Abstract
An enzyme of cephalosporin-acid synthetase produced by the E. coli strain VKPM B-10182 has specificity for the synthesis of β-lactam antibiotics of the cephalosporin acids class (cefazolin, cefalotin, cefezole etc.). A comparison of the previously determined genomic sequence of E. coli VKPM B-10182 with a genome of the parent E. coli strain ATCC 9637 was performed. Multiple mutations indicating the long selection history of the strain were detected, including mutations in the genes of RNase and β-lactamases that could enhance the level of enzyme synthesis and reduce the degree of degradation of the synthesized cephalosporin acids. The CASA gene—a direct homolog of the penicillin G-acylase gene—was identified by bioinformatics methods. The homology of the gene was confirmed by gene cloning and the expression and determination of its enzymatic activity in the reaction of cefazolin synthesis. The CASA gene was isolated and cloned into the original expression vector, resulting in an effective E. coli BL21(DE3) pMD0107 strain producing CASA.
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Volpato, G., Rodrigues, R.C., and Fernandez-Lafuente, R., Curr. Med. Chem., 2010, vol. 17, no. 32, pp. 3855–3873.
The, Mycota., in X Industrial Applications, 2nd. ed., Esser, K. and Hofrichter, M., Eds., Berlin: SpringerVerlag, vol. 5, pp. 101–121.
AL, BlumJ.K. and Bommarius, A.S., in Flickinger M.C. Encyclopedia Industrial Biotechnology: Bioprocess, Bioseparation, and Cell Technology (Kingfisher Guide), Flickinger, M.C., Ed., Hoboken, N.J.: John Wiley Sons, Inc., 2010, pp. 1–32.
Sklyarenko, A.V., Kurochkina, V.B., and Egorov, A.M., in New Res. Biotechnol. Biol. Med., Egorov, A.M. and Zaikov, G., Eds., Nova Sci. Publ., 2006, vol. 8, pp. 73–86.
Kurochkina, V.B. and Sklyarenko, A.V., Antibiot. Khimioter., 2005, vol. 50, no. 5-6, pp. 39–58.
Grulich, M., Stepanek, V., and Kyslik, P., Biotechnol. Adv., 2013, vol. 31, no. 8, pp. 1458–1472.
Srirangan, K., Orr, V., Akawi, L., Westbrook, A., Moo Young, M., and Chou, C.P., Biotechnol. Adv., 2013, vol. 31, no. 8, pp. 1319–1332.
Kurochkina, V.B., Sklyarenko, A.V., Berezina, O.V., and Yarotskii, S.V., Appl. Biochem. Microbiol., 2013, vol. 49, pp. 672–694.
RF Patent no. RU2381273, 2010.
RF Patent no. RU2535893, 2014.
RF Patent no. RU2420581, 2011.
Kurochkina, V.B. and Nys, P.S., Antibiot. Khimioter., 1999, vol. 44, no. 5, pp. 12–16.
Nys, P.S. and Kurochkina, V.B., Appl. Biochem. Biotechnol., 2000, vol. 88, nos 1-3, pp. 221–230.
RF Patent no. RU2210596, 2000.
Shiloach, J. and Bauer, S., Biotechnol. Bioeng., 1975, vol. 17, no. 2, pp. 227–239.
Nagata, S., J. Biosci. Bioeng., 2001, vol. 92, no. 4, pp. 324–329.
Gleiser, I. and Bauer, S., Biotechnol. Bioeng., 1981, vol. 23, no. 5, pp. 1015–1021.
Mardanov, A.V., Eldarov, M.A., Sklyarenko, A.V., Dumina, M.V., Beletsky, A.V., Yarotsky, S.V., and Ravin, N.V., Genome Announc, 2014, vol. 2, no. 6. doi:10.1128/genomeA.01222-14.
Moriya, Y., Itoh, M., Okuda, S., Yoshizawa, A.C., and Kanehisa, M., Nucleic Acids Res., 2007, vol. 35, pp. W182–W185.
Altschul, S.F., Madden, T.L., Schaffer, A.A., Zhang, J., Zhang, Z., Miller, W., and Lipman, D.J., Nucleic Acid Res., 1997, vol. 25, no. 17, pp. 3389–3402.
Khatuntseva, S.A., Eldarov, M.A., Redo, V.A., and Skryabin, K.G., J. Biotechnol., 2008, vol. 133, no. 1, pp. 250–256.
Nys, P.S., Savitskaya, E.M., and Kolygina, T.S., Antibiotiki, 1973, vol. 17, pp. 270–276.
Balasingham, K., Warburton, D., Dunnill, P., and Lilly, M.D., Biochim. Biophys. Acta—Enzymol., 1972, vol. 276, pp. 250–256.
Laemmli, U., K:, Nature, 1970, vol. 227, no. 5259, pp. 680–685.
Keseler, I.M., Mackie, A., Peralta-Gil, M., Santos-Zavaleta, A., Gama-Castro, S., Bonavides-Martinez, C., Fulcher, C., Huerta, A.M., Kothari, A., Krummenacker, M., Latendresse, M., Muniz-Rascado, L., Ong, Q., Paley, S., Schroder, I., Shearer, A.G., Subhraveti, P., Travers, M., Weerasinghe, D., Weiss, V., Collado-Vides, J., Gunsalus, R.P., Paulsen, I., and Karp, P.D., Nucleic Acids Res., 2013, no. 41 (Database issue), pp. D605–D612.
Park, C.B., Lee, S.B., and Ryu, D.D., J. Mol. Catal., 2000, vol. 9, nos. 4–6, pp. 275–281.
Velez, A.M., Silva, A.J., Luperni, HortaA.C., Sargo, C.R., Campani, G., Goncalves, SilvaG., de Lima, Camargo., Giordano, R., and Zangirolami, T.C., BMC Biotechnol., 2014, vol. 14, no. 1, p. 6. doi:10.1186/1472-6750-14-6.
Viegas, S.C., Schmidt, D., Kasche, V., Arraiano, C.M., and Ignatova, Z., FEBS Lett., 2005, vol. 579, no. 22, pp. 5069–5073.
Xu, Y., Rosenkranz, S., Weng, C.-L., Scharer, J.M., Moo-Young, M., and Chou, C.P., Appl. Microbiol. Biotechnol., 2006, vol. 72, no. 3, pp. 529–536.
Narayanan, N., Hsieh, M.-Y., Xu, Y., and Chou, C.P., Biotechnol. Prog., 2006, vol. 22, no. 3, pp. 617–625.
Narayanan, N., Xu, Y., and Chou, C.P., Biotechnol. Prog., 2006, vol. 22, no. 6, pp. 1518–1523.
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Original Russian Text © M.A. Eldarov, A.V. Sklyarenko, A.V. Mardanov, A.V. Beletsky, A.A. Zhgun, M.V. Dumina, N.V. Medvedeva, D.E. Satarova, N.V. Ravin, S.V. Yarockii, 2015, published in Prikladnaya Biokhimiya i Mikrobiologiya, 2015, Vol. 51, No. 5, pp. 465–471.
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Eldarov, M.A., Sklyarenko, A.V., Mardanov, A.V. et al. Cephalosporin-acid synthetase of Escherichia coli strain VKPM B-10182: Genomic context, gene identification, producer strain production. Appl Biochem Microbiol 51, 505–510 (2015). https://doi.org/10.1134/S0003683815050051
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DOI: https://doi.org/10.1134/S0003683815050051