Author Report for: Yan YNo contact information in database for Yan Y
Hu Z, Jiao L, Xie X, Xu L, Yan J, Yang M, Yan Y Ref: Int J Mol Sci, 24:8924, 2023 : PubMed ESTHER: Hu_2023_Int.J.Mol.Sci_24_8924 PubMedSearch: Hu 2023 Int.J.Mol.Sci 24 8924 PubMedID: 37240270 Gene_locus related to this paper: psefs-c3kbe9 Abstract The search for and characterization of new lipases with excellent properties has always been urgent and is of great importance to meet industrial needs. In this study, a new lipase, lipB, from Pseudomonas fluorescens SBW25, belonging to the lipase subfamily I.3, was cloned and expressed in Bacillus subtilis WB800N. Enzymatic properties studies of recombinant LipB found that it exhibited the highest activity towards p-nitrophenyl caprylate at 40 degreesC and pH 8.0, retaining 73% of its original activity after incubation at 70 degreesC for 6 h. In addition, Ca(2+), Mg(2+), and Ba(2+) strongly enhanced the activity of LipB, while Cu(2+), Zn(2+), Mn(2+), and CTAB showed an inhibiting effect. The LipB also displayed noticeable tolerance to organic solvents, especially acetonitrile, isopropanol, acetone, and DMSO. Moreover, LipB was applied to the enrichment of polyunsaturated fatty acids from fish oil. After hydrolyzing for 24 h, it could increase the contents of polyunsaturated fatty acids from 43.16% to 72.18%, consisting of 5.75% eicosapentaenoic acid, 19.57% docosapentaenoic acid, and 46.86% docosahexaenoic acid, respectively. The properties of LipB render it great potential in industrial applications, especially in health food production. Title: Alteration of Chain-Length Selectivity and Thermostability of Rhizopus oryzae Lipase via Virtual Saturation Mutagenesis Coupled with Disulfide Bond Design Huang J, Dai S, Chen X, Xu L, Yan J, Yang M, Yan Y Ref: Applied Environmental Microbiology, :e0187822, 2023 : PubMed ESTHER: Huang_2023_Appl.Environ.Microbiol__e0187822 PubMedSearch: Huang 2023 Appl.Environ.Microbiol e0187822 PubMedID: 36602359 Gene_locus related to this paper: rhidl-lipas Abstract Rhizopus oryzae lipase (ROL) is one of the most important enzymes used in the food, biofuel, and pharmaceutical industries. However, the highly demanding conditions of industrial processes can reduce its stability and activity. To seek a feasible method to improve both the catalytic activity and the thermostability of this lipase, first, the structure of ROL was divided into catalytic and noncatalytic regions by identifying critical amino acids in the crevice-like binding pocket. Second, a mutant screening library aimed at improvement of ROL catalytic performance by virtual saturation mutagenesis of residues in the catalytic region was constructed based on Rosetta's Cartesian_ddg protocol. A double mutant, E265V/S267W (with an E-to-V change at residue 265 and an S-to-W change at residue 267), with markedly improved catalytic activity toward diverse chain-length fatty acid esters was identified. Then, computational design of disulfide bonds was conducted for the noncatalytic amino acids of E265V/S267W, and two potential disulfide bonds, S61C-S115C and E190C-E238C, were identified as candidates. Experimental data validated that the variant E265V/S267W/S61C-S115C/E190C-E238C had superior stability, with an increase of 8.5 degreesC in the melting temperature and a half-life of 31.7 min at 60 degreesC, 4.2-fold longer than that of the wild-type enzyme. Moreover, the variant improved the lipase activity toward five 4-nitrophenyl esters by 1.5 to 3.8 times, exhibiting a potential to modify the catalytic efficiency. IMPORTANCE Rhizopus oryzae lipase (ROL) is very attractive in biotechnology and industry as a safe and environmentally friendly biocatalyst. Functional expression of ROL in Escherichia coli facilitates effective high-throughput screening for positive variants. This work highlights a method to improve both selectivity and thermostability based on a combination of virtual saturation mutagenesis in the substrate pocket and disulfide bond prediction in the noncatalytic region. Using the method, ROL thermostability and activity to diverse 4-nitrophenyl esters could be substantially improved. The strategy of rational introduction of multiple mutations in different functional domains of the enzyme is a great prospect in the modification of biocatalysts. Title: De Novo Computational Design of a Lipase with Hydrolysis Activity towards Middle-Chained Fatty Acid Esters Huang J, Xie X, Zheng Z, Ye L, Wang P, Xu L, Wu Y, Yan J, Yang M, Yan Y Ref: Int J Mol Sci, 24:8581, 2023 : PubMed ESTHER: Huang_2023_Int.J.Mol.Sci_24_8581 PubMedSearch: Huang 2023 Int.J.Mol.Sci 24 8581 PubMedID: 37239928 Gene_locus related to this paper: 9zzzz-1a8uD1 Abstract Innovations in biocatalysts provide great prospects for intolerant environments or novel reactions. Due to the limited catalytic capacity and the long-term and labor-intensive characteristics of mining enzymes with the desired functions, de novo enzyme design was developed to obtain industrial application candidates in a rapid and convenient way. Here, based on the catalytic mechanisms and the known structures of proteins, we proposed a computational protein design strategy combining de novo enzyme design and laboratory-directed evolution. Starting with the theozyme constructed using a quantum-mechanical approach, the theoretical enzyme-skeleton combinations were assembled and optimized via the Rosetta "inside-out" protocol. A small number of designed sequences were experimentally screened using SDS-PAGE, mass spectrometry and a qualitative activity assay in which the designed enzyme 1a8uD(1) exhibited a measurable hydrolysis activity of 24.25 +/- 0.57 U/g towards p-nitrophenyl octanoate. To improve the activity of the designed enzyme, molecular dynamics simulations and the RosettaDesign application were utilized to further optimize the substrate binding mode and amino acid sequence, thus keeping the residues of theozyme intact. The redesigned lipase 1a8uD(1)-M8 displayed enhanced hydrolysis activity towards p-nitrophenyl octanoate-3.34 times higher than that of 1a8uD(1). Meanwhile, the natural skeleton protein (PDB entry 1a8u) did not display any hydrolysis activity, confirming that the hydrolysis abilities of the designed 1a8uD(1) and the redesigned 1a8uD(1)-M8 were devised from scratch. More importantly, the designed 1a8uD(1)-M8 was also able to hydrolyze the natural middle-chained substrate (glycerol trioctanoate), for which the activity was 27.67 +/- 0.69 U/g. This study indicates that the strategy employed here has great potential to generate novel enzymes exhibiting the desired reactions. Title: Immobilization of Rhizomucor miehei lipase on magnetic multiwalled carbon nanotubes towards the synthesis of structured lipids rich in sn-2 palmitic acid and sn-1,3 oleic acid (OPO) for infant formula use Ghide MK, Li K, Wang J, Abdulmalek SA, Yan Y Ref: Food Chem, 390:133171, 2022 : PubMed ESTHER: Ghide_2022_Food.Chem_390_133171 PubMedSearch: Ghide 2022 Food.Chem 390 133171 PubMedID: 35551020 Abstract Nowadays, breast milk is considered as the ideal food for infants owing to the most common oleic acid-palmitic acid-oleic acid (OA-PA-OA) fatty acid distribution of the human milk fat (HMF). This study reports the synthesis of 1,3-dioleoyl-2-palmotoylglycerol (OPO)-rich human milk fat substitutes in a two-step enzymatic acidolysis reaction with Rhizomucor miehei lipase (RML) immobilized on magnetic multi-walled carbon nanotubes(mMWCNTs). The immobilized RML (RML-mMWCNTs) showed better thermal and pH stability, convenient recovery and reusability than the free soluble form. Under optimized reaction conditions (1:8 tripalmitin (PPP)/OA, 10%wt. enzyme, 50 degreesC, 5 h), PA content at the sn-2 position and OA incorporation at the sn-1,3 positions reached 93.46% and 59.54%, respectively. Comparison tests have also showed that RML-mMWCNTs has better catalytic activity and reusability than the commercial lipase Lipozyme RM IM. The results suggest that RML-mMWCNTs is a promising biocatalyst for the synthesis of OPO-rich TAGs with potential use in infant formulas. Title: Glimepiride Use is Associated with Reduced Cardiovascular Mortality in Patients with Type 2 Diabetes and Chronic Heart Failure: A Prospective Cohort Study He W, Yuan G, Han Y, Yan Y, Li G, Zhao C, Shen J, Jiang X, Chen C and Wang DW <1 more author(s)> He W, Yuan G, Han Y, Yan Y, Li G, Zhao C, Shen J, Jiang X, Chen C, Ni L, Wang DW (- 1) Ref: Eur J Prev Cardiol, :, 2022 : PubMed ESTHER: He_2022_Eur.J.Prev.Cardiol__ PubMedSearch: He 2022 Eur.J.Prev.Cardiol PubMedID: 36573717 Abstract BACKGROUND: Glimepiride has good cardiovascular safety. However, whether glimepiride benefits clinical cardiovascular outcomes is unclear. METHODS: A total of 21,451 inpatients with type 2 diabetes (T2D) and chronic heart failure (CHF) were analyzed, including 638 who received glimepiride treatment and 20,813 who did not. Propensity score matching yielded 509 pairs (glimepiride and non-glimepiride groups), and both groups were followed up. Kaplan-Meier and Cox regression analyses were used to compare all-cause mortality, cardiovascular mortality, hospitalizations and emergency visits for heart failure, and hospitalizations for acute myocardial infarction or stroke. RESULTS: During follow-up, the all-cause mortality (adjusted hazard ratio [HR], 0.47; 95% confidence interval [CI], 0.35-0.63; P < 0.001), cardiovascular mortality (adjusted HR, 0.34; 95% CI, 0.24-0.48; P < 0.001), and number of hospitalizations and emergency visits for heart failure (adjusted HR, 0.42; 95% CI, 0.36-0.50; P < 0.001) and hospitalizations for acute myocardial infarction or stroke (adjusted HR, 0.53; 95% CI, 0.38-0.73; P < 0.001) were significantly lower in the glimepiride group; the conclusion remained similar in all subgroups. Furthermore, high-dose glimepiride use (2-4mg/day) was associated with lower cardiovascular mortality than low-dose (1mg/day) (adjusted HR, 0.55; 95% CI, 0.31-0.99; P = 0.047). Glimepiride exhibited good molecular docking with soluble epoxide hydrolase (sEH) and increased the level epoxyeicosatrienoic acid (EET). CONCLUSIONS: Long-term continuous glimepiride use is associated with better survival, fewer hospitalizations and emergency visits for heart failure, and fewer hospitalizations for acute myocardial infarction or stroke in patients with T2D and CHF. High-dose glimepiride has greater cardiovascular protective advantages than low-dose glimepiride. The cardiovascular protective effect of glimepiride may be related to the EET level increase through sEH inhibition. Title: Employing Engineered Enolase Promoter for Efficient Expression of Thermomyces lanuginosus Lipase in Yarrowia lipolytica via a Self-Excisable Vector Jiao L, Li W, Li Y, Zhou Q, Zhu M, Zhao G, Zhang H, Yan Y Ref: Int J Mol Sci, 24:, 2022 : PubMed ESTHER: Jiao_2022_Int.J.Mol.Sci_24_ PubMedSearch: Jiao 2022 Int.J.Mol.Sci 24 PubMedID: 36614159 Abstract Yarrowia lipolytica is progressively being employed as a workhouse for recombinant protein expression. Here, we expanded the molecular toolbox by engineering the enolase promoter (pENO) and developed a new self-excisable vector, and based on this, a combined strategy was employed to enhance the expression of Thermomyces lanuginosus lipase (TLL) in Y. lipolytica. The strength of 11 truncated enolase promoters of different length was first identified using eGFP as a reporter. Seven of the truncated promoters were selected to examine their ability for driving TLL expression. Then, a series of enolase promoters with higher activities were developed by upstream fusing of different copies of UAS1B, and the recombinant strain Po1f/hp16e(100)-tll harboring the optimal promoter hp16e(100) obtained a TLL activity of 447 U/mL. Additionally, a new self-excisable vector was developed based on a Cre/loxP recombination system, which achieved efficient markerless integration in Y. lipolytica. Subsequently, strains harboring one to four copies of the tll gene were constructed using this tool, with the three-copy strain Po1f/3tll showing the highest activity of 579 U/mL. The activity of Po1f/3tll was then increased to 720 U/mL by optimizing the shaking flask fermentation parameters. Moreover, the folding-related proteins Hac1, Pdi, and Kar2 were employed to further enhance TLL expression, and the TLL activity of the optimal recombinant strain Po1f/3tll-hac1-pdi-kar2 reached 1197 U/mL. By using this combined strategy, TLL activity was enhanced by approximately 39.9-fold compared to the initial strain. Thus, the new vector and the combined strategy could be a useful tool to engineer Y. lipolytica for high-level expression of heterologous protein. Title: Short-term prognostic factors for hepatitis B virus-related acute-on-chronic liver failure Ye QX, Huang JF, Xu ZJ, Yan YY, Yan Y, Liu LG Ref: World J Clin Cases, 10:8186, 2022 : PubMed ESTHER: Ye_2022_World.J.Clin.Cases_10_8186 PubMedSearch: Ye 2022 World.J.Clin.Cases 10 8186 PubMedID: 36159528 Abstract BACKGROUND: Acute-on-chronic liver failure (ACLF) is the abrupt exacerbation of declined hepatic function in patients with chronic liver disease. AIM: To explore the independent predictors of short-term prognosis in patients with hepatitis B virus (HBV)-related ACLF and to establish a predictive short-term prognosis model for HBV-related ACLF. METHODS: From January 2016 to December 2019, 207 patients with HBV-related ACLF attending the 910th Hospital of Chinese People's Liberation Army were continuously included in this retrospective study. Patients were stratified based on their survival status 3 mo after diagnosis. Information was collected regarding gender and age; coagulation function in terms of prothrombin time and international normalized ratio (INR); hematological profile in terms of neutrophil-to-lymphocyte ratio (NLR) and platelet count (PLT); blood biochemistry in terms of alanine aminotransferase, aspartate aminotransferase, total bilirubin (Tbil), albumin, cholinesterase, blood urea nitrogen (BUN), creatinine, blood glucose, and sodium (Na); tumor markers including alpha-fetoprotein (AFP) and Golgi protein 73 (GP73); virological indicators including HBV-DNA, HBsAg, HBeAg, Anti-HBe, and Anti-HBc; and complications including hepatic encephalopathy, hepatorenal syndrome, spontaneous peritonitis, gastrointestinal bleeding, and pulmonary infection. RESULTS: There were 157 and 50 patients in the survival and death categories, respectively. Univariate analysis revealed significant differences in age, PLT, Tbil, BUN, NLR, HBsAg, AFP, GP73, INR, stage of liver failure, classification of liver failure, and incidence of complications (pulmonary infection, hepatic encephalopathy, spontaneous bacterial peritonitis, and upper gastrointestinal bleeding) between the two groups (P < 0.05). GP73 [hazard ratio (HR): 1.009, 95% confidence interval (CI): 1.005-1.013, P = 0.000], middle stage of liver failure (HR: 5.056, 95%CI: 1.792-14.269, P = 0.002), late stage of liver failure (HR: 22.335, 95%CI: 8.544-58.388, P = 0.000), pulmonary infection (HR: 2.056, 95%CI: 1.145-3.690, P = 0.016), hepatorenal syndrome (HR: 6.847, 95%CI: 1.930-24.291, P = 0.003), and HBsAg (HR: 0.690, 95%CI: 0.524-0.908, P = 0.008) were independent risk factors for short-term prognosis in patients with HBV-related ACLF. Following binary logistics regression analysis, we arrived at the following formula for predicting short-term prognosis: Logit(P) = Ln(P/1-P) = 0.013 x (GP73 ng/mL) + 1.907 x (middle stage of liver failure) + 4.146 x (late stage of liver failure) + 0.734 x (pulmonary infection) + 22.320 x (hepatorenal syndrome) - 0.529 x (HBsAg) - 5.224. The predictive efficacy of the GP73-ACLF score was significantly better than that of the Model for End-Stage Liver Disease (MELD) and MELD-Na score models (P < 0.05). CONCLUSION: The stage of liver failure, presence of GP73, pulmonary infection, hepatorenal syndrome, and HBsAg are independent predictors of short-term prognosis in patients with HBV-related ACLF, and the GP73-ACLF model has good predictive value among these patients. Title: Novel AP2238-clorgiline hybrids as multi-target agents for the treatment of Alzheimer's disease: Design, synthesis, and biological evaluation Zhong G, Guo J, Pang C, Su D, Tang C, Jing L, Zhang F, He P, Yan Y and Jiang N <2 more author(s)> Zhong G, Guo J, Pang C, Su D, Tang C, Jing L, Zhang F, He P, Yan Y, Chen Z, Liu J, Jiang N (- 2) Ref: Bioorg Chem, 130:106224, 2022 : PubMed ESTHER: Zhong_2022_Bioorg.Chem_130_106224 PubMedSearch: Zhong 2022 Bioorg.Chem 130 106224 PubMedID: 36332315 Abstract Cholinesterase and monoamine oxidase are potential targets for the therapy of Alzheimer's disease. A series of novel AP2238-clorgiline hybrids as multi-target agents were designed, synthesized and investigated in vitro for their inhibition of cholinesterases and monoamine oxidases. Many compounds displayed balanced and good inhibitory activity against AChE, BuChE and MAO-B with an obvious selective inhibitory effect on MAO-B. Among them, Compound 5l showed the most balanced potency to inhibit ChEs (eeAChE: IC(50) = 4.03 +/- 0.03 microM, eqBuChE: IC(50) = 5.64 +/- 0.53 microM; hAChE: IC(50) = 8.30 +/- 0.04 microM, hBuChE: IC(50) = 1.91 +/- 0.06 microM) and hMAO-B (IC(50) = 3.29 +/- 0.09 microM). Molecular modeling and kinetic studies showed that 5l was a mixed inhibitor for both AChE and BuChE, and a competitive MAO-B inhibitor. Compound 5l exhibited no toxicity to PC12 and BV-2 cells at 12.5 microM and no acute toxicity at a dosage of 2500 mg/kg. Moreover, 5l can improve the memory function of mice with scopolamine-induced memory impairment and have an excellent ability to cross the blood-brain barrier. Overall, these findings suggested that compound 5l could be deemed as a promising, balanced multi-target drug candidate against Alzheimer's disease. Title: Pancreatic Cancer Cell-Derived Exosomes Promote Lymphangiogenesis by Downregulating ABHD11-AS1 Expression Zhou X, Zhong F, Yan Y, Wu S, Wang H, Liu J, Li F, Cui D, Xu M Ref: Cancers (Basel), 14:, 2022 : PubMed ESTHER: Zhou_2022_Cancers.(Basel)_14_ PubMedSearch: Zhou 2022 Cancers.(Basel) 14 PubMedID: 36230535 Gene_locus related to this paper: human-ABHD11 Abstract Research on pancreatic cancer microbiomes has attracted attention in recent years. The current view is that enriched microbial communities in pancreatic cancer tissues may affect pancreatic cancer metastasis, including lymph node (LN) metastasis. Similar to carriers of genetic information between cells, such as DNA, mRNA, protein, and non-coding RNA, exosomes are of great importance in early LN metastasis in tumors, including pancreatic cancer. Our previous study showed that the long non-coding RNA ABHD11-AS1 was highly expressed in tissues of patients with pancreatic cancer, and was correlated with patient survival time. However, the role of ABHD11-AS1 in pancreatic cancer LN metastasis has rarely been studied. Hence, in this paper we confirmed that exosomes derived from pancreatic cancer cells could promote lymphangiogenesis in vitro and in vivo, and that the mechanism was related to the downregulation of ABHD11-AS1 expression in lymphatic endothelial cells, and to the enhancement of their ability to proliferate, migrate, and form tubes. These findings preliminarily show a new mechanism by which pancreatic cancer cells regulate peripheral lymphangiogenesis, providing a new therapeutic strategy for inhibiting LN metastasis in pancreatic cancer. Title: A Polyketide Cyclase That Forms Medium-Ring Lactones Gao DW, Jamieson CS, Wang G, Yan Y, Zhou J, Houk KN, Tang Y Ref: Journal of the American Chemical Society, 143:80, 2021 : PubMed ESTHER: Gao_2021_J.Am.Chem.Soc_143_80 PubMedSearch: Gao 2021 J.Am.Chem.Soc 143 80 PubMedID: 33351624 Gene_locus related to this paper: beab2-j4wat9 Abstract Medium-ring lactones are synthetically challenging due to unfavorable energetics involved in cyclization. We have discovered a thioesterase enzyme DcsB, from the decarestrictine C1 (1) biosynthetic pathway, that efficiently performs medium-ring lactonizations. DcsB shows broad substrate promiscuity toward linear substrates that vary in lengths and substituents, and is a potential biocatalyst for lactonization. X-ray crystal structure and computational analyses provide insights into the molecular basis of catalysis. Title: Developmental neurotoxicity of antimony (Sb) in the early life stages of zebrafish Xia S, Zhu X, Yan Y, Zhang T, Chen G, Lei D, Wang G Ref: Ecotoxicology & Environmental Safety, 218:112308, 2021 : PubMed ESTHER: Xia_2021_Ecotoxicol.Environ.Saf_218_112308 PubMedSearch: Xia 2021 Ecotoxicol.Environ.Saf 218 112308 PubMedID: 33975224 Abstract Accumulating studies have revealed the toxicity of antimony (Sb) to soil-dwelling and aquatic organisms at the individual level. However, little is known about the neurotoxic effects of antimony and its underlying mechanisms. To assess this issue, we investigated the neurotoxicity of antimony (0, 200, 400, 600 and 800 mg/L) in zebrafish embryos. After exposure, zebrafish embryos showed abnormal phenotypes such as a shortened body length, morphological malformations, and weakened heart function. Behavioral experiments indicated that antimony caused neurotoxicity in zebrafish embryos, manifested in a decreased spontaneous movement frequency, delayed response to touch, and reduced movement distance. We also showed that antimony caused a decrease in acetylcholinesterase (AChE) levels in zebrafish embryos, along with decreased expression of neurofunctional markers such as gfap, nestin, mbp, and shha. Additionally, antimony significantly increased reactive oxygen species levels and significantly reduced glutathione (GSH) and superoxide dismutase (SOD) activity. In summary, our findings indicated that antimony can induce developmental toxicity and neurotoxicity in zebrash embryos by affecting neurotransmitter systems and oxidative stress, thus altering behavior. These outcomes will advance our understanding of antimony-induced neurotoxicity, environmental problems, and health hazards. Title: Enzyme-Responsive Aqueous Two-Phase Systems in a Cationic-Anionic Surfactant Mixture Xiao X, Qiao Y, Xu Z, Wu T, Wu Y, Ling Z, Yan Y, Huang J Ref: Langmuir, :, 2021 : PubMed ESTHER: Xiao_2021_Langmuir__ PubMedSearch: Xiao 2021 Langmuir PubMedID: 34714092 Abstract Enzyme-instructed self-assembly is an increasingly attractive topic owing to its broad applications in biomaterials and biomedicine. In this work, we report an approach to construct enzyme-responsive aqueous surfactant two-phase (ASTP) systems serving as enzyme substrates by using a cationic surfactant (myristoylcholine chloride) and a series of anionic surfactants. Driven by the hydrophobic interaction and electrostatic attraction, self-assemblies of cationic-anionic surfactant mixtures result in biphasic systems containing condensed lamellar structures and coexisting dilute solutions, which turn into homogeneous aqueous phases in the presence of hydrolase (cholinesterase). The enzyme-sensitive ASTP systems reported in this work highlight potential applications in the active control of biomolecular enrichment/release and visual detection of cholinesterase. Title: A highly contiguous genome assembly of a polyphagous predatory mite Stratiolaelaps scimitus (Womersley) (Acari: Laelapidae) Yan Y, Zhang N, Liu C, Wu X, Liu K, Yin Z, Zhou X, Xie L Ref: Genome Biol Evol, :, 2021 : PubMed ESTHER: Yan_2021_Genome.Biol.Evol__ PubMedSearch: Yan 2021 Genome.Biol.Evol PubMedID: 33528489 Abstract As a polyphagous soil-dwelling predatory mite, Stratiolaelaps scimitus (Womersley) (Acari: Laelapidae), formerly known as Stratiolaelaps miles (Berlese), is native to the Northern hemisphere and preys on soil invertebrates, including fungus gnats, springtails, thrips nymphs, nematodes, and other species of mites. Already mass-produced and commercialized in North America and Europe, S. scimitus is now introduced in China as a biocontrol agent for field crop. The introduction, however, can lead to unexpected genetic changes within populations of biological control agents, which might decrease the efficacy of pest management or increase the risks to local environments. To better understand the genetic basis of its biology and behavior, we sequenced and assembled the draft genome of S. scimitus using the PacBio Sequel platform II. We generated -150 x (64.81 Gb) PacBio long reads with an average read length of 12.60 kb. Reads longer than 5 kb were assembled into contigs, resulting in the final assembly of 158 contigs with a N50 length of 7.66 Mb, and captured 93.1% of the BUSCO gene set (n = 1,066). We identified 16.39% (69.91 Mb) repetitive elements, 1,686 non-coding RNAs, and 13,305 protein-coding genes, which represented 95.8% BUSCO completeness. Combining analyses of genome family evolution and function enrichment of gene ontology and pathway, a total of 135 families experienced significant expansions, which were mainly involved in digestion, detoxification, immunity and venom. Major expansions of the detoxification enzymes, i.e., P450s and carboxylesterases, suggest a possible genetic mechanism underlying polyphagy and ecological adaptions. Our high-quality genome assembly and annotation provide new insights on the evolutionary biology, soil ecology and biological control for predaceous mites. Title: A De Novo Designed Esterase with p-Nitrophenyl Acetate Hydrolysis Activity Li G, Xu L, Zhang H, Liu J, Yan J, Yan Y Ref: Molecules, 25:4658, 2020 : PubMed ESTHER: Li_2020_Molecules_25_4658 PubMedSearch: Li 2020 Molecules 25 4658 PubMedID: 33066055 Gene_locus related to this paper: aspor-cutas, fusso-cutas, hevbr-hnl Abstract Esterases are a large family of enzymes with wide applications in the industry. However, all esterases originated from natural sources, limiting their use in harsh environments or newly- emerged reactions. In this study, we designed a new esterase to develop a new protocol to satisfy the needs for better biocatalysts. The ideal spatial conformation of the serine catalytic triad and the oxygen anion hole at the substrate-binding site was constructed by quantum mechanical calculation. The catalytic triad and oxygen anion holes were then embedded in the protein scaffold using the new enzyme protocol in Rosetta 3. The design results were subsequently evaluated, and optimized designs were used for expression and purification. The designed esterase had significant lytic activities towards p-nitrophenyl acetate, which was confirmed by point mutations. Thus, this study developed a new protocol to obtain novel enzymes that may be useful in unforgiving environments or novel reactions. Title: Design, synthesis and biological evaluation of novel carboline-cinnamic acid hybrids as multifunctional agents for treatment of Alzheimer's disease Liao Q, Li Q, Zhao Y, Jiang P, Yan Y, Sun H, Liu W, Feng F, Qu W Ref: Bioorg Chem, 99:103844, 2020 : PubMed ESTHER: Liao_2020_Bioorg.Chem_99_103844 PubMedSearch: Liao 2020 Bioorg.Chem 99 103844 PubMedID: 32325336 Abstract Alzheimer's disease (AD) is a complex neurodegenerative disease with multiple pathological features. Multifunctional compounds able to simultaneously interact with several pathological components have been considered as a solution to treat the complex pathologies of neurodegenerative diseases. beta-carboline and cinnamic acid have been extensively studied for their widespread biological effects in treatment of AD, further application is limited due to its poor solubility and high toxicity. Herein, a series of carboline-cinnamic acid hybrids was designed and synthesized to obtain new multifunctional molecules with low toxicity and good physicochemical properties. In particular, e3 and e12 exhibited significant inhibition of Abeta aggregation (inhibitory rate at 25 M: 65% and 72% respectively), moderate BuChE inhibition, excellent neuroprotective effects and low neurotoxicity. Furthermore, in the AD mice model, e3 and e12 could restore learning and memory function to a comparable level to that of the control and did not exhibit any acute toxicity in vivo at a relatively high dose of 600 mg/kg. Thus, these new compounds can be further studied as multifunctional molecules for AD. Title: Edaravone at high concentrations attenuates cognitive dysfunctions induced by abdominal surgery under general anesthesia in aged mice Zhou Y, Wu X, Ye L, Bai Y, Zhang H, Xuan Z, Feng Y, Zhang P, Chen Y and Cui W <2 more author(s)> Zhou Y, Wu X, Ye L, Bai Y, Zhang H, Xuan Z, Feng Y, Zhang P, Chen Y, Yan Y, Zhu B, Cui W (- 2) Ref: Metabolic Brain Disease, :, 2020 : PubMed ESTHER: Zhou_2020_Metab.Brain.Dis__ PubMedSearch: Zhou 2020 Metab.Brain.Dis PubMedID: 31916204 Abstract Postoperative cognitive dysfunction (POCD) is a common neurological disease affecting the elderly patients after surgery. Unfortunately, no effective treatment for this disease has been discovered. Edaravone, a clinical-used free radical scavenger, at 3 mg/kg has been reported to prevent neuroinflammation induced by the combination of surgery and lipopolysaccharide in adult rodents. However, we found that edaravone at such low concentration could not inhibit POCD in aged mice. Instead, edaravone at 33.2 mg/kg significantly prevented recognition and spatial cognitive dysfunctions in 14 month aged mice after abdominal surgery under general anesthesia with isoflurane. Furthermore, edaravone significantly prevented the increase of tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta) and interleukin-6 (IL-6) induced by abdominal surgery in aged mice. Edaravone could also decrease glial fibrillary acidic protein (GFAP) and ionized calcium binding adaptor molecule-1 (Iba-1) positive areas in the hippocampal