Chow J

References (21)

Title : An archaeal lid-containing feruloyl esterase degrades polyethylene terephthalate - Perez-Garcia_2023_Commun.Chem_6_193
Author(s) : Perez-Garcia P , Chow J , Costanzi E , Gurschke M , Dittrich J , Dierkes RF , Molitor R , Applegate V , Feuerriegel G , Tete P , Danso D , Thies S , Schumacher J , Pfleger C , Jaeger KE , Gohlke H , Smits SHJ , Schmitz RA , Streit WR
Ref : Commun Chem , 6 :193 , 2023
Abstract : Polyethylene terephthalate (PET) is a commodity polymer known to globally contaminate marine and terrestrial environments. Today, around 80 bacterial and fungal PET-active enzymes (PETases) are known, originating from four bacterial and two fungal phyla. In contrast, no archaeal enzyme had been identified to degrade PET. Here we report on the structural and biochemical characterization of PET46 (RLI42440.1), an archaeal promiscuous feruloyl esterase exhibiting degradation activity on semi-crystalline PET powder comparable to IsPETase and LCC (wildtypes), and higher activity on bis-, and mono-(2-hydroxyethyl) terephthalate (BHET and MHET). The enzyme, found by a sequence-based metagenome search, is derived from a non-cultivated, deep-sea Candidatus Bathyarchaeota archaeon. Biochemical characterization demonstrated that PET46 is a promiscuous, heat-adapted hydrolase. Its crystal structure was solved at a resolution of 1.71 A. It shares the core alpha/beta-hydrolase fold with bacterial PETases, but contains a unique lid common in feruloyl esterases, which is involved in substrate binding. Thus, our study widens the currently known diversity of PET-hydrolyzing enzymes, by demonstrating PET depolymerization by a plant cell wall-degrading esterase.
ESTHER : Perez-Garcia_2023_Commun.Chem_6_193
PubMedSearch : Perez-Garcia_2023_Commun.Chem_6_193
PubMedID: 37697032
Gene_locus related to this paper: 9arch-PETcan211 , 9cren-PETcan204 , 9arch-PET46

Title : The metagenome-derived esterase PET40 is highly promiscuous and hydrolyses polyethylene terephthalate (PET) - Zhang_2023_Febs.j__
Author(s) : Zhang H , Dierkes RF , Perez-Garcia P , Costanzi E , Dittrich J , Cea PA , Gurschke M , Applegate V , Partus K , Schmeisser C , Pfleger C , Gohlke H , Smits SHJ , Chow J , Streit WR
Ref : Febs J , : , 2023
Abstract : Polyethylene terephthalate (PET) is a widely used synthetic polymer and known to contaminate marine and terrestrial ecosystems. Only few PET-active microorganisms and enzymes (PETases) are currently known and it is debated whether degradation activity for PET originates from promiscuous enzymes with broad substrate spectra that primarily act on natural polymers or other bulky substrates, or whether microorganisms evolved their genetic makeup to accepting PET as a carbon source. Here, we present a predicted diene lactone hydrolase designated PET40, which acts on a broad spectrum of substrates, including PET. It is the first esterase with activity on PET from a GC-rich Gram-positive Amycolatopsis species belonging to the Pseudonocardiaceae (Actinobacteria). It is highly conserved within the genera Amycolatopsis and Streptomyces. PET40 was identified by sequence-based metagenome search using a PETase-specific Hidden Markov Model (HMM). Besides acting on PET, PET40 has a versatile substrate spectrum, hydrolyzing delta-lactones, beta-lactam antibiotics, the polyester-polyurethane Impranil(a) DLN, and various para-nitrophenyl (pNP) ester substrates. Molecular docking suggests that the PET degradative activity is likely a result of the promiscuity of PET40, as potential binding modes were found for substrates encompassing mono(2-hydroxyethyl) terephthalate (MHET), bis(2-hydroxyethyl) terephthalate (BHET), and a PET trimer (PET(3) ). We also solved the crystal structure of the inactive PET40 variant S178A to 1.60 A resolution. PET40 is active throughout a wide pH (pH 4-10) and temperature range (4-65 degreesC) and remarkably stable in the presence of 5% sodium dodecyl sulfate (SDS), making it a promising enzyme as a starting point for further investigations and optimization approaches.
ESTHER : Zhang_2023_Febs.j__
PubMedSearch : Zhang_2023_Febs.j__
PubMedID: 37549040
Gene_locus related to this paper: 9pseu-PET40

Title : The PET-Degrading Potential of Global Metagenomes: From In Silico Mining to Active Enzymes - Chow_2023_Methods.Mol.Biol_2555_139
Author(s) : Chow J , Perez-Garcia P , Dierkes RF , Zhang H , Streit WR
Ref : Methods Mol Biol , 2555 :139 , 2023
Abstract : Against the background of the steadily increasing amount of plastic waste in the sea and on land, it is more important than ever to find ways out of this situation. In recent years, microorganisms have been discovered that are capable of degrading artificial polymers such as polyethylene terephthalate (PET). Even if the turnover rates of the enzymes responsible for this reaction may be too low to solve the global plastic pollution problem, it is still of great societal interest to find microorganisms that are able to degrade the polymer. The corresponding enzymes, PET esterases (PETases) can be used in biotechnological processes and could contribute to a resource-saving circular economy. In this chapter, we present a sequence-based in silico screening method to find new PETases in metagenomic datasets. This method can easily be adapted to find other enzyme classes. We also list a number of assays that can be used to test the enzymes for activity on PET as well as other substrates.
ESTHER : Chow_2023_Methods.Mol.Biol_2555_139
PubMedSearch : Chow_2023_Methods.Mol.Biol_2555_139
PubMedID: 36306084

