文献类型：期刊文献

英文题名：Molecular cloning and characterization of a cytochrome P450 taxoid 9a-hydroxylase in Ginkgo biloba cells

作者：Zhang, Nan[1] Han, Zhentai[2] Sun, Guiling[3] Hoffman, Angela[4] Wilson, Iain W.[5] Yang, Yanfang[1] Gao, Qian[2] Wu, Jianqiang[3] Xie, Dan[6,7] Dai, Jungui[6,7] Qiu, Deyou[1]

通信作者：Qiu, DY[1]|[a000598d4c276c5f9aea6]邱德有;

机构：[1]Chinese Acad Forestry, State Key Lab Tree Genet & Breeding, Res Inst Forestry, Beijing 100091, Peoples R China;[2]Chinese Acad Forestry, Key Lab Forest Ecol Environm, Minist Forestry, Res Inst Forest Ecol Environm & Protect, Beijing 100091, Peoples R China;[3]Chinese Acad Sci, Key Lab Econ Plants & Biotechnol, Kunming Inst Bot, Kunming 650201, Peoples R China;[4]Univ Portland, Dept Chem, Portland, OR 97203 USA;[5]CSIRO Plant Ind, Canberra, ACT 2001, Australia;[6]Chinese Acad Med Sci, State Key Lab Bioact Subst & Funct Nat Med, Inst Mat Med, Beijing 100050, Peoples R China;[7]Peking Union Med Coll, Beijing 100050, Peoples R China

年份：2014

卷号：443

期号：3

起止页码：938-943

外文期刊名：BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000331415000026)】；

基金：This work was supported by a grant for the National non-profit Research Institutions of Chinese Academy of Forestry (CAFYBB2012042).

语种：英文

外文关键词：Taxol; Taxoid 9 alpha-hydroxylase; Ginkgo biloba; CYP450; Cell-free protein synthesis system; LC-MS

摘要：Taxol is a well-known effective anticancer compound. Due to the inability to synthesize sufficient quantities of taxol to satisfy commercial demand, a biotechnological approach for a large-scale cell or cell-free system for its production is highly desirable. Several important genes in taxol biosynthesis are currently still unknown and have been shown to be difficult to isolate directly from Taxus, including the gene encoding taxoid 9 alpha-hydroxylase. Ginkgo biloba suspension cells exhibit taxoid hydroxylation activity and provides an alternate means of identifying genes encoding enzymes with taxoid 9 alpha-hydroxylation activity. Through analysis of high throughput RNA sequencing data from G. biloba, we identified two candidate genes with high similarity to Taxus CYP450s. Using in vitro cell-free protein synthesis assays and LC-MS analysis, we show that one candidate that belongs to the CYP716B, a subfamily whose biochemical functions have not been previously studied, possessed 9 alpha-hydroxylation activity. This work will aid future identification of the taxoid 9 alpha-hydroxylase gene from Taxus sp. (C) 2013 Elsevier Inc. All rights reserved.