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Periplasmic Export of Bile Salt Hydrolase in Escherichia coli by the Twin-Arginine Signal Peptides

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Abstract

Bile salt hydrolase (BSH, EC 3.5.1.24) is considered as an ideal way with lower cost and less side effects to release the risk of coronary heart disease caused by hypercholesterolemia. As bile salt hydrolase from Lactobacillus plantarum BBE7 could not be efficiently exported by PelB signal peptide of the general secretory (Sec) pathway, three twin-arginine signal peptides from twin-arginine translocation (Tat) pathway were synthesized, fused with bsh gene, inserted into expression vectors pET-20b(+) and pET-22b(+), and transformed into four different Escherichia coli hosts, respectively. Among the 24 recombinant bacteria obtained, E. coli BL21 (DE3) pLysS (pET-20b(+)-dmsA-bsh) showed the highest BSH activity in periplasmic fraction, which was further increased to 1.21 ± 0.03 U/mL by orthogonal experimental design. And, signal peptide dimethyl sulfoxide reductase subunit DmsA (DMSA) had the best activity of exported BSH. More importantly, the presence of BSH in the periplasm had proven to be caused by the export rather than cell leakage. For the first time, we report the periplasmic expression of BSH by signal peptides from the Tat pathway. This will lay a solid foundation for the purification and biochemical characterization of BSH from the supernatant, and strategies adopted here could be used for the periplasmic expression of other proteins in E. coli.

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Acknowledgments

This project was financially supported by Synergetic Innovation Center of Food Safety and Nutrition, the National Natural Science Foundation of China (No. 31100064), the National High Technology Research and Development Program of China (863 Program, No. 2011AA100905), Program for Changjiang Scholars and Innovative Research Team in University (No. IRT1135), the Major State Basic Research Development Program of China (973 Program, 2012CB720802, 2012CB720806), and the Natural Science Foundation of Jiangsu Province (No. BK2012553).

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Authors and Affiliations

  1. College of Chemical Engineering and Material Science, Tianjin University of Science & Technology, Tianjin, 300457, China

    Zixing Dong

  2. Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, China

    Juan Zhang, Guocheng Du, Jian Chen & Huazhong Li

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

    Guocheng Du & Jian Chen

  4. Department of Food Science and Biotechnology, Faculty of Agriculture and Life Sciences, Kangwon National University, Chuncheon, 200-701, South Korea

    Byonghoon Lee

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  1. Zixing Dong
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  2. Juan Zhang
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  3. Guocheng Du
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Correspondence to Juan Zhang or Byonghoon Lee.

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Dong, Z., Zhang, J., Du, G. et al. Periplasmic Export of Bile Salt Hydrolase in Escherichia coli by the Twin-Arginine Signal Peptides. Appl Biochem Biotechnol 177, 458–471 (2015). https://doi.org/10.1007/s12010-015-1755-3

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  • DOI: https://doi.org/10.1007/s12010-015-1755-3

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