白藜芦醇
异源表达
生物合成
生物化学
酶
ATP合酶
化学
植保素
大肠杆菌
酶动力学
基因
重组DNA
活动站点
作者
Huili Guo,Luo Zai-qi,Yadong Yang,Mingfeng Yang,Heshu Lü,Chunmei Liu,Jing Yang,You‐Nian Wang,Lanqing Ma
出处
期刊:PubMed
[National Institutes of Health]
日期:2014-10-01
卷期号:30 (10): 1622-33
被引量:3
摘要
Resveratrol is a natural phytoalexin with special pharmacological and health functions. Stilbene synthase (STS) is a key and rate-limiting enzyme in the biosynthesis of resveratrol that is present only in a limited number of plants. The content of resveratrol from Polygonum cuspidatum is more than 1000 times higher than grapes and peanuts. We speculate that the catalytic ability of different STS may be one of the reasons causing differences in the content of resveratrol. To verify the above speculation, Vitis vinifera stilbene synthase gene (VvSTS) was amplified according to overlap PCR protocol with genomic DNA as template. VvSTS and PcSTS (PcPKS5) were analyzed through heterologous expression in Escherichia coli. The expression products were purified with Ni-NTA sepharose affinity chromatography and desalted through PD-10 column. The molecular weight of the two fusion proteins was about 43 kDa. Enzyme reaction and product analysis showed that the two products were resveratrol. The enzyme kinetic analysis showed that the catalyze efficiency (Kcat/Km) of PcPKS5 was 2.4 times of the VvSTS. Our findings confirms that STS from certain plants has much higher catalytic capability.
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