Extracellular production of human cystatin S and cystatin SA by Bacillus subtilis

Shunichi Akiba, Yasuhiro Hayashi, Yoshihiro Hakamada, Keiji Endo, Katsutoshi Ara*, Shuji Kawai, Eiichi Saitoh

*この研究の対応する著者

研究成果: Article査読

3 被引用数 (Scopus)

抄録

We herein describe the development of a Bacillus subtilis system that can be used to produce large quantities of recombinant (r-) human salivary cystatins, a cysteine protease inhibitor of family 2 in the cystatin superfamily. The B. subtilis that lacked the alkaline protease E gene (ΔaprE type mutant strain) was prepared by homologous recombination. The cDNA fragments coding for mature cystatins (S and SA) were ligated in frame to the DNA segment for the signal peptide of endoglucanase in the pHSP-US plasmid vector that was then use to transform the ΔaprE type mutant strain of B. subtilis. The transformants carrying the expression vectors were cultivated in 5-L jar fermenters for 3 days at 30 °C. Both r-cystatin S and r-cystatin SA were successfully expressed and secreted into the culture broth, and were purified using a fast performance liquid chromatography system. The first use of ΔaprE type mutant strain of B. subtilis made it possible to obtain a high yield of secreted protein, which makes this system an improvement over expression in Escherichia coli. We conclude that this system has high utility for expression of commercial quantities of secreted proteins.

本文言語English
ページ(範囲)203-210
ページ数8
ジャーナルProtein Expression and Purification
49
2
DOI
出版ステータスPublished - 2006 10
外部発表はい

ASJC Scopus subject areas

  • バイオテクノロジー

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