酯酶基因Est2的异源表达及酶学性质研究  

Heterologous Expression of Esterase Gene Est2 and Study of Its Enzymatic Properties

作　　者：邓名 杨云娟 DENG Ming;YANG Yun-juan(Engineering Research Center of Sustainable Development and Utilization of Biomass Energy of the Ministry of Education,School of Life Sciences,Yunnan Normal University,Kunming 650500,China)

机构地区：[1]云南师范大学生命科学学院,生物能源持续开发利用教育部工程研究中心,云南昆明650500

出　　处：《云南师范大学学报（自然科学版）》2021年第2期51-58,共8页Journal of Yunnan Normal University:Natural Sciences Edition

基　　金：国家自然科学基金资助项目(31960023);云南省中青年学术和技术带头人后备人才资助项目(2018HB059).

摘　　要：对来自α/β水解酶超家族的酯酶基因Est2进行了克隆、异源表达及纯化,然后对其酶学性质进行了研究.研究结果显示:酯酶Est2的最适底物为对硝基苯乙酸酯(p-NPC_(2)),最适反应温度为50℃,最适pH为8.0;Est2具有较好的pH稳定性,在pH 5.0~10.0范围内,其相对酶活性保持在50%以上;在40℃条件下具有最好的温度稳定性,在40℃耐受1 h之后仍有大于60%的酶活性;金属离子Fe^(2+)和有机试剂甲醇对酯酶Est2的酶活性具有激活作用.酯酶Est2在不同pH缓冲液中的稳定性以及对于Fe^(2+)和有机试剂较好的耐受性,暗示其具有较好的潜在工业应用价值.In this study,an esterase gene named Est2 fromα/βhydrolase superfamily was cloned,heterologously expressed and purified.Using the purified enzyme Est2,its enzymatic activity was determined.The results showed that maximum enzymatic activity of Est2 was recorded at 50℃and pH 8.0 with p-nitrophenol acetate(p-NPC_(2))as substrate.Est2 retained more than 50%of its original activity when incubated in the pH range of 5.0 to 10.0,exhibiting good pH stability.When incubated Est2 at 40℃for one hour,more than 60%of its original activity was left.The metal ion Fe^(2+)and organic reagent methanol can activate the activity of Est2.The pH stability of Est2 and its good tolerance to Fe^(2+)and organic reagent methanol indicate that Est2 has good potential industrial application value.

关 键 词：酯酶 酶学性质 基因克隆 异源表达 

分 类 号：Q78[生物学—分子生物学]