不同培养条件对大肠杆菌工程菌产β-胡萝卜素的影响  

Effects of Different Culture Conditions on Production of β-carotene by Engineering Escherichia coli

作　　者：杜桂彩[1] 刘敏[2] 赵瑜[1] 李荣贵[1] 

机构地区：[1]青岛大学生物系,山东省天然色素重点实验室,山东青岛266071 [2]山东师范大学逆境植物研究所,山东济南250014

出　　处：《食品科学》2008年第7期272-276,共5页Food Science

基　　金：青岛市自然科学基金项目(05-1-JC-91)

摘　　要：将克隆了噬夏孢欧文氏菌类胡萝卜素合成相关基因crtE、crtB、crtI的重组质粒pET-15bcrtEIB和crtY的重组质粒pACYC-184crtY共转化E.coliBL21(DE3)构建工程菌。研究了碳源、氮源、金属离子、培养温度、光照、pH值、诱导时间等参数对工程菌生长及色素累积的影响,确定了合适的培养条件:培养基为改良LB培养基(蛋白胨10g/L、酵母提取物10g/L、可溶性淀粉5g/L、MgCl20.04g/L、FeCl30.01g/L、NaCl10g/L,pH5.8),光照,起始培养温度为37℃,培养至OD600为0.6时加入IPTG至终浓度为0.5mmol/L,诱导温度降至28℃;诱导时间为12h。发酵完成后工程菌的生物量为7.32gdw/L,β-胡萝卜素的最高含量可达5.11mg/gdw。Two expression vectors, pET-15bcrtEIB in which crtE, crtB, crtI of Erwinia uredovora were cloned and pACYC-184crtY in which crtY was cloned, were transformed into E.coli BL21（DE3） simultaneously to construct engineering bacterium. The engineering bacterium could accumulate β-carotene when induced by IPTG. Effects of different culture conditions such as carbon sources, nitrogen sources, metal ions, culture temperature, light, medium pH and induction time on bacterial growth and β-carotene accumulation were investigated. The optimal culturing conditions obtained are as follows： Modified LB broth （trypton 10 g/L, yeast extract 10 g/L, soluble starch 5 g/L, MgCl2 0.04 g/L, FeCl3 0.01 g/L and NaCl 10 g/L as well as pH 5.8） as medium and in light, the engineering bacterium firstly is shake-cultured at 37 ℃ to OD600 0.6 of culture liquid, subsequently IPTG added to final concentration 0.5 mmol/L, and then further cultured for 14 h at 28 ℃. The engineering bacterium could grow to 7.32 g dw/ L in LB medium and accumulate β-carotene to 5.11 mg/g dw under optimal culture conditions.

关 键 词：Β-胡萝卜素 工程菌 培养条件 

分 类 号：TS201.3[轻工技术与工程—食品科学]