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产重组别藻蓝蛋白类荧光蛋白的重组大肠杆菌发酵条件的优化
张伟杰1,2, 杨雨1,2, 关翔宇3, 葛宝胜4, 陈华新1, 李富超1, 秦松1
1.中国科学院海洋研究所;2.中国科学院研究生院;3.中国海洋大学;4.中国石油大学
摘要:
对重组别藻蓝蛋白类荧光蛋白基因工程菌在装有150 mL液体培养基的250 mL摇瓶中的发酵条件进行优化。对起始pH值、接种量、诱导起始时间、诱导温度和时长以及IPTG浓度对重组荧光蛋白表达量进行了分析。结果表明,在装有150 mL液体培养基的250 mL摇瓶中,在起始pH值为7.5, 6 %接种量,培养温度为37℃,培养时长为3~5 h,诱导温度为22℃,IPTG浓度为0.2 mmol/L,诱导时长为7~8 h的条件下发酵重组别藻蓝蛋白类荧光蛋白,其表达量最高。
关键词:  重组别藻蓝蛋白类荧光蛋白  发酵  优化
DOI:
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基金项目:中国科学院知识创新工程项目(KZCX2-YW-209,KZCX2-YW-216);国家863计划项目(2006AA090303);青岛市科技计划项目(06-2-2-12-JCH)
Optimization of fermentation conditions for producing allophycocyanin alike recombinant fluorescent protein in Escherichia coli
ZHANG Wei-jie,YANG Yu,GUAN Xiang-yu,GE Bao-sheng,CHEN Hua-xin,LI Fu-chao,QIN Song
Abstract:
In present study, fermentation conditions of allophycocyanin alike recombinant fluorescent protein engineering bacteria in 250 mL shaking flask with 150 mL medium were optimized. Affections of inoculation volume, initial acidity basicity, starting time, temperature and duration of inducement, and IPTG cencentration on allophycocyanin alike recombinant fluorescent protein produce were analyzed. The result showed that the highest output of recombinant fluorescent protein could be achieved under the following conditions: 6% inoculum volume, initial medium pH7.5, incubation temperature 37℃, incubation duration 3~5 h, induction temperature 22℃, IPTG concentration 0.2 mmol/L, and induction duration 7~8 h. All these conditions provided a reference for a large scale and high density fermentation of allophycocyanin alike recombinant fluorescent protein engineering bacteria.
Key words:  allophycocyanin alike recombinant fluorescent protein  fermentation  optimization
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