可降解磺胺嘧啶的重组细菌漆酶诱导表达和酶学性质研究  

Induced expression and enzymatic properties of recombinant bacterial laccase degrading sulfadiazine

作　　者：王婧涵 庞立[2] 田红 詹依雅 易永真 夏菠[1] 刘霞[1] WANG Jing-Han;PANG Li;TIAN Hong;ZHAN Yi-Ya;YI Yong-Zhen;XIA Bo;LIU Xia(College of Food Science and Technology,Hunan Agricultural University,Changsha 410128,China;College of Horticulture,Hunan Agricultural University,Changsha 410128,China)

机构地区：[1]湖南农业大学食品科学技术学院,长沙410128 [2]湖南农业大学园艺学院,长沙410128

出　　处：《食品安全质量检测学报》2025年第6期232-238,共7页Journal of Food Safety and Quality

基　　金：湖南省自然科学基金项目(2022JJ30299,2022JJ30290)。

摘　　要：目的 探究重组细菌漆酶诱导表达水平及酶学特性,提高细菌漆酶对磺胺嘧啶抗生素的降解效率。方法 本研究通过单因素实验探讨了诱导温度、诱导时间以及异丙基-β-D-硫代半乳糖苷(isopropylβ-D-thiogalactopyranoside,IPTG)浓度对酶表达量的影响。并通过p H和温度两方面对重组细菌漆酶的酶学性质进行探究。结果 24 h重组细菌漆酶对磺胺嘧啶的降解率达50%。重组细菌漆酶在大肠杆菌BL21 (DE3)中的最适表达条件为:诱导温度16℃,诱导时间16 h,IPTG浓度0.2 mmol/L。最适p H为3,在p H为7时,稳定性最好,6 h酶活达到150%;最适温度为80℃,在温度为70℃时热稳定性最好,1 h提高到120%,半衰期约为6 h。结论 本研究为细菌漆酶在磺胺嘧啶抗生素降解中的应用提供了实验基础,并为未来研究其在工业中的潜在应用奠定了理论依据。Objective To investigate the induced expression level and enzymatic properties of recombinant bacterial laccase and to improve the degradation efficiency of sulfadiazine antibiotics by bacterial laccase.Methods In this study,the effects of induction temperature,induction time,and concentration of isopropylβ-D-thiogalactopyranoside(IPTG)on enzyme expression were investigated by one-way experiments.The enzymatic properties of recombinant bacterial laccase were also investigated by both pH and temperature.Results The degradation of sulfadiazine by recombinant bacterial laccase at 24 h was 50%. The optimal conditions for the expression of recombinant bacterial laccase in Escherichia coli BL21 (DE3) were as follows: Induction temperature of 16 ℃, induction time of 16 h, and IPTG concentration of 0.2 mmol/L. The optimal pH was 3, and the stability was best at pH 7, with the enzyme activity reaching 150% in 6 h. The optimal temperature was 80 ℃, and the thermal stability was best at 70 ℃, with the enzyme activity increasing to 120% in 1 h. The half-life was about 6 h. Conclusion This study provides an experimental basis for the application of bacterial laccase in the degradation of sulfadiazine antibiotics and lays down a theoretical basis for the future study of its potential application in the industry.

关 键 词：细菌漆酶 磺胺嘧啶 抗生素降解 异源表达 酶学性质 蛋白浓度 

分 类 号：S85[农业科学—兽医学]