在Cibotium barometz中三合成酶的功能性表征
Zhongju Ji1, Baolian Fan1, Yidu Chen1
1Guangdong Provincial Key Laboratory of Translational Cancer Research of Chinese Medicines, Joint International Research Laboratory of Translational Cancer Research of Chinese Medicines, International Institute for Translational Chinese Medicine, School of Pharmaceutical Sciences, Guangzhou University of Chinese Medicine, Guangzhou, China.
Synthetic and systems biotechnology
|July 7, 2023
概括
研究人员确定了三种Cibotium barometz三合成酶基因 (CbTSs),参与生产各种三烯. 这些在根茎中高度表达的基因使用修改的酵母系统进行了表征,揭示了它们在生物合成中的特定作用.
科学领域:
- 植物生物化学 植物生物化学
- 分子生物学分子生物学
- 代谢学 代谢学 代谢学
背景情况:
- 一种有价值的药用,Cibotium barometz,可以产生各种生物活性三烯.
- 目前,C. barometz中这些三二烯的生物合成途径尚不清楚.
- 了解三烯生物合成对于其药用应用至关重要.
研究的目的:
- 为了确定负责C. barometz.中三烯生物合成的候选基因.
- 阐明各种三烯生产背后的酶机制.
- 描述已识别的C. barometz三合成酶 (CbTSs) 的功能.
主要方法:
- 在C. barometz根茎和叶子中进行De novo转录组测序.
- 鉴定和分离了三个候选CbTS基因.
- 一个过度生产素和氧素的Saccharomyces cerevisiae底盘的建造.
- 在工程酵母中表达CbTS1,CbTS2和CbTS3的异质表达.
- 对CbTS基因的遗传学分析.
主要成果:
- 鉴定了三种CbTS基因 (CbTS1,CbTS2,CbTS3),发现它们在C. barometz根茎中表达很高.
- 酵母中的异质表达产生了环甲醇 (CbTS1),达马拉丁烯 (CbTS2) 和双烯 (CbTS3).
- 遗传学分析将CbTS1分类为氧化二烯环酶,CbTS2/CbTS3分类为二烯环酶.
结论:
- 这项研究成功地确定并功能性地表征了参与C. barometz三烯生物合成的三个关键酶.
- 这些发现提供了关于C. barometz.中观察到的多样化的三二烯配置文件的酶基础的见解.
- 这项研究为未来在这个重要的药用植物中生产三烯的代谢工程奠定了基础.
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