Metabolism of phenolic compounds catalyzed by Tomato CYP736A61

IF 3.4 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Enzyme and Microbial Technology Pub Date : 2024-03-07 DOI:10.1016/j.enzmictec.2024.110425

Thanh Dat Mai , Hyun Min Kim , Seo Young Park , Sang Hoon Ma , Ju Hui Do , Won Choi , Hye Min Jang , Hyeon Bae Hwang , Eun Gyeong Song , Jae Sung Shim , Young Hee Joung

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Abstract

Cytochrome P450s (CYPs) regulate plant growth and stress responses by producing diverse primary and secondary metabolites. However, the function of many plant CYPs remains unknown because, despite their structural similarity, predicting the enzymatic activity of CYPs is difficult. In this study, one member of the CYP736A subfamily (CYP736A61) from tomatoes was isolated and characterized its enzymatic functions. CYP736A61 was successfully expressed in Escherichia coli through co-expression with molecular chaperones. The purified CYP736A61 showed hydroxylation activity toward 7-ethoxycoumarin, producing 7-hydroxycoumarin or 3-hydroxy 7-ethoxycoumarin. Further substrate screening revealed that dihydrochalcone and stilbene derivates (resveratrol and polydatin) are the substrates of CYP736A61. CYP736A61 also mediated the hydroxylation of resveratrol and polydatin, albeit with low activity. Importantly, CYP736A61 mediated the cleavage of resveratrol and polydatin as well as pinostilbene and pterostilbene. Interestingly, CY736A61 also converted phloretin to naringenin chalcone. These results suggest that CYP736A61 is a novel CYP enzyme with stilbene cleavage activity.

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番茄 CYP736A61 催化的酚类化合物代谢

细胞色素 P450s（CYPs）通过产生多种初级和次级代谢产物来调节植物的生长和胁迫反应。然而，许多植物 CYPs 的功能仍然未知，因为尽管它们的结构相似，但预测 CYPs 的酶活性却很困难。本研究从番茄中分离出了 CYP736A 亚家族的一个成员（CYP736A61），并鉴定了其酶功能。通过与分子伴侣共同表达，CYP736A61 成功地在大肠杆菌中表达。纯化的 CYP736A61 对 7-乙氧基香豆素具有羟化活性，可产生 7-羟基香豆素或 3-羟基 7-乙氧基香豆素。进一步的底物筛选显示，二氢查尔酮和二苯乙烯衍生物（白藜芦醇和多靛酚）是 CYP736A61 的底物。CYP736A61 还介导了白藜芦醇和多靛红的羟基化，尽管活性较低。重要的是，CYP736A61 介导了白藜芦醇和多靛红以及松芪和紫檀芪的裂解。有趣的是，CYP736A61 还能将白藜芦醇转化为柚皮苷查尔酮。这些结果表明，CYP736A61 是一种具有链烯裂解活性的新型 CYP 酶。

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来源期刊

Enzyme and Microbial Technology 生物-生物工程与应用微生物

CiteScore

7.60

自引率

5.90%

发文量

142

审稿时长

38 days

期刊介绍： Enzyme and Microbial Technology is an international, peer-reviewed journal publishing original research and reviews, of biotechnological significance and novelty, on basic and applied aspects of the science and technology of processes involving the use of enzymes, micro-organisms, animal cells and plant cells. We especially encourage submissions on: Biocatalysis and the use of Directed Evolution in Synthetic Biology and Biotechnology Biotechnological Production of New Bioactive Molecules, Biomaterials, Biopharmaceuticals, and Biofuels New Imaging Techniques and Biosensors, especially as applicable to Healthcare and Systems Biology New Biotechnological Approaches in Genomics, Proteomics and Metabolomics Metabolic Engineering, Biomolecular Engineering and Nanobiotechnology Manuscripts which report isolation, purification, immobilization or utilization of organisms or enzymes which are already well-described in the literature are not suitable for publication in EMT, unless their primary purpose is to report significant new findings or approaches which are of broad biotechnological importance. Similarly, manuscripts which report optimization studies on well-established processes are inappropriate. EMT does not accept papers dealing with mathematical modeling unless they report significant, new experimental data.