不同连接肽在硫酸软骨素酶 ABC I重组表达中的应用研究.

李晔; 陈振娅; 王瑞丽; 沈荣; 危晴; 陈亮

Chondroitinase ABC I (ChSase ABC I) is a kind of lyases that can degrade glycosaminoglycans such as chondroitin sulfate, chondroitin and hyaluronic acid into oligosaccharides and unsaturated disaccharides. The ChSase ABC I gene was linked to maltose binging protein (MBP) gene using two linker peptides including FFFFF and RRRRR, respectively, cloned into pMAL-c2X vector, and successfully expressed in Escherichia coli BL21 (DE3). The results showed that the enzymatic activity and specific enzyme activity of the recombinase MBP-FFFFF-ChSase ABC I were 716.5 IU/L fermentation broth and 1.15 IU/mg protein, respectively, and the recombinase MBP-RRRRR-ChSase ABC I were 741.0 IU/L fermentation broth and 1.13 IU/mg protein, respectively. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) results showed that the molecular weights of the recombinases MBP-FFFFF-ChSase ABC I and MBP-RRRRR-ChSase ABC I were 130 kDa, and they were soluble proteins.

MOLECULAR weights; CHONDROITIN sulfates; LYASES; DISACCHARIDES; GENETIC vectors

China Brewing, 2020, Vol 39, Issue 9, p121

0254-5071

Academic Journal

10.11882/j.issn.0254-5071.2020.09.023