人血管抑素基因工程菌发酵条件的初步研究  

Studies on Fermentation Conditions of Genetically Engineered Escherichial Coli for Recombinant Human Angiostatin

作　　者：卢文菊[1] 罗进贤[2] 何建国[2] 张添元[2] 

机构地区：[1]广州医学院检验系,广州510182 [2]中山大学生命科学学院,广州510275

出　　处：《广州医学院学报》2000年第3期1-3,共3页Academic Journal of Guangzhou Medical College

基　　金：国家自然科学基金! (396 70 0 13);广东省自然科学基金! (96 0 0 52 )

摘　　要：目的 :探讨发酵条件对大肠杆菌工程菌表达重组人血管抑素 (rhAGN)的影响。方法 :应用摇瓶和发酵罐发酵试验观察不同诱导时机和诱导时间以及 pH、通气量等对rhAGN表达量的影响。结果 :(1)工程菌E .coliDH5α(pBVA2 )在 30℃培养至A60 0nm 为 0 .3～ 0 .5时于 4 2℃诱导4hrhAGN表达量最高 ,约为 2 12mg L ;(2 ) 15L发酵罐发酵参数为 pH7.2 ,搅拌转速 50 0r min ,通气量 4～ 8L min ,溶氧 50 %时 ,rhAGN表达量提高到 2 76mg L。结论 :确定了最适诱导时机和诱导时间 ,通过调节 pH和改善通气 ,初步形成了一套高表达rhAGN的发酵工艺。Objective:To investigate the effects of fermentation conditions on the production of recombinant human angiostatin (rhAGN)by genetically engineered E.coli .Methods:The effects of different cell growth phases for induction,different inducing period,pH and air flow on rhAGN production were observed using shaking flask and fermentor respectively.Results:When induced at 42℃ for 4 hours after the cultivation of E.coli DH5α(pBVA2) in shaking flask reached to A 600nm ≈0.3～0.5 at 30℃,the yield of rhAGN reached to the highest level of 212mg/ml. When cultivated in a 15l fermentor at 30℃ ,with pH of 7.2,rotation rate of 500r/min,air flow of 4～6L/min and soluble oxygen saturation of 50%,for the engineered bacteria to reach to A 600nm ≈0.5 and then induced at 42℃ for another 4 hours,the yield of rhAGN was 276mg/L.Conclusion:The optimal growth phase for induction and the inducing period were determined.Through the effective controll of the technical parameters,the yield of rhAGN in a fermentor was increased to a higher level than that in a shaking flask.

关 键 词：人血管抑素 基因表达 大肠杆菌 发酵 

分 类 号：R394.8[医药卫生—医学遗传学] Q786[医药卫生—基础医学]