Overexpression, purification and characterization of a recombinant secretary catalase from Bacillus subtilis.
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 approximately 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.
Shi
Xunlong
X
Department of Drug Biosynthesis, School of Pharmacy, Fudan University, Shanghai 200032, PR China.
Feng
Meiqing
M
Zhao
Yujie
Y
Guo
Xin
X
Zhou
Pei
P
D001412:Bacillus subtilis / Q000502:physiology*; D002374:Catalase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D004789:Enzyme Activation; D004795:Enzyme Stability; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism
D011994:Recombinant Proteins; D002374:Catalase