一株葛根素羟基化真菌的筛选及鉴定

Screening and identification of a fungal strain functioning in hydroxylation of puerarin

【目的】葛根素是一种常用的抗心血管药物,主要从葛根中提取,本文从土壤中分离转化葛根素的菌株,并对转化产物进行了鉴定。【方法】用马铃薯培养基从土壤中分离真菌,采用静息细胞转化法,通过高效液相色谱法检测转化效果,用有机溶剂萃取法和半制备高效液相色谱分离纯化得到转化产物,质谱和核磁共振分析转化产物。【结果】从土壤中分离到186株真菌,其中1株具有较高的转化活性,命名为NT-01,经形态学和ITS序列系统发育分析鉴定为粘帚霉菌(Gliocladium sp.)。质谱分析转化产物分子离子峰为433,而底物分子离子峰为417,表明转化产物为葛根素羟基化物,核磁共振分析羟基位于葛根素B-环的3′-位碳原子上。【结论】真菌NT-01菌株能转化葛根素形成3′-羟基葛根素。

[Objective] Puerarin is a Chinese herbal medicine. We isolated a fungus from soil which could convert puerarin to 3＇- hydroxypuerarin. [ Methods] Strains were cultivated on potato dextrose agar（PDA）. The biotransformation product was extracted with an organic solvent and detected on high performance liquid chromatography（HPLC） . [ Results] A total of 186 fungal strains were isolated from soil, and strain NT-01 was determined to transform puerarin into 3＇-bydroxypuerarin. The product was identified as 3＇-hydroxypuerarin by MS and 1H-NMR and 13 C-NMR. The morphology characteristic of strain NT-01 was consistent with Gliocladium deliguescens. Phylogenic analysis on sequences showed that strain NT-01 was clustered together with G. deliguescens and G. viride with high boostrap supporting, so that strain NT-01 was identified as Gliocladium sp. [ Conclusion] Gliocladium strain NT-01 can convert puerarin to 3＇-hydroxypuerarin.