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Cloning and expression of variants of the glutaryl-7-aminocephalosporic acid acylase of the bacterium Brevundimonas diminuta in Escherichia coli cells

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Abstract

The gene coding for glutaryl-7-aminocephalosporic acid acylase (Gl7ACA acylase) of the bacterium Brevundimonas diminuta (BrdGl7ACA), a commercial enzyme widely used in modern biocatalytic technologies for manufacture of β-lactam antibiotics, was cloned. Efficient expression systems for producing a “native” recombinant BrdGl7ACA and its analogs modified by attaching affinity groups—the chitin-binding domain of chitinases A1 and hexahistidine sequence—were designed. It was demonstrated that both the recombinant hybrid proteins and the native Gl7ACA acylase produced in E. coli cells underwent a correct autoproteolytic processing with generation of functionally active enzymes and could be isolated with a high yield using one-step affinity chromatography.

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Authors and Affiliations

  1. Bioengineering Center, Russian Academy of Sciences, Moscow, 117312, Russia

    S. A. Khatuntseva, M. A. El’darov, S. A. Lopatin, O. A. Zeinalov & K. G. Skryabin

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  1. S. A. Khatuntseva
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  2. M. A. El’darov
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  3. S. A. Lopatin
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  4. O. A. Zeinalov
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  5. K. G. Skryabin
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Original Russian Text © S.A. Khatuntseva, M.A. El’darov, S.A. Lopatin, O.A. Zeinalov, K.G. Skryabin, 2007, published in Prikladnaya Biokhimiya i Mikrobiologiya 2007, Vol. 43, No. 4, pp. 462–470.

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Khatuntseva, S.A., El’darov, M.A., Lopatin, S.A. et al. Cloning and expression of variants of the glutaryl-7-aminocephalosporic acid acylase of the bacterium Brevundimonas diminuta in Escherichia coli cells. Appl Biochem Microbiol 43, 414–421 (2007). https://doi.org/10.1134/S0003683807040102

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  • DOI: https://doi.org/10.1134/S0003683807040102

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