Biotechnology and Bioprocess Engineering:BBE

The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
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Optimization of Staphylokinase Production in Bacillus subtilis Using Inducible and Constitutive Promoters

  • Kim, June-Hyung (School of Chemical Engineering and The Institute of Molecular Biology and Genetics, Seoul National University) ;
  • Wong, Sui-Lam (Department of Biological Sciences, Division of Cellular and Microbial biology, University of Calgary) ;
  • Kim, Byung-Gee (School of Chemical Engineering and The Institute of Molecular Biology and Genetics, Seoul National University)
  • Published : 2001.05.01
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Abstract

Staphylokinase (SAK) was produced in B. subtilis using two different promoter systems, i.e. the P43 and sacB promoters. To maximize SAK expression in B. subtilis, fermentation control strategies for each promoter were examined. SAK, under P43, a vegetative promoter transcribed mainly by $\sigma$(sup)B containing RNA polymerase, was overexpressed at low dissolved oxygen (D.O.) levels, suggesting that the sigB operon is somewhat affected by the energy charge of the cells. The expression of SAK at the 10% D.O. level was three times higher than that at the 50% D.O. level. In the case of sacB, a sucrose-inducible promoter, sucrose feeding was used to control the induction period and induction strength. Since sucrose is hydrolyzed by two sucrose hydrolyzing enzymes in the cell and culture broth, the control strategy was based on replenishing the loss of sucrose in the culture. With continuous feeding of sucrose, WB700 (pSAKBQ), which contains the SAK gene under sacB promoter, yielded ca. 35% more SAK than the batch culture. These results present efficient promoter-dependent control strategies in B. subtilis host system for foreign protein expression.

Keywords

References

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