高粱3-脱氧花青素对糖苷水解酶活性的抑制机制  

Inhibitory Activity and Mechanism of 3-Deoxyanthocyanidins in Sorghum on Glycoside Hydrolase

作　　者：张旭[1] 宋爽[1] 王璐璐 仵雁北 王静[1,2] ZHANG Xu;SONG Shuang;WANG Lulu;WU Yanbei;WANG Jing(School of Food and Health,Beijing Technology and Business University,Beijing 100048,China;China-Canada Joint Lab of Food Nutrition and Health,Beijing Technology and Business University,Beijing 100048,China)

机构地区：[1]北京工商大学食品与健康学院,北京100048 [2]北京工商大学中国-加拿大食品营养与健康联合实验室,北京100048

出　　处：《食品科学技术学报》2025年第2期29-41,共13页Journal of Food Science and Technology

基　　金：国家自然科学基金资助项目(32101950)。

摘　　要：高粱3-脱氧花青素(3-DAS)对α-淀粉酶与α-葡萄糖苷酶活性具有抑制作用,但其作用机制尚不明确。考察了不同高粱品种间3-DAS的酶抑制活性差异,以确定对α-淀粉酶和α-葡萄糖苷酶抑制作用最强的高粱3-DAS提取物;采用多种光谱学技术及酶动力学分析法,进一步探究3-DAS与两种糖苷水解酶的相互作用机制。结果表明,红茅粱6号提取物中的3-DAS含量最高且对酶的活性抑制作用最强,但其对α-淀粉酶和α-葡萄糖苷酶的作用特性略有不同,3-DAS以非竞争性方式抑制α-淀粉酶,而以竞争性为主的混合型方式抑制α-葡萄糖苷酶。高粱3-DAS与酶结合后,可导致α-淀粉酶和α-葡萄糖苷酶在200 nm附近的蛋白质骨架特征吸收峰发生红移,同时引起两种酶的内在荧光分别发生静态猝灭和静态-动态混合猝灭,并使酶的色氨酸和酪氨酸残基周围环境的疏水性升高。高粱3-DAS能减少α-淀粉酶和α-葡萄糖苷酶的α-螺旋结构含量,从而改变酶构象。高粱3-DAS与两种糖苷水解酶间的相互作用力均以静电作用力为主,结合过程为自发进行的放热反应。研究结果表明,高粱3-DAS具有作为α-淀粉酶和α-葡萄糖苷酶抑制剂的潜在价值,研究结果旨在为高粱及其副产物的高值化应用提供基础数据支撑。Sorghum 3-deoxyanthocyanins(3-DAS)can inhibit the activities of α-amylase and α-glucosidase,but the mechanism is still unclear.The differences in the enzyme inhibitory activities of 3-DAS among different sorghum varieties were investigated to determine the sorghum 3-DAS extract with the strongest inhibitory effect on α-amylase and α-glucosidase.A variety of spectroscopic techniques and enzyme kinetic analysis were used to further explore the interaction mechanism between 3-DAS and the two glycoside hydrolases.The results showed that 3-DAS from Hong Mao Grain No.6 extract had the highest content and the strongest enzyme inhibitory activity,but its action characteristics for α-amylase and α-glucosidase were slightly different.3-DAS inhibitedα-amylase in a non-competitive manner,but inhibited α-glucosidase in a competitive mixed manner.The binding of sorghum 3-DAS to the enzyme resulted in a red shift of the characteristic absorption peaks of the protein cytoskeleton around 200 nm forα-amylase and α-glucosidase,a static quenching and a static-dynamic mixed quenching of the intrinsic fluorescence of the two enzymes,respectively,and an increase in the hydrophobicity of the environment around the tryptophan and tyrosine residues of the enzymes.Sorghum 3-DAS could reduce the content of α-helix structure of α-amylase and α-glucosidase,thereby changing the conformation of the enzyme.The interaction between sorghum 3-DAS and the two glycoside hydrolases was dominated by electrostatic force,and the binding process was a spontaneous exothermic reaction.The results showed that sorghum 3-DAS had potential value as an inhibitor of α-amylase and α-glucosidase,and the results aimed to provide basic data support for high-value application of sorghum and its by-products.

关 键 词：高粱 3-脱氧花青素 Α-淀粉酶 Α-葡萄糖苷酶 酶活性 抑制机理 

分 类 号：TS201.4[轻工技术与工程—食品科学]