天津科技大学学报2017，Vol.32Issue(6)：8-14,7.DOI:10.13364/j.issn.1672-6510.20160142
黑曲霉ATCC1015催化16α,17α–环氧黄体酮11α–羟基化及相关P450基因诱导表达
11α-Hydroxylation of 16α,17α-Epoxy Progesterone by Aspergillus niger ATCC1015 and Induction Expression of Relevant Cytochromes P450 Genes
摘要
Abstract
The activity of Aspergillus niger ATCC1015 to biotransform 16α,17α-epoxy progesterone was investigated for rational engineering ofAspergillus niger.The transformation product was determined as 11α-hydroxy-16α,17α-epoxy pro-gesterone based on analyses of TLC,HPLC and NMR spectroscopy.In addition,the 11α-hydroxylation activities ofA.niger ATCC1015 on 16α,17α-epoxy progesterone were shown to be induced by substrate 16α,17α-epoxy progesterone.Given that fungal steroid hydroxylases are members of the cytochrome P450 family,57 CYP genes were selected as potential can-didates of the target hydroxylase genes.Quantitative Real-time PCR showed that expression of 2 of 57 CYP genes named AnA100 and AnA154 were highly induced by 16α,17α-epoxy progesterone treatment.RecombinantS.cerevisiae stains pYES2-AnA100 and pYES2-AnA154 were constructed with genes AnA100 and AnA154 respectively.The results of the steroidal transformation showed that recombinantS.cerevisiaepYES2-AnA100 can transform 16α,17α-epoxy progesterone to 11α-hydroxylation of 16α,17α-epoxyprogesterone.关键词
黑曲霉ATCC1015/11α–羟基化/16α,17α–环氧黄体酮/实时荧光定量PCRKey words
Aspergillus nigerATCC1015/11α-hydroxylase/16α/17α-epoxy progesterone/Real-time PCR分类
生物科学引用本文复制引用
林本凤,职亚飞,刘晓光,路福平..黑曲霉ATCC1015催化16α,17α–环氧黄体酮11α–羟基化及相关P450基因诱导表达[J].天津科技大学学报,2017,32(6):8-14,7.基金项目
国家高技术研究发展计划(863计划)资助项目(2011AA02A211) （863计划）
