摘要
咖啡酸及其衍生物是传统中药材紫草的重要水溶性有效成分,具有抗菌、抗氧化、保护心脑血管、抗肝纤维化和保肝、抗病毒及抗肿瘤等多种生物学活性。由于该类化合物经苯丙烷途径合成,而UDP-葡萄糖基转移酶(UDP-glycosyltransferases,UGTs)对苯丙烷途径的调控发挥重要作用,因此推测紫草细胞中存在糖基转移酶参与咖啡酸或/和其衍生物的修饰。该研究通过挖掘新疆紫草Arnebia euchroma转录组数据克隆获得15条糖基转移酶候选基因全长,并通过原核表达检测了对咖啡酸及迷迭香酸的体外活性。结果表明AeUGT_01,AeUGT_02,AeUGT_03,AeUGT_04,AeUGT_10可催化咖啡酸或/和迷迭香酸生成1~3种糖基化产物。这些糖基转移酶处于不同的系统进化分支,与同分支其他糖基转移酶具有底物或催化功能的相似性。因此推测糖基转移酶不同分支通过独立进化不断扩张而产生了趋同进化现象,糖基转移酶的底物宽泛性是这一现象产生的基础。上述工作为进一步研究新疆紫草咖啡酸及迷迭香酸糖基转移酶体内功能奠定了基础,有助于明确咖啡酸衍生物的形成及调控机制;也为咖啡酸及迷迭香酸的糖基化修饰提供了工具酶。
Caffeic acid and its oligomers are the main water-soluble active constituents of the traditional Chinese medicine(TCM)Arnebiae Radix.These compounds possess multiple biological activities such as antimicrobial,antioxidant,cardiovascular protective,liver protective,anti-liver fibrosis,antiviral and anticancer activities.The phenylpropanoid pathway in plants is responsible for the biosynthesis of caffeic acid and its oligomers.Glycosylation can change phenylpropanoid solubility,stability and toxic potential,as well as influencing compartmentalization and biological activity.In view of the important role played by de-glycosylation in the regulation of phenylpropanoid homeostasis,the biosynthesis of caffeic acid and its oligomers are supposed to be under the control of relative UDP-glycosyltransferases(UGTs).Through the data mining of Arnebia euchroma transcriptome,we cloned 15 full-length putative UGT genes.After recombinant expression using the prokaryotic system,the crude enzyme solution of the putative UGTs was examined for the glycosylation activities towards caffeic acid and rosmarinic acid in vitro.AeUGT_01,AeUGT_02,AeUGT_03,AeUGT_04 and AeUGT_10 were able to glycosylate caffeic acid and/or rosmarinic acid resulting in different mono-and/or di-glycosylated products in the UPLC-MS analyses.The characterized UGTs were distantly related to each other and divided into different clades of the phylogenetic tree.Based on the observation that each characterized UGT exhibited substrate or catalytic similarity with the members in their own clade,we supposed the glycosylation abilities towards caffeic acid and/or rosmarinic acid were evolved independently in different clades.The identification of caffeic acid and rosmarinic acid UGTs from A.euchroma could lead to deeper understanding of the caffeic acid oligomers biosynthesis and its regulation.Furthermore,these UGTs might be used for regiospecific glycosylation of caffeic acid and rosmarinic acid to produce bioactive compounds for potential therapeutic applications.
作者
王瑞杉
王升
梁玖雯
李锬
周利
詹志来
万修福
康传志
郭兰萍
WANG Rui-shan;WANG Sheng;LIANG Jiu-wen;LI Tan;ZHOU Li;ZHAN Zhi-lai;WAN Xiu-fu;KANG Chuan-zhi;GUO Lan-ping(State Key Laboratory Breeding Base of Dao-di Herbs,National Resource Center for Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China;Guangdong Provincial Research Center on Good Agricultural Practice&Comprehensive Agricultural Development Engineering Technology of Cantonese Medicinal Materials,College of Traditional Chinese Medicine,Guangdong Pharmaceutical University,Guangzhou 510006,China)
出处
《中国中药杂志》
CAS
CSCD
北大核心
2021年第1期86-93,共8页
China Journal of Chinese Materia Medica
基金
国家自然科学基金项目(81603239,81703648)
中央级公益性科研院所基本科研业务费专项(ZZXT202005,ZZ13-YQ-092,ZZXT201901,ZZ13-YQ-084)
国家重点研发计划项目(2017YFC1700701,2017YFC1701405)。
关键词
新疆紫草
糖基转移酶
咖啡酸
迷迭香酸
系统进化分析
Arnebia euchroma
glycosyltransferase
caffeic acid
rosmarinic acid
phylogenetic analysis
