Identification and recombinant production of a flavonoid glucosyltransferase with broad substrate specificity from Vaccinium corymbosum

IF 16.4 1区化学 Q1 CHEMISTRY, MULTIDISCIPLINARY

Accounts of Chemical Research Pub Date : 2024-02-26 DOI:10.1007/s13562-024-00876-2

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Abstract

Glucosyltransferases (GTs) are enzymes that use UDP-glucose to glucosylate wide variety of substrates, including the aglycones of anthocyanins. Anthocyanins are glycosylated polyphenolic plant pigments possessing potential health benefits to humans. The berries of Vaccinium species plants are rich in anthocyanins. Although the flavonoid content of bilberries is well characterized, the enzymes responsible for carrying out anthocyanin modifications are not thoroughly studied. In this study, a predicted sequence of an anthocyanin glucosyltransferase was identified from the genomic data of Vaccinium corymbosum. The codon-optimized gene sequence of the protein was integrated into the genome of P. pastoris. Constitutive expression in yeast extract-peptone-dextrose based media gave satisfactory amount of recombinant protein. The enzyme activity assays revealed that the V. corymbosum GT transferred glucosyl moieties to up to three positions of diverse flavonoids, such as naringenin, kaempferol, eriodictyol and cyanidin 3-O-glucoside, being therefore a rather unique enzyme among GTs described so far. The enzyme preferred cyanidin 3-O-glucoside, peonidin 3-O-glucoside and eriodictyol as substrates. This enzyme could find application in biotechnological production of glucosylated flavonoids.

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鉴定并重组生产具有广泛底物特异性的越橘类黄酮葡萄糖基转移酶

摘要糖基转移酶（GT）是一种利用 UDP-葡萄糖对包括花青素苷元在内的多种底物进行糖基化的酶。花青素是糖基化的多酚植物色素，对人类健康具有潜在的益处。越橘属植物的浆果富含花青素。虽然山桑子的类黄酮含量已被充分描述，但对负责进行花青素修饰的酶却没有进行深入研究。本研究从蔓越橘的基因组数据中确定了花青素葡萄糖基转移酶的预测序列。该蛋白的密码子优化基因序列被整合到 P. pastoris 的基因组中。在以酵母提取物-蛋白胨-葡萄糖为基础的培养基中，重组蛋白的表达量令人满意。酶活性测定显示，V. corymbosum GT 能将葡萄糖基转移到多种黄酮类化合物的三个位置，如柚皮苷、山柰酚、麦饭石酚和青花素 3-O-葡萄糖苷。该酶偏爱青花素 3-O-葡萄糖苷、芍药苷 3-O-葡萄糖苷和二碘酪醇作为底物。这种酶可用于葡萄糖基黄酮类化合物的生物技术生产。

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来源期刊

Accounts of Chemical Research 化学-化学综合

CiteScore

31.40

自引率

1.10%

发文量

312

审稿时长

2 months

期刊介绍： Accounts of Chemical Research presents short, concise and critical articles offering easy-to-read overviews of basic research and applications in all areas of chemistry and biochemistry. These short reviews focus on research from the author’s own laboratory and are designed to teach the reader about a research project. In addition, Accounts of Chemical Research publishes commentaries that give an informed opinion on a current research problem. Special Issues online are devoted to a single topic of unusual activity and significance. Accounts of Chemical Research replaces the traditional article abstract with an article "Conspectus." These entries synopsize the research affording the reader a closer look at the content and significance of an article. Through this provision of a more detailed description of the article contents, the Conspectus enhances the article's discoverability by search engines and the exposure for the research.