摘要
戊二酰 7 氨基头孢烷酸 (GL 7ACA)酰化酶能够催化GL 7ACA分解生成 7 ACA ,后者是工业半合成生产头孢类抗菌素所需的重要前体。为了准确地检测GL 7ACA酰化酶及其突变体的表达 ,本研究通过构建一系列质粒载体 ,建立了两个简便有效地测定GL 7ACA酰化酶基因acy表达量的系统 ,从而可对酶的比活力进行定量。我们将两个报告基因 ,即儿茶酚双加氧酶基因 (xylE)和 β 半乳糖苷酶基因 (lacZ)分别置于acy基因的下游 ,使之与acy基因共用一个启动子 ,进行串联表达 ,各自构成一个多顺反子系统。实验证明 ,基因融合后的儿茶酚双加氧酶或 β 半乳糖苷酶的活力可以间接反映acy的表达量。
Glutaryl-7-amino cephalosporanic acid (GL-7ACA)acylase catalyzes the conversion of GL-7ACA to 7-amino cephalosporanic acid (7-ACA).The product 7-ACA is a starting compound for semi-synthetic cephalosporin antibiotics in industry.In order to detect the expression and spesific activity of protein-engineered GL-7ACA acylase accurately,two useful detective systems for its expression has been established,in which reporter genes %xylE% and %lacZ% were fused to the downstream the GL-7ACA acylase gene %acy% respectively and the activity of catechol dioxygenase or β-galactosidase could indicate the amount of acy expression.
出处
《生物工程学报》
CAS
CSCD
北大核心
2001年第6期673-677,共5页
Chinese Journal of Biotechnology
基金
国家"8 63"计划资助项目 ( 10 3 13 0 2 0 1)~~
