食品工业科技2025，Vol.46Issue(2)：25-34,10.DOI:10.13386/j.issn1002-0306.2024040241
三种芸香糖苷酶的底物特异性和酶学特征
Substrate Specificity and Enzymatic Characteristics of Three Rutinosidases
摘要
Abstract
Quercetin,naringenin,and hesperetin are the hydrolysis products of rutinoside flavonoids,known for their multiple biological activities and potential applications.To explore the characteristics of different rutinosidases in preparing quercetin,naringenin,and hesperetin,the rutinosidase AnRut from Aspergillus niger CBS 513.88,and αRβD Ⅰ and αRβD Ⅱfrom Acremonium sp.DSM 24697 were selected for comparative studies on substrate specificity and enzymatic properties.The results showed that,all three rutinosidases could only hydrolyze flavonoid compounds with α-1,6-linked rutinosides,but had no effect on flavonoid compounds with α-1,2-linked neohesperidosides.AnRut primarily hydrolyzed 3-O-linked rutin,αRβD Ⅰ mainly hydrolyzed 7-O-linked narirutin and hesperidin,while αRβD Ⅱ showed no significant difference in hydrolytic activity towards both types of substrates.Molecular docking results indicated that there were distinct binding modes within the three rutinosidases with rutin,narirutin,and hesperidin,and the substrate specificities of the three rutinosidases were influenced with variations in their interactions with the glycoside structures rutinoside flavonoids.The optimal temperature for AnRut was 50 ℃,and the optimal pH was 4.0.Additionally,10 mmol/L β-ME and DTT significantly enhanced AnRut's enzymatic activity,increasing the relative activity to 223％ and 242％ of the wild type,respectively.The optimum temperature and pH of αRβD Ⅰ was 70 ℃ and 4.0,demonstrating efficient hydrolysis of narirutin and hesperidin under acidic conditions.αRβD Ⅱ,meanwhile,had an optimal temperature of 40 ℃ and an optimal pH of 6.0,indicating its suitability for hydrolyzing rutinoside flavonoids under neutral conditions.This study would provide experimental and theoretical references for the preparation of quercetin,naringenin,and hesperetin using rutinosidases and lay the groundwork for future research on the structure-activity relationship of rutinosidases.关键词
芸香糖苷酶/芸香糖苷类黄酮/底物特异性/酶学特征/分子对接Key words
rutinosidase/rutinoside flavonoids/substrate specificity/enzymatic characteristics/molecular docking分类
生物科学引用本文复制引用
肖枫伟,孙江,叶家影,李利君,倪辉..三种芸香糖苷酶的底物特异性和酶学特征[J].食品工业科技,2025,46(2):25-34,10.基金项目
国家自然科学基金项目(32170056). （32170056）
