酰胺水解酶基因工程菌的构建及其对棘白菌素B的转化

Construction of ECB deacylase genetically engineered strain and transformation of ECB

棘白菌素B_0(ECB)去侧链母核为重要抗真菌药物阿尼芬净的半合成前体。本实验室从保存的犹他游动放线菌(Actinoplanes utahensis)SIPI-A.2001基因组中克隆到ECB酰胺水解酶及其上下游基因,并将其构建入表达质粒pTGV2,通过接合转移的方法将此表达质粒导入到变铅青链霉菌(Streptomyces lividans)TK24中,构建可以水解ECB底物生成其母核的基因工程菌,静息细胞转化法生成ECB母核。HPLC检测表明,构建的基因工程菌可以有效水解ECB侧链,转化率达53.2%。

Echinocandin B0 （ECB）, a side-chain cleavaged nucleus, is a semi-synthetic precursor of significant antifungal anidulafungin. A fragment containing the ECB deacylase encoding region and its flank genes was amplified from Actinoplanes utahensis SIPI-A. 2001, and linked into the overexpression vector pTGV2. Then the plasmid was transformed into Streptornyces lizriclans TK24. This genetically engineered strain could transform ECB into ECB nucleus. HPLC analysis showed that the genetically engineered strain could effectively cleave ECB side chain, and the conversion rate was as high as 53.2%.