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Halophilic β-lactamase as a new solubility- and folding-enhancing tag protein: production of native human interleukin 1α and human neutrophil α-defensin

  • Applied Genetics and Molecular Biotechnology
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Abstract

The amino acid composition of halophilic enzymes is characterized by an abundant content of acidic amino acid, which confers to the halophilic enzymes extensive negative charges at neutral pH and high aqueous solubility. This negative charge prevents protein aggregation when denatured and thereby leads to highly efficient protein refolding. β-Lactamase from periplasmic space of moderate halophile (BLA), a typical halophilic enzyme, can be readily expressed as a native, active form in Escherichia coli cytoplasm. Similar to other halophilic enzymes, BLA is soluble upon denaturation by heat or urea treatments and, hence, can be efficiently refolded. Such high solubility and refolding efficiency make BLA a potential fusion partner for expression of aggregation-prone heterologous proteins to be expressed in E. coli. Here, we succeeded in the soluble expression of several “difficult-to-express” proteins as a BLA fusion protein and verified biological activities of human interleukin 1α and human neutrophil α-defensin, HNP-1.

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Acknowledgements

This study was supported by the Salt Science Research Foundation (Program for Research on Halophilic Organisms), and Grant in Aid for Science Research (20580372) from MEXT Japan.

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Authors and Affiliations

  1. Applied and Molecular Microbiology, Faculty of Agriculture, Kagoshima University, 1-21-24 Korimoto, Kagoshima, 890-0065, Japan

    Hiroko Tokunaga, Shoko Saito, Kazuki Sakai, Rui Yamaguchi & Masao Tokunaga

  2. Epoch Medical International Inc. (EMI), K-I build., 1-6-9 Hiranomachi, Chuo-ku, Osaka, 541-0046, Japan

    Shoko Saito & Iwao Katsuyama

  3. Department of Molecular Health Sciences, Graduate School of Pharmaceutical Sciences, Nagoya City University, Nagoya, 467-8603, Japan

    Tomohiro Arakawa & Kikuo Onozaki

  4. Alliance Protein Laboratory, 3957 Corte Cancion, Thousand Oaks, CA, 91360, USA

    Tsutomu Arakawa

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  1. Hiroko Tokunaga
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  2. Shoko Saito
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  3. Kazuki Sakai
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  4. Rui Yamaguchi
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  5. Iwao Katsuyama
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  6. Tomohiro Arakawa
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  8. Tsutomu Arakawa
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  9. Masao Tokunaga
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Correspondence to Masao Tokunaga.

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Tokunaga, H., Saito, S., Sakai, K. et al. Halophilic β-lactamase as a new solubility- and folding-enhancing tag protein: production of native human interleukin 1α and human neutrophil α-defensin. Appl Microbiol Biotechnol 86, 649–658 (2010). https://doi.org/10.1007/s00253-009-2325-9

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  • DOI: https://doi.org/10.1007/s00253-009-2325-9

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