Title : An Ultra-Sensitive Comamonas thiooxidans Biosensor for the Rapid Detection of Enzymatic Polyethylene Terephthalate (PET) Degradation - Dierkes_2022_Appl.Environ.Microbiol__e0160322
Author(s) : Dierkes RF , Wypych A , Perez-Garcia P , Danso D , Chow J , Streit WR
Ref : Applied Environmental Microbiology , :e0160322 , 2022
Abstract : Polyethylene terephthalate (PET) is a prevalent synthetic polymer that is known to contaminate marine and terrestrial environments. Currently, only a limited number of PET-active microorganisms and enzymes (PETases) are known. This is in part linked to the lack of highly sensitive function-based screening assays for PET-active enzymes. Here, we report on the construction of a fluorescent biosensor based on Comamonas thiooxidans strain S23. C. thiooxidans S23 transports and metabolizes TPA, one of the main breakdown products of PET, using a specific tripartite tricarboxylate transporter (TTT) and various mono- and dioxygenases encoded in its genome in a conserved operon ranging from tphC-tphA1. TphR, an IclR-type transcriptional regulator is found upstream of the tphC-tphA1 cluster where TPA induces transcription of tphC-tphA1 up to 88-fold in exponentially growing cells. In the present study, we show that the C. thiooxidans S23 wild-type strain, carrying the sfGFP gene fused to the tphC promoter, senses TPA at concentrations as low as 10 microM. Moreover, a deletion mutant lacking the catabolic genes involved in TPA degradation thphA2-A1 (deltatphA2A3BA1) is up to 10,000-fold more sensitive and detects TPA concentrations in the nanomolar range. This is, to our knowledge, the most sensitive reporter strain for TPA and we demonstrate that it can be used for the detection of enzymatic PET breakdown products. IMPORTANCE Plastics and microplastics accumulate in all ecological niches. The construction of more sensitive biosensors allows to monitor and screen potential PET degradation in natural environments and industrial samples. These strains will also be a valuable tool for functional screenings of novel PETase candidates and variants or monitoring of PET recycling processes using biocatalysts. Thereby they help us to enrich the known biodiversity and efficiency of PET degrading organisms and enzymes and understand their contribution to environmental plastic degradation.
ESTHER : Dierkes_2022_Appl.Environ.Microbiol__e0160322
PubMedSearch : Dierkes_2022_Appl.Environ.Microbiol__e0160322
PubMedID: 36507653

Title : Plastics degradation by hydrolytic enzymes: The plastics-active enzymes database-PAZy - Buchholz_2022_Proteins_90_1443
Author(s) : Buchholz PCF , Feuerriegel G , Zhang H , Perez-Garcia P , Nover LL , Chow J , Streit WR , Pleiss J
Ref : Proteins , 90 :1443 , 2022
Abstract : Petroleum-based plastics are durable and accumulate in all ecological niches. Knowledge on enzymatic degradation is sparse. Today, less than 50 verified plastics-active enzymes are known. First examples of enzymes acting on the polymers polyethylene terephthalate (PET) and polyurethane (PUR) have been reported together with a detailed biochemical and structural description. Furthermore, very few polyamide (PA) oligomer active enzymes are known. In this article, the current known enzymes acting on the synthetic polymers PET and PUR are briefly summarized, their published activity data were collected and integrated into a comprehensive open access database. The Plastics-Active Enzymes Database (PAZy) represents an inventory of known and experimentally verified enzymes that act on synthetic fossil fuel-based polymers. Almost 3000 homologs of PET-active enzymes were identified by profile hidden Markov models. Over 2000 homologs of PUR-active enzymes were identified by BLAST. Based on multiple sequence alignments, conservation analysis identified the most conserved amino acids, and sequence motifs for PET- and PUR-active enzymes were derived.
ESTHER : Buchholz_2022_Proteins_90_1443
PubMedSearch : Buchholz_2022_Proteins_90_1443
PubMedID: 35175626

Title : Investigation of the halophilic PET hydrolase PET6 from Vibrio gazogenes - Weigert_2022_Protein.Sci__e4500
Author(s) : Weigert S , Perez-Garcia P , Gisdon FJ , Gagsteiger A , Schweinshaut K , Ullmann GM , Chow J , Streit WR , Hcker B
Ref : Protein Science , :e4500 , 2022
Abstract : The handling of plastic waste and the associated ubiquitous occurrence of microplastic poses one of the biggest challenges of our time. Recent investigations of plastic degrading enzymes have opened new prospects for biological microplastic decomposition as well as recycling applications. For polyethylene terephthalate, in particular, several natural and engineered enzymes are known to have such promising properties. From a previous study that identified new PETase candidates by homology search, we chose the candidate PET6 from the globally distributed, halophilic organism Vibrio gazogenes for further investigation. By mapping the occurrence of Vibrios containing PET6 homologs we demonstrated their ubiquitous prevalence in the pangenome of several Vibrio strains. The biochemical characterization of PET6 showed that PET6 has a comparatively lower activity than other enzymes but also revealed a superior turnover at very high salt concentrations. The crystal structure of PET6 provides structural insights into this adaptation to saline environments. By grafting only a few beneficial mutations from other PET degrading enzymes onto PET6, we increased the activity up to three-fold, demonstrating the evolutionary potential of the enzyme. MD simulations of the variant helped rationalize the mutational effects of those mutants and elucidate the interaction of the enzyme with a PET substrate. With tremendous amounts of plastic waste in the Ocean and the prevalence of Vibrio gazogenes in marine biofilms and estuarine marshes, our findings suggest that Vibrio and the PET6 enzyme are worthy subjects to study the PET degradation in marine environments. This article is protected by copyright. All rights reserved.
ESTHER : Weigert_2022_Protein.Sci__e4500
PubMedSearch : Weigert_2022_Protein.Sci__e4500
PubMedID: 36336469
Gene_locus related to this paper: vibga-a0a1m5fok3

