Abstract 3β,7α,15α-trihydroxy-5-androsten-17-one (7α,15α-diOH-DHEA), a key precursor of the novel oral contraceptive “Yasmin”, can be obtained by dihydroxylation in the 7α and 15α positions of dehydroepiandrosterone (DHEA) by Colletotrichum lini CGMCC 6051. However, the commercial-scale application of 7α,15α-diOH-DHEA is limited by the poor dihydroxylation efficiency and its low yield. This research investigated different organic solvents to improve the dihydroxylation efficiency of DHEA by C. lini CGMCC 6051. Results showed that ethylene glycol was the optimum organic solvent. With the addition of 0.5% (v/v) ethylene glycol, the molar yield of 7α,15α-diOH-DHEA increased from 26.0% to 79.4% at 12 g/L DHEA, and that of the intermediate product 7α-OH-DHEA decreased to 10.5%. We then investigated the related functional mechanism of ethylene glycol on DHEA bioconversion. Results of two-dimensional electrophoresis proteomics analysis demonstrated that the expression levels of cytochrome P450 and 6-phosphogluconate dehydrogenase were increased upon the addition of ethylene glycol. Further studies suggested that ethylene glycol played critical roles in stabilizing the pH of the transformation system and generating co-enzyme NADPH during DHEA bioconversion. Hence, the addition of ethylene glycol induced the expression level of the key hydroxylase cytochrome P450 and increased the concentration of NADPH, which aided the dihydroxylation of DHEA in C. lini CGMCC6051.

中文翻译：

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乙二醇对炭疽菌DHEA二羟基化的影响：增加细胞色素P450和6-磷酸葡萄糖酸脱氢酶的表达，促进NADPH的产生

摘要 3β,7α,15α-trihydroxy-5-androsten-17-one (7α,15α-diOH-DHEA) 是新型口服避孕药“Yasmin”的关键前体，可通过其 7α 和 15α 位的二羟基化获得。脱氢表雄酮 (DHEA) by Colletotrichum lini CGMCC 6051。然而，7α,15α-diOH-DHEA 的商业规模应用受到二羟基化效率差和产率低的限制。本研究研究了不同的有机溶剂，以提高 C. lini CGMCC 6051 对 DHEA 的二羟基化效率。结果表明，乙二醇是最佳的有机溶剂。添加0.5% (v/v)乙二醇后，12 g/L DHEA时7α,15α-diOH-DHEA的摩尔收率从26.0%增加到79.4%，中间产物7α-OH-DHEA的摩尔收率从26.0%增加到79.4%下降到 10.5%。然后我们研究了乙二醇对 DHEA 生物转化的相关功能机制。二维电泳蛋白质组学分析结果表明，加入乙二醇后细胞色素P450和6-磷酸葡萄糖酸脱氢酶的表达水平增加。进一步的研究表明，乙二醇在稳定转化系统的 pH 值和在 DHEA 生物转化过程中产生辅酶 NADPH 方面发挥了关键作用。因此，乙二醇的添加诱导了关键羟化酶细胞色素 P450 的表达水平并增加了 NADPH 的浓度，这有助于 C.lini CGMCC6051 中 DHEA 的二羟基化。二维电泳蛋白质组学分析结果表明，加入乙二醇后细胞色素P450和6-磷酸葡萄糖酸脱氢酶的表达水平增加。进一步的研究表明，乙二醇在稳定转化系统的 pH 值和在 DHEA 生物转化过程中产生辅酶 NADPH 方面发挥了关键作用。因此，乙二醇的添加诱导了关键羟化酶细胞色素 P450 的表达水平并增加了 NADPH 的浓度，这有助于 C.lini CGMCC6051 中 DHEA 的二羟基化。二维电泳蛋白质组学分析结果表明，加入乙二醇后细胞色素P450和6-磷酸葡萄糖酸脱氢酶的表达水平增加。进一步的研究表明，乙二醇在稳定转化系统的 pH 值和在 DHEA 生物转化过程中产生辅酶 NADPH 方面发挥了关键作用。因此，乙二醇的添加诱导了关键羟化酶细胞色素 P450 的表达水平并增加了 NADPH 的浓度，这有助于 C.lini CGMCC6051 中 DHEA 的二羟基化。