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Zarubina S.A.,Moscow State University | Uporov I.V.,Moscow State University | Fedorchuk E.A.,Moscow State University | Fedorchuk V.V.,Moscow State University | And 3 more authors.
Acta Naturae | Year: 2013

Alpha-amino acid ester hydrolase (EC 3.1.1.43, AEH) is a promising biocatalyst for the production of semi-synthetic β-lactam antibiotics, penicillins and cephalosporins. The AEH gene from Xanthomonas rubrilineans (XrAEH) was recently cloned in this laboratory. The three-dimensional structure of XrAEH was simulated using the homology modeling method for rational design experiments. The analysis of the active site was performed, and its structure was specified. The key amino acid residues in the active site - the catalytic triad (Ser175, His341 and Asp308), oxyanion hole (Tyr83 and Tyr176), and carboxylate cluster (carboxylate groups of Asp209, Glu310 and Asp311) - were identified. It was shown that the optimal configuration of residues in the active site occurs with a negative net charge -1 in the carboxylate cluster. Docking of different substrates in the AEH active site was carried out, which allowed us to obtain structures of XrAEH complexes with the ampicillin, amoxicillin, cephalexin, D-phenylglycine, and 4-hydroxy-D-phenylglycine methyl ester. Modeling of XrAEH enzyme complexes with various substrates was used to show the structures for whose synthesis this enzyme will show the highest efficiency. © 2013 Park-media, Ltd. Source


Borshchevskaya L.N.,State Research Institute for Genetics and Selection of Industrial Microorganisms GosNIIgenetika | Kalinina A.N.,State Research Institute for Genetics and Selection of Industrial Microorganisms GosNIIgenetika | Sineokii S.P.,State Research Institute for Genetics and Selection of Industrial Microorganisms GosNIIgenetika
Applied Biochemistry and Microbiology | Year: 2013

A method for the taxonomic identification of seven closely related bacterial species of the Bacillus subtilis group (B. subtilis, B. amyloliquefaciens, B. licheniformis, B. vallismortis, B. atrophaeus, B. sonorensis, and B. mojavensis) using specific primers selected on the basis of the gyrA gene sequences was developed. The effectiveness of this method both for the identification of pure cultures of type strains of this group and for the precise species identification of collection and industrial bacterial strains was demonstrated. The principal possibility of using this method for detecting B. subtilis group bacteria in mixed cultures was shown. © 2013 Pleiades Publishing, Inc. Source

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