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- Title
硫酸软骨素ABC I酶的融合表达及传感器应用研究.
- Authors
李晔; 金丽华; 李栋; 辛秀兰; 陈振娅
- Abstract
Chondroitinase (ChSase) is a class of enzymes that can degrade chondroitin sulfate (CS) to small molecular polysaccharides, which can be classified as ChSase ABC, ChSase AC, ChSase B and ChSase C according to the kind of substrates. In this study, the ChSase ABC I derived from Proteus vulgaris KCTC 2579 was integrated and expressed with maltose-binding protein (MBP). The recombinant vector pMAL-c2x-ChSase ABC I was constructed and successfully expressed in Escherichia coli BL21 (DE3). The maltose-binding protein-chondroitinase (MBP-ChSase) ABC I enzyme was immobilized on a polyaniline carrier to prepare a biosensor, which was detected by cyclic voltammetry. The results showed that the enzyme activity and specific activity of MBP-ChSase ABC I were 3 180 IU/L fermentation broth and 76 IU/mg protein, respectively. Using chondroitin sulfate A (CS A) as the reaction substrate of the enzyme, under the optimized conditions, the peak current of the reaction had a linear relationship with the concentration of CS A of 0.1 nmol/L-0.1 μmol/L. The biosensor reacted quickly to the substrate, with high sensitivity, and could perform effective detection.
- Subjects
CHONDROITIN sulfates; CYCLIC voltammetry; ESCHERICHIA coli; CHONDROITINASE; POLYSACCHARIDES
- Publication
China Brewing, 2020, Vol 39, Issue 11, p153
- ISSN
0254-5071
- Publication type
Article
- DOI
10.11882/j.issn.0254-5071.2020.11.029