regions of surgery mice, suggesting that edaravone might inhibit surgery-induced over-activation of microglia and astrocytes. Moreover, edaravone substantially increased the expression of PSD-95 and pSer9-glycogen synthase kinase-3beta (pSer9-GSK3beta) as demonstrated by Western blotting assay. Furthermore, the activity of acetylcholinesterase (AChE) is decreased in the mice in edaravone group. All these results suggested that edaravone at high concentrations could inhibit surgery-induced cognitive impairments in aged animals, possibly via the attenuation of neuroinflammation, the increase of synaptic proteins, and the elevation of cholinergic transmission, providing a further support that edaravone might be developed as a treatment of POCD. Title: Metal-Organic Frameworks Conjugated Lipase with Enhanced Bio-catalytic Activity and Stability Zou B, Zhang L, Xia J, Wang P, Yan Y, Wang X, Adesanya IO Ref: Appl Biochem Biotechnol, :, 2020 : PubMed ESTHER: Zou_2020_Appl.Biochem.Biotechnol__ PubMedSearch: Zou 2020 Appl.Biochem.Biotechnol PubMedID: 32323142 Abstract Covalent immobilization of lipase onto a solid carrier is an effective way to enhance stability. Immobilization inhibits the activity of lipase due to decreased flexibility of enzyme structure via the covalent bond. In this study, monomer of the metal-organic frameworks (MOFs) material ZIF-8 (2-methyl imidazole-4-carboxylic acid) was innovatively used as a chemical modifier of Candida nrugosa lipase (CRL). The circular dichroism spectra results show that the CRL molecule was altered by chemical modification and thus its catalytic activity was 1.3 times higher than that of the free CRL. The modified CRL molecule was further immobilized in the "skeleton" of ZIF-8 through the monomer while in situ forming the cell skeleton of the MOFs, which prevent the active center from being destroyed. The results show that conjugation of chemical modification and immobilized enzymes ensure that there was no obvious reduction in the activity of CRL after immobilization and the stability of CRL was improved. Especially, the organic solvent stability of the modified immobilization CRL in isopropanol was significantly improved and retained more than 148% of its activity. Title: Enhancing bio-catalytic activity and stability of lipase nanogel by functional ionic liquids modification Zou B, Yan Y, Xia J, Zhang L, Adesanya IO Ref: Colloids Surf B Biointerfaces, 195:111275, 2020 : PubMed ESTHER: Zou_2020_Colloids.Surf.B.Biointerfaces_195_111275 PubMedSearch: Zou 2020 Colloids.Surf.B.Biointerfaces 195 111275 PubMedID: 32739774 Abstract A novel integrated lipase nanogel based on functional ionic liquid modification and polymerization immobilization with improved stability was designed. Characterization before and after modification and polymerization was conducted using infrared spectroscopy, Circular dichroism spectroscopy, fluorescence spectroscopy, and scanning electron microscopy. It was shown that the modification of the ionic liquid influenced the catalytic behavior of lipase significantly due to the changed structure and surface properties of lipase. The enzymatic properties, including acid-base stability, thermal stability, organic solvents stability, and storage stability of CRL nanogel, were investigated in the p-nitrophenyl palmitate hydrolysis reaction (CRL, Lipase from Candida Rugosa). The results indicated that CRL nanogel has a better pH, heat, and organic solvent tolerance after immobilization. After seven weeks of storage, the natural CRL gradually lost its enzymatic activity, and only 17.5+/-1.7 % of the catalytic activity remained, the residual activity of CRL nanogel was 97.3+/-1.8 %. It was indicated that the novel CRL nanogel was an excellent biocatalyst. Title: Musa balbisiana genome reveals subgenome evolution and functional divergence Wang Z, Miao H, Liu J, Xu B, Yao X, Xu C, Zhao S, Fang X, Jia C and Jin Z <30 more author(s)> Wang Z, Miao H, Liu J, Xu B, Yao X, Xu C, Zhao S, Fang X, Jia C, Wang J, Zhang J, Li J, Xu Y, Ma W, Wu Z, Yu L, Yang Y, Liu C, Guo Y, Sun S, Baurens FC, Martin G, Salmon F, Garsmeur O, Yahiaoui N, Hervouet C, Rouard M, Laboureau N, Habas R, Ricci S, Peng M, Guo A, Xie J, Li Y, Ding Z, Yan Y, Tie W, D'Hont A, Hu W, Jin Z (- 30) Ref: Nat Plants, 5:810, 2019 : PubMed ESTHER: Wang_2019_Nat.Plants_5_810 PubMedSearch: Wang 2019 Nat.Plants 5 810 PubMedID: 31308504 Gene_locus related to this paper: musam-m0tuu7, musam-a0a804kav5 Abstract Banana cultivars (Musa ssp.) are diploid, triploid and tetraploid hybrids derived from Musa acuminata and Musa balbisiana. We presented a high-quality draft genome assembly of M. balbisiana with 430 Mb (87%) assembled into 11 chromosomes. We identified that the recent divergence of M. acuminata (A-genome) and M. balbisiana (B-genome) occurred after lineage-specific whole-genome duplication, and that the B-genome may be more sensitive to the fractionation process compared to the A-genome. Homoeologous exchanges occurred frequently between A- and B-subgenomes in allopolyploids. Genomic variation within progenitors resulted in functional divergence of subgenomes. Global homoeologue expression dominance occurred between subgenomes of the allotriploid. Gene families related to ethylene biosynthesis and starch metabolism exhibited significant expansion at the pathway level and wide homoeologue expression dominance in the B-subgenome of the allotriploid. The independent origin of 1-aminocyclopropane-1-carboxylic acid oxidase (ACO) homoeologue gene pairs and tandem duplication-driven expansion of ACO genes in the B-subgenome contributed to rapid and major ethylene production post-harvest in allotriploid banana fruits. The findings of this study provide greater context for understanding fruit biology, and aid the development of tools for breeding optimal banana cultivars. Title: Excipient-free nanodispersion of 7-ethyl-10-hydroxycamptothecin exerts potent therapeutic effects against pancreatic cancer cell lines and patient-derived xenografts Zhang L, Zhou J, Yan Y, Zhou X, Zhou Q, Du R, Hu S, Ge W, Huang Y and Wang W <5 more author(s)> Zhang L, Zhou J, Yan Y, Zhou X, Zhou Q, Du R, Hu S, Ge W, Huang Y, Xu H, Kong Y, Zheng H, Ding Y, Shen Y, Wang W (- 5) Ref: Cancer Letters, 465:36, 2019 : PubMed ESTHER: Zhang_2019_Cancer.Lett_465_36 PubMedSearch: Zhang 2019 Cancer.Lett 465 36 PubMedID: 31479691 Abstract Irinotecan (CPT-11) is an anti-tumor drug and formulated as nanomedicines to reduce side effects and improve efficacy. In vivo, CPT-11 must be hydrolyzed by carboxylesterase to its active form 7-ethyl-10-hydroxycamptothecin (SN-38) to exert anti-tumor activity, but the lack of this enzyme in humans causes inefficient generation of SN-38. Thus, direct delivery of SN-38, not relying on carboxylesterase, will potentially achieve higher efficacy. However, it is difficult to effectively formulate SN-38 using current excipients due to its hydrophobicity and tendency to crystallize. Herein, we report the nanodispersion of SN-38 with its amphiphilic prodrug, CPT-11, as an effective treatment for pancreatic cancer (PC). SN-38 and CPT-11 formed stable nanoparticles without any other excipients, and showed potent cytotoxicity against PC cells in vitro, slowed tumor growth in vivo, namely subcutaneously and orthotopically xenografted mice, with minimal adverse effects, and prolonged their overall survival. Even in clinically-relevant patient-derived xenograft (PDX) models, the nanodispersion showed greater anti-tumor efficacy than CPT-11. Importantly, the nanodispersion directly released SN-38, resulting in carboxylesterase-independent anti-tumor activity, in contrast to carboxylesterase-dependent CPT-11. These characteristics may enable the excipient-free nanodispersion to exert potent therapeutic effects in patients. Title: Excellent Degradation Performance of a Versatile Phthalic Acid Esters-Degrading Bacterium and Catalytic Mechanism of Monoalkyl Phthalate Hydrolase Fan S, Wang J, Yan Y, Jia Y Ref: Int J Mol Sci, 19:, 2018 : PubMed ESTHER: Fan_2018_Int.J.Mol.Sci_19_ PubMedSearch: Fan 2018 Int.J.Mol.Sci 19 PubMedID: 30231475 Gene_locus related to this paper: 9acto-q2mhh5 Abstract Despites lots of characterized microorganisms that are capable of degrading phthalic acid esters (PAEs), there are few isolated strains with high activity towards PAEs under a broad range of environmental conditions. In this study, Gordonia sp. YC-JH1 had advantages over its counterparts in terms of di(2-ethylhexyl) phthalate (DEHP) degradation performance. It possessed an excellent degradation ability in the range of 20(-)50 degrees C, pH 5.0(-)12.0, or 0(-)8% NaCl with the optimal degradation condition 40 degrees C and pH 10.0. Therefore, strain YC-JH1 appeared suitable for bioremediation application at various conditions. Metabolites analysis revealed that DEHP was sequentially hydrolyzed by strain YC-JH1 to mono(2-ethylhexyl) phthalate (MEHP) and phthalic acid (PA). The hydrolase MphG1 from strain YC-JH1 hydrolyzed monoethyl phthalate (MEP), mono-n-butyl phthalate (MBP), mono-n-hexyl phthalate (MHP), and MEHP to PA. According to molecular docking and molecular dynamics simulation between MphG1 and monoalkyl phthalates (MAPs), some key residues were detected, including the catalytic triad (S125-H291-D259) and the residues R126 and F54 potentially binding substrates. The mutation of these residues accounted for the reduced activity. Together, the mechanism of MphG1 catalyzing MAPs was elucidated, and would shed insights into catalytic mechanism of more hydrolases. Title: A novel chlorpyrifos hydrolase CPD from Paracoccus sp. TRP: Molecular cloning, characterization and catalytic mechanism Fan S, Li K, Yan Y, Wang J, Qiao C, Yang T, Jia Y, Zhao B Ref: Electronic Journal of Biotechnology, 31:10, 2018 : PubMed ESTHER: Fan_2018_Electron.J.Biotechnol_31_10 PubMedSearch: Fan 2018 Electron.J.Biotechnol 31 10 Gene_locus related to this paper: 9rhob-a0a1x7ll67 Abstract Background: Biodegradation is a reliable approach for efficiently eliminating persistent pollutants such as chlorpyrifos. Despite many bacteria or fungi isolated from contaminated environment and capable of degrading chlorpyrifos, limited enzymes responsible for its degradation have been identified, let alone the catalytic mechanism of the enzymes. Results: In present study, the gene cpd encoding a chlorpyrifos hydrolase was cloned by analysis of genomic sequence of Paracoccus sp. TRP. Phylogenetic analysis and BLAST indicated that CPD was a novel member of organophosphate hydrolases. The purified CPD enzyme, with conserved catalytic triad (Ser155-Asp251-His281) and motif Gly-Asp-Ser-Ala-Gly, was significantly inhibited by PMSF, a serine modifier. Molecular docking between CPD and chlorpyrifos showed that Ser155 was adjacent to chlorpyrifos, which indicated that Ser155 may be the active amino acid involved in chlorpyrifos degradation. This speculation was confirmed by site-directed mutagenesis of Ser155Ala accounting for the decreased activity of CPD towards chlorpyrifos. According to the key role of Ser155 in chlorpyrifos degradation and molecular docking conformation, the nucleophilic catalytic mechanism for chlorpyrifos degradation by CPD was proposed. Conclusion: The novel enzyme CPD was capable of hydrolyze chlorpyrifos and Ser155 played key role during degradation of chlorpyrifos. Title: Efficient Heterologous Production of Rhizopus oryzae Lipase via Optimization of Multiple Expression-Related Helper Proteins Jiao L, Zhou Q, Su Z, Yan Y Ref: Int J Mol Sci, 19:, 2018 : PubMed ESTHER: Jiao_2018_Int.J.Mol.Sci_19_ PubMedSearch: Jiao 2018 Int.J.Mol.Sci 19 PubMedID: 30373304 Abstract This study is dedicated to efficiently produce Rhizopus oryzae lipase (ROL) by optimizing the expression of multiple expression-related helper proteins in Pichia pastoris. A series of engineered strains harboring different copy numbers of the ROL gene and different copies of the chaperone Pdi gene were first constructed to examine the influence of Pdi gene copy number on ROL production. The results showed that multiple copies of Pdi gene did not significantly improve ROL expression. Then, the effect of the co-overexpression of 10 expression-related helper proteins on ROL secretion was investigated by screening 20 colonies of each transformants. The data from shaking-flask fermentation suggested that Ssa4, Bmh2, Sso2, Pdi, Bip, Hac1, and VHb had positive effects on ROL expression. Subsequently, Ssa4, Bmh2, and Sso2, which all participate in vesicular trafficking and strongly promote ROL expression, were combined to further improve ROL production level. ROL activity of the screened strain GS115/5ROL-Ssa4-Sso2-Bmh2 4# attained 5230 U/mL. Furthermore, when the helper proteins Pdi, Bip, Hac1, and VHb were individually co-expressed with ROL in the strain GS115/5ROL-Ssa4-Sso2-Bmh2 4#, lipase activity increased to 5650 U/mL in the strain GS115/5ROL-Ssa4-Sso2-Bmh2-VHb 9#. Additionally, the maximum ROL activity of 41,700 U/mL was achieved in a 3 L bioreactor for high-density fermentation via a sorbitolmethanol co-feeding strategy, reaching almost twofold the value of the initial strain GS115/pAOalpha-5ROL 11#. Thus, the strategies in this study significantly increased ROL expression level, which is of great potential for the large-scale production of ROL in P. pastoris. Title: Enhancing the Thermostability of Rhizomucor miehei Lipase with a Limited Screening Library by Rational-Design Point Mutations and Disulfide Bonds Li G, Fang X, Su F, Chen Y, Xu L, Yan Y Ref: Applied Environmental Microbiology, 84:, 2018 : PubMed ESTHER: Li_2018_Appl.Environ.Microbiol_84_ PubMedSearch: Li 2018 Appl.Environ.Microbiol 84 PubMedID: 29101200 Gene_locus related to this paper: rhimi-lipas Abstract Rhizomucor miehei lipase (RML), as a kind of eukaryotic protein catalyst, plays an important role in the food, organic chemical, and biofuel industries. However, RML retains its catalytic activity below 50 degrees C, which limits its industrial applications at higher temperatures. Soluble expression of this eukaryotic protein in Escherichia coli not only helps to screen for thermostable mutants quickly but also provides the opportunity to develop rapid and effective ways to enhance the thermal stability of eukaryotic proteins. Therefore, in this study, RML was engineered using multiple computational design methods, followed by filtration via conservation analysis and functional region assessment. We successfully obtained a limited screening library (only 36 candidates) to validate thermostable single point mutants, among which 24 of the candidates showed higher thermostability and 13 point mutations resulted in an apparent melting temperature ([Formula: see text]) of at least 1 degrees C higher. Furthermore, both of the two disulfide bonds predicted from four rational-design algorithms were further