Pietruszka J

References (4)

Title : Discovery of new acetylcholinesterase inhibitors for Alzheimer's disease: virtual screening and in vitro characterisation - David_2021_J.Enzyme.Inhib.Med.Chem_36_491
Author(s) : David B , Schneider P , Schafer P , Pietruszka J , Gohlke H
Ref : J Enzyme Inhib Med Chem , 36 :491 , 2021
Abstract : For more than two decades, the development of potent acetylcholinesterase (AChE) inhibitors has been an ongoing task to treat dementia associated with Alzheimer's disease and improve the pharmacokinetic properties of existing drugs. In the present study, we used three docking-based virtual screening approaches to screen both ZINC15 and MolPort databases for synthetic analogs of physostigmine and donepezil, two highly potent AChE inhibitors. We characterised the in vitro inhibitory concentration of 11 compounds, ranging from 14 to 985 microM. The most potent of these compounds, S-I 26, showed a fivefold improved inhibitory concentration in comparison to rivastigmine. Moderate inhibitors carrying novel scaffolds were identified and could be improved for the development of new classes of AChE inhibitors.
ESTHER : David_2021_J.Enzyme.Inhib.Med.Chem_36_491
PubMedSearch : David_2021_J.Enzyme.Inhib.Med.Chem_36_491
PubMedID: 33478277

Title : Identification of novel esterase-active enzymes from hot environments by use of the host bacterium Thermus thermophilus - Leis_2015_Front.Microbiol_6_275
Author(s) : Leis B , Angelov A , Mientus M , Li H , Pham VT , Lauinger B , Bongen P , Pietruszka J , Goncalves LG , Santos H , Liebl W
Ref : Front Microbiol , 6 :275 , 2015
Abstract : Functional metagenomic screening strategies, which are independent of known sequence information, can lead to the identification of truly novel genes and enzymes. Since E. coli has been used exhaustively for this purpose as a host, it is important to establish alternative expression hosts and to use them for functional metagenomic screening for new enzymes. In this study we show that Thermus thermophilus HB27 is an excellent screening host and can be used as an alternative provider of truly novel biocatalysts. In a previous study we constructed mutant strain BL03 with multiple markerless deletions in genes for major extra- and intracellular lipolytic activities. This esterase-diminished strain was no longer able to grow on defined minimal medium supplemented with tributyrin as the sole carbon source and could be used as a host to screen for metagenomic DNA fragments that could complement growth on tributyrin. Several thousand single fosmid clones from thermophilic metagenomic libraries from heated compost and hot spring water samples were subjected to a comparative screening for esterase activity in both T. thermophilus strain BL03 and E. coli EPI300. We scored a greater number of active esterase clones in the thermophilic bacterium than in the mesophilic E. coli. From several thousand functionally screened clones only two thermostable alpha/beta-fold hydrolase enzymes with high amino acid sequence similarity to already characterized enzymes were identifiable in E. coli. In contrast, five further fosmids were found that conferred lipolytic activities in T. thermophilus only. Four open reading frames (ORFs) were found which did not share significant similarity to known esterase enzymes but contained the conserved GXSXG motif regularly found in lipolytic enzymes. Two of the genes were expressed in both hosts and the novel thermophilic esterases, which based on their primary structures could not be assigned to known esterase or lipase families, were purified and preliminarily characterized. Our work underscores the benefit of using additional screening hosts other than E. coli for the identification of novel biocatalysts with industrial relevance.
ESTHER : Leis_2015_Front.Microbiol_6_275
PubMedSearch : Leis_2015_Front.Microbiol_6_275
PubMedID: 25904908

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 : 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 3.1.1.3) 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