嘌呤降解酶的反向虚拟筛选、重组表达及活性表征  

Inverse Virtual Screening,Recombinant Expression,and Activity Characterization of Purine Degrading Enzymes

作　　者：刘瑜 张鑫鑫 王睿 孟祥红[1] LIU Yu;ZHANG Xinxin;WANG Rui;MENG Xianghong(College of Food Science and Engineering,Ocean University of China,Qingdao 266404,China)

机构地区：[1]中国海洋大学食品科学与工程学院,山东青岛266404

出　　处：《食品工业科技》2025年第5期145-152,共8页Science and Technology of Food Industry

基　　金：“十四五”国家重点研发计划重点专项(2021YFD21005)。

摘　　要：为开发可用于食品领域的新型嘌呤降解酶,本研究采用反向虚拟筛选技术和分子对接技术,筛选具有潜在嘌呤降解活性的酶,对该酶进行克隆表达、纯化及活性验证,并探究其降解嘌呤的最优反应条件。结果表明,由反向虚拟筛选平台PharmMapper筛选得到的二氢乳清酸酶和腺嘌呤、鸟嘌呤、次黄嘌呤和黄嘌呤的对接结合能分别为-7.3、-6.9、-7.6、-8.8 kcal/mol,对接结果最好。经克隆表达获得分子量为41 kDa的二氢乳清酸酶,其催化鸟嘌呤降解的最适条件为pH8.0,25℃,在Zn^(2+)(0.5 mmol/L)催化下,可降解46.1%的鸟嘌呤。因此,本研究发现二氢乳清酸酶具有一定的鸟嘌呤降解活性,并确定了其降解鸟嘌呤的最佳反应条件,为其在食品嘌呤降解中的应用奠定基础。To develop a novel purine degrading enzyme that can be used in the food industry,this study aimed to screen an enzyme with potential purine degradation activity using inverse virtual screening technology and molecular docking technology,clone express,purify,and validate the activity of this enzyme,and explore the optimal reaction conditions for its degradation of purines.The results showed that the docking binding energies of dihydroorotase screened by the inverse virtual screening platform PharmMapper with adenine,guanine,hypoxanthine,and xanthine were-7.3,-6.9,-7.6,and-8.8 kcal/mol,respectively,which had the optimum docking result.The dihydroorotase with a molecular weight of 41 kDa was obtained with cloning expression.The optimal conditions for catalyzing the degradation of guanine were pH8.0,25℃,and under the catalysis of Zn^(2+)(0.5 mmol/L),guanine could be degraded at a ratio of 46.1%.Therefore,it found that dihydroorotase had the degradation activity for guanine,and the optimal reaction conditions for guanine degradation were determined,which laid foundation for the application in degradation of food purine.

关 键 词：反向虚拟筛选 分子对接 克隆表达 鸟嘌呤 降解 

分 类 号：TS201.2[轻工技术与工程—食品科学]