introduced and found to stabilize RML. The most stable mutant, with T18K/T22I/E230I/S56C-N63C/V189C-D238C mutations, exhibited a 14.3 degrees C-higher [Formula: see text] and a 12.5-fold increase in half-life at 70 degrees C. The catalytic efficiency of the engineered lipase was 39% higher than that of the wild type. The results demonstrate that rationally designed point mutations and disulfide bonds can effectively reduce the number of screened clones to enhance the thermostability of RML.IMPORTANCER. miehei lipase, whose structure is well established, can be widely applied in diverse chemical processes. Soluble expression of R. miehei lipase in E. coli provides an opportunity to explore efficient methods for enhancing eukaryotic protein thermostability. This study highlights a strategy that combines computational algorithms to predict single point mutations and disulfide bonds in RML without losing catalytic activity. Through this strategy, an RML variant with greatly enhanced thermostability was obtained. This study provides a competitive alternative for wild-type RML in practical applications and further a rapid and effective strategy for thermostability engineering. Title: Modification-free carbon dots as turn-on fluorescence probe for detection of organophosphorus pesticides Lin B, Yan Y, Guo M, Cao Y, Yu Y, Zhang T, Huang Y, Wu D Ref: Food Chem, 245:1176, 2018 : PubMed ESTHER: Lin_2018_Food.Chem_245_1176 PubMedSearch: Lin 2018 Food.Chem 245 1176 PubMedID: 29287338 Abstract It is important to detect pesticides residues due to the concern over food safety. In this work, the burning ash of waste paper was used as carbon source to synthesize carbon dots (C-dots). The fluorescence of obtained C-dots could been turn off by Fe(3+) which was from Fe(2+) oxidized by H2O2, organophosphorus pesticides could effectively inhibit the production of H2O2 by destroying the acetylcholinesterase activity, so the fluorescence of C-dots hold turning on in the presence of organophosphorus pesticides. Based on above principle that the fluorescence intensity of C-dots was proportional to the pesticides concentration, take chlorpyrifos for example, a universal method for pesticides detection was established. The linear range was 0.01-1.0mug/mL with detection limit of 3ng/mL. The method was reliable and sensitive to actual samples. The imaging of chlorpyrifos on cabbages leaves indicated this method could be used for visualization detection of organophosphorus pesticides in vegetables. Title: Discovery and identification of O, O-diethyl O-(4-(5-phenyl-4, 5-dihydroisoxazol-3-yl) phenyl) phosphorothioate (XP-1408) as a novel mode of action of organophosphorus insecticides Zeng Z, Yan Y, Wang B, Liu N, Xu H Ref: Sci Rep, 7:3617, 2017 : PubMed ESTHER: Zeng_2017_Sci.Rep_7_3617 PubMedSearch: Zeng 2017 Sci.Rep 7 3617 PubMedID: 28620187 Abstract Organophosphorus (OP) insecticides play an important role in pest control. Many OP insecticides have been removed from the market because of their high toxicity to humans. We designed and synthesized a new OP insecticide with the goal of providing a low cost, and less toxic insecticide. The mode of action of O, O-diethyl O-(4-(5-phenyl-4, 5-dihydroisoxazol-3-yl) phenyl) phosphorothioate (XP-1408) was studied in Drosophila melanogaster. Bioassays showed that XP-1408 at a concentration of 50 mg/L delayed larval development. Molecular docking into Drosophila acetylcholinesterase (AChE) and voltage-gated sodium channels suggested that XP-1408 fitted into their active sites and could be inhibitory. Whole-cell patch clamp recordings indicated that XP-1408 exhibited synergistic effects involving the inhibition of cholinergic synaptic transmission and blockage of voltage-gated potassium (Kv) channels and sodium (Nav) channels. In conclusion, the multiple actions of XP-1408 rendered it as a lead compound for formulating OP insecticides with a novel mode of action. Title: Fabrication of Propeller-Shaped Supra-amphiphile for Construction of Enzyme-Responsive Fluorescent Vesicles Li J, Liu K, Han Y, Tang BZ, Huang J, Yan Y Ref: ACS Appl Mater Interfaces, 8:27987, 2016 : PubMed ESTHER: Li_2016_ACS.Appl.Mater.Interfaces_8_27987 PubMedSearch: Li 2016 ACS.Appl.Mater.Interfaces 8 27987 PubMedID: 27668305 Abstract Propeller-shaped molecules have been recognized to display fantastic AIE (aggregation induced emission), but they can hardly self-assemble into nanostructures. Herein, we for the first time report that ionic complexation between a water-soluble tetrapheneyl derivative and an enzyme substrate in aqueous media produces a propeller-shaped supra-amphiphile that self-assembles into enzyme responsive fluorescent vesicles. The supra-amphiphile was fabricated upon complexation between a water-soluble propeller-shaped AIE luminogen TPE-BPA and myristoylcholine chloride (MChCl) in aqueous media. MChCl filled in the intramolecular voids of propeller-shaped TPE-BPA upon supra-amphiphile formation, which endows the supra-amphiphile superior self-assembling ability to the component molecules thus leading to the formation of fluorescent vesicles. Because MChCl is the substrate of cholinesterases, the vesicles dissemble in the presence of cholinesterases, and the fluorescent intensity can be correlated to the level of enzymes. The resulting fluorescent vesicles may be used to recognize the site of Alzheimer's disease, to encapsulate the enzyme inhibitor, and to release the inhibitor at the disease site. Title: Overexpression of Candida rugosa lipase Lip1 via combined strategies in Pichia pastoris Li X, Liu Z, Wang G, Pan D, Jiao L, Yan Y Ref: Enzyme Microb Technol, 82:115, 2016 : PubMed ESTHER: Li_2016_Enzyme.Microb.Technol_82_115 PubMedSearch: Li 2016 Enzyme.Microb.Technol 82 115 PubMedID: 26672457 Abstract In this study, combined strategies were employed to heterologously overexpress Candida rugosa lipase Lip1 (CRL1) in a Pichia pastoris system. The LIP1 gene was systematically codon-optimized and synthesized in vitro. The Lip1 activity of a recombinant strain harboring three copies of the codon-optimized LIP1 gene reached 1200 U/mL in a shake flask culture. Higher lipase activity, 1450 U/mL, was obtained using a five copy number construct. Co-expressing one copy of the ERO1p and BiP chaperones with Lip1p, the CRL1 lipase yield further reached 1758 U/mL, which was significantly higher than that achieved by expressing Lip1p alone or only co-expressing one molecular chaperone. When cultivated in a 3 L fermenter under optimal conditions, the recombinant strain GS115/87-ZA-ERO1p-BiP #7, expressing the molecular chaperones Ero1p and BiP, produced 13,490 U/mL of lipase activity at 130 h, which was greater than the 11,400 U/mL of activity for the recombinant strain GS115/pAO815-alpha-mCRL1 #87, which did not express a molecular chaperone. This study indicates that a strategy of combining codon optimization with co-expression of molecular chaperones has great potential for the industrial-scale production of pure CRL1. Title: Enhanced performance of lipase via microcapsulation and its application in biodiesel preparation Su F, Li G, Fan Y, Yan Y Ref: Sci Rep, 6:29670, 2016 : PubMed ESTHER: Su_2016_Sci.Rep_6_29670 PubMedSearch: Su 2016 Sci.Rep 6 29670 PubMedID: 27424490 Abstract In the present study, a surface-active enzyme, lipase was immobilized in polyethyleneimine (PEI) microcapsules and then modified with oxidized multiwall carbon nanotubes (MWCNTs). The resulting lipase microcapsules exhibited higher activity and stability, since the activity of microcapsules was 21.9 folds than that of the free counterpart. Numerous interfaces which were created in polycondensation enhanced the performance of lipases. Illustrated by confocal laser scanning microscope (CLSM), it was found that microcapsules, whose barrier properties against molecules with diameter >4.6 nm, were with a semipermeable and porous membrane structure. The lipases preferred to locate in the wall of the microcapsules. The oxidized multiwall carbon nanotubes (MWCNTs) were further added to modify microcapsules, resulting in even higher activity. The nanocomposites were examined by scanning electron microscope (SEM) and zeta-potential analyzer. The results indicated the superior catalytic performances were attributed to the augmented interface and decreased positive charge. Finally, the MWCNTs modified microcapsules were utilized in producing biodiesel with a 97.15% yield and retained nearly 90% yield after running 10 cycles. This approach of microcapsulation will be highly beneficial for preparing various bio-active microcapsules with excellent catalytic performance. Title: Silibinin inhibits acetylcholinesterase activity and amyloid beta peptide aggregation: a dual-target drug for the treatment of Alzheimer's disease Duan S, Guan X, Lin R, Liu X, Yan Y, Zhang T, Chen X, Huang J, Sun X and Gu H <4 more author(s)> Duan S, Guan X, Lin R, Liu X, Yan Y, Zhang T, Chen X, Huang J, Sun X, Li Q, Fang S, Xu J, Yao Z, Gu H (- 4) Ref: Neurobiology of Aging, 36:1792, 2015 : PubMed ESTHER: Duan_2015_Neurobiol.Aging_36_1792 PubMedSearch: Duan 2015 Neurobiol.Aging 36 1792 PubMedID: 25771396 Abstract Alzheimer's disease (AD) is characterized by amyloid beta (Abeta) peptide aggregation and cholinergic neurodegeneration. Therefore, in this paper, we examined silibinin, a flavonoid extracted from Silybum marianum, to determine its potential as a dual inhibitor of acetylcholinesterase (AChE) and Abeta peptide aggregation for AD treatment. To achieve this, we used molecular docking and molecular dynamics simulations to examine the affinity of silibinin with Abeta and AChE in silico. Next, we used circular dichroism and transmission electron microscopy to study the anti-Abeta aggregation capability of silibinin in vitro. Moreover, a Morris Water Maze test, enzyme-linked immunosorbent assay, immunohistochemistry, 5-bromo-2-deoxyuridine double labeling, and a gene gun experiment were performed on silibinin-treated APP/PS1 transgenic mice. In molecular dynamics simulations, silibinin interacted with Abeta and AChE to form different stable complexes. After the administration of silibinin, AChE activity and Abeta aggregations were down-regulated, and the quantity of AChE also decreased. In addition, silibinin-treated APP/PS1 transgenic mice had greater scores in the Morris Water Maze. Moreover, silibinin could increase the number of newly generated microglia, astrocytes, neurons, and neuronal precursor cells. Taken together, these data suggest that silibinin could act as a dual inhibitor of AChE and Abeta peptide aggregation, therefore suggesting a therapeutic strategy for AD treatment. Title: Protein-Coated Microcrystals from Candida rugosa Lipase: Its Immobilization, Characterization, and Application in Resolution of Racemic Ibuprofen Huang S, Li X, Xu L, Ke C, Zhang R, Yan Y Ref: Appl Biochem Biotechnol, 177:36, 2015 : PubMed ESTHER: Huang_2015_Appl.Biochem.Biotechnol_177_36 PubMedSearch: Huang 2015 Appl.Biochem.Biotechnol 177 36 PubMedID: 26137875 Abstract In this study, an economical heterogeneous biocatalyst, protein-coated microcrystals (PCMCs), was prepared from a commercial Candida rugosa lipase (CRL) and used for catalyzing esterification of (R, S)-ibuprofen enantiomers with isooctanol in isooctane. The main variables controlling the process (precipitating solvents, pH, saturated K2SO4 solution, and water content) were optimized via single-factorial experiments. Under optimum conditions, the enantiomeric excess of active S(+)-ibuprofen and total conversion rate were 97.34 and 49.83 %, respectively, and the corresponding enzyme (PCMC-CRL) activity attained 387.29 mumol/min/g protein, a 5.78-fold enhancement over the free lipase powder. Additionally, the thermostability, organic-solvent tolerance, and operational stability of PCMC-CRL were greatly improved as compared to the free enzyme. Fourier transform infrared (FTIR) spectroscopy was employed to reveal the correlation between conformation and enzyme activity enhancement. Moreover, the PCMC-CRL retained most of its original activity following use in more than 15 successive batches, suggesting that it exhibits adequate operational stability. These results indicate that PCMC-CRL is of great potential use in industrial applications. Title: Probing role of key residues in the divergent evolution of Yarrowia lipolytica lipase 2 and Aspergillus niger eruloyl esterase A Wang G, Liu Z, Xu L, Zhang H, Yan Y Ref: Microbiol Res, 178:27, 2015 : PubMed ESTHER: Wang_2015_Microbiol.Res_178_27 PubMedSearch: Wang 2015 Microbiol.Res 178 27 PubMedID: 26302844 Abstract Yarrowia lipolytica lipase 2 (YLLip2) and Aspergillus niger feruloyl esterase A (AnFaeA) are enzymes of similar structures but with different functions. They are both classified into the same homologous family in Lipase Engineering Database (LED). The major difference between the two enzymes is that YLLip2 exhibits interfacial activity while AnFaeA does not. In order to better understand the interfacial activation mechanisms of YLLip2, structure guided site-directed mutagenesis were performed, mutants were constructed, kinetics parameters and lipase properties were detected. Mutant enzymes showed enhanced catalytic efficiency towards p-nitrophenyl butyrin (pNPB) but their catalytic efficiency decreased towards p-nitrophenyl palmitate (pNPP), their catalysis behavior was more close to feruloyl esterase. Moreover, the mutant enzymes exhibited enhanced thermostability compared with their wild type. These results indicate that I100 and F129 are probably cut-off point of divergent functions between the two enzymes during evolution. Title: Characterizing LipR from Pseudomonas sp. R0-14 and Applying in Enrichment of Polyunsaturated Fatty Acids from Algal Oil Yang W, Xu L, Zhang H, Yan Y Ref: J Microbiol Biotechnol, 25:1880, 2015 : PubMed ESTHER: Yang_2015_J.Microbiol.Biotechnol_25_1880 PubMedSearch: Yang 2015 J.Microbiol.Biotechnol 25 1880 PubMedID: 26215266 Abstract In this study, Pseudomonas R0-14, which was isolated from Arctic soil samples, showed a clear halo when grown on M9 medium agarose plates containing olive oil-rhodamine B as substrate, suggesting that it expressed putative lipase(s). A putative lipase gene, lipR, was cloned from R0-14 by genome walking and Touchdown PCR. lipR encodes a 562-amino-acid polypeptide showing a typical alpha/beta hydrolase structure with a catalytic triad consisting of Ser153-Asp202-His260 and one alpha-helical lid (residues 103-113). A phylogenetic analysis revealed that LipR belongs to the lipase subfamily I.3. LipR was successfully expressed in Escherichia coli, purified, and biochemically characterized. Recombinant LipR exhibited its maximum activity towards p-nitrophenyl butyrate at pH 8.5 and 60 degrees C with a Km of 0.37 mM and a kcat of 6.42 s(-1). It retained over 90% of its original