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Surface display of the thermophilic lipase Tm1350 on the spore of Bacillus subtilis by the CotB anchor protein

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Abstract

Lipases expressed in microbial hosts have great commercial value, but their applications are restricted by the high costs of production and harsh conditions used in industrial processes, such as high temperature and alkaline environment. In this study, an Escherichia coli–Bacillus subtilis shuttle vector (pHS-cotB-Tm1350) was constructed for the spore surface display of the lipase Tm1350 from hyperthermophilic bacterium Thermotoga maritima MSB8. Successful display of the CotB-Tm1350 fusion protein on spore surface was confirmed by Western blot analysis and activity measurements. The optimal catalytic temperature and pH of the spore surface-displayed Tm1350 were 80 °C and 9, respectively, which were higher than non-immobilized Tm1350 (70 °C and pH 7.5). Analysis of thermal and pH stability showed that spore surface-displayed Tm1350 retained 81 or 70 % of its original activity after 8 h of incubation at pH 8 or pH 9 (70 °C), which were 18 % higher than the retained activity of the non-immobilized Tm1350 under the same conditions. Meanwhile, recycling experiments showed that the recombinant spores could be used for up to three reaction cycles without a significant decrease in the catalytic rate (84 %). These results suggested that enzyme display on the surface of the B. subtilis spore could serve as an effective approach for enzyme immobilization, which has potential applications in the harsh biochemical industry.

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Acknowledgments

This study was supported by the National Key Basic Research Program of China (973 Program, No. 2011CBA00800), the Open Funding Project of National Key Laboratory of Biochemical Engineering, and the Key Agriculture Support Project of Jiangsu Province, China (No. BE2013400).

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Authors and Affiliations

  1. Institute of Life Sciences, Jiangsu University, Zhenjiang, 212013, China

    Huayou Chen, Rui Tian, Zhong Ni, Qing Zhang, Tianxi Zhang, Zhi Chen, Keping Chen & Shengli Yang

  2. National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing, 10090, China

    Huayou Chen

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  1. Huayou Chen
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  2. Rui Tian
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  3. Zhong Ni
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  4. Qing Zhang
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  5. Tianxi Zhang
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  6. Zhi Chen
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  7. Keping Chen
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  8. Shengli Yang
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Correspondence to Huayou Chen.

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Communicated by F. Robb.

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Chen, H., Tian, R., Ni, Z. et al. Surface display of the thermophilic lipase Tm1350 on the spore of Bacillus subtilis by the CotB anchor protein. Extremophiles 19, 799–808 (2015). https://doi.org/10.1007/s00792-015-0755-0

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  • DOI: https://doi.org/10.1007/s00792-015-0755-0

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