Title : Exploring the global metagenome for plastic-degrading enzymes - Perez-Garcia_2021_Methods.Enzymol_648_137
Author(s) : Perez-Garcia P , Danso D , Zhang H , Chow J , Streit WR
Ref : Methods Enzymol , 648 :137 , 2021
Abstract : Plastics are extensively used in our daily life, but they are also a major pollutant of our biosphere accumulating in both the ocean and the land. In the recent years, few enzymes and microorganisms have been discovered with the ability to degrade even fewer synthetic polymers. Nevertheless, more active species and enzymes need to be discovered and described in order to gain more knowledge about protein adaptation to the degradation of not-naturally-occurring polymers. Within this chapter, we focus on efficient methods to identify novel polyethylene terephthalate-degrading enzymes (PETases) from culturable and non-culturable microorganisms by a combination of sequence- and function-based screening. This protocol can be adapted to discover other plastic hydrolases and in general for other enzymes, for which not many characterized specimens are yet available.
ESTHER : Perez-Garcia_2021_Methods.Enzymol_648_137
PubMedSearch : Perez-Garcia_2021_Methods.Enzymol_648_137
PubMedID: 33579401

Title : Plastics degradation by hydrolytic enzymes: the Plastics-Active Enzymes Database - PAZy - Buchholz_2021_Authorea__
Author(s) : Buchholz PCF , Zhang H , Perez-Garcia P , Nover LL , Chow J , Streit WR , Pleiss J
Ref : Authorea , : , 2021
Abstract : Petroleum based plastics are durable and accumulate in all ecological niches. Knowledge on enzymatic degradation is sparse. Today, less than 50 verified plastics-active enzymes are known. First examples of enzymes acting on the polymers polyethylene terephthalate (PET) and polyurethane (PUR) have been reported together with a detailed biochemical and structural description. Further, very few polyamide (PA) oligomer active enzymes are known. In this paper, the current known enzymes acting on the synthetic polymers PET and PUR are briefly summarized, their published activity data were collected and integrated into a comprehensive open access database. The Plastics-Active Enzymes Database (PAZy) represents an inventory of known and experimentally verified plastics-active enzymes. Almost 3000 homologues of PET-active enzymes were identified by profile hidden Markov models. Over 2000 homologues of PUR-active enzymes were identified by BLAST. Based on multiple sequence alignments, conservation analysis identified the most conserved amino acids, and sequence motifs for PET- and PUR-active enzymes were derived.
ESTHER : Buchholz_2021_Authorea__
PubMedSearch : Buchholz_2021_Authorea__

Title : The abundance of mRNA transcripts of bacteroidetal polyethylene terephthalate (PET) esterase genes may indicate a role in marine plastic degradation - Zhang_2021_ResearchSquare__
Author(s) : Zhang H , Dierkes R , Perez-Garcia P , Weigert S , Sternagel S , Hallam S , Schott T , Juergens K , Vollstedt C , Chibani C , Danso D , Buchholz PCF , Pleiss J , Almeida A , Hocker B , Schmitz R , Chow J , Streit WR
Ref : ResearchSquare , : , 2021
Abstract : Polyethylene terephthalate (PET) is an important synthetic polymer accumulating in nature 2 and recent studies have identified microorganisms capable of degrading PET. While the majority of 3 known PET hydrolases originate from the Actinobacteria and Proteobacteria, here we describe the 4 first functional PET-active enzymes from the Bacteroidetes phylum. Using a PETase-specific 5 Hidden-Markov-Model (HMM)-based search algorithm we identified two promiscuous and cold6 active esterases derived from Aequorivita sp. (PET27) and Chryseobacterium jeonii (PET30) acting 7 on PET foil and powder. Notably, one of the enzymes (PET30) was able to hydrolyze PET at 8 temperatures between 4 - 30 C with a similar turnover rate compared to the well-known Ideonella 9 sakaiensis enzyme (IsPETase). 10 PET27 and PET30 homologues were detected in metagenomes encompassing a wide range 11 of different global climate zones. Additional transcript abundance mapping of marine samples imply 12 that these promiscuous enzymes and source organisms may play a role in the long-term 13 degradation of microplastic particles and fibers.
ESTHER : Zhang_2021_ResearchSquare__
PubMedSearch : Zhang_2021_ResearchSquare__
Gene_locus related to this paper: flutr-f2ie04 , 9flao-a0a0c1f4u8 , 9flao-kjj39608 , 9flao-a0a1m6f5v0 , 9flao-a0a330mq60

