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Design of an expression vector and its application to heterologous protein expression in Bacillus subtilis

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Abstract

An expression vector, pUBEX, was constructed for extracellular production of heterologous proteins in Bacillus subtilis using a polyhistidine tag on the C-terminal sequence, providing an efficient and easy purification of the protein. A CII protein, a member of Bowman–Birk protease inhibitors, which was expressed as an inactive protein in Escherichia coli, was successfully expressed in Bacillus subtilis using the pUBEX vector and was purified to 6.4 mg l−1 by the immobilized metal affinity chromatography.

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References

  • Back SJ, Jung KH, Kim H, Kim SF (1997) Expression and secretion of carboxymethyl cellulose in Bacillus subtilis by Lactobacillus casei lactate dehydrogenase gene promoter. Biotechnol. Lett. 19: 27-29.

    Google Scholar 

  • Baek JM, Song JC, Choi YD, Kim SI (1994) Nucleotide sequence homology of cDNAs encoding soybean Bowman-Birk type proteinase inhibitor and is isoinhibitors. Biosci. Biotechnol. Biochem. 58: 843-846.

    Google Scholar 

  • Bellini AV, Gali G, Fascetti E, Frascotti G, Branduzzi P, Lucchese G, Grandi G (1991) Production process of recombinant IL-1 from Bacillus subtilis: comparison between intracellular and extracellular expression. J. Biotechnol. 18: 177-192.

    Google Scholar 

  • Kyhse-Andersen J (1984) Electroblotting of multiple gels: a simple apparatus without buffer tank for rapid transfer of protein from polyacrylamide to nitrocellulose. J. Biochem. Biophys. Meth. 10: 203-209.

    Google Scholar 

  • Lam KHE, Chow KC, Wong WKR (1998) Construction of an ef-ficient Bacillus subtilis system for extracellular production of heterologous proteins. J. Biotechnol. 63: 167-177.

    Google Scholar 

  • Sadaie Y, Kada T (1983) Formation of competent Bacillus subtilis cells. J. Bacteriol. 153: 813-821.

    Google Scholar 

  • Wong SL (1995) Advances in the use of Bacillus subtilis for the expression and secretion of heterologous proteins. Curr. Opin. Biotechnol. 6: 517-522.

    Google Scholar 

  • Wu SC, Yeung JC, Hwang PM, Wong SL (2002) Design, production, and characterization of an engineered biotin ligase (BirA) and its application for affinity purification of staphylokinase pro1874 duced from Bacillus subtilis via secretion. Protein. Expr. Purif. 24: 357-365.

    Google Scholar 

  • Wu XC, Lee W, Tran L, Wong SL (1991) Engineering a Bacillus subtilis expression-secretion system with a strain deficient in six extracellular proteases. J. Bacteriol. 173: 4952-4958.

    Google Scholar 

  • Wu XC, Ng SC, Near RI, Wong SL (1993) Efficient production of a functional single-chain antidigoxin antibody fragment via an engineered Bacillus subtilis expression-secretion system. Biotechnology 11: 71-76.

    Google Scholar 

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So, MY., Pan, CH., Rhee, S. et al. Design of an expression vector and its application to heterologous protein expression in Bacillus subtilis . Biotechnology Letters 24, 1869–1874 (2002). https://doi.org/10.1023/A:1020909619931

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  • DOI: https://doi.org/10.1023/A:1020909619931

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