黄体酮C11α,C17α位双羟化菌株的筛选及发酵优化

A screen for strains producing 11α,17α-dihydroxy progesterone and optimization of fermentation conditions

皮质激素在临床上应用广泛,其中11α,17α-二羟基黄体酮对皮质激素生产至关重要,其生物制备方法主要以17α-羟基黄体酮为底物通过微生物转化生成.而以黄体酮为底物,实现11α,17α-二羟基黄体酮的高效生产更具有经济效应.通过筛选实验室菌种库中能够转化黄体酮的菌株,发现赤霉菌CA3-1对黄体酮有转化能力,其转化产物经HPLC、MS和NMR鉴定为11α,17α-二羟基黄体酮.进一步通过单因素实验和正交实验对菌株发酵培养基组分进行优化,确定最适发酵培养基:25 g/L乳糖、15 g/L酵母粉、0.4 g/L硫酸铁、15 g/L玉米浆.当底物投料量为0.5 g/L,转化72 h时,底物转化率为82.2%,11α,17α-二羟基黄体酮的浓度为43.5 mg/L,较优化前产物浓度30.2 mg/L提高44.0%.本研究成功筛选获得可以双羟基化黄体酮的菌株,并通过单因素实验和正交实验优化发酵培养基提高了11α,17α-二羟基黄体酮的浓度,为11α,17α-二羟基黄体酮的工业生产提供了新的思路.

Corticosteroids are widely used in clinical practice.11α,17α-dihydroxy progesterone serves as a crucial intermediate in corticosteroid synthesis.The prevailing biological method for its preparation involves microbial biosynthesis using 17α-hydroxy progesterone as a substrate.However,achieving efficient production of 11α,17α-dihydroxy progesterone using progesterone as substrate is more cost-effective.We screened our laboratory library for strains capable of modifying progesterone,and found Gibberella intermedia CA3-1to proficient in utilizing progesterone.HPLC,MS,and NMR analyses confirmed that the product was indeed 11α,17α-dihydroxy progesterone.We subsequently optimized fermentation medium components through single-factor experiments and orthogonal experiments to determine the optimal composition:lactose(25 g/L),yeast powder(15 g/L),Fe2(SO4)3(0.4 g/L),and corn steep liquor(15 g/L).With a substrate feeding amount of 0.5 g/L and a 72 h fermentation time,we achieved a substrate conversion rate of 82.2%and a final concentration of 11α,17α-dihydroxy progesterone of 43.5 mg/L compared to the pre-optimization concentration of 30.2 mg/L representing a significant 44%improvement.We successfully identified a novel strain capable of hydroxylating progesterone through effective screening methods,and enhanced its yield of 11α,17α-dihydroxy progesterone by optimizing the fermentation medium using single-factor and orthogonal testing.This discovery offers a promising approach for the industrial production of 11α,17α-dihydroxy progesterone.