葡萄糖氧化酶基因在解脂耶氏酵母中的表达及发酵优化  

Expression of Glucose Oxidase Gene in Yarrowia lipolytica and Fermentation Optimization

作　　者：刘晓筱[1,2] 张娟 陈坚[1,2] 堵国成[1,2] LIU Xiaoxiao;ZHANG Juan;CHEN Jian;DU Guocheng(Key Laboratory of Industrial Biotechnology,Ministry of Education,Jiangnan University,Wuxi 214122,China;School of Biotechnology,Jiangnan University,Wuxi 214122,China)

机构地区：[1]江南大学工业生物技术教育部重点实验室,江苏无锡214122 [2]江南大学生物工程学院,江苏无锡214122

出　　处：《食品与生物技术学报》2018年第12期1233-1241,共9页Journal of Food Science and Biotechnology

基　　金：国家自然科学基金项目(31470160);国家高技术研究发展计划项目(2011AA100905);中国博士后科学基金特别资助项目(114957)

摘　　要：葡萄糖氧化酶(β-D-glucose:oxygen 1-oxidoreductase;EC 1.1.3.4,简称GOD)是生物领域中至关重要的工具酶之一,被广泛应用于食品工业、饲料工业、纺织、医药等行业中。作者构建了一株重组解脂耶氏酵母Yarrowia lipolytica 1-28,分泌GOD,并进一步运用单因素实验与正交实验在摇瓶水平对其发酵培养基进行了优化。实验结果显示,最佳发酵培养基配方为:甘油20 g/L,酵母膏2.64 g/L,氯化铵2.64 g/L,无水硫酸镁0.13 g/L,磷酸二氢钾0.32 g/L,维生素B13.34×10-4g/L,初始pH值6.0。优化后GOD发酵产量达到11.0 U/mL,较初始发酵培养基GOD产量提高了72%。在此基础上进行3 L罐发酵,重组菌在甘油补料30 g/L,pH为5.0时,28℃发酵190h,发酵液酶活达到81.6 U/mL。Glucose oxidase(β-D-glucose:oxygen 1-oxidoreductase;EC 1.1.3.4,GOD) has gained considerable commercial importance in the last few years due to its multitude of applications in the chemical,food,pharmaceutical,beverage,biotechnology and other industries.In this study,the fermentation medium of the recombinant Yarrowia lipolytica strain 1-28 that was constructed to secrete GOD was optimized through single-factor and orthogonal tests.The optimal fermentation medium was consisted of 20 g/L glycerol,2.64 g/L yeast extract,2.64 g/L NH4Cl,0.13 g/L MgSO4,0.32 g/L KH2PO4,3.34×10^-4 g/L Vitamin B1 at pH 6.0.The yield of GOD corresponded to 11.0 U/mL when cultured in the optimized medium,which was 72% higher than that at initial conditions.When fermented in the 3 L tank at 28℃ for 190 h,the enzyme activity of GOD reached 81.6 U/mL at pH 5.0 and glycerol feeding at the concentration of 30 g/L.

关 键 词：解脂耶氏酵母 葡萄糖氧化酶 发酵优化 

分 类 号：TQ925[轻工技术与工程—发酵工程]