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Expression, purification and thermostability of MBP-chondroitinase ABC I from Proteus vulgaris

Chen, Zhenya, Li, Ye, Yuan, Qipeng
International journal of biological macromolecules 2015 v.72 pp. 6-10
Escherichia coli, Proteus vulgaris, chondroitin sulfate, enzyme activity, fermentation, oligosaccharides, rapid methods, thermal stability
Chondroitinase ABC I (ChSase ABC I) which can degrade chondroitin sulfate (CS) and other glycosaminoglycan to oligosaccharide or unsaturated disaccharide, was fusionally expressed with maltose-binding protein (MBP) in Escherichia coli BL21(DE3) (E. coli BL21(DE3)) and purified for the first time in this study. The result showed that the productivity of recombinant MBP-ChSase ABC I was 3180IU/(L fermentation liquor) with CS A as substrate, and the productivity might be the highest level when compared to the reported ones. The specific activity of recombinant MBP-ChSase ABC I was 76IU/(mg protein) after purification. The Vmax, Km and kcat were 18.7±0.3μmol/Ls, 73.1±4.1μmol/L and 586.7±10.8s−1, respectively. Enzyme activity of the purified enzyme remained about 78% after 210min when the enzyme incubated at 30°C.This study introduces a rapid method for highly expressing ChSase ABC I, and the method could be adopted in the process of industrial production. Furthermore the investigation of thermostability might lead to an important guide in clinical treatment.