Acta NaturaeActa NaturaeActa Naturae2075-8251Acta Naturae Ltd1078610.32607/20758251-2009-1-3-94-98ArticlesСтатьиResearch ArticleMutation of Residue βF71 of Escherichia coli Penicillin Acylase Results in Enhanced Enantioselectivity and Improved Catalytic PropertiesMutation of Residue βF71 of Escherichia coli Penicillin Acylase Results in Enhanced Enantioselectivity and Improved Catalytic PropertiesShapovalovaI VAlkemaW BLJamskovaO Vde VriesEGurandaD TJanssenD BŠvedasV Kvytas@belozersky.msu.ruBelozersky Institute of Physicochemical Biology and Faculty of Bioengineering and Bioinformatics, Lomonosov Moscow State UniversityDepartment of Biochemistry, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen1512200913NO3 (2009)№3 (2009)949817012020Copyright ©; 2009, Shapovalova I.V., Alkema W.B., Jamskova O.V., de Vries E., Guranda D.T., Janssen D.B., Švedas V.K.Copyright ©; 2009, Shapovalova I.V., Alkema W.B., Jamskova O.V., de Vries E., Guranda D.T., Janssen D.B., Švedas V.K.2009Shapovalova I.V., Alkema W.B., Jamskova O.V., de Vries E., Guranda D.T., Janssen D.B., Švedas V.K.Shapovalova I.V., Alkema W.B., Jamskova O.V., de Vries E., Guranda D.T., Janssen D.B., Švedas V.K.https://creativecommons.org/licenses/by/4.0https://actanaturae.ru/2075-8251/article/view/10786

Residue phenylalanine 71 of the β-chain of penicillin acylase from E. coli is involved in substrate binding and chiral discrimination of its enantiomers. Different amino acid residues have been introduced at position βF71, and the mutants were studied with respect to their enantioselectivity and substrate specificity. Some mutants demonstrated remarkably improved catalytic activity. Moreover, mutation of βF71 residue allowed to enhance penicillin acylase enantioselectivity. The catalytic activity to the specific substrates was improved up to 36 times, most notably for K, R, and L mutants. Increased activity to a D-phenylglycine derivative – a valuable specificity improvement for biocatalytic synthesis of new penicillins and cephalosporins – was shown for βF71R and βF71L mutants. The synthetic capacity of penicillin acylase with 6-aminopenicillanic acid as an external nucleophile was especially sensitive to mutation of the β71 residue in contrast to the synthesis with 7-aminodeacetoxycephalosporanic acid.

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