Title : The Bacteroidetes Aequorivita sp. and Kaistella jeonii Produce Promiscuous Esterases With PET-Hydrolyzing Activity - Zhang_2022_Front.Microbiol_12_803896
Author(s) : Zhang H , Perez-Garcia P , Dierkes RF , Applegate V , Schumacher J , Chibani CM , Sternagel S , Preuss L , Weigert S , Schmeisser C , Danso D , Pleiss J , Almeida A , Hocker B , Hallam SJ , Schmitz RA , Smits SHJ , Chow J , Streit WR
Ref : Front Microbiol , 12 :803896 , 2021
Abstract : Certain members of the Actinobacteria and Proteobacteria are known to degrade polyethylene terephthalate (PET). Here, we describe the first functional PET-active enzymes from the Bacteroidetes phylum. Using a PETase-specific Hidden-Markov-Model- (HMM-) based search algorithm, we identified several PETase candidates from Flavobacteriaceae and Porphyromonadaceae. Among them, two promiscuous and cold-active esterases derived from Aequorivita sp. (PET27) and Kaistella jeonii (PET30) showed depolymerizing activity on polycaprolactone (PCL), amorphous PET foil and on the polyester polyurethane Impranil((a)) DLN. PET27 is a 37.8 kDa enzyme that released an average of 174.4 nmol terephthalic acid (TPA) after 120 h at 30 degreesC from a 7 mg PET foil platelet in a 200 microl reaction volume, 38-times more than PET30 (37.4 kDa) released under the same conditions. The crystal structure of PET30 without its C-terminal Por-domain (PET30deltaPorC) was solved at 2.1 A and displays high structural similarity to the IsPETase. PET30 shows a Phe-Met-Tyr substrate binding motif, which seems to be a unique feature, as IsPETase, LCC and PET2 all contain Tyr-Met-Trp binding residues, while PET27 possesses a Phe-Met-Trp motif that is identical to Cut190. Microscopic analyses showed that K. jeonii cells are indeed able to bind on and colonize PET surfaces after a few days of incubation. Homologs of PET27 and PET30 were detected in metagenomes, predominantly aquatic habitats, encompassing a wide range of different global climate zones and suggesting a hitherto unknown influence of this bacterial phylum on man-made polymer degradation.
ESTHER : Zhang_2022_Front.Microbiol_12_803896
PubMedSearch : Zhang_2022_Front.Microbiol_12_803896
PubMedID: 35069509
Gene_locus related to this paper: flutr-f2ie04 , 9flao-a0a0c1f4u8 , 9flao-kjj39608 , 9flao-a0a330mq60

Title : Plastics: Environmental and Biotechnological Perspectives on Microbial Degradation - Danso_2019_Appl.Environ.Microbiol_85_e01095
Author(s) : Danso D , Chow J , Streit WR
Ref : Applied Environmental Microbiology , 85 : , 2019
Abstract : Plastics are widely used in the global economy, and each year, at least 350 to 400 million tons are being produced. Due to poor recycling and low circular use, millions of tons accumulate annually in terrestrial or marine environments. Today it has become clear that plastic causes adverse effects in all ecosystems and that microplastics are of particular concern to our health. Therefore, recent microbial research has addressed the question of if and to what extent microorganisms can degrade plastics in the environment. This review summarizes current knowledge on microbial plastic degradation. Enzymes available act mainly on the high-molecular-weight polymers of polyethylene terephthalate (PET) and ester-based polyurethane (PUR). Unfortunately, the best PUR- and PET-active enzymes and microorganisms known still have moderate turnover rates. While many reports describing microbial communities degrading chemical additives have been published, no enzymes acting on the high-molecular-weight polymers polystyrene, polyamide, polyvinylchloride, polypropylene, ether-based polyurethane, and polyethylene are known. Together, these polymers comprise more than 80% of annual plastic production. Thus, further research is needed to significantly increase the diversity of enzymes and microorganisms acting on these polymers. This can be achieved by tapping into the global metagenomes of noncultivated microorganisms and dark matter proteins. Only then can novel biocatalysts and organisms be delivered that allow rapid degradation, recycling, or value-added use of the vast majority of most human-made polymers.
ESTHER : Danso_2019_Appl.Environ.Microbiol_85_e01095
PubMedSearch : Danso_2019_Appl.Environ.Microbiol_85_e01095
PubMedID: 31324632

Title : The Thaumarchaeon N. gargensis carries functional bioABD genes and has a promiscuous E. coli DeltabioH-complementing esterase EstN1 - Chow_2018_Sci.Rep_8_13823
Author(s) : Chow J , Danso D , Ferrer M , Streit WR
Ref : Sci Rep , 8 :13823 , 2018
Abstract : Biotin is an essential cofactor required for carboxylation and decarboxylation reactions in all domains of life. While biotin biosynthesis in most Bacteria and Eukarya is well studied, the complete pathway for this vitamer in Archaea is still not known. Detailed genome searches indicated the presence of possible bio gene clusters only in Methanococcales and Thaumarchaeota. Therefore, we analysed the functionality of the predicted genes bioA, bioB, bioD and bioF in the Thaumarchaeon Nitrososphaera gargensis Ga2.9 which are essential for the later steps of biotin synthesis. In complementation tests, the gene cluster-encoded N. gargensis bioABD genes except bioF restored growth of corresponding E. coli Rosetta-gami 2 (DE3) deletion mutants. To find out how biotin biosynthesis is initiated, we searched the genome for a possible bioH analogue encoding a pimeloyl-ACP-methylester carboxylesterase. The respective amino acid sequence of the ORF estN1 showed weak conserved domain similarity to this class of enzymes (e-value 3.70e(-42)). Remarkably, EstN1 is a promiscuous carboxylesterase that complements E. coli DeltabioH and Mesorhizobium loti DeltabioZ mutants for growth on biotin-free minimal medium. Additional 3D-structural models support the hypothesis that EstN1 is a BioH analogue. Thus, this is the first report providing experimental evidence that Archaea carry functional bio genes.
ESTHER : Chow_2018_Sci.Rep_8_13823
PubMedSearch : Chow_2018_Sci.Rep_8_13823
PubMedID: 30218044

