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OALib Journal期刊
ISSN: 2333-9721
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顺丁烯二酸异构酶工程菌发酵条件优化
DOI: 10.3724/SP.J.1145.2011.00558, PP. 558-562
Keywords: 顺丁烯二酸异构酶,发酵条件优化,重组工程菌,正交试验设计,酶活力
Abstract:
为提高重组大肠杆菌中顺丁烯二酸异构酶表达量，通过正交试验设计，对工程菌的生长条件和目的蛋白可溶表达条件进行优化.采用250ml三角瓶中装有50ml（amp100mgl-1）的培养基，分别研究培养基中葡萄糖、蛋白胨、酵母浸粉的浓度，培养基ph值以及摇床转速、装液量、接种量等对蛋白可溶表达量的影响.确定顺丁烯二酸异构酶工程菌最优化培养基为：蛋白胨20gl-1、酵母浸粉2.5gl-1、k2hpo4?3h2o3.0gl-1、kh2po41.5gl-1、nacl6gl-1、mgso43gl-1，培养基ph调至6.5.确定顺丁烯二酸异构酶工程菌可溶性表达最优条件为：37℃下培养至d600nm值为1.0时，添加终浓度为0.05mmoll-1的iptg进行诱导，诱导温度37℃，摇床转速220rmin-1，装液量20%，接种量5%，诱导时长为6h.利用bioflo415发酵罐以最优化的培养基和发酵条件对该工程菌进行了3批发酵实验，与摇瓶实验相比，顺丁烯二酸异构酶的表达量提高了近1.5倍，单位发酵液的酶活力由46uml-1发酵液提高到78uml-1发酵液.以上数据为顺丁烯二酸异构酶重组工程菌的中试发酵奠定了基础.图7表2参17
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