5-ketoreductase from Streptomyces bingchengensis: overexpression and preliminary characterization
5-ketoreductase from Streptomyces bingchengensis: overexpression and preliminary characterization
复制标题
丙城链霉菌中的 5-酮还原酶：过表达和初步表征
DOI：
10.1007/s10529-010-0320-y
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发表时间：
2010-06
影响因子：
2.7
通讯作者：
Xiang, Wen-Sheng
中科院分区：
文献类型：
--
作者：
Wang, Xiang-Jing;Wang, Cheng-Qin;Sun, Xiao-Lin;Xiang, Wen-Sheng
为了阐明从5-氧球霉素A3和A4的生物转化到Bingchengensis链霉菌中的Milbemycins A3和A4，使用PCR克隆了C5-酮还原酶基因（MILF），并使用PCR与来自同源核苷酸序列设计的特异性引物。 c
To elucidate the biotransformation from 5-oxomilbemycins A3 and A4 to milbemycins A3 and A4 in Streptomyces bingchengensis, the C5-ketoreductase gene (milF) was cloned using PCR with the specific primer designed from homologous nucleotide sequences. The C5-ketoreductase (MilF) was heterologously expressed in E. coli BL21 (DE3) as a His-tagged fusion protein. The characterization and biotransformation function of purified MilF was verified by in vitro enzyme assay. MilF is an NADPH-dependent reductase. The biotransformation products, analyzed by LC-APCI/MS, were identified as milbemycin A3 and milbemycin A4. MilF is thus present in Streptomyces bingchengensis and can transform 5-oxomilbemycins A3 and A4 to milbemycins A3 and A4. These findings are significant for understanding the biosynthetic pathway of milbemycins in Streptomyces bingchengensis and pave the way to obtain a producer strain of 5-oxomilbemycins directly by targeted milF disruption.
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DOI：
--
发表时间：
2000
期刊：
--
影响因子：
--
作者：
T. Kieser
通讯作者：
T. Kieser
DOI：
--
发表时间：
2001-01
期刊：
--
影响因子：
--
作者：
J. Sambrook;E. Fritsch;T. Maniatis
通讯作者：
J. Sambrook;E. Fritsch;T. Maniatis
影响因子：
3.3
作者：
Hu, ZH;Reid, R;Gramajo, H
通讯作者：
Gramajo, H
DOI：
--
发表时间：
1993
期刊：
--
影响因子：
--
作者：
J. Ide
通讯作者：
J. Ide
影响因子：
3.3
作者：
Wang, Xiang-Jing;Guo, Suo-Lian;Xiang, Wen-Sheng
通讯作者：
Xiang, Wen-Sheng