| 重组变形链球菌表面蛋白PAc发酵工艺研究 |
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| 作者姓名： | Jin J Li YH Fan MW |
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| 作者单位： | 1. 杭州口腔医院,牙体牙髓病科,浙江,杭州,310006;武汉大学口腔生物医学工程教育部重点实验室,湖北,武汉,430079
2. 武汉大学口腔生物医学工程教育部重点实验室,湖北,武汉,430079 |
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| 基金项目： | 国家自然科学基金(30600707)~~ |
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| 摘 要： | 目的:研究表达重组变形链球菌表面蛋白(rPAc)的工程菌pET20b(+)-AP/BL21(DE3)plysS的发酵条件。方法:通过摇瓶培养,进行发酵条件优化,在5L发酵罐中发酵pET20b(+)-AP/BL21(DE3)plysS工程菌,并对发酵产物进行SDS-PAGE、Western印迹分析鉴定。结果:确立了pET20b(+)-AP/BL21(DE3)plysS工程菌的高密度发酵工艺,即LB-2培养基,初始pH值为7.2,溶氧控制为30%以上,补料为10%甘油、5%酵母粉和5%蛋白胨。目的蛋白呈可溶性表达,表达量占菌体总蛋白的45%以上,菌体密度达到44。Western印迹分析显示,rPAc蛋白和抗PAc抗体有良好的结合活性。结论:通过发酵工艺优化,提高了rPAc蛋白的可溶性表达量,为进一步规模化研究和生产奠定了基础。
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| 关 键 词： | 发酵 纯化 发酵优化 |
Study on the fermentation conditions of recombinant Streptococcus mutans surface protein |
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| Jin J,Li YH,Fan MW.Study on the fermentation conditions of recombinant Streptococcus mutans surface protein[J].Shanghai Journal of Stomatology,2011,20(5):459-463. |
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| Authors: | Jin Jie Li Yu-hong Fan Ming-wen |
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| Institution: | JIN Jie1,2,LI Yu-hong2,FAN Ming-wen2.(1.Department of Operative Dentistry and Endodontics,Stomatological Hospital of Hangzhou.Hangzhou 310006,Zhejiang Province,2.Key Laboratory for Oral Biomedical Engineering of Ministry of Education,Wuhan University.Wuhan 430079,Hubei Province,China) |
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| Abstract: | PURPOSE: To develop the best fermentation process on pET20b(+)-AP/BL21(DE3)plysS expressing recombinant Streptococcus mutans surface protein(rPAc).METHODS: The strains were activated and then cultured in orbital shakers.Pilot test of flask shaking culture was done to get optimized culture conditions,including pH value for culture,type of culture medium,range of culture volume and type of feeding medium.According to the optimal fermentation condition of flask shaking culture,fermentation was carried out with... |
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| Keywords: | Fermentation Purification Fermentation optimization |
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