activity after incubation at 50 degrees C for 12 h. In addition, LipR was activated by Ca(2+), Mg(2+), Ba(2+), and Sr(2+), while strongly inhibited by Cu(2+), Zn(2+), Mn(2+), and ethylenediaminetetraacetic acid. Moreover, it showed a certain tolerance to organic solvents, including acetonitrile, isopropanol, acetone, methanol, and tert-butanol. When algal oil was hydrolyzed by LipR for 24 h, there was an enrichment of n-3 long-chain polyunsaturated fatty acids, including eicosapentaenoic acid (1.22%, 1.65-fold), docosapentaenoic acid (21.24%, 2.04-fold), and docosahexaenoic acid (36.98%, 1.33-fold), and even a certain amount of diacylglycerols was also produced. As a result, LipR has great prospect in industrial applications, especially in food and/or cosmetics applications. Title: A novel eurythermic and thermostale lipase LipM from Pseudomonas moraviensis M9 and its application in the partial hydrolysis of algal oil Yang W, Cao H, Xu L, Zhang H, Yan Y Ref: BMC Biotechnol, 15:94, 2015 : PubMed ESTHER: Yang_2015_BMC.Biotechnol_15_94 PubMedSearch: Yang 2015 BMC.Biotechnol 15 94 PubMedID: 26463643 Abstract BACKGROUND: Lipases are regularly used in biotechnology to catalyse the hydrolysis of triglycerides and the synthesis of esters. Microbial lipases in particular have been widely used in a variety of industrial applications. However, the current commercial microbial lipases cannot meet industrial demand due to rapid inactivation under harsh conditions. Therefore, in order to identify more stable enzymes, we isolated novel eurythermic and thermostable lipase(s) from Pseudomonas moraviensis M9. METHODS: Cloning of lipM was based on Touchdown PCR and genome walking, and then recombinant LipM was purified by guanidine hydrochloride and the nickel-nitrilotriacetic acid resins affinity chromatography. Finally, the hydrolysis of algal oil by LipM was analyzed by gas chromatograph-mass spectrometer, thin layer chromatography and gas chromatograph. RESULTS: The lipM gene was first cloned from Pseudomonas moraviensis M9 via Touchdown PCR and genome walking. Sequence analysis reveals that LipM is a member of subfamily I.3 of lipases, and the predicted amino acid sequences of LipM has 82 % identity to lipase LipT from Pseudomonas mandelii JR-1, and 54 % identity to lipase PML from Pseudomonas sp. MIS38 and lipase Lip I.3 from Pseudomonas sp. CR-611. LipM was expressed in Escherichia coli, purified from inclusion bodies, and further biochemically characterized. Purified LipM differed significantly from previously reported subfamily I.3 lipases, and was eurythermic between 10 degreesC-95 degreesC. LipM activity was enhanced by Ca(2+), Sr(2+), Mn(2+), and Ba(2+), but sharply inhibited by Cu(2+), Zn(2+), Co(2+), Ni(2+), and EDTA. Compared with other lipases, LipM exhibited medium tolerance to methanol, ethanol, and isopropanol. When applied for hydrolysis of algal oil, LipM could enrich 65.88 % polyunsaturated fatty acids, which include 1.25 % eicosapentaenoic acid, 17.61 % docosapentaenoic acid, and 47.02 % docosahexaenoic acid with derivative glycerides containing 32.46 % diacylglycerols. CONCLUSIONS: A novel eurythermic I.3 subfamily lipase with high tolerance and stability was identified from Pseudomonas moraviensis and biochemically characterized. It will not only improve our understanding of subfamily I.3 lipases, but also provides an ideal biocatalyst for the enrichment of polyunsaturated fatty acids. Pseudomonas moraviensis have been investigated as a potential resource of lipases. Title: Omarigliptin (MK-3102): a novel long-acting DPP-4 inhibitor for once-weekly treatment of type 2 diabetes Biftu T, Sinha-Roy R, Chen P, Qian X, Feng D, Kuethe JT, Scapin G, Gao YD, Yan Y and Weber AE <19 more author(s)> Biftu T, Sinha-Roy R, Chen P, Qian X, Feng D, Kuethe JT, Scapin G, Gao YD, Yan Y, Krueger D, Bak A, Eiermann G, He J, Cox J, Hicks J, Lyons K, He H, Salituro G, Tong S, Patel S, Doss G, Petrov A, Wu J, Xu SS, Sewall C, Zhang X, Zhang B, Thornberry NA, Weber AE (- 19) Ref: Journal of Medicinal Chemistry, 57:3205, 2014 : PubMed ESTHER: Biftu_2014_J.Med.Chem_57_3205 PubMedSearch: Biftu 2014 J.Med.Chem 57 3205 PubMedID: 24660890 Gene_locus related to this paper: human-DPP4 Inhibitor(s) related to this paper: Omarigliptin Abstract In our effort to discover DPP-4 inhibitors with added benefits over currently commercially available DPP-4 inhibitors, MK-3102 (omarigliptin), was identified as a potent and selective dipeptidyl peptidase 4 (DPP-4) inhibitor with an excellent pharmacokinetic profile amenable for once-weekly human dosing and selected as a clinical development candidate. This manuscript summarizes the mechanism of action, scientific rationale, medicinal chemistry, pharmacokinetic properties, and human efficacy data for omarigliptin, which is currently in phase 3 clinical development. Title: Biochemical Characterization of a Carboxylesterase from the Archaeon Pyrobaculum sp. 1860 and a Rational Explanation of Its Substrate Specificity and Thermostability Shao H, Xu L, Yan Y Ref: Int J Mol Sci, 15:16885, 2014 : PubMed ESTHER: Shao_2014_Int.J.Mol.Sci_15_16885 PubMedSearch: Shao 2014 Int.J.Mol.Sci 15 16885 PubMedID: 25250909 Abstract In this work, genome mining was used to identify esterase/lipase genes in the archaeon Pyrobaculum sp. 1860. A gene was cloned and functionally expressed in Escherichia coli as His-tagged protein. The recombinant enzyme (rP186_1588) was verified by western blotting and peptide mass fingerprinting. Biochemical characterization revealed that rP186_1588 exhibited optimum activity at pH 9.0 and 80 degrees C towards p-nitrophenyl acetate (Km: 0.35 mM, kcat: 11.65 s-1). Interestingly, the purified rP186_1588 exhibited high thermostability retaining 70% relative activity after incubation at 90 degrees C for 6 h. Circular dichroism results indicated that rP186_1588 showed slight structure alteration from 60 to 90 degrees C. Structural modeling showed P186_1588 possessed a typical alpha/beta hydrolase's fold with the catalytic triad consisting of Ser97, Asp147 and His172, and was further confirmed by site-directed mutagenesis. Comparative molecular simulations at different temperatures (300, 353, 373 and 473 K) revealed that its thermostability was associated with its conformational rigidity. The binding free energy analysis by MM-PBSA method revealed that the van der Waals interaction played a major role in p-NP ester binding for P186_1588. Our data provide insights into the molecular structures of this archaeal esterase, and may help to its further protein engineering for industrial applications. Title: Aromatic Amino Acid Mutagenesis at the Substrate Binding Pocket of Yarrowia lipolytica Lipase Lip2 Affects Its Activity and Thermostability Wang G, Liu Z, Xu L, Yan Y Ref: ScientificWorldJournal, 2014:382581, 2014 : PubMed ESTHER: Wang_2014_ScientificWorldJournal_2014_382581 PubMedSearch: Wang 2014 ScientificWorldJournal 2014 382581 PubMedID: 25197700 Abstract The lipase2 from Yarrowia lipolytica (YLLip2) is a yeast lipase exhibiting high homologous to filamentous fungal lipase family. Though its crystal structure has been resolved, its structure-function relationship has rarely been reported. By contrast, there are two amino acid residues (V94 and I100) with significant difference in the substrate binding pocket of YLLip2; they were subjected to site-directed mutagenesis (SDM) to introduce aromatic amino acid mutations. Two mutants (V94W and I100F) were created. The enzymatic properties of the mutant lipases were detected and compared with the wild-type. The activities of mutant enzymes dropped to some extent towards p-nitrophenyl palmitate (pNPC16) and their optimum temperature was 35 degrees C, which was 5 degrees C lower than that of the wild-type. However, the thermostability of I100F increased 22.44% after incubation for 1 h at 40 degrees C and its optimum substrate shifted from p-nitrophenyl laurate (pNPC12) to p-nitrophenyl caprate (pNPC10). The above results demonstrated that the two substituted amino acid residuals have close relationship with such enzymatic properties as thermostability and substrate selectivity. Title: N-terminal transmembrane domain of lipase LipA from Pseudomonas protegens Pf-5: A must for its efficient folding into an active conformation Zha D, Zhang H, Xu L, Yan Y Ref: Biochimie, 105:165, 2014 : PubMed ESTHER: Zha_2014_Biochimie_105_165 PubMedSearch: Zha 2014 Biochimie 105 165 PubMedID: 25038570 Gene_locus related to this paper: psef5-q4kj24 Abstract LipA from Pseudomonas protegens Pf-5 has been proven not to be secreted into the extracytoplasmic space, proposing that it is a membrane protein in virtue of its N-terminal transmembrane domain predicted by the TMHMM 2.0. However, LipA was confirmed to be an intracellular protein through determining the effects of lipA deletion or overexpression on the lipase activities in the whole-cell, lysis supernatant and lysis pellet, even through its transmembrane domain being able to make heterologous LacZ locate on the cytoplasmic membrane via construction of beta-galactosidase reporter strains. Subsequently, lipase activity assays showed that the transmembrane domain played an indispensable role for the catalytic function of LipA through construction of the markerless deletion mutant of transmembrane domain sequence of lipA and the expression and purification of LipA and LipADeltaTMD. To further investigate why the transmembrane domain lost its membrane localization function and significantly affected the catalytic function of LipA, the 3D structures of LipA and LipADeltaTMD were constructed. The results indicated that the transmembrane domain, located in the interior of LipA, helped the alpha-helical lid to form an open conformation by the mediation of alpha5 helix. It seems to act as a kind of intramolecular chaperone like the beta-roll motif of subfamily I.3 lipases, which is novel and is the first to notify the intramolecular chaperone of a subfamily I.1 lipase. Title: Assessment of activities and conformation of lipases treated with sub- and supercritical carbon dioxide Chen D, Peng C, Zhang H, Yan Y Ref: Appl Biochem Biotechnol, 169:2189, 2013 : PubMed ESTHER: Chen_2013_Appl.Biochem.Biotechnol_169_2189 PubMedSearch: Chen 2013 Appl.Biochem.Biotechnol 169 2189 PubMedID: 23417391 Abstract In order to illustrate the underlining mechanism of the effect of high pressure on lipases from different resources, the influence of compressed carbon dioxide treatment on the esterification activities and conformation of the three lipases Candida rugosa lipase (CRL), Pseudomonas fluorescens lipase, and Rhizopus oryzae lipase was investigated in the present work. The results showed that the lipases activities were significantly enhanced in most of high-pressure treatments, except the pressure had a negative effect on CRL activity in supercritical condition. Mild depressurization rate could remain the lipase's activity by protecting its rigid structure under supercritical fluid. Conformational analysis by Fourier transform-infrared spectrometry and fluorescence emission spectra revealed that the variances of lipase activity after high-pressure treatment were correlated with the changes of its alpha-helix content and fluorescence intensity. Additionally, transesterification catalyzed by three lipases in supercritical carbon dioxide were conducted, and 87.2 % biodiesel conversion was obtained by CRL after 3 h, resulting in a great reduction of reaction time. Title: Improving activity and enantioselectivity of lipase via immobilization on macroporous resin for resolution of racemic 1- phenylethanol in non-aqueous medium Li X, Huang S, Xu L, Yan Y Ref: BMC Biotechnol, 13:92, 2013 : PubMed ESTHER: Li_2013_BMC.Biotechnol_13_92 PubMedSearch: Li 2013 BMC.Biotechnol 13 92 PubMedID: 24168516 Gene_locus related to this paper: burce-lipaa Abstract BACKGROUND: Burkholderia cepacia lipase (BCL) has been proved to be capable of resolution reactions. However, its free form usually exhibits low stability, bad resistance and no reusability, which restrict its further industrial applications. Therefore, it is of great importance to improve the catalytic performance of free lipase in non-aqueous medium. Title: Conformation and Catalytic Properties Studies of Candida rugosa Lip7 via Enantioselective Esterification of Ibuprofen in Organic Solvents and Ionic Liquids Li X, Huang S, Xu L, Yan Y Ref: ScientificWorldJournal, 2013:364730, 2013 : PubMed ESTHER: Li_2013_ScientificWorldJournal_2013_364730 PubMedSearch: Li 2013 ScientificWorldJournal 2013 364730 PubMedID: 24381516 Abstract Enantioselective esterification of ibuprofen was conducted to evaluate the enzyme activity and ees of lipase from Candida rugosa (CRL7) in ten conventional organic solvents and three ionic liquids. Different alcohols were tested for selecting the most suitable acyl acceptor due to the fact that the structure of alcohols (branch and length of carbon chains; location of -OH functional group) could affect the enzyme activity and ees. The results of alcohol and solvent selection revealed that 1-isooctanol and isooctane were the best substrate and reaction medium, respectively, because of the highest enzyme activity and ees. Compared with the control, conformational studies via FT-IR indicate that the variations of CRL7's secondary structure elements are probably responsible for the differences of enzyme activity and ees in the organic solvents and ionic liquids. Moreover, the effects of reaction parameters, such as molar ratio, water content, temperature, and reaction time, in the selected reaction medium, were also examined. Title: A novel oriented immobilized lipase on magnetic nanoparticles in reverse micelles system and its application in the enrichment of polyunsaturated fatty acids Liu T, Zhao Y, Wang X, Li X, Yan Y Ref: Bioresour Technol, 132C:99, 2013 : PubMed ESTHER: Liu_2013_Bioresour.Technol_132C_99 PubMedSearch: Liu 2013 Bioresour.Technol 132C 99 PubMedID: 23395761 Abstract A novel oriented immobilized lipase was derived from Yarrowia lipolytica lipase LIP2 covalently immobilized on functionalized Fe(3)O(4) magnetic nanoparticles (MNPs) in reverse micelles system (RMS). The activity recovery reached 382% compared with 29% in aqueous phase, and further ran up to 1425% under optimum conditions. (3-Aminopropyl) triethoxysilane (APTES) coated Fe(3)O(4) nanoparticles were characterized by Fourier transform infrared (FT-IR) and X-ray diffraction (XRD). A significant alteration in the secondary structure of the lipase in RMS with a 15.5% increase of alpha-helix content and a 12.5% decrease of beta-sheet content was detected by circular dichroism (CD). The immobilized lipase was employed to enrich polyunsaturated fatty acids in fish oil, a 90% increase of DHA content was obtained after 12h, and after 20 cycles of successive usage, it still