• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Journal of Biotechnology >Discovery of a steroid 11 alpha-hydroxylase from Rhizopus oryzae and its biotechnological application
【24h】

Discovery of a steroid 11 alpha-hydroxylase from Rhizopus oryzae and its biotechnological application

机译：米根霉甾体11α-羟化酶的发现及其生物技术应用

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

To overcome the chemically laborious stereo- and regioselective hydroxylation steps in the pharmaceutical production of corticosteroids and progestogens, certain fungal species, e.g. Rhizopus spp. and Aspergillus spp., are employed to perform the 11 alpha-hydroxylation of the steroid skeleton, thereby significantly simplifying steroid drug production. Here we report for the first time the identification and expression of a fungal ha-steroid hydroxylase, CYP509C12. The newly identified cytochrome P450, which is one of the 48 putative CYP genes in Rhizopus oryzae, was induced in the fungus by progesterone. By functionally expressing CYP509C12 in recombinant fission yeast, we were able to determine that its substrate spectrum includes progesterone as well as testosterone, 11-deoxycorticosterone, and 11-deoxycortisol, with the hydroxylations taking place predominantly at 11 alpha and 6 beta positions of the steroid ring system. To increase the 11a-hydroxylation activity of CYP509C12 in recombinant fission yeast, its natural redox partner, the R. oryzae NAD(P)H-dependent reductase, was coexpressed. The coexpression improved electron transfer to CYP509C12 and thus an increase in productivity from 246 to 300 mu M hydroxy Pg d(-1) was observed, as well as a 7-fold increase of rate of hydroxyprogesterone formation within the linear phase of transformation. This newly developed strain displayed total bioconversion of progesterone into 11 alpha-hydroxyprogesterone and small amounts of 6 beta-hydroxyprogesterone within the first 6h of incubation with progesterone as substrate, hence demonstrating its potential for biotechnological application
机译：为了克服在皮质类固醇和孕激素的药物生产中化学费力的立体和区域选择性羟基化步骤，某些真菌物种例如根霉曲霉属和曲霉属（Aspergillus spp。）被用于进行甾体骨架的11α-羟基化，从而显着简化了甾体药物的生产。在这里，我们首次报告真菌ha-类固醇羟化酶CYP509C12的鉴定和表达。新鉴定的细胞色素P450是米黄根霉中48个推定的CYP基因之一，是由孕酮在真菌中诱导的。通过在重组裂变酵母中功能性表达CYP509C12，我们能够确定其底物谱包括黄体酮以及睾丸激素，11-脱氧皮质酮和11-脱氧皮质醇，羟化反应主要发生在甾体的11 alpha和6 beta位置铃声系统。为了增加CYP509C12在重组裂变酵母中的11a-羟基化活性，共表达了其天然氧化还原伴侣米曲霉NAD（P）H依赖性还原酶。共表达改善了向CYP509C12的电子转移，因此观察到生产率从246增加到300μM羟基Pg d（-1），并且在线性转化相中羟基孕酮形成的速率增加了7倍。该新开发的菌株在以孕酮为底物孵育的最初6小时内显示出孕酮完全生物转化为11种α-羟基孕酮和少量6β-羟基孕酮，因此证明了其在生物技术应用中的潜力

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号