17α-羟基黄体酮C11α-羟化菌株的筛选及工艺优化

Screening and Fermentation Optimization of 17α-hydroxyprogesterone C11α-hydroxylation Strain

11α,17α-二羟基黄体酮(11α,17α-dihydroxy progesterone)作为甾体激素类药物的重要中间体,其生物合成主要以17α-羟基黄体酮作为底物转化生成。为探究不同微生物对17α-羟基黄体酮的转化能力,以11株具有甾体羟化能力的菌株为研究对象,通过全细胞生物转化实验,筛选得到了一株转化能力最强的菌株亚麻刺盘孢SF-307。通过单因素实验和正交设计进行菌株发酵培养基组分优化,确定了最适发酵培养基:15 g/L可溶性淀粉、1.8 g/L氯化铵、0.6 g/L氯化镁、3 g/L玉米浆。优化培养基后的亚麻刺盘孢SF-307转化产物唯一,当底物投料量为0.5 g/L,添加1%(V/V)乙醇进行助溶,转化56 h时,底物转化率为93.2%,11α,17α-二羟基黄体酮的最高浓度为224.1 mg/L,较优化前提高61.1%。上述结果表明:亚麻刺盘孢SF-307作为一株全新的17α-羟基黄体酮羟化菌株,发酵优化可显著增强17α-羟基黄体酮转化产物的选择性,缩短转化周期,对11α,17α-二羟基黄体酮的工业生产意义重大。

11α,17α-dihydroxy progesterone is an important intermediate of steroid hormone drugs and its biosynthesis is mainly produced by microbial transformation of 17α-hydroxyprogesterone.In order to explore the transformation ability of different microorganisms to 17α-hydroxyprogesterone,11 strains with steroid hydroxylation ability were selected.Through the whole-cell biotransformation experiment,Colletotrichum lini SF-307 with the strongest transformation ability was obtained.Then,the optimal composition of the fermentation medium was determined by a single-factor experiment and orthogonal design.The most suitable fermentation medium was determined:15 g/L soluble starch,1.8 g/L ammonium chloride,0.6 g/L magnesium chloride,and 3 g/L corn pulp.After optimization,the only product 11α,17α-dihydroxy progesterone was produced by C.lini SF-307.When the substrate was fed at 0.5 g/L with the addition of 1%(V/V)ethanol for co-solubilization,the substrate conversion was 93.2%and the highest concentration was 224.1 mg/L at 56 h,which was increased by 61.1%compared to the original.The results showed that C.lini SF-307 was a new strain of 17α-hydroxyprogesterone hydroxylation.The fermentation optimization can significantly enhance selectivity of 17α-hydroxyprogesterone conversion products and shorten the transformation period.This study is of great significance to the industrial production of 11α,17α-dihydroxy progesterone.