remained over 80% of relative hydrolysis degree, which shows a good recyclability. Title: Molecular cloning and characterization of a newly isolated pyrethroid-degrading esterase gene from a genomic library of Ochrobactrum anthropi YZ-1 Ruan Z, Zhai Y, Song J, Shi Y, Li K, Zhao B, Yan Y Ref: PLoS ONE, 8:e77329, 2013 : PubMed ESTHER: Ruan_2013_PLoS.One_8_e77329 PubMedSearch: Ruan 2013 PLoS.One 8 e77329 PubMedID: 24155944 Gene_locus related to this paper: 9rhiz-c4wm13 Abstract A novel pyrethroid-degrading esterase gene pytY was isolated from the genomic library of Ochrobactrum anthropi YZ-1. It possesses an open reading frame (ORF) of 897 bp. Blast search showed that its deduced amino acid sequence shares moderate identities (30% to 46%) with most homologous esterases. Phylogenetic analysis revealed that PytY is a member of the esterase VI family. pytY showed very low sequence similarity compared with reported pyrethroid-degrading genes. PytY was expressed, purified, and characterized. Enzyme assay revealed that PytY is a broad-spectrum degrading enzyme that can degrade various pyrethroids. It is a new pyrethroid-degrading gene and enriches genetic resource. Kinetic constants of Km and Vmax were 2.34 mmol.L(-1) and 56.33 nmol min(-1), respectively, with lambda-cyhalothrin as substrate. PytY displayed good degrading ability and stability over a broad range of temperature and pH. The optimal temperature and pH were of 35 degrees C and 7.5. No cofactors were required for enzyme activity. The results highlighted the potential use of PytY in the elimination of pyrethroid residuals from contaminated environments. Title: Isolation and characterization of a thermostable esterase from a metagenomic library Shao H, Xu L, Yan Y Ref: J Ind Microbiol Biotechnol, 40:1211, 2013 : PubMed ESTHER: Shao_2013_J.Ind.Microbiol.Biotechnol_40_1211 PubMedSearch: Shao 2013 J.Ind.Microbiol.Biotechnol 40 1211 PubMedID: 23934105 Gene_locus related to this paper: 9bact-s5tby8 Abstract A novel esterase gene was isolated by functional screening of a metagenomic library prepared from an activated sludge sample. The gene (est-XG2) consists of 1,506 bp with GC content of 74.8 %, and encodes a protein of 501 amino acids with a molecular mass of 53 kDa. Sequence alignment revealed that Est-XG2 shows a maximum amino acid identity (47 %) with the carboxylesterase from Thermaerobacter marianensis DSM 12885 (YP_004101478). The catalytic triad of Est-XG2 was predicted to be Ser192-Glu313-His412 with Ser192 in a conserved pentapeptide (GXSXG), and further confirmed by site-directed mutagenesis. Phylogenetic analysis suggested Est-XG2 belongs to the bacterial lipase/esterase family VII. The recombinant Est-XG2, expressed and purified from Escherichia coli, preferred to hydrolyze short and medium length p-nitrophenyl esters with the best substrate being p-nitrophenyl acetate (K m and k cat of 0.33 mM and 36.21 s(-1), respectively). The purified enzyme also had the ability to cleave sterically hindered esters of tertiary alcohols. Biochemical characterization of Est-XG2 revealed that it is a thermophilic esterase that exhibits optimum activity at pH 8.5 and 70 degrees C. Est-XG2 had moderate tolerance to organic solvents and surfactants. The unique properties of Est-XG2, high thermostability and stability in the presence of organic solvents, may render it a potential candidate for industrial applications. Title: Extinction bursts in rats trained to self-administer nicotine or food in 1-h daily sessions Pushparaj A, Pryslawsky Y, Forget B, Yan Y, Le Foll B Ref: Am J Transl Res, 4:422, 2012 : PubMed ESTHER: Pushparaj_2012_Am.J.Transl.Res_4_422 PubMedSearch: Pushparaj 2012 Am.J.Transl.Res 4 422 PubMedID: 23145210 Abstract Extinction bursts are characterized by a temporary increase in responding when drug access is withheld from rats trained to self-administer drugs of abuse. Thus far, one study has examined extinction bursts for nicotine self-administration using a 23-h access paradigm [1]. Here we examined extinction bursts using previously published and unpublished data in which rats were trained to self-administer nicotine (0.03mg/kg/infusion) or food pellets (as a comparator) in 1-h sessions under an FR5 schedule of reinforcement followed by 1-h extinction sessions. Analysis of response rates during nicotine self-administration (NSA) was indicative of a loading phase, as response rates were significantly higher at the beginning of the session, which was not observed for food self-administration. At the start of extinction for both food and nicotine, although sessional response rates did not increase, there was an increase in response rate during the first 5-min of the first extinction session relative to self-administration. This transient extinction burst following nicotine was observed in a minority of subjects and correlated with the number of nicotine infusions obtained during self-administration. This transient extinction burst following food was observed in all subjects. Nicotine and food produce more transient extinction bursts compared to other drugs of abuse and only for a minority of animals in the case of nicotine. This study supports the presence of a loading phase in rats trained to self-administer nicotine in 1-r daily sessions and the presence of a transient extinction burst. Title: Constitutive expression of Yarrowia lipolytica lipase LIP2 in Pichia pastoris using GAP as promoter Wang X, Sun Y, Ke F, Zhao H, Liu T, Xu L, Liu Y, Yan Y Ref: Appl Biochem Biotechnol, 166:1355, 2012 : PubMed ESTHER: Wang_2012_Appl.Biochem.Biotechnol_166_1355 PubMedSearch: Wang 2012 Appl.Biochem.Biotechnol 166 1355 PubMedID: 22246727 Abstract A gene encoding Yarrowia lipolytica lipase LIP2 (YlLIP2) was cloned into a constitutive expression vector pGAPZalphaA and electrotransformed into the Pichia pastoris X-33 strain. The high-yield clones obtained by high copy and enzyme activity screening were chosen as the host strains for shaking flask and fermentor culture. The results showed that glucose was the optimum carbon source for YlLIP2 production, and the maximum hydrolytic activity of recombinant YlLIP2 reached 1,315 U/ml under the flask culture at 28 degrees C, pH 7.0, for 48 h. The fed-batch fermentation was carried out in 3- and 10-l bioreactors by continuously feeding glucose into the growing medium for achieving high cell density and YlLIP2 yields. The maximum hydrolytic activity of YlLIP2 and cell density obtained in the 3-l bioreactor were 10,300 U/ml and 116 g dry cell weight (DCW)/l, respectively. The peak hydrolytic activity of YlLIP2 and cell density were further improved in the 10-l fermentor where the values respectively attained were 13,500 U/ml and 120 g DCW/l. The total protein concentration in the supernatant reached 3.3 g/l and the cell viability remained approximately 99% after 80 h of culture. Furthermore, the recombinant YlLIP2 produced in P. pastoris pGAP and pAOX1 systems have similar content of sugar (about 12%) and biochemical characteristics. The above results suggest that the GAP promoter-derived expression system of P. pastoris is effective for the expression of YlLIP2 by high cell density culture and is probably an alternative to the conventional AOX1 promoter expression system in large-scale production of industrial lipases. Title: Nicotine-taking and nicotine-seeking in C57Bl/6J mice without prior operant training or food restriction Yan Y, Pushparaj A, Gamaleddin I, Steiner RC, Picciotto MR, Roder J, Le Foll B Ref: Behavioural Brain Research, 230:34, 2012 : PubMed ESTHER: Yan_2012_Behav.Brain.Res_230_34 PubMedSearch: Yan 2012 Behav.Brain.Res 230 34 PubMedID: 22326373 Abstract The ability to examine genetically engineered mice in a chronic intravenous (IV) nicotine self-administration paradigm will be a powerful tool for investigating the contribution of specific genes to nicotine reinforcement and more importantly, to relapse behavior. Here we describe a reliable model of nicotine-taking and -seeking behavior in male C57BL/6J mice without prior operant training or food restriction. Mice were allowed to self-administer either nicotine (0.03mg/kg/infusion) or saline in 2-h daily sessions under fixed ratio 1 (FR1) followed by FR2 schedules of reinforcement. In the nicotine group, a dose-response curve was measured after the nose-poke behavior stabilized. Subsequently, nose-poke behavior was extinguished and ability of cue presentations, priming injections of nicotine, or intermittent footshock to reinstate responding was assessed in both groups. C57BL/6J mice given access to nicotine exhibited high levels of nose-poke behavior and self-administered a high number of infusions as compared to mice given access to saline. After this acquisition phase, changing the unit-dose of nicotine resulted in a flat dose-response curve for nicotine-taking and subsequently reinstatement of nicotine-seeking behavior was achieved by both nicotine-associated light cue presentation and intermittent footshock. Nicotine priming injections only triggered significant reinstatement on the second consecutive day of priming. In contrast, mice previously trained to self-administer saline did not increase their responding under those conditions. These results demonstrate the ability to produce nicotine-taking and nicotine-seeking behavior in naive C57BL/6J mice without both prior operant training and food restriction. Future work will utilize these models to evaluate nicotine-taking and relapsing behavior in genetically-altered mice. Title: Genome sequence and transcriptome analysis of the radioresistant bacterium Deinococcus gobiensis: insights into the extreme environmental adaptations Yuan M, Chen M, Zhang W, Lu W, Wang J, Yang M, Zhao P, Tang R, Li X and Lin M <8 more author(s)> Yuan M, Chen M, Zhang W, Lu W, Wang J, Yang M, Zhao P, Tang R, Li X, Hao Y, Zhou Z, Zhan Y, Yu H, Teng C, Yan Y, Ping S, Wang Y, Lin M (- 8) Ref: PLoS ONE, 7:e34458, 2012 : PubMed ESTHER: Yuan_2012_PLoS.ONE_7_E34458 PubMedSearch: Yuan 2012 PLoS.ONE 7 E34458 PubMedID: 22470573 Gene_locus related to this paper: deigi-h8gtt4, deigi-h8h0j9 Abstract The desert is an excellent model for studying evolution under extreme environments. We present here the complete genome and ultraviolet (UV) radiation-induced transcriptome of Deinococcus gobiensis I-0, which was isolated from the cold Gobi desert and shows higher tolerance to gamma radiation and UV light than all other known microorganisms. Nearly half of the genes in the genome encode proteins of unknown function, suggesting that the extreme resistance phenotype may be attributed to unknown genes and pathways. D. gobiensis also contains a surprisingly large number of horizontally acquired genes and predicted mobile elements of different classes, which is indicative of adaptation to extreme environments through genomic plasticity. High-resolution RNA-Seq transcriptome analyses indicated that 30 regulatory proteins, including several well-known regulators and uncharacterized protein kinases, and 13 noncoding RNAs were induced immediately after UV irradiation. Particularly interesting is the UV irradiation induction of the phrB and recB genes involved in photoreactivation and recombinational repair, respectively. These proteins likely include key players in the immediate global transcriptional response to UV irradiation. Our results help to explain the exceptional ability of D. gobiensis to withstand environmental extremes of the Gobi desert, and highlight the metabolic features of this organism that have biotechnological potential. Title: Molecular cloning, purification and biochemical characterization of a novel pyrethroid-hydrolyzing carboxylesterase gene from Ochrobactrum anthropi YZ-1 Zhai Y, Li K, Song J, Shi Y, Yan Y Ref: J Hazard Mater, 221-222:206, 2012 : PubMed ESTHER: Zhai_2012_J.Hazard.Mater_221-222_206 PubMedSearch: Zhai 2012 J.Hazard.Mater 221-222 206 PubMedID: 22579404 Gene_locus related to this paper: 9hyph-h2esq9 Abstract Strain YZ-1 was isolated from activated sludge and identified as Ochrobactrum anthropi. This strain was capable of degrading pyrethroids pesticides, suggesting the presence of degrading enzymes. In the present study, a novel esterase gene pytZ was cloned from the genomic library of YZ-1 successfully. The pytZ contained an open reading frame of 606bp encoding a pyrethroid-hydrolyzing carboxylesterase. Deduced amino acid sequence showed moderate identities (39-59%) with most homologous carboxylesterase, except a putative carboxylesterase from O. anthropi ATCC 49188 with the highest identity of 85%. Phylogenetic analysis revealed that PytZ belonged to esterase VI family. The gene pytZ showed no any sequence similarity with reported pyrethroid-hydrolyzing genes and was a new pyrethroid-degrading gene. PytZ was expressed in Escherichia coli BL21 (DE3) and purified using Ni-NTA Fast Start. PytZ was able to degrade various pyrethroids. The optimal temperature and pH were 35 degrees C and 7.5. This enzyme was very stable over a wide range of temperature and pH. No cofactors were required for enzyme activity. Broad substrate specificity, high enzyme activity, and the favorable stability make the PytZ a potential candidate for the detoxification of pyrethroid residues in biotechnological application. Title: Lipase-coated K2SO4 micro-crystals: preparation, characterization, and application in biodiesel production using various oil feedstocks Zheng J, Xu L, Liu Y, Zhang X, Yan Y Ref: Bioresour Technol, 110:224, 2012 : PubMed ESTHER: Zheng_2012_Bioresour.Technol_110_224 PubMedSearch: Zheng 2012 Bioresour.Technol 110 224 PubMedID: 22330591 Abstract This study investigated the preparation and characteristics of protein-coated microcrystals (PCMCs) from Pseudomonas cepacia lipase (PS) and K(2)SO(4), and their application in biodiesel synthesis, via single factorial experiments and response surface methodology (RSM), the optimized PCMC-PS exhibited high activity and stability; the optimal temperature was 60 degrees C (which gave 99.83% conversion), although fairly high activity was exhibited after incubation at different temperatures (25-70 degrees C). The organic solvents stability of the PCMC-PS was improved, and it significantly reduced ethanol toxicity. Circular dichroism (CD) analysis revealed the correlation between the conformation and the enzyme activity. The morphology of the PCMC-PS was also confirmed via scanning electron microscopy (SEM). When catalyzed by PCMC-PS, above 83% biodiesel yield was obtained for most of the seven oils tested. The PCMC-PS (washed with hexane) activity remained relatively stable after eight batch reactions, with only a 15.73% reduction in the conversion (from 99.02% to 83.29%). Title: Biodiesel synthesis and conformation of lipase from Burkholderia cepacia in room temperature ionic liquids and organic solvents Liu Y, Chen D, Yan Y, Peng