大豆血红蛋白在马克斯克鲁维酵母中的重组表达及条件优化  

Recombinant Expression and Optimization Conditions of Soybean Hemoglobin in K.marxianus

作　　者：田甜[1,2] 鲁心怡 曾俊源 周峻岗 余垚[1,2] 吕红 TIAN Tian;LU Xinyi;ZENG Junyuan;ZHOU Jungang;YU Yao;LU Hong(State Key Laboratory of Genetic Engineering,Fudan University,Shanghai 200438,China;Shanghai Engineering Research Center of Industrial Microorganisms,School of Life Sciences,Fudan University,Shanghai 200438,China)

机构地区：[1]复旦大学遗传工程国家重点实验室,上海200438 [2]复旦大学生命科学学院上海工业菌株技术研究中心,上海200438

出　　处：《复旦学报（自然科学版）》2024年第6期744-754,共11页Journal of Fudan University：Natural Science

基　　金：国家重点研发计划(2021YFA0910601)、上海市科委工程中心项目(19DZ2282100)。

摘　　要：本研究在马克斯克鲁维酵母中测试了不同来源血红蛋白表达情况,选择初始表达量最高的豆血红蛋白A(LBA)作为研究对象。探究了融合标签蛋白以及改变培养基条件对LBA表达的影响,结果显示培养基最佳pH为5.5,额外补充60μmol/L氯化血红素使LBA可溶表达量提升了21.1%,额外补充5μmol/L Fe^(2+)使LBA可溶表达量提升了56.5%。最后在5 L发酵罐分批补料发酵中额外补充5μmol/L Fe ^(2+)发酵72 h LBA产量最高,为1.97 g/L,较对照提高了18.7%。In this study,different sources of hemoglobin expression were assessed in Kluyveromyces marxianus,with leghemoglobin A(LBA),exhibiting the highest initial expression levels,chosen for investigation.The impact of fusion tag proteins and altered culture conditions on LBA expression was explored,revealing an optimal pH of 5.5 in the culture medium.The addition of 60μmol/L hemin chloride led to a 21.1%increase in soluble LBA expression,while supplementing with 5μmol/L iron ions resulted in a 56.5%increase.Lastly,in a 5 L fermenter with batch feeding,supplementing with 5μmol/L iron ions for 72 hours yielded the highest LBA production at 1.97 g/L,representing an 18.7%increase compared to the control.

关 键 词：豆血红蛋白 马克斯克鲁维酵母 血红素 蛋白表达 

分 类 号：Q36[生物学—遗传学]