| 摘要: |
| 探究重组大肠杆菌产尿素酶B（urease B subunit, UreB）的高密度发酵条件。通过实验室摇瓶和30 L发酵罐对UreB基因工程菌的发酵条件进行优化。结果表明：30 L发酵罐中以TB培养基为发酵培养基，接种量为5%，发酵温度为37 ℃，pH为6.8，溶氧量为30%左右，培养至2 h开始恒速流加50%甘油，4 h流加50%酵母提取物和50%胰蛋白胨，并加入终浓度为0.5 mmol/L的异丙基β-D-硫代半乳糖苷（isopropyl β-D-thiogalactoside，IPTG)，诱导表达4 h，结束发酵,所得菌体干物质约为25.7 g/L，UreB表达量为31.4%。此工艺可以提高UreB的产量。 |
| 关键词: 尿素酶B亚单位 高密度发酵 摇瓶发酵 |
| DOI：10.14188/j.ajsh.2019.03.012 |
| 分类号:Q935 |
| 基金项目:四川省科技厅人才创新计划 2017RZ0083四川省科技厅人才创新计划(2017RZ0083); |
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| Optimization of high density fermentation conditions for UreB production from recombinant Escherichia coli |
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SUN Lijin1,ZHANG Zhongbao2,Li Hao2,CHEN Yongmei2,ZHANG Zhi1,LI Zaixin1
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1.Department of Biological Engineering, Zigong 643000, Sichuan, China;2.School of Chemical Engineering, Sichuan University of Science & Engineering, Zigong 643000, Sichuan, China
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| Abstract: |
| To explore the high density fermentation conditions of recombinant E.coli strain for expression of urease B subunit (UreB), the recombinant Escherichia coli BL21 (DE3) expressing virulence protein UreB was used for fermentation in 30-liter fermenter. Results indicated that: TB medium was used as the basal medium, the inoculation size was 5%, fermentation temperature was 37 °C, the pH was 6.8, the dissolved oxygen content was about 30%, 50% glycerol was added at a constant speed after fermentation for 2 h, 50% yeast extract and 50% tryptone were added at 4 h, then, IPTG with a final concentration of 0.5 mmol/L was added followed by incubation for 4 h.The dry weight of recombinant E.coli and expression rate of UreB reached 25.7g/L and 31.4%, respectively. Our results demonstrated that this process increases the yield of UreB. |
| Key words: UreB high density fermentation shake flask fermentation |
