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Molecular chaperones facilitate the soluble expression of N-acyl-d-amino acid amidohydrolases in Escherichia coli

  • Original Paper
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Journal of Industrial Microbiology and Biotechnology

Abstract

The overproduction of d-aminoacylase (d-ANase, 233.8 U/mg), N-acyl-d-glutamate amidohydrolase (d-AGase, 38.1 U/mg) or N-acyl-d-aspartate amidohydrolase (d-AAase, 6.2 U/mg) in Escherichia coli is accompanied by aggregation of the overproduced protein. To facilitate the expression of active enzymes, the molecular chaperones GroEL-GroES (GroELS), DnaK-DnaJ-GrpE (DnaKJE), trigger factor (TF), GroELS and DnaKJE or GroELS and TF were coexpressed with the enzymes. d-ANase (313.3 U/mg) and d-AGase (95.8 U/mg) were overproduced in an active form at levels 1.3- and 1.8-fold higher, respectively, upon co-expression of GroELS and TF. An E. coli strain expressing the d-AAase gene simultaneously with the TF gene exhibited a 4.3-fold enhancement in d-AAase activity (32.0 U/mg) compared with control E. coli expressing the d-AAase gene alone.

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Authors and Affiliations

  1. Department of Applied Chemistry, Faculty of Engineering, Oita University, Dannoharu, Oita, 870-1192, Japan

    Kazuaki Yoshimune, Yoko Ninomiya & Mitsuaki Moriguchi

  2. Department of Bioscience and Biotechnology, Faculty of Science and Engineering, Ritsumeikan University, Nojihigashi, Kusatsu, 525-8577, Japan

    Mamoru Wakayama

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  1. Kazuaki Yoshimune
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  2. Yoko Ninomiya
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  3. Mamoru Wakayama
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  4. Mitsuaki Moriguchi
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Correspondence to Mitsuaki Moriguchi.

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Yoshimune, K., Ninomiya, Y., Wakayama, M. et al. Molecular chaperones facilitate the soluble expression of N-acyl-d-amino acid amidohydrolases in Escherichia coli. J IND MICROBIOL BIOTECHNOL 31, 421–426 (2004). https://doi.org/10.1007/s10295-004-0163-4

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  • DOI: https://doi.org/10.1007/s10295-004-0163-4

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