Title : Determinants and prediction of esterase substrate promiscuity patterns - Martinez-Martinez_2018_ACS.Chem.Biol_13_225
Author(s) : Martinez-Martinez M , Coscolin C , Santiago G , Chow J , Stogios PJ , Bargiela R , Gertler C , Navarro-Fernandez J , Bollinger A , Thies S , Mendez-Garcia C , Popovic A , Brown G , Chernikova TN , Garcia-Moyano A , Bjergah GE , Perez-Garcia P , Hai T , Del Pozo MV , Stokke R , Steen IH , Cui H , Xu X , Nocek BP , Alcaide M , Distaso M , Mesa V , Pelaez AI , Sanchez J , Buchholz PCF , Pleiss J , Fernandez-Guerra A , Glockner FO , Golyshina OV , Yakimov MM , Savchenko A , Jaeger KE , Yakunin AF , Streit WR , Golyshin PN , Guallar V , Ferrer M
Ref : ACS Chemical Biology , 13 :225 , 2018
Abstract : Esterases receive special attention because their wide distribution in biological systems and environments and their importance for physiology and chemical synthesis. The prediction of esterases substrate promiscuity level from sequence data and the molecular reasons why certain such enzymes are more promiscuous than others, remain to be elucidated. This limits the surveillance of the sequence space for esterases potentially leading to new versatile biocatalysts and new insights into their role in cellular function. Here we performed an extensive analysis of the substrate spectra of 145 phylogenetically and environmentally diverse microbial esterases, when tested with 96 diverse esters. We determined the primary factors shaping their substrate range by analyzing substrate range patterns in combination with structural analysis and protein-ligand simulations. We found a structural parameter that helps ranking (classifying) promiscuity level of esterases from sequence data at 94% accuracy. This parameter, the active site effective volume, exemplifies the topology of the catalytic environment by measuring the active site cavity volume corrected by the relative solvent accessible surface area (SASA) of the catalytic triad. Sequences encoding esterases with active site effective volumes (cavity volume/SASA) above a threshold show greater substrate spectra, which can be further extended in combination with phylogenetic data. This measure provides also a valuable tool for interrogating substrates capable of being converted. This measure, found to be transferred to phosphatases of the haloalkanoic acid dehalogenase superfamily and possibly other enzymatic systems, represents a powerful tool for low-cost bioprospecting for esterases with broad substrate ranges, in large scale sequence datasets.
ESTHER : Martinez-Martinez_2018_ACS.Chem.Biol_13_225
PubMedSearch : Martinez-Martinez_2018_ACS.Chem.Biol_13_225
PubMedID: 29182315
Gene_locus related to this paper: 9zzzz-a0a2k8jn75 , 9zzzz-a0a2k8jt94 , 9zzzz-a0a0g3fj44 , 9zzzz-a0a0g3fh10 , 9zzzz-a0a0g3fh03 , 9bact-a0a1s5qkj8 , 9zzzz-a0a0g3feh5 , 9zzzz-a0a0g3fkz4 , 9zzzz-a0a0g3fh07 , 9zzzz-a0a0g3fh34 , 9zzzz-a0a0g3fh31 , 9bact-KY458167 , alcbs-q0vqa3 , 9bact-a0a1s5qki8 , 9zzzz-a0a0g3feq8 , 9zzzz-a0a0g3feh8 , 9zzzz-a0a0g3fh19 , 9bact-KY203037 , 9bact-a0a1s5ql22 , 9bact-a0a1s5qm34 , 9bact-KY203034 , 9bact-r9qzg0 , 9bact-a0a1s5qly8 , 9zzzz-a0a0g3fkz8 , 9zzzz-a0a0g3feg9 , 9zzzz-KY203033 , 9zzzz-a0a0g3fes4 , 9zzzz-a0a0g3fh42 , 9bact-a0a1s5qlx2 , 9zzzz-KY483651 , 9bact-a0a1s5qmh4 , 9zzzz-KY203032 , 9zzzz-EH87 , 9zzzz-a0a0g3fei1 , 9zzzz-a0a0g3fet2 , 9zzzz-KY483647 , 9zzzz-EH82 , 9zzzz-a0a0g3fe15 , 9bact-KY203031 , 9bact-t1w006 , 9zzzz-a0a0g3fet6 , 9bact-KY458164 , geoth-g8myf3 , 9bact-a0a1s5ql04 , 9gamm-a0a1y0ihk7 , 9bact-a0a1s5qly6 , 9bact-a0a1s5qkg4 , 9bact-a0a1s5qkm4 , 9gamm-s5tv80 , 9gamm-a0a0c4zhg2 , 9zzzz-t1b379 , 9gamm-KY483646 , 9bact-KY458160 , 9zzzz-a0a0g3fj57 , 9gamm-s5t8349 , 9arch-KY203036 , 9bact-KY458168 , 9zzzz-a0a0g3fes0 , 9zzzz-t1be47 , 9bact-KY458159 , 9zzzz-a0a0g3fh39 , 9bact-t1vzd5 , 9prot-EH41 , 9bact-Lip114 , alcbs-q0vt77 , 9bact-a0a1s5qke6 , 9bact-a0a1s5qkf3 , 9prot-SRP030024 , 9gamm-s5t532 , 9bact-a0a1s5qkl2 , 9bact-a0a1s5qkk8 , 9zzzz-KY203030 , 9zzzz-t1d4I7 , 9prot-KY019260 , 9bact-a0a1s5qm38 , 9arch-KY458161 , 9prot-KY010302 , 9zzzz-a0a0g3fl25 , 9actn-KY010298 , 9gamm-s5u059 , 9bact-a0a1s5qmi7 , 9bact-KY010297 , 9bact-KY483642 , 9bact-a0a1s5qkj1 , 9bact-KY010299 , 9bact-KY483648 , alcbs-q0vtl7 , 9bact-a0a1s5qf1 , 9bact-a0a1s5qkg0 , 9bact-a0a0h4tgu6 , 9bact-MilE3 , 9bact-LAE6 , 9alte-MGS-MT1 , 9bact-r9qzf7 , 9gamm-k0c6t6 , alcbs-q0vl36 , alcbs-q0vlq1 , alcbs-q0vq49 , bacsu-pnbae , canar-LipB , canan-lipasA , geost-lipas , marav-a1u5n0 , pseps-i7k8x5 , staep-GEHD , symth-q67mr3 , altma-s5cfn7 , cycsp-k0c2b8 , alcbs-q0vlk5 , 9bact-k7qe48 , 9bact-MGS-M1 , 9bact-MGS-M2 , 9bact-a0a0b5kns5 , 9zzzz-a0a0g3fej4 , 9zzzz-a0a0g3fj60 , 9zzzz-a0a0g3fej0 , 9zzzz-a0a0g3fj64 , 9bact-a0a0b5kc16 , 9zzzz-a0a0g3feg6 , 9zzzz-a0a0g3feu6

