Functional characterization of squalene epoxidase and NADPH-cytochrome P450 reductase in Dioscorea zingiberensis

被引：19
作者：
Song, Wei [1 ]
Yan, Shan [1 ]
Li, Yi [1 ]
Feng, Shan [1 ]
Zhang, Jia-jiao [1 ]
Li, Jia-ru [1 ]
机构：
[1] Wuhan Univ, Coll Life Sci, State Key Lab Hybrid Rice, Wuhan 430072, Hubei, Peoples R China
来源：
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS | 2019年 / 509卷 / 03期
基金：
中国国家自然科学基金;
关键词：
Dioscorea zingiberensis; Squalene epoxidase; NADPH-Cytochrome P450 reductase; Enzyme activity; SUBCELLULAR-LOCALIZATION; BIOSYNTHESIS; SAPONIN; TRITERPENE; CLONING; EXPRESSION; DIOSGENIN; ROOT;
D O I：
10.1016/j.bbrc.2019.01.010
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
Dioscorea zingiberensis is a perennial medicinal herb rich in a variety of pharmaceutical steroidal saponins. Squalene epoxidase (SE) is the key enzyme in the biosynthesis pathways of triterpenoids and sterols, and catalyzes the epoxidation of squalene in coordination with NADPH-cytochrome P450 reductase (CPR). In this study, we cloned DzSE and DzCPR gene sequences from D. zingiberensis leaves, encoding proteins with 514 and 692 amino acids, respectively. Recombinant proteins were successfully expressed in vitro, and enzymatic analysis indicated that, when SE and CPR were incubated with the substrates squalene and NADPH, 2,3-oxidosqualene was formed as the product. Subcellular localization revealed that both the DzSE and DzCPR proteins are localized to the endoplasmic reticulum. The changes in transcription of DzSE and DzCPR were similar in several tissues. DzSE expression was enhanced in a time-dependent manner after methyl jasmonate (MeJA) treatments, while DzCPR expression was not inducible. (C) 2019 Elsevier Inc. All rights reserved.
引用
收藏
页码：822 / 827
页数：6
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