摘要
penDE基因编码40k的酰基-CoA:6-APA酰基转移酶(IAT),可催化产黄青霉中青霉素生物合成途径的最后一步酶反应。利用RT-PCR法从产黄青霉中扩增得1.1kbp的不含内含子的penDE基因,并将其克隆至大肠杆菌-链霉菌穿梭质粒pUWL201,使penDE基因位于ermEp*启动子的下游,再将其转化带小棒链霉菌,获得了能够表达IAT的转化子。PDAB法和抗菌活性测定表明,所表达的IAT蛋白对6-氨基青霉烷酸和苯乙酰CoA有明显的活性。
Acyl-CoA：6-aminopenicillanic acid acyltransferase （AAT）, encoded by penDE, catalyze the final step of the penicillin biosynthesis pathway in Penicillium chrysogenum. The 1.1kbp intronless penDE was amplified from P. chrysogenum total RNA by RT-PCR, and cloned into a shuttle vector pUWL201. The recombinant plasmid in which penDE was then located immediately downstream from the ermEp＊ promoter, was then transformed into S. clavuligerus and successfully produced the AAT. The PDAB and antimierobial assay indicated that the AAT possessed high activities with 6-APA and phenylacyl-CoA as the substrates.
出处
《中国医药工业杂志》
CAS
CSCD
北大核心
2007年第1期15-18,共4页
Chinese Journal of Pharmaceuticals
