分散泛菌DJL-B醇脱氢酶的重组表达与固定化研究  

Recombinant Expression and Immobilization of Pantoea dispersa DJL-B Alcohol Dehydrogenase

作　　者：田红 庞立[2] 杨文韬 陈晓 王婧涵 夏菠[1] 蒋立文[1] TIAN Hong;PANG Li;YANG Wentao;CHEN Xiao;WANG Jinghan;XIA Bo;JIANG Liwen(College of Food Science and Technology,Hunan Agricultural University,Changsha 410128,China;Horticulture College,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,湖南长沙410128 [2]湖南农业大学园艺学院,湖南长沙410128

出　　处：《食品工业科技》2025年第9期176-184,共9页Science and Technology of Food Industry

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

摘　　要：为了提高重组醇脱氢酶在大肠杆菌BL21(DE3)中的表达水平,本研究通过单因素实验探讨了诱导温度、诱导时间以及IPTG浓度对酶表达量的影响。并在此基础上,进一步通过海藻酸钠包埋法研究了重组醇脱氢酶的固定化,以提升其对香叶醇转化的应用价值。结果表明,重组醇脱氢酶在大肠杆菌BL21(DE3)中的最适表达条件为:诱导温度28℃,诱导时间20 h,IPTG浓度0.1 mmol/L。固定化的条件为:固定化温度20℃、海藻酸钠浓度3.5%、加酶量500μL。同时固定化酶表现出良好的机械稳定性,经重复使用5次后,相对酶活性仍保持在60%左右,显示出较好的重复使用性。研究结果为工业上利用固定化酶催化法制备香叶醛提供了重要的理论参考,具有良好的应用前景。To enhance the expression level of recombinant alcohol dehydrogenase in E.coli BL21(DE3),this study investigated the effects of induction temperature,induction time,and IPTG concentration on enzyme expression through one-way experiments.Additionally,the immobilization of recombinant alcohol dehydrogenase using sodium alginate embedding was examined to improve its application in geraniol conversion.The results indicated that the optimal expression conditions for recombinant alcohol dehydrogenase in E.coli BL21(DE3)were an induction temperature of 28℃,an induction time of 20 hours,and an IPTG concentration of 0.1 mmol/L.Furthermore,the conditions for immobilization were:immobilization temperature of 20℃,sodium alginate concentration of 3.5%,and enzyme addition of 500μL.Meanwhile,the immobilized enzyme showed good mechanical stability,and the relative enzyme activity was still maintained at about 60%after 5 times of reuse,demonstrating good reusability.The results of the study provide an important theoretical reference for the industrial preparation of geranial using immobilized enzyme catalysis,which has good application prospects.

关 键 词：醇脱氢酶 异源表达 固定化 香叶醛 生物转化 

分 类 号：Q55[生物学—生物化学]