作　　者： ; ; ; (李岩）; (吴同山）; (王进军）;

机构地区： 华南农业大学食品学院

出　　处： 《食品与发酵工业》 2013年第6期28-33,共6页

摘　　要： 为获得含猪脂联素球状结构域gAd基因的重组乳酸乳球菌的高效表达,对其表达条件和培养基配方进行了优化。首先采用单因素分析法确定重组乳球菌最佳诱导剂浓度、诱导时间点(OD600)和诱导后培养温度,以及培养基中的乳糖浓度、氮源浓度、Na2HPO4浓度和MgSO4浓度对表达量的影响;之后通过正交试验,确定重组乳酸乳球菌的最佳表达条件。结果显示,其最佳诱导剂nisin浓度为30ng/mL、诱导的时间点是OD600≈0.4、诱导后培养温度为25℃;最佳培养基配方是:乳糖添加量为7%、大豆蛋白胨为2.7%、酵母膏为1.3%、Na2HPO4浓度为0.25%、MgSO4浓度为0.02%。优化后,脂联素的表达量为29.4%的总蛋白含量。 In this paper we studied the induction conditions and cultural media of recombinant Lactococcus lactis NZ9000-gAd for expression of porcine adiponectin protein. The aim of this study was to obtain maximum yield of pro- tein gAd. Firstly, the amount of nisin added for induction, the time point of induction （cell density of the culture） , and the temperature for induction in improved M17G culture medium were optimized through single factor experi- ments. Then the concentrations of lactose, nitrogen source, N%HPO4 and MgSO4 in basic medium were also deter- mined through single factor experiments. Finally, the optimal induction and cultural media were determined though orthogonal experiment. The optimized conditions of induced expression were obtained. The amount of nisin was $0ng/ mL, the time point of induction was OD600 ≈0.4, and the culture temperature after induction was 25℃. The optimal cultural medium contained 7% lactose, 2.7% peptone, 1.3% yeast extract, 0.25% Na2 HPO4 , and 0.02% MgSO4. In optimization conditions the amount of expressed protein-gAd could account for 29.4% of total protein.

关 键 词： 脂联素 球状结构域 乳酸乳球菌 正交试验 表达优化

领　　域： [轻工技术与工程] [轻工技术与工程]