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Secretory expression of the rat aryl sulfotransferases IV with improved catalytic efficiency by molecular engineering.

3 Biotech. 2019; 
ZhouZhengxiong,LiQing,XuRuirui,WangBingbing,DuGuocheng,Kang
Products/Services Used Details Operation
Bacterial Expression System … Codon-optimized rat AST IV gene for expression in E. coli was synthesized by Genscript (Nanjing, Jiangsu, China), and inserted into pColdIII plasmid (Takara Bio Inc., Shiga, Japan) after digestion with BamHI and XhoI to generate pColdIII-AST IV … Get A Quote

摘要

The rat aryl sulfotransferases IV (AST IV) has been used to catalyze 3'-phosphoadenosine-5'-phosphate (PAP) into the sulfuryl group donor 3'-phosphoadenosine-5'-phosphosulfate (PAPS) in biotechnological production of glycosaminoglycans. The performance of native AST IV is not satisfying due to the lower catalytic activity with substrate PAP. In the present study, we achieved secretory expression of the AST IV and improved its catalytic efficiency by molecular engineering. Fusion with signal peptides Cex, YebF and PelB allow for secretory expression of AST IV and the secreted AST IV yield reached 4.21 ± 0.23 U/mL, 8.67 ± 0.34 U/mL and 21.35 ± 0.87 U/mL, respectively. Modification of Pel... More

关键词

AST IV,Glycosaminoglycans,Molecular evolution,PAPS,Sulfa