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OALib Journal期刊
ISSN: 2333-9721
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溶氧反馈分批补料高密度培养枯草杆菌谷氨酰胺合成酶工程菌
, PP. 96-103
Keywords: 谷氨酰胺合成酶,重组大肠杆菌,分批补料培养,高密度培养
Abstract:
为了建立枯草杆菌谷氨酰胺合成酶基因重组工程菌bl21/pet28b-glna的高密度培养工艺,首先以摇瓶培养为基础,对影响工程菌生长及目的蛋白表达的因素如培养基、ph值、诱导剂浓度及时间等进行优化,再扩大至biotech-5bg(5l)自动玻璃发酵罐培养,利用溶氧反馈补料策略进行分批补料,并在培养过程中保持20%~50%的溶解氧,7.0~7.5的ph,以及1.0g/l乳糖诱导12h,成功地进行了工程菌的高密度培养,最终细菌干重达到48.1g/l,目的蛋白的表达量占菌体蛋白总量的55%,粗提物酶比活为10.35u/mg,整个补料过程细菌比生长率稳定的控制在0.1h-1左右.
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