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Comparative Analysis of Bacterial Expression Systems for Keratinase Production

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Abstract

To explore a better expression system for the production of keratinase, the keratinase gene from Bacillus licheniformis BBE11-1 was expressed in Escherichia coli, Bacillus subtilis, and Pichia pastoris. The corresponding recombinant keratinases were named ker E, ker B, and ker P, respectively. All recombinant keratinases had an optimal pH at 10 although the pH stability of ker E and ker P was higher than that of ker E. The optimal temperature and thermostability of ker P were enhanced compared with those of ker E and ker B. The recombinant keratinases were inhibited by Mn2+ but experienced little influence from other metal ions. Furthermore, all recombinant keratinases could retain almost 80 % activity after treatment with 1 M hydrogen peroxide for 5 h. Under optimized conditions in a 3-L fermenter, the maximum keratinase activities obtained from recombinant B. subtilis and P. pastoris were 3,010 and 1,050 U/mL, respectively. This maximum keratinase activity from B. subtilis is the highest activity ever reported for any bacterial strain. These results indicate that B. subtilis is the ideal host for keratinase production, with potential applications in textile processing and feed supplements.

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Acknowledgments

This project was financially supported by the National High Technology Research and Development Program of China (863 Program, 2011AA100905), the Program for Changjiang Scholars and Innovative Research Team in University (No. IRT1135), the China Postdoctoral Science Foundation (2013M540538),the 111 project (111-2-06), and the Natural Science Foundation of Jiangsu Province (BK2012553).

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Authors and Affiliations

  1. Synergetic Innovation Center of Food Safety and Nutrition, 214122, Wuxi, China

    Baihong Liu & Lei Gu

  2. Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, 214122, Wuxi, China

    Juan Zhang & Xiangru Liao

  3. National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, 214122, Wuxi, China

    Jian Chen

  4. The Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, Jiangnan University, 214122, Wuxi, China

    Guocheng Du

  5. School of Biotechnology, Jiangnan University, 214122, Wuxi, China

    Baihong Liu, Juan Zhang, Lei Gu, Guocheng Du, Jian Chen & Xiangru Liao

Authors
  1. Baihong Liu
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  2. Juan Zhang
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  3. Lei Gu
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  4. Guocheng Du
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  5. Jian Chen
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  6. Xiangru Liao
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Correspondence to Juan Zhang or Xiangru Liao.

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Liu, B., Zhang, J., Gu, L. et al. Comparative Analysis of Bacterial Expression Systems for Keratinase Production. Appl Biochem Biotechnol 173, 1222–1235 (2014). https://doi.org/10.1007/s12010-014-0925-z

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  • DOI: https://doi.org/10.1007/s12010-014-0925-z

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