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Novel strategy for expression of authentic and bioactive human basic fibroblast growth factor in Bacillus subtilis

  • Applied genetics and molecular biotechnology
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Abstract

Inteins, also known as “protein introns,” have been found to be present in many microbial species and widely employed for the expression and purification of recombinant proteins in Escherichia coli. However, interestingly, until now there has not been much information on the identification and application of inteins to protein expression in Bacillus subtilis. In this article, for the first time, despite the likelihood of absence of inteins in B. subtilis, this bacterium was shown to be able to facilitate auto-catalytic cleavages of fusions formed between inteins and recombinant proteins. Employing a construct expressing the intein, Ssp DnaB, (DnaB), which was fused at its N-terminus with the cellulose-binding domain (CellBD) of an endoglucanase encoded by the cenA gene of Cellulomonas fimi, the construct was demonstrated to be capable of mediating intracellular expression of basic fibroblast growth factor (bFGF), followed by auto-processing of the CellBD-DnaB-bFGF fusion to result in bFGF possessing the 146-residue authentic structure. The mentioned fusion was shown to result in a high yield of 84 mg l−1 of biologically active bFGF. Future work in improving the growth of B. subtilis may enable the use of this bacterium, working in cooperation with inteins, to result in a new platform for efficient expression of valuable proteins.

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Acknowledgements

We thank Biosciences Central Research Facility at HKUST for their mass spectrometry services.

Funding

This study was funded by RGC project: GRF16101515 and Research Contracts: 13142580CLIL07W011 and 1617219-0.

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Authors and Affiliations

  1. Division of Life Science, The Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong, China

    Xiuhua Hu, Cheuk Yin Nelson Lai, T. Sivakumar, Hao Wang, K. L. Ng, C. C. Lam & W. K. R. Wong

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  1. Xiuhua Hu
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  2. Cheuk Yin Nelson Lai
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  3. T. Sivakumar
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  4. Hao Wang
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  5. K. L. Ng
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  6. C. C. Lam
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  7. W. K. R. Wong
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Correspondence to W. K. R. Wong.

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Hu, X., Lai, C.Y.N., Sivakumar, T. et al. Novel strategy for expression of authentic and bioactive human basic fibroblast growth factor in Bacillus subtilis. Appl Microbiol Biotechnol 102, 7061–7069 (2018). https://doi.org/10.1007/s00253-018-9176-1

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