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Identification of new secreted proteins and secretion of heterologous amylase by C. glutamicum

  • Applied Genetics and Molecular Biotechnology
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Abstract

In this study, secreted Corynebacterium glutamicum proteins were investigated by two-dimensional gel electrophoresis. Around 100 spots observed in the pH range 4.5–5.5 had molecular masses that varied from 10 to 50 kDa. Upon N-terminal amino acid sequence analysis by Edman degradation, two of them were hits to two hypothetical proteins encoded by cgR_1176 and cgR_2070 on C. glutamicum R genome, respectively. Active-form α-amylase derived from Geobacillus stearothermophilus was successfully secreted by using the predicted cgR_1176 and cgR_2070 signal sequences, indicating that these hypothetical proteins were secreted proteins. Analysis using a disruption mutant of the twin-arginine translocation (Tat) export pathway machinery of C. glutamicum suggested that one is Tat pathway dependent secretion while the other is independent of the pathway. Our results demonstrate that C. glutamicum can secrete exoproteins by using its own signal sequences, indicating its potential as a host for protein productions.

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Acknowledgments

We wish to thank Dr. C. Omumasaba (internal) for critical reading of the manuscript and helpful comments. This study was funded by New Energy and Industrial Technology Development Organization (NEDO).

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Authors and Affiliations

  1. Molecular Microbiology and Biotechnology Group, Research Institute of Innovative Technology for the Earth (RITE), 9-2, Kizugawadai, Kizugawa City, Kyoto, 619-0292, Japan

    Nobuaki Suzuki, Keiro Watanabe, Naoko Okibe, Yoshiki Tsuchida, Masayuki Inui & Hideaki Yukawa

  2. Honda R&D Co., Ltd., 1-4-1 Chuo Wako-shi, Saitama, 351-0193, Japan

    Yoshiki Tsuchida

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  1. Nobuaki Suzuki
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  2. Keiro Watanabe
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  3. Naoko Okibe
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  4. Yoshiki Tsuchida
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  5. Masayuki Inui
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  6. Hideaki Yukawa
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Correspondence to Hideaki Yukawa.

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Suzuki, N., Watanabe, K., Okibe, N. et al. Identification of new secreted proteins and secretion of heterologous amylase by C. glutamicum . Appl Microbiol Biotechnol 82, 491–500 (2009). https://doi.org/10.1007/s00253-008-1786-6

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  • DOI: https://doi.org/10.1007/s00253-008-1786-6

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