Title : New insights into the function and global distribution of polyethylene terephthalate (PET) degrading bacteria and enzymes in marine and terrestrial metagenomes - Danso_2018_Appl.Environ.Microbiol_84_e2773
Author(s) : Danso D , Schmeisser C , Chow J , Zimmermann W , Wei R , Leggewie C , Li X , Hazen T , Streit WR
Ref : Applied Environmental Microbiology , 84 :e2773 , 2018
Abstract : Polyethylene terephthalate (PET) is one of the most important synthetic polymers used nowadays. Unfortunately, the polymers accumulate in nature and until now, no highly active enzymes are known that can degrade it at high velocity. Enzymes involved in PET degradation are mainly alpha/beta-hydrolases like cutinases and related enzymes (E.C. 3.1.-). Currently, only a small number of such enzymes are well characterized. Within this work, a search algorithm was developed that identified 504 possible PET hydrolase candidate genes from various databases. A further global search that comprised more than 16 GB of sequence information within 108 marine and 25 terrestrial metagenomes obtained from the IMG data base detected 349 putative PET hydrolases. Heterologous expression of four such candidate enzymes verified the function of these enzymes and confirmed the usefulness of the developed search algorithm. Thereby, two novel and thermostable enzymes with high potential for downstream application were in part characterized. Clustering of 504 novel enzyme candidates based on amino acid similarities indicated that PET hydrolases mainly occur in the phylum of Actinobacteria, Proteobacteria and Bacteroidetes Within the Proteobacteria, the Beta-, Delta- and Gammaproteobacteria were the main hosts. Remarkably enough, in the marine environment, bacteria affiliated with the phylum of the Bacteroidetes appear to be the main host of PET hydrolase genes rather than Actinobacteria or Proteobacteria as observed for the terrestrial metagenomes. Our data further imply that PET hydrolases are truly rare enzymes. The highest occurrence of 1.5 hits/Mb was observed in a sample site containing crude oil.IMPORTANCE Polyethylene terephthalate (PET) accumulates in our environment without significant microbial conversion. Although few PET hydrolases are already known it is still unknown how frequent they appear and which main bacterial phyla they are affiliated with. In this study, deep sequence mining of protein databases and metagenomes demonstrated that PET hydrolases indeed are occurring at very low frequencies in the environment. Further it was possible to link them to phyla which were previously unknown to harbor such enzymes. This work contributes novel knowledge to the phylogenetic relationship, the recent evolution and the global distribution of PET hydrolases. Finally, we describe biochemical traits of four novel PET hydrolases.
ESTHER : Danso_2018_Appl.Environ.Microbiol_84_e2773
PubMedSearch : Danso_2018_Appl.Environ.Microbiol_84_e2773
PubMedID: 29427431
Gene_locus related to this paper: 9burk-PET10 , 9burk-PET11 , 9gamm-a0a0d4l7e6 , 9alte-n6vy44 , 9zzzz-a0a0f9x315 , deiml-e8u721 , olean-r4ykl9 , vibga-a0a1z2siq1 , 9burk-a0a0g3bi90 , 9bact-c3ryl0 , 9actn-h6wx58 , idesa-peth , 9bact-g9by57 , acide-PBSA , morsp-lip1

Title : Evolution of novel wood decay mechanisms in Agaricales revealed by the genome sequences of Fistulina hepatica and Cylindrobasidium torrendii - Floudas_2015_Fungal.Genet.Biol_76_78
Author(s) : Floudas D , Held BW , Riley R , Nagy LG , Koehler G , Ransdell AS , Younus H , Chow J , Chiniquy J , Lipzen A , Tritt A , Sun H , Haridas S , LaButti K , Ohm RA , Kues U , Blanchette RA , Grigoriev IV , Minto RE , Hibbett DS
Ref : Fungal Genet Biol , 76 :78 , 2015
Abstract : Wood decay mechanisms in Agaricomycotina have been traditionally separated in two categories termed white and brown rot. Recently the accuracy of such a dichotomy has been questioned. Here, we present the genome sequences of the white-rot fungus Cylindrobasidium torrendii and the brown-rot fungus Fistulina hepatica both members of Agaricales, combining comparative genomics and wood decay experiments. C. torrendii is closely related to the white-rot root pathogen Armillaria mellea, while F. hepatica is related to Schizophyllum commune, which has been reported to cause white rot. Our results suggest that C. torrendii and S. commune are intermediate between white-rot and brown-rot fungi, but at the same time they show characteristics of decay that resembles soft rot. Both species cause weak wood decay and degrade all wood components but leave the middle lamella intact. Their gene content related to lignin degradation is reduced, similar to brown-rot fungi, but both have maintained a rich array of genes related to carbohydrate degradation, similar to white-rot fungi. These characteristics appear to have evolved from white-rot ancestors with stronger ligninolytic ability. F. hepatica shows characteristics of brown rot both in terms of wood decay genes found in its genome and the decay that it causes. However, genes related to cellulose degradation are still present, which is a plesiomorphic characteristic shared with its white-rot ancestors. Four wood degradation-related genes, homologs of which are frequently lost in brown-rot fungi, show signs of pseudogenization in the genome of F. hepatica. These results suggest that transition toward a brown-rot lifestyle could be an ongoing process in F. hepatica. Our results reinforce the idea that wood decay mechanisms are more diverse than initially thought and that the dichotomous separation of wood decay mechanisms in Agaricomycotina into white rot and brown rot should be revisited.
ESTHER : Floudas_2015_Fungal.Genet.Biol_76_78
PubMedSearch : Floudas_2015_Fungal.Genet.Biol_76_78
PubMedID: 25683379
Gene_locus related to this paper: 9agar-a0a0d6zyq5 , 9agar-a0a0d7a2p9 , 9agar-a0a0d7a2v2 , 9agar-a0a0d7abt2 , 9agar-a0a0d7acd3 , 9agar-a0a0d7acx0 , 9agar-a0a0d7acx9 , 9agar-a0a0d7adg2 , 9agar-a0a0d7a6d0 , 9agar-a0a0d7aen7 , 9agar-a0a0d7aez7 , 9agar-a0a0d7ahq5 , 9agar-a0a0d7akr6 , 9agar-a0a0d7al29 , 9agar-a0a0d7an16 , 9agar-a0a0d7ann7 , 9agar-a0a0d7anv1 , 9homo-a0a0d7atv2 , 9homo-a0a0d7ay28 , 9homo-a0a0d7ayz7 , 9homo-a0a0d7b1w8 , 9homo-a0a0d7b2p0 , 9homo-a0a0d7b4n4 , 9homo-a0a0d7b624 , 9homo-a0a0d7b7r3 , 9homo-a0a0d7b7w3 , 9homo-a0a0d7bac5 , 9homo-a0a0d7bav7 , 9homo-a0a0d7bbx7 , 9homo-a0a0d7bdn7 , 9homo-a0a0d7bgj9 , 9homo-a0a0d7biw2 , 9homo-a0a0d7bqi1 , 9homo-a0a0d7bv80 , 9agar-a0a0d7b6f6 , 9agar-a0a0d7b976 , 9agar-a0a0d7aeu9 , 9agar-a0a0d7ag53 , 9agar-a0a0d7b8a5

