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Cloning, sequencing, expression and characterization of the manganese superoxide dismutase gene from Vibrio alginolyticus
Author(s) -
Shyu YuChiau,
Chiu ChiChien,
Lin FuPang
Publication year - 1999
Publication title -
iubmb life
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.132
H-Index - 113
eISSN - 1521-6551
pISSN - 1521-6543
DOI - 10.1080/15216549900201893
Subject(s) - vibrio alginolyticus , escherichia coli , superoxide dismutase , open reading frame , biochemistry , microbiology and biotechnology , gene , peptide sequence , amino acid , molecular cloning , recombinant dna , biology , chemistry , cloning (programming) , dismutase , homology (biology) , expression vector , enzyme , vibrio , bacteria , genetics , computer science , programming language
The sodA gene coding for manganese superoxide dismutase from the marine microorganism Vibrio alginolyticus was cloned, sequenced and over‐expressed in Escherichia coli using the pET20b (+) expression vector. The full‐length gene was consisted of 603bp open reading frame, which encoded a polypeptide of 201 amino acid residues, with a calculated molecular weight of 22672Da. The deduced amino acid sequence of the sodA showed considerable homology to other Mn‐SODs. The recombinant enzyme was efficiently purified from crude E. coli cell lysate by the metal ion affinity chromatography. The recombinant VAMn‐SOD resisted thermo‐denaturation up to 60°C and was insensitive to inhibitors such as H2O2, NaN3 and diethyldithiocarbamic acid.