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Functional Analysis of an Uridine Diphosphate Glycosyltransferase Involved in the Biosynthesis of Polyphenolic Glucoside in Tea Plants (Camellia sinensis)
Journal of Agricultural and Food Chemistry ( IF 5.7 ) Pub Date : 2017-12-11 00:00:00 , DOI: 10.1021/acs.jafc.7b04969 Xuecheng Zhao 1 , Xinlong Dai 1 , Liping Gao 1 , Lina Guo 1 , Juhua Zhuang 1 , Yajun Liu 1 , Xiubing Ma 1 , Rui Wang 1 , Tao Xia 1 , Yunsheng Wang 1
Journal of Agricultural and Food Chemistry ( IF 5.7 ) Pub Date : 2017-12-11 00:00:00 , DOI: 10.1021/acs.jafc.7b04969 Xuecheng Zhao 1 , Xinlong Dai 1 , Liping Gao 1 , Lina Guo 1 , Juhua Zhuang 1 , Yajun Liu 1 , Xiubing Ma 1 , Rui Wang 1 , Tao Xia 1 , Yunsheng Wang 1
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Polyphenols are one of the largest groups of compounds that confer benefits to the health of plants and humans. Flavonol glycosides are a major ingredient of polyphenols in Camellia sinensis. Flavonol-3-O-glycosides are characteristic astringent taste compounds in tea infusion. A polyphenolic glycosyltransferase (CsUGT72AM1) belonging to cluster IIIb was isolated from the tea plant. The full-length cDNA of CsUGT72AM1 is 1416 bp. It encodes 472 amino acids with a calculated molecular mass of 50.92 kDa and an isoelectric point of 5.21. The recombinant CsUGT72AM1 protein was expressed in Escherichia coli and exhibited catalytic activity toward multiple flavonoids and coniferyl aldehyde. The enzyme assay indicated that rCsUGT72AM1 could perform glycosidation of flavonols or coniferyl aldehyde in vitro to form 3-O-glucoside or 4-O-glucoside, respectively. Interestingly, this enzyme also had activities and performed multisite glycosidation toward flavanones. The consistent products were confirmed to be naringenin-7-O-glucoside and -4′-O-glucoside by the nuclear magnetism assay. In addition, in the enzyme assay with cyanidin as the substrate, the results suggested that the glycosylated activity of CsUGT72AM1 was remarkably inhibited by a high concentration of anthocyanins. The above results indicate that CsUGT72AM1 may be involved in the metabolism of flavonol, flavanone, anthocyanin, and lignin.
中文翻译：
的尿苷二磷酸葡萄糖基转移酶在茶树中多酚类葡萄糖苷的生物合成相关的功能分析（茶树）
多酚是赋予植物和人类健康益处的最大化合物之一。黄酮苷是山茶中多酚的主要成分。黄酮醇-3- O-糖苷是茶浸剂中特征性的涩味化合物。从茶树中分离出属于簇IIIb的多酚糖基转移酶（CsUGT72AM1）。CsUGT72AM1的全长cDNA为1416 bp。它编码472个氨基酸，计算的分子量为50.92 kDa，等电点为5.21。重组Cs UGT72AM1蛋白在大肠杆菌中表达并表现出对多种类黄酮和松柏基醛的催化活性。酶测定表明，r Cs UGT72AM1可以在体外进行黄酮醇或松柏树醛的糖苷化反应，分别形成3- O-葡萄糖苷或4- O-葡萄糖苷。有趣的是，这种酶也具有活性，并能对黄烷酮进行多位糖基化。一贯的产品被证实为柚皮素-7- Ø葡萄糖苷和-4' - Ø糖苷通过核磁检测。另外，在以花青素为底物的酶分析中，结果表明Cs的糖基化活性。UGT72AM1被高浓度的花青素显着抑制。以上结果表明Cs UGT72AM1可能参与了黄酮醇，黄烷酮，花色苷和木质素的代谢。
更新日期：2017-12-11
中文翻译：
的尿苷二磷酸葡萄糖基转移酶在茶树中多酚类葡萄糖苷的生物合成相关的功能分析（茶树）
多酚是赋予植物和人类健康益处的最大化合物之一。黄酮苷是山茶中多酚的主要成分。黄酮醇-3- O-糖苷是茶浸剂中特征性的涩味化合物。从茶树中分离出属于簇IIIb的多酚糖基转移酶（CsUGT72AM1）。CsUGT72AM1的全长cDNA为1416 bp。它编码472个氨基酸，计算的分子量为50.92 kDa，等电点为5.21。重组Cs UGT72AM1蛋白在大肠杆菌中表达并表现出对多种类黄酮和松柏基醛的催化活性。酶测定表明，r Cs UGT72AM1可以在体外进行黄酮醇或松柏树醛的糖苷化反应，分别形成3- O-葡萄糖苷或4- O-葡萄糖苷。有趣的是，这种酶也具有活性，并能对黄烷酮进行多位糖基化。一贯的产品被证实为柚皮素-7- Ø葡萄糖苷和-4' - Ø糖苷通过核磁检测。另外，在以花青素为底物的酶分析中，结果表明Cs的糖基化活性。UGT72AM1被高浓度的花青素显着抑制。以上结果表明Cs UGT72AM1可能参与了黄酮醇，黄烷酮，花色苷和木质素的代谢。
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