C, Xu L Ref: Bioresour Technol, 102:10414, 2011 : PubMed ESTHER: Liu_2011_Bioresour.Technol_102_10414 PubMedSearch: Liu 2011 Bioresour.Technol 102 10414 PubMedID: 21955878 Abstract Biodiesel synthesis and conformation of Burkholderia cepacia lipase (BCL) were studied in 19 different room temperature ionic liquids (RTLLs) with a range of cation and anion structures. Overall, anion selection had a greater influence on biodiesel conversion than cation choice. RTILs containing Tf2N- and PF6- anions were suitable reaction media, while RTIL of [OmPy][BF4] was the best reaction medium with a biodiesel yield of 82.2+/-1.2%. RTILs with strong water miscible properties showed very low biodiesel yields. Conformational analysis by FT-IR revealed that higher biodiesel conversion in RTILs was correlated with a low tendency in alpha-helix content of BCL. An ultrasound-assisted biocatalysis process in RTILs was used to improve mass transfer rate, leading to 83% reduction of the reaction time for biodiesel production. Title: Complete genome sequence of the nitrogen-fixing and rhizosphere-associated bacterium Pseudomonas stutzeri strain DSM4166 Yu H, Yuan M, Lu W, Yang J, Dai S, Li Q, Yang Z, Dong J, Sun L and Lin M <8 more author(s)> Yu H, Yuan M, Lu W, Yang J, Dai S, Li Q, Yang Z, Dong J, Sun L, Deng Z, Zhang W, Chen M, Ping S, Han Y, Zhan Y, Yan Y, Jin Q, Lin M (- 8) Ref: Journal of Bacteriology, 193:3422, 2011 : PubMed ESTHER: Yu_2011_J.Bacteriol_193_3422 PubMedSearch: Yu 2011 J.Bacteriol 193 3422 PubMedID: 21515765 Gene_locus related to this paper: pseu5-a4vfn0, pseu5-a4vj02, pseu5-a4vrl9, pseut-f8h720, pseu5-a4vkl7, pseu5-a4vgd4, pseu5-a4vr33 Abstract We present here the analysis of the whole-genome sequence of Pseudomonas stutzeri strain DSM4166, a diazotrophic isolate from the rhizosphere of a Sorghum nutans cultivar. To our knowledge, this is the second genome to be sequenced for P. stutzeri. The availability and analysis of the genome provide insight into the evolution of the nitrogen fixation property and identification of rhizosphere competence traits required in interactions with host plants. Title: Genome sequence of Acinetobacter calcoaceticus PHEA-2, isolated from industry wastewater Zhan Y, Yan Y, Zhang W, Yu H, Chen M, Lu W, Ping S, Peng Z, Yuan M and Lin M <2 more author(s)> Zhan Y, Yan Y, Zhang W, Yu H, Chen M, Lu W, Ping S, Peng Z, Yuan M, Zhou Z, Elmerich C, Lin M (- 2) Ref: Journal of Bacteriology, 193:2672, 2011 : PubMed ESTHER: Zhan_2011_J.Bacteriol_193_2672 PubMedSearch: Zhan 2011 J.Bacteriol 193 2672 PubMedID: 21441526 Gene_locus related to this paper: aciad-q6fc40, aciba-d0c992, aciba-q76hj1, acibt-a3m5r6, acibt-a3m5t3, acibt-a3m5x2, acica-d0ryi9, acica-d0ryx6, acicp-f0kgu1, acicp-f0kh68, acicp-f0khy0, acicp-f0kj61, aciba-w3svn7, aciba-a0a009wzt4 Abstract Genome analysis of Acinetobacter calcoaceticus PHEA-2 was undertaken because of the importance of this bacterium for bioremediation of phenol-polluted water and because of the close phylogenetic relationship of this species with the human pathogen Acinetobacter baumannii. To our knowledge, this is the first strain of A. calcoaceticus whose genome has been sequenced. Title: [Heterologous expression and characterization of Yarrowia lipolytica lipase 4 and lipase 5 in Pichia pastoris] Zhao H, Xiao X, Xu L, Liu Y, Yan Y Ref: Wei Sheng Wu Xue Bao, 51:1374, 2011 : PubMed ESTHER: Zhao_2011_Wei.Sheng.Wu.Xue.Bao_51_1374 PubMedSearch: Zhao 2011 Wei.Sheng.Wu.Xue.Bao 51 1374 PubMedID: 22233059 Abstract OBJECTIVE: To clone cDNA sequences of lipase 4 (LIP4) and lipase 5 (LIPS), analyze gene structures and express them in Pichia pastoris so as to investigate their enzymatic characteristics. METHODS: We first cloned cDNA sequences of LIP4 and LIP5 by reverse transcription PCR and analyzed their gene structures by SignalP 3.0. Then, intracellular expression vectors pPIC3. 5K-Lip4, pPIC3. 5K-Lip5 and inducible secretion vectors pPIC9K-Lip4, pPIC9K-Lip5 were constructed. All vectors were transformed into Pichia pastoris GS115 by electroporation, resulting in a series of engineered strains. After fermentation and NTA-Ni resin purification, the enzymatic properties of LIP4 and LIP5 were examined. RESULTS: The cloned cDNA sequences revealed that there was no intron in both of Lip4 and Lip5. The two lipases both contained catalytic triads and conserved GHSLG motifs. Their optimal substrate, pH, temperature were respectively pNP-caprylate (C8), 7.0 and 40 degrees C. The activities of LIP4 and LIPS were 10.16 U/mg and 5.1 U/mg, respectively. It was found that LIP4 was more sensitive to the variations of pH and temperature than LIP5. LIP4 and LIP5 could both be stimulated by Ca2+, besides LIPS could also be activated by Mg2+. They were both strongly inhibited by Hg2+, Phenylmethanesulfonyl fluoride (PMSF) and Dithiothreitol (DTT). CONCLUSION: The cloning of Lip4 and Lip5, expression in P. pastoris and characterization of their properties would offer a solid basis for their large-scale production and future application. In addition, the results also enriched the data for a systematic research on the lipase gene family of Y. lipolytica. Title: Optimization of lipase-catalyzed transesterification of lard for biodiesel production using response surface methodology Huang Y, Zheng H, Yan Y Ref: Appl Biochem Biotechnol, 160:504, 2010 : PubMed ESTHER: Huang_2010_Appl.Biochem.Biotechnol_160_504 PubMedSearch: Huang 2010 Appl.Biochem.Biotechnol 160 504 PubMedID: 18931953 Abstract Biodiesel, an alternative diesel fuel made from renewable biological resources, has become more and more attractive recently. Combined use of two immobilized lipases with complementary position specificity instead of one lipase is a potential way to significantly reduce cost of lipase-catalyzed biodiesel production. In this study, the process of biodiesel production from lard catalyzed by the combined use of Novozym435 (non-specific) and Lipozyme TLIM (1,3-specific) was optimized by response surface methodology. The optimal reaction conditions were 0.04 of amount of lipase/oil (w/w), 0.49 of proportion of Novozym435/total lipases (w/w), 0.55 of quantity of tert-butanol/oil (v/v), 5.12 of quantity of methanol/oil (mol/mol), and 20 h of reaction time, by which 97.2% of methyl ester (ME) yield was attained, very close to the predicted value (97.6%). This optimal reaction condition could be true of other similar reactions with plant and animal oil resources; their ME yield could be higher than 95%. The lipases regenerated by washing with organic solvent after each reaction cycle could be continuously reused for 20 cycles without any loss of activity, exhibiting very high manipulation stability. Title: Surface display of active lipase in Saccharomyces cerevisiae using Cwp2 as an anchor protein Liu W, Zhao H, Jia B, Xu L, Yan Y Ref: Biotechnol Lett, 32:255, 2010 : PubMed ESTHER: Liu_2010_Biotechnol.Lett_32_255 PubMedSearch: Liu 2010 Biotechnol.Lett 32 255 PubMedID: 19821073 Abstract Lipase Lip2 from Yarrowia lipolytica was displayed on the cell surface of Saccharomyces cerevisiae using Cwp2 as an anchor protein. Successful display of the lipase on the cell surface was confirmed by immunofluorescence microscopy and halo assay. The length of linker sequences was further examined to confirm that the correct conformation of Lip2 was maintained. The results showed that the displayed Lip2 exhibited the highest activity at 7.6 +/- 0.4 U/g (dry cell) when using (G(4)S)(3) sequence as the linker, with an optimal temperature and pH at 40 degrees C and pH 8.0. The displayed lipase did not lose any activity after being treated with 0.1% Triton X-100 and 0.1% Tween 80 for 30 min, and it retained 92% of its original activity after incubation in 10% DMSO for 30 min. It also exhibited better thermostability than free Lip2 as reported previously. Title: Esterification activity and conformation studies of Burkholderia cepacia lipase in conventional organic solvents, ionic liquids and their co-solvent mixture media Pan S, Liu X, Xie Y, Yi Y, Li C, Yan Y, Liu Y Ref: Bioresour Technol, 101:9822, 2010 : PubMed ESTHER: Pan_2010_Bioresour.Technol_101_9822 PubMedSearch: Pan 2010 Bioresour.Technol 101 9822 PubMedID: 20713309 Abstract In this work, experiments were carried out to evaluate the esterification activity and conformation of lipase from Burkholderia cepacia in the selected conventional organic solvents, ionic liquids and their co-solvent mixture media. The results revealed that the activity of esterification of B. cepacia lipase was mostly highest in co-solvent mixture of ionic liquids-organic solvents, followed by conventional organic solvents and ionic liquids. Hence, co-solvent mixture was a high-effective strategy to enhance the activity of B. cepacia lipase for non-aqueous enzymology reaction. Conformational studies via circular dichroism spectroscopy indicated that the secondary structure of B. cepacia lipase was variant in the above-mentioned media, especially the content of alpha-helix, which was probably responsible for lipase activity difference. Title: Cloning of a novel lipase gene, lipJ08, from Candida rugosa and expression in Pichia pastoris by codon optimization Xu L, Jiang X, Yang J, Liu Y, Yan Y Ref: Biotechnol Lett, 32:269, 2010 : PubMed ESTHER: Xu_2010_Biotechnol.Lett_32_269 PubMedSearch: Xu 2010 Biotechnol.Lett 32 269 PubMedID: 19841868 Abstract A novel lipase gene, lipJ08, was cloned from Candida rugosa ATCC14830, along with the already reported five lipase genes (lip1-lip5). Nucleotide sequencing indicated that the lipJ08 gene contains a 1650 bp open reading frame (ORF) without introns. The deduced amino acid sequence corresponds to 534 amino acid residues, including a putative signal sequence of 15 amino acid residues. Seventeen of the non-universal serine codons (CTG) of lipJ08 were converted into universal serine codons (TCT) by PCR-based mutagenesis. The native and codon-optimized lipJ08 genes were expressed in Pichia pastoris. The hydrolytic activity of the recombinant LIPJ08 was 4.7 U/ml, whereas the activity of the recombinant wild-type lipase could not be detected. Title: lip2, a novel lipase gene cloned from Aspergillus niger exhibits enzymatic characteristics distinct from its previously identified family member Yang J, Sun J, Yan Y Ref: Biotechnol Lett, 32:951, 2010 : PubMed ESTHER: Yang_2010_Biotechnol.Lett_32_951 PubMedSearch: Yang 2010 Biotechnol.Lett 32 951 PubMedID: 20213520 Abstract We have cloned a novel lipase gene, lip2, from Aspergillus niger and expressed it in Escherichia coli. Upon purification of the recombinant Lip2 protein, its properties were characterized. In comparison with a previously identified lipase Lip1, both enzymes are acid lipases (optimal pH <6.5), Ca(2+)-dependent and PMSF-sensitive, but have different molecular weights (35 and 43 kDa), optimal substrate spectra (C10 and C8), optimal reaction temperatures (45 and 50 degrees C) and thermal stability. Circular dichroism spectroscopy revealed that Lip2 contains a typical Ca(2+)-active site. This first report on the cloning of the Lip2 gene and its enzymatic characteristics may greatly facilitate its potential industrial application. Title: [Homologous expression of Burkholderia cepacia G63 lipase gene based on T7 RNA polymerase expression system] Jia B, Yang J, Yan Y Ref: Sheng Wu Gong Cheng Xue Bao, 25:215, 2009 : PubMed ESTHER: Jia_2009_Sheng.Wu.Gong.Cheng.Xue.Bao_25_215 PubMedSearch: Jia 2009 Sheng.Wu.Gong.Cheng.Xue.Bao 25 215 PubMedID: 19459326 Abstract In order to realize over-expression of Burkholderia cepacia (B. cepacia) lipase, we introduced the widely used T7 RAN polymerase expression system into B. cepacia G63 to over-express the lipase gene. By using PCR technique, we amplified the T7 RNA polymerase gene (T7 RNAP) from the BL21 (DE3) and cloned it into the suicide plasmid pJQ200SK. After that, we flanked T7 RNAP with two 500 bp homologous fragments and integrated it into the genomes of B. cepacia by tri-parental mating, so that T7 RNAP was under-controlled by lipase gene (lipA) promoter. Then, we cloned the lipA and its partner gene lipB into the vector pUCPCM and pBBR22b both or separately. Therefore, we got 7 expression plasmids pBBR22blipAB, pBBR22blipA, pUCPCMlipAB, pUCPCMlipA, pUCPCMdeltalipAlipB, pUCPCMdeltalipA, pUCPCMdeltalipB, and then electroporated them into B. cepacia containing T7 RNA. After shake flask culture, we found B. cepacia containing pUCPCMlipAB produced the most quantity of lipase, and lipase activity was up to 607.2 U/mg, 2.8-folds higher than that of the wild strain. Moreover, lipase activities of all engineering strains except the one containing pUCPCMdeltalipB were enhanced to some extent. The specific activities of wild type B. cepacia and B. cepacia containing pUCPCMlipAB were respectively 29 984 U/mg and 30 875 U/mg after ammonium sulfate precipitation and gel filtration chromatography. The T7 RNA polymerase expression system could effectively enhanced lipase expression in B. cepacia, and secretion signal PelB and ribosome-binding site may promote lipase expression in engineering strain. Title: Aspergillus niger lipase: Heterologous expression in Pichia pastoris, molecular modeling prediction and the importance of the hinge domains at both sides of the lid domain to interfacial activation Shu Z, Duan M, Yang J, Xu L, Yan Y Ref: Biotechnol Prog, 25:409, 2009 : PubMed ESTHER: Shu_2009_Biotechnol.Prog_25_409 PubMedSearch: Shu 2009 Biotechnol.Prog 25 409 PubMedID: 19248178 Abstract Aspergillus niger lipase (ANL) is an important biocatalyst in the food processing industry. However, there is no report of its detailed three-dimensional structure because of difficulties in crystallization. In this article, based on experimental data and bioinformational analysis results, the structural features of ANL were simulated. Firstly, two recombinant ANLs expressed in Pichia pastoris were purified to homogeneity and their corresponding secondary structure compositions were determined by circular dichroism spectra. Secondly, the primary structure, the secondary structure and the three-dimensional structure of ANL were modeled by comparison with homologous lipases with known three-dimensional structures using the BioEdit software, lipase engineering database (http://www.led.uni-stuttgart.de/), PSIPRED server and SwissModel server. The predicted molecular structure of ANL presented typical features of the alpha/beta hydrolase fold including positioning of the putative catalytic triad residues and the GXSXG signature motif. Comparison of the predicted three-dimensional structure of ANL with the X-ray three-dimensional structure of A. niger feruloyl esterase showed that the functional difference of interfacial activation between lipase and esterase was concerned with the difference in position of the lid. Our three-dimensional model of ANL helps to