为了对石斛碱生物合成途径进一步解析,对石斛碱生物合成途径研究进展、5 种药用萜类生物 合成的 CYP450 酶功能、CYP450 酶研究手段三个方面进行文献综述。 通过综述石斛碱生物合成研究进展, 提出 CYP450 酶在石斛碱合成途径中羟基化中间体的作用;总结对药用萜类青蒿酸、丹参酮、鼠尾草酸、人参 皂苷、甘草甜素等合成途径中 CYP450 酶的功能研究与鉴定,发现 CYP450 酶在萜类化合物中的主要功能为 羟基化、酮基化等氧化功能;总结 CYP450 酶的研究手段:一方面基因的筛选可以通过数据库检索或植物组 织、多因子诱导表达差异筛选,另一方面基因的表达和功能鉴定通过基因工程的多种手段,转入原核或真核 宿主细胞中表达出酶蛋白,然后加入底物鉴定其催化活性。 最后,展望下一步石斛碱生物合成 CYP450 酶的 挖掘,认为可以通过已有的转录组中筛选出差异 CYP450 酶基因并进行蛋白表达,然后通过上游酶催化或从 植物中分离纯化的方式获得底物,从酶蛋白与底物的催化挖掘与石斛碱合成相关 CYP450 酶类。
In order to further analyze the biosynthetic pathway of dendrobine the literature review was conducted on the research progress of dendrobine biosynthesis pathway the function of CYP450 enzymes in biosynthesis of five medicinal terpenoids and the research methods of CYP450 enzymes. By summarizing the research progress of dendrobine biosynthesis the role of CYP450 enzyme in the hydroxylation intermediate of dendrobine biosynthesis was put forward. This paper summarized the research and identification of CYP450 enzyme functions in the synthetic pathways of medicinal terpenoids artemisinic acid tanshinone carnosic acid ginsenoside and glycyrrhizin and found that the main functions of CYP450 enzyme in terpenoids were oxidation functions such as hydroxylation and ketonization. This paper summarizes the research methods of CYP450 enzyme on one hand the screening of genes can be performed by database search or differential screening of plant tissues and multi-factor induced expression on the other hand the expression and function identification of genes are transferred into prokaryotic or eukaryotic host cells to express the enzyme protein through various means of genetic engineering and then the substrate is added to identify its catalytic activity. Finally looking forward to the next step in the mining of dendrobine biosynthetic CYP450 enzymes it is believed that the differential CYP450 enzyme genes can be screened from the existing transcriptome and protein expression and then the substrate can be obtained by upstream enzyme catalysis or separation and purification from plants and the CYP450 enzymes related to dendrobium alkaloids synthesis can be discovered by the catalysis of enzyme proteins and substrates.
覃宏婷, 龚道勇, 李 标.石斛碱生物合成途径及萜类化合物CYP450酶的研究进展[J].重庆工商大学学报（自然科学版）,2022,39(6):1-13
QIN Hong-ting, GONG Dao-yong, LI Biao. Research Progress on Dendrobium Biosynthesis Pathway and Terpenoid CYP450 Enzymes[J]. Journal of Chongqing Technology and Business University(Natural Science Edition）,2022,39(6):1-13