Title : A novel thermoalkalostable esterase from Acidicaldus sp. strain USBA-GBX-499 with enantioselectivity isolated from an acidic hot springs of Colombian Andes - Lopez_2014_Appl.Microbiol.Biotechnol_98_8603
Author(s) : Lopez G , Chow J , Bongen P , Lauinger B , Pietruszka J , Streit WR , Baena S
Ref : Applied Microbiology & Biotechnology , 98 :8603 , 2014
Abstract : Several thermo- and mesoacidophilic bacterial strains that revealed high lipolytic activity were isolated from water samples derived from acidic hot springs in Los Nevados National Natural Park (Colombia). A novel lipolytic enzyme named 499EST was obtained from the thermoacidophilic alpha-Proteobacterium Acidicaldus USBA-GBX-499. The gene estA encoded a 313-amino-acid protein named 499EST. The deduced amino acid sequence showed the highest identity (58 %) with a putative alpha/beta hydrolase from Acidiphilium sp. (ZP_08632277.1). Sequence alignments and phylogenetic analysis indicated that 499EST is a new member of the bacterial esterase/lipase family IV. The esterase reveals its optimum catalytic activity at 55 degrees C and pH 9.0. Kinetic studies showed that 499EST preferentially hydrolyzed middle-length acyl chains (C6-C8), especially p-nitrophenyl (p-NP) caproate (C6). Its thermostability and activity were strongly enhanced by adding 6 mM FeCl3. High stability in the presence of water-miscible solvents such as dimethyl sulfoxide and glycerol was observed. This enzyme also exhibits stability under harsh environmental conditions and enantioselectivity towards naproxen and ibuprofen esters, yielding the medically relevant (S)-enantiomers. In conclusion, according to our knowledge, 499EST is the first thermoalkalostable esterase derived from a Gram-negative thermoacidophilic bacterium.
ESTHER : Lopez_2014_Appl.Microbiol.Biotechnol_98_8603
PubMedSearch : Lopez_2014_Appl.Microbiol.Biotechnol_98_8603
PubMedID: 24818691
Gene_locus related to this paper: 9prot-a0a068lg40

Title : Cloning, expression, purification and preliminary X-ray analysis of EstN2, a novel archaeal alpha\/beta-hydrolase from Candidatus Nitrososphaera gargensis - Kaljunen_2014_Acta.Crystallogr.F.Struct.Biol.Commun_70_1394
Author(s) : Kaljunen H , Chow J , Streit WR , Mueller-Dieckmann J
Ref : Acta Crystallographica F Struct Biol Commun , 70 :1394 , 2014
Abstract : EstN2 is a novel alpha/beta-hydrolase originating from the ammonia-oxidizing thaumarchaeon Candidatus Nitrososphaera gargensis. The genome of the organism was sequenced and genes conferring putative lipolytic activity were amplified and cloned into Escherichia coli as a heterologous host. Through function-based screening, esterase and lipase activity was detected. A recombinant enzyme designated EstN2 was successfully expressed, purified and crystallized. The crystals belonged to space group I2, with one molecule per asymmetric unit, and diffracted X-rays to 1.5 A resolution.
ESTHER : Kaljunen_2014_Acta.Crystallogr.F.Struct.Biol.Commun_70_1394
PubMedSearch : Kaljunen_2014_Acta.Crystallogr.F.Struct.Biol.Commun_70_1394
PubMedID: 25286947
Gene_locus related to this paper: nitgg-k0im51