modify lipase structure by protein engineering, which will further expand the scope of application of ANL. Title: Cloning and expression of Pseudomonas fluorescens 26-2 lipase gene in Pichia pastoris and characterizing for transesterification Yang J, Zhang B, Yan Y Ref: Appl Biochem Biotechnol, 159:355, 2009 : PubMed ESTHER: Yang_2009_Appl.Biochem.Biotechnol_159_355 PubMedSearch: Yang 2009 Appl.Biochem.Biotechnol 159 355 PubMedID: 19005622 Abstract Pseudomonas lipases are important biocatalysts widely used in a variety of industrial fields. An extracellular lipase gene lipA with 1,854-bp open reading frame was cloned from Pseudomonas fluorescens 26-2. The multialignment assay of the putative amino acid and the secondary structure prediction revealed this enzyme could be classified into the lipolytic subfamily I.3 and secreted via adenosine-triphosphate-binding cassette pathway. The lipA gene was integrated into Pichia pastoris GS115, and the methanol-inducible recombinants with Mut(S) and Mut(+) phenotypes were acquired. The characteristics and the transesterification capacity shown by this enzyme suggested it is a useful biocatalyst for biodiesel preparation. Title: [Cloning, expression and characterization of a novel lipase gene lipB from Aspergillus niger F044] Yang J, Zhang Z, Liu L, Yan Y Ref: Wei Sheng Wu Xue Bao, 49:1095, 2009 : PubMed ESTHER: Yang_2009_Wei.Sheng.Wu.Xue.Bao_49_1095 PubMedSearch: Yang 2009 Wei.Sheng.Wu.Xue.Bao 49 1095 PubMedID: 19835173 Abstract OBJECTIVE: We cloned, expressed and characterized a novel lipase gene lipB from Aspergillus niger F044, to facilitate the large scale production and application of that enzyme. METHOD: We cloned lipB gene and the cDNA sequence by PCR and RT-PCR, and then cloned the open reading frame of lipB into pET28a vector and expressed by isopropyl beta-D-1-thiogalactopyranoside (IPTG) induction. After Ni-agarose purification, the characteristics were determined and the conformation change was checked by circular dichroism methods. RESULTS: The novel lipase genes cDNA of lipB were cloned from Aspergillus niger F044 (GenBank: FJ536287, FJ536288) and expressed in Escherichia coli. The molecular weight of LipB was about 43 kDa. The optimal substrate of this enzyme is 4-nitrophenyl octanoate (pNPC-C8) with Km = 5.98 mmol/L. The optimal temperature and pH was 50 degrees C and pH 6.0. The enzyme was stable below 40 degrees C. After incubated at 60 degrees C for 1 h, only 18.8% activity remained. After treated by 2 mmol/L Ca2+ for 1 h, the activity improved 2.6-fold. CONCLUSION: Enzymatic characteristics of LipB determined showed this enzyme might have potential in industrial applications. Title: [Directed evolution of lipase of Bacillus pumilus YZ02 by error-prone PCR] Huang Y, Cai Y, Yang J, Yan Y Ref: Sheng Wu Gong Cheng Xue Bao, 24:445, 2008 : PubMed ESTHER: Huang_2008_Sheng.Wu.Gong.Cheng.Xue.Bao_24_445 PubMedSearch: Huang 2008 Sheng.Wu.Gong.Cheng.Xue.Bao 24 445 PubMedID: 18589821 Abstract Random mutagenesis on Bacillus pumilus lipase YZ02 gene was conducted by using error-prone PCR strategy. Through two cycles of directed evolution, two optimum mutants BpL1-7 and BpL2-1369 with lipase activity improved 2 folds and 6 folds respectively were screened. The sequence of BpL2-1369 lipase gene showed that four nucleotides substitution, T61C, C147T, A334G and T371A have occurred, and three of them caused amino acid changes. Thus, amine acid Ser21 was changed into Pro21, Arg112 to Gly112, and Leu124 to His124. According to the 3D structure of Bacillus pumilus lipase mimicked by SWISS-MODEL Repository, three mutated amino acids were located at the third amino acid of the first alpha-helix, the turn between the fourth and fifth beta fold, and the first amino acid of the fifth beta fold, respectively. The BpL and BpL2-1369 genes were ligated into pET28a vector, and transferred into E. coli BL21 (DE3). After induced by IPTG the lipases were purified and characterized. The results showed that the specific activity of the evolved lipase was 1.31-fold than that of the wild lipase, and the Km decreased from 8.24 mmol/L to 7.17 mmol/L. The pH stability of the evolved lipase was better than wild lipase when pH>8.0. Title: Lipase-catalyzed biodiesel production with methyl acetate as acyl acceptor Huang Y, Yan Y Ref: Z Naturforsch C, 63:297, 2008 : PubMed ESTHER: Huang_2008_Z.Naturforsch.C_63_297 PubMedSearch: Huang 2008 Z.Naturforsch.C 63 297 PubMedID: 18533477 Abstract Biodiesel is an alternative diesel fuel made from renewable biological resources. During the process of biodiesel production, lipase-catalyzed transesterification is a crucial step. However, current techniques using methanol as acyl acceptor have lower enzymatic activity; this limits the application of such techniques in large-scale biodiesel production. Furthermore, the lipid feedstock of currently available techniques is limited. In this paper, the technique of lipase-catalyzed transesterification of five different oils for biodiesel production with methyl acetate as acyl acceptor was investigated, and the transesterification reaction conditions were optimized. The operation stability of lipase under the obtained optimal conditions was further examined. The results showed that under optimal transesterification conditions, both plant oils and animal fats led to high yields of methyl ester: cotton-seed oil, 98%; rapeseed oil, 95%; soybean oil, 91%; tea-seed oil, 92%; and lard, 95%. Crude and refined cottonseed oil or lard made no significant difference in yields of methyl ester. No loss of enzymatic activity was detected for lipase after being repeatedly used for 40 cycles (ca. 800 h), which indicates that the operational stability of lipase was fairly good under these conditions. Our results suggest that cotton-seed oil, rape-seed oil and lard might substitute soybean oil as suitable lipid feedstock for biodiesel production. Our results also show that our technique is fit for various lipid feedstocks both from plants and animals, and presents a very promising way for the large-scale biodiesel production. Title: [Cell surface display of Yarrowia lipolytica lipase Lip2 in Saccharomyces cerevisiae with a-agglutinin as carrier protein] Liu W, Xu L, Zhao H, Yang J, Yan Y Ref: Wei Sheng Wu Xue Bao, 48:1543, 2008 : PubMed ESTHER: Liu_2008_Wei.Sheng.Wu.Xue.Bao_48_1543 PubMedSearch: Liu 2008 Wei.Sheng.Wu.Xue.Bao 48 1543 PubMedID: 19149173 Abstract OBJECTIVE: In order to display extracellular.lipase Lip2 from Yarrowia lipolytica on the surface of yeast Saccharomyces cerevisiae for whole cell catalysts. METHODS: The mature Lip2 encoding fragment was amplified from Yarrowia lipolytica total DNA, and was inserted into the 3'terminal of AGA2 to give the plasmid pCTLIP2 for surface display of Lip2. Olive oil, tributyrin and p-nitrophenyl palmitate (pNPP) were used as substrates to measure lipase activity. Moreover, the characterization of displayed lipase and its free form was analyzed. RESULTS: The surface displayed lipase was confirmed to be active towards olive oil, tributyrin and p-nitrophenyl palmitate (pNPP), and reached its highest expression level at 182 U/g dry cell after induced by galactose for 72h. The optimum temperature of cell surface displayed Lip2 was 40 degrees C After incubated at 50 degrees C for 4h, the surface displayed lipase retained 23.2% of its full activity, improved a little compared to free Lip2. The surface displayed lipase showed a preference to medium-chain and long-chain fatty acids p-nitrophenyl esters (C8-C16). CONCLUSION: The cell surface display system based on a-agglutinin is an effective system for displaying Lip2, and the whole cell EBY100-pCTLIP2 will be probably suited to a different range of applications. Title: Characterization of ST-4821 complex, a unique Neisseria meningitidis clone Peng J, Yang L, Yang F, Yang J, Yan Y, Nie H, Zhang X, Xiong Z, Jiang Y and Jin Q <9 more author(s)> Peng J, Yang L, Yang F, Yang J, Yan Y, Nie H, Zhang X, Xiong Z, Jiang Y, Cheng F, Xu X, Chen S, Sun L, Li W, Shen Y, Shao Z, Liang X, Xu J, Jin Q (- 9) Ref: Genomics, 91:78, 2008 : PubMed ESTHER: Peng_2008_Genomics_91_78 PubMedSearch: Peng 2008 Genomics 91 78 PubMedID: 18031983 Gene_locus related to this paper: neigo-pip, neima-metx, neimb-q9k0t9, neime-ESD, neime-NMA2216, neime-NMB0276, neime-NMB1877 Abstract Ten outbreaks of a new serogroup C meningococcal disease emerged during 2003-2005 in China. The multilocus sequence typing results indicated that unique sequence type 4821 clone meningococci were responsible for these outbreaks. Herein, we determined the entire genomic DNA sequence of serogroup C isolate 053442, which belongs to ST-4821. Comparison of 053442 gene contents with other meningococcal genomes shows that they have similar characteristics, including thousands of repetitive elements and simple sequence repeats, numerous phase-variable genes, and similar virulence-related factors. However, many strain-specific regions were found in each genome. We also present the results of a genomic comparison of 28 ST-4821 complex isolates that were isolated from different serogroups using comparative genomic hybridization analysis. Genome comparison between the newly emerged hyperinvasive isolates belonging to different serogroups will further our understanding of their respective pathogenetic mechanisms. Title: [Cloning and overexpression of lipase gene from Geotrichum candidum Y162] Yan J, Yang J, Xu L, Yan Y Ref: Wei Sheng Wu Xue Bao, 48:184, 2008 : PubMed ESTHER: Yan_2008_Wei.Sheng.Wu.Xue.Bao_48_184 PubMedSearch: Yan 2008 Wei.Sheng.Wu.Xue.Bao 48 184 PubMedID: 18437999 Abstract By means of bioinformatics, we aligned nucleotide sequence of reported lipase gene from Geotrichum. Primers were designed based on the conservative nucleotide sequence, and the lipase gene of G. candidum Y162 was cloned for the first time in China. Nucleotide sequencing revealed that the open reading frame has 1692 nucleotides without any introns, encoding 563 amino acid residues including a signal sequence of 19 amino acid residues, which is 86% identical to lipase I of G. fermentans. Subsequently, we cloned the lipase gene into expression vector pPIC9K, and then transformed into Pichia pastoris GS115. Cultures of recombined P. pastoris accumulated active enzyme in the supernatant to levels of 55 U/mL after induction for 96 hours in shake flasks. The purified lipase exhibited maximum activity at 50 degrees C and pH 8.0, and was stable between pH 6.0 and 10.0 and below 60 degrees C. Lipase activity was compatible with the presence of organic solvents such as methanol, n-heptane, hexane, cyclohexane, glycerol, benzene and diethyl ether. Lipase showed hydrolysis preference for triacylglycerol substrates containing cis-9 unsaturated fatty acid. The results suggest that the lipase could be a candidate for industrial applications. Title: Genome dynamics and diversity of Shigella species, the etiologic agents of bacillary dysentery Yang F, Yang J, Zhang X, Chen L, Jiang Y, Yan Y, Tang X, Wang J, Xiong Z and Jin Q <17 more author(s)> Yang F, Yang J, Zhang X, Chen L, Jiang Y, Yan Y, Tang X, Wang J, Xiong Z, Dong J, Xue Y, Zhu Y, Xu X, Sun L, Chen S, Nie H, Peng J, Xu J, Wang Y, Yuan Z, Wen Y, Yao Z, Shen Y, Qiang B, Hou Y, Yu J, Jin Q (- 17) Ref: Nucleic Acids Research, 33:6445, 2005 : PubMed ESTHER: Yang_2005_Nucleic.Acids.Res_33_6445 PubMedSearch: Yang 2005 Nucleic.Acids.Res 33 6445 PubMedID: 16275786 Gene_locus related to this paper: ecoli-yeiG, shidy-IROD, shidy-q67dv1, shifl-AES, shifl-BIOH, shifl-entf, shifl-FES, shifl-PLDB, shifl-PTRB, shifl-S2753, shifl-SF1334, shifl-SF1808, shifl-SF3046, shifl-SF3908, shifl-yafa, shifl-YBFF, shifl-YCDJ, shifl-ycfp, shifl-YCJY, shifl-YFBB, shifl-YHET, shifl-YJFP, shifl-YPFH, shiss-yaim, shiss-yeiG, shiss-yqia Abstract The Shigella bacteria cause bacillary dysentery, which remains a significant threat to public health. The genus status and species classification appear no longer valid, as compelling evidence indicates that Shigella, as well as enteroinvasive Escherichia coli, are derived from multiple origins of E.coli and form a single pathovar. Nevertheless, Shigella dysenteriae serotype 1 causes deadly epidemics but Shigella boydii is restricted to the Indian subcontinent, while Shigella flexneri and Shigella sonnei are prevalent in developing and developed countries respectively. To begin to explain these distinctive epidemiological and pathological features at the genome level, we have carried out comparative genomics on four representative strains. Each of the Shigella genomes includes a virulence plasmid that encodes conserved primary virulence determinants. The Shigella chromosomes share most of their genes with that of E.coli K12 strain MG1655, but each has over 200 pseudogenes, 300 approximately 700 copies of insertion sequence (IS) elements, and numerous deletions, insertions, translocations and inversions. There is extensive diversity of putative virulence genes, mostly acquired via bacteriophage-mediated lateral gene transfer. Hence, via convergent evolution involving gain and loss of functions, through bacteriophage-mediated gene acquisition, IS-mediated DNA rearrangements and formation of pseudogenes, the Shigella spp. became highly specific human pathogens with variable epidemiological and pathological features. Title: Efficient water removal in lipase-catalyzed esterifications using a low-boiling-point azeotrope Yan Y, Bornscheuer UT, Schmid RD Ref: Biotechnol Bioeng, 78:31, 2002 : PubMed ESTHER: Yan_2002_Biotechnol.Bioeng_78_31 PubMedSearch: Yan 2002 Biotechnol.Bioeng 78 31 PubMedID: 11857278 Abstract High conversions in lipase-catalyzed syntheses of esters from free acyl donors and an alcohol requires efficient removal of water preferentially at temperatures compatible to enzyme activity. Using a lipase B from Candida antarctica (CAL-B)-mediated synthesis of sugar fatty-acid esters, we show that a mixture of ethyl methylketone (EMK) and hexane (best ratio: 4:1, vo/vo) allows efficient removal of water generated during esterification. Azeotropic distillation of the solvent mixture (composition: 26% EMK, 55% hexane, 19% water) takes place at 59 degrees C, which closely matches the optimum temperature reported for CAL-B. Water is then removed from the azeotrope by membrane vapor permeation. In case of glucose stearate, 93% yield was achieved after 48 h using an equimolar ratio of glucose and stearic acid. CAL-B could be reused for seven reaction cycles, with 86% residual activity after 14 d total reaction time at 59 degrees C. A decrease in fatty-acid chain length as well as increasing temperatures (75 degrees C) resulted in lower conversions. In addition, immobilization of CAL-B on a magnetic polypropylene carrier (EP 100) facilitated separation of the biocatalyst. | |||||||
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