Overexpression, purification and characterization of a recombinant secretary catalase from Bacillus subtilis

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• 作者：Xunlong Shi ; Meiqing Feng ; Yujie Zhao ; Xin Guo and Pei Zhou
• 关键词：Bacillus subtilis ; Catalase ; Expression ; katA ; Secretary
• 刊名：Biotechnology Letters
• 出版年：2008
• 出版时间：January, 2008
• 年：2008
• 卷：30
• 期：1
• 页码：181-186
• 全文大小：259.9 KB
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Life Sciences
  Microbiology
  Biotechnology
  Applied Microbiology
  Biochemistry
  
• 出版者：Springer Netherlands
• ISSN：1573-6776

文摘

A recombinant Bacillus subtilis strain (KN25) was generated for the large-scale preparation of catalase. The B. subtilis katA gene encoding for catalase was cloned into the shuttle vector PRB374, downstream of the constitutively active vegII promoter, followed by transformation of the B. subtilis strain WB600 with the plasmid. The transformant strain, KN25 secretes high levels (3,500 U/ml) of catalase, which facilitates its purification. Three simple purification steps yielded nearly homogeneous catalase, with ∼70% recovery. The purified recombinant catalase has a specific activity of 34,600 U/mg under optimal conditions, and is more resistant to acidic conditions than bovine liver catalase.