TRANSFORMATION OF 4-ANDROSTEN-3,17-DIONE BY GROWING CULTURES AND CELL-EXTRACTS OF CLOSTRIDIUM - PARAPUTRIFICUM
TRANSFORMATION OF 4-ANDROSTEN-3,17-DIONE BY GROWING CULTURES AND CELL-EXTRACTS OF CLOSTRIDIUM - PARAPUTRIFICUM
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DOI：
10.1016/0005-2760(79)90066-3
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发表时间：
1979-01-01
期刊：
影响因子：
--
通讯作者：
FINEGOLD, SM
中科院分区：
文献类型：
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作者：
GLASS, TL;WHEELER, LA;FINEGOLD, SM
培养C. paraputrificum将4-雄甾烯-3，17-二酮转化为3 α-羟基-5 β-雄甾烷-17-酮与5 β-雄甾烷-3，17-二酮作为中间体。将1.5mM甲萘醌加入对数期培养物中，雄甾烯-3，17-二酮导致该类固醇部分再氧化为4-雄甾烯-3，17-二酮。该处理还导致培养物生长的短暂抑制。生长的恢复伴随着4-雄烯-3，17-二酮完全还原为5 β-雄烯。雄甾烷-3，17-二酮C. paraputrificum能够进行这些还原性转化的情况下添加的辅因子。Sephadex G-25处理的提取物需要NADH或NADPH进行这些反应。黄素核苷酸，FAD（-lus NADH或NADPH）或FMN（加上NADH）对4-雄甾烯-3，17-二酮还原为5 β-脱氢酶具有高度刺激性。雄甾烷-3，17-二酮NADH是用于还原C4-C5双键的优选还原吡啶核苷酸，而时程测量表明NADPH是用于还原3-酮基的优选供体。
Growing cultures of C. paraputrificum transformed 4-androsten-3,17-dione to 3.alpha.-hydroxy-5.beta.-androstan-17-one in a sequential manner with 5.beta.-androstan-3,17-dione as an intermediate. The addition of 1.5 mM menadione to log-phase cultures which had formed 5.beta.-androstan-3,17-dione resulted in a partial reoxidation of this steroid to 4-androsten-3,17-dione. This treatment also resulted in transient inhibition of culture growth. Resumption of growth was accompanied by complete reduction of 4-androsten-3,17-dione to 5.beta.-androstan-3,17-dione. Cell extracts of C. paraputrificum were capable of carrying out these reductive transformations in the absence of added cofactors. Sephadex G-25 treated extracts required NADH or NADPH for these reactions. A flavin nucleotide, either FAD (-lus NADH or NADPH) or FMN (plus NADH) was highly stimulatory for 4-androsten-3,17-dione reduction to 5.beta.-androstan-3,17-dione. NADH was the preferred reduced pyridine nucleotide for reduction of the C4-C5 double bond, while time-course measurements suggested that NADPH was the preferred donor for reduction of the 3-keto group.