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Enhanced production of heterologous proteins via engineering the cell surface of Bacillus licheniformis

  • Genetics and Molecular Biology of Industrial Organisms - Original Paper
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

Cell surface engineering was proven as the efficient strategy for enhanced production of target metabolites. In this study, we want to improve the yield of target protein by engineering cell surface in Bacillus licheniformis. First, our results confirmed that deletions of d-alanyl-lipoteichoic acid synthetase gene dltD, cardiolipin synthase gene clsA and CDP-diacylglycerol-serine O-phosphatidyltransferase gene pssA were not conducive to cell growth, and the biomass of gene deletion strains were, respectively, decreased by 10.54 ± 1.43%, 14.17 ± 1.51%, and 17.55 ± 1.28%, while the concentrations of total extracellular proteins were improved, due to the increases of cell surface net negative charge and cell membrane permeability. In addition, the activities of target proteins, nattokinase, and α-amylase were also improved significantly in gene deletion strains. Furthermore, the triplicate gene (dltD, clsA, and pssA) deletion strain was constructed, which further led to the 45.71 ± 2.43% increase of cell surface net negative charge and 26.45 ± 2.31% increase of cell membrane permeability, and the activities of nattokinase and α-amylase reached 37.15 ± 0.89 FU/mL and 305.3 ± 8.4 U/mL, increased by 46.09 ± 3.51% and 96.34 ± 7.24%, respectively. Taken together, our results confirmed that cell surface engineering via deleting dltD, clsA, and pssA is an efficient strategy for enhanced production of target proteins, and this research provided a promising host strain of B. licheniformis for efficient protein expression.

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Acknowledgements

This study was funded by the National Key Research and Development Program of China (2018YFA0900303, 2015CB150505), the Technical Innovation Special Fund of Hubei Province (No. 2018ACA149), China Postdoctoral Science Foundation (2018M642802), and the Science and Technology Project of Hubei Tobacco Company (027Y2019-018).

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Authors and Affiliations

  1. State Key Laboratory of Biocatalysis and Enzyme Engineering, Environmental Microbial Technology Center of Hubei Province, College of Life Sciences, Hubei University, 368 Youyi Avenue, Wuchang District, Wuhan, 430062, Hubei, People’s Republic of China

    Fei Mo, Dongbo Cai, Penghui He, Fan Yang, Yaozhong Chen, Xin Ma & Shouwen Chen

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  1. Fei Mo
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  6. Xin Ma
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Contributions

XM and SC designed and supervised the study. FM, DC, PH, FY, and YC performed the experiments. FM, DC, PH, XM, and SC analyzed the data and wrote the manuscript. All authors read and approved the final manuscript.

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Correspondence to Xin Ma or Shouwen Chen.

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Mo, F., Cai, D., He, P. et al. Enhanced production of heterologous proteins via engineering the cell surface of Bacillus licheniformis. J Ind Microbiol Biotechnol 46, 1745–1755 (2019). https://doi.org/10.1007/s10295-019-02229-8

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