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将可催化合成西格列汀的转氨酶基因克隆到表达载体p GEX-6p-1上,并转入大肠杆菌E.coli中表达,获得高效表达重组转氨酶的菌株。分别考察基因在不同大肠杆菌[E.coli W3110、E.coli BL21(DE3)、E.coli Rosetta(DE3)和E.coli Rosetta-gami2(DE3)]中的表达情况,结果显示,转氨酶在大肠杆菌BL21(DE3)中的表达量较高,而且多为可溶性表达。所得重组转氨酶(粗酶)比活力也较高,为0.32 u/mg。
The transaminase gene, which can catalyze the synthesis of sitagliptin, was cloned into the expression vector pGEX-6p-1 and transformed into E.coli for expression. Recombinant transaminase was efficiently expressed. The gene expression in E. coli W3110, E. coli BL21 (DE3), E. coli Rosetta (DE3) and E. coli Rosetta-gami2 (DE3) were investigated respectively. The results showed that transaminases were expressed in the large intestine Bacillus BL21 (DE3) expression in the higher, but mostly soluble expression. The resulting recombinant aminotransferase (crude enzyme) also has a higher specific activity of 0.32 u / mg.