Title : Genome sequencing provides insight into the reproductive biology, nutritional mode and ploidy of the fern pathogen Mixia osmundae - Toome_2014_New.Phytol_202_554
Author(s) : Toome M , Ohm RA , Riley RW , James TY , Lazarus KL , Henrissat B , Albu S , Boyd A , Chow J , Clum A , Heller G , Lipzen A , Nolan M , Sandor L , Zvenigorodsky N , Grigoriev IV , Spatafora JW , Aime MC
Ref : New Phytol , 202 :554 , 2014
Abstract : Mixia osmundae (Basidiomycota, Pucciniomycotina) represents a monotypic class containing an unusual fern pathogen with incompletely understood biology. We sequenced and analyzed the genome of M. osmundae, focusing on genes that may provide some insight into its mode of pathogenicity and reproductive biology. Mixia osmundae has the smallest plant pathogenic basidiomycete genome sequenced to date, at 13.6 Mb, with very few repeats, high gene density, and relatively few significant gene family gains. The genome shows that the yeast state of M. osmundae is haploid and the lack of segregation of mating genes suggests that the spores produced on Osmunda spp. fronds are probably asexual. However, our finding of a complete complement of mating and meiosis genes suggests the capacity to undergo sexual reproduction. Analyses of carbohydrate active enzymes suggest that this fungus is a biotroph with the ability to break down several plant cell wall components. Analyses of publicly available sequence data show that other Mixia members may exist on other plant hosts and with a broader distribution than previously known.
ESTHER : Toome_2014_New.Phytol_202_554
PubMedSearch : Toome_2014_New.Phytol_202_554
PubMedID: 24372469
Gene_locus related to this paper: mixos-g7dyk5 , mixos-g7eay5

Title : Complete Genome Sequence of Geobacillus sp. Strain GHH01, a Thermophilic Lipase-Secreting Bacterium - Wiegand_2013_Genome.Announc_1_e0009213
Author(s) : Wiegand S , Rabausch U , Chow J , Daniel R , Streit WR , Liesegang H
Ref : Genome Announc , 1 :e0009213 , 2013
Abstract : Geobacillus sp. strain GHH01 was isolated during a screening for producers of extracellular thermostable lipases. The completely sequenced and annotated 3.6-Mb genome encodes 3,478 proteins. The strain is genetically equipped to utilize a broad range of different substrates and might develop natural competence.
ESTHER : Wiegand_2013_Genome.Announc_1_e0009213
PubMedSearch : Wiegand_2013_Genome.Announc_1_e0009213
PubMedID: 23618712
Gene_locus related to this paper: geoka-q5l3h0 , geosc-d7d055 , geotn-a4isp0 , geos2-a0a0e0tby6

Title : The metagenome-derived enzymes LipS and LipT increase the diversity of known lipases - Chow_2012_PLoS.One_7_e47665
Author(s) : Chow J , Kovacic F , Dall Antonia Y , Krauss U , Fersini F , Schmeisser C , Lauinger B , Bongen P , Pietruszka J , Schmidt M , Menyes I , Bornscheuer UT , Eckstein M , Thum O , Liese A , Mueller-Dieckmann J , Jaeger KE , Streit WR
Ref : PLoS ONE , 7 :e47665 , 2012
Abstract : Triacylglycerol lipases (EC catalyze both hydrolysis and synthesis reactions with a broad spectrum of substrates rendering them especially suitable for many biotechnological applications. Most lipases used today originate from mesophilic organisms and are susceptible to thermal denaturation whereas only few possess high thermotolerance. Here, we report on the identification and characterization of two novel thermostable bacterial lipases identified by functional metagenomic screenings. Metagenomic libraries were constructed from enrichment cultures maintained at 65 to 75 degrees C and screened resulting in the identification of initially 10 clones with lipolytic activities. Subsequently, two ORFs were identified encoding lipases, LipS and LipT. Comparative sequence analyses suggested that both enzymes are members of novel lipase families. LipS is a 30.2 kDa protein and revealed a half-life of 48 h at 70 degrees C. The lipT gene encoded for a multimeric enzyme with a half-life of 3 h at 70 degrees C. LipS had an optimum temperature at 70 degrees C and LipT at 75 degrees C. Both enzymes catalyzed hydrolysis of long-chain (C(12) and C(14)) fatty acid esters and additionally hydrolyzed a number of industry-relevant substrates. LipS was highly specific for (R)-ibuprofen-phenyl ester with an enantiomeric excess (ee) of 99%. Furthermore, LipS was able to synthesize 1-propyl laurate and 1-tetradecyl myristate at 70 degrees C with rates similar to those of the lipase CalB from Candida antarctica. LipS represents the first example of a thermostable metagenome-derived lipase with significant synthesis activities. Its X-ray structure was solved with a resolution of 1.99 A revealing an unusually compact lid structure.
ESTHER : Chow_2012_PLoS.One_7_e47665
PubMedSearch : Chow_2012_PLoS.One_7_e47665
PubMedID: 23112831
Gene_locus related to this paper: symth-q67mr3 , 9bact-k7qe48

Title : Cloning, expression, purification and preliminary X-ray analysis of a putative metagenome-derived lipase - Fersini_2012_Acta.Crystallogr.Sect.F.Struct.Biol.Cryst.Commun_68_923
Author(s) : Fersini F , Dall'Antonia Y , Chow J , Streit WR , Mueller-Dieckmann J
Ref : Acta Crystallographica Sect F Struct Biol Cryst Commun , 68 :923 , 2012
Abstract : LipS is a novel thermostable putative lipase that was isolated from a metagenomic library using functional screening methods. The corresponding gene shows high similarity to that encoding a putative but uncharacterized esterase from Symbiobacterium thermophilum IAM14863 (99% nucleotide-sequence similarity). Two different constructs of the recombinant lipase were crystallized. Crystals belonging to space group P4(2)2(1)2 diffracted X-ray radiation to 2.8 A resolution and crystals belonging to space group P4 diffracted to 2.0 A resolution. The most probable content of their asymmetric units were two molecules (P4(2)2(1)2) and four or five molecules (P4), respectively.
ESTHER : Fersini_2012_Acta.Crystallogr.Sect.F.Struct.Biol.Cryst.Commun_68_923
PubMedSearch : Fersini_2012_Acta.Crystallogr.Sect.F.Struct.Biol.Cryst.Commun_68_923
PubMedID: 22869123
Gene_locus related to this paper: symth-q67mr3