不同连接肽在硫酸软骨素酶 ABC I重组表达中的应用研究

硫酸软骨素酶ABC I（ChSase ABC I）是一类能够将硫酸软骨素、软骨素、透明质酸等糖胺多糖降解为寡糖及不饱和二糖的裂解酶。该研究将硫酸软骨素酶ABC I基因与麦芽糖结合蛋白（MBP）基因分别用FFFFF和RRRRR两种连接肽连接,并克隆到pMAL-c2X载体上,在大肠杆菌（Escherichia coli）BL21(DE3)高效表达。结果表明,重组酶MBP-FFFFF-ChSase ABC I的酶活和比酶活分别为716.5 IU/L发酵液和1.15 IU/mg蛋白,重组酶MBP-RRRRR-ChSase ABC I的酶活和比酶活分别为741.0 IU/L发酵液和1.13 IU/mg蛋白。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）结果表明,重组酶MBP-FFFFF-ChSase ABC I和MBP-RRRRR-ChSase ABC I的分子质量均为130 kDa,并且都为可溶性蛋白。

Application of different linker peptides in Chondroitinase ABC I recombinant expression

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.