重组巴斯德毕赤酵母发酵生产几丁质酶的条件优化研究

The Research on Optimal Culture Condition of Recombintant Chitinase Production in Pichia pastoris

摘要:

研究了利用重组巴斯德毕赤酵母诱导表达重组几丁质酶的条件。在摇瓶水平上研究了诱导时间、pH、甲醇流加量、油酸等因素对重组几丁质酶表达的影响。结果发现诱导108 h蛋白表达量最高；偏酸性环境不利于蛋白表达，维持在pH5.5～6.0最佳；甲醇最佳诱导浓度为1%；添加0.05%的油酸有助于提高蛋白表达量。在此基础上通过正交试验设计优化了培养基配方，在优化条件下，蛋白表达量达171.99mg/L,酶活达49.58U/mL。

Abstract:

The fermentation condition of a recombinant strain to produce chitinase in Pichia pastoris was studied in this paper.By the shaking flask,the influences of inducing time, pH,methanol,oleic acid on the expression of chitinase were investigated.Results showed that the highest productivity of chitinase was obtained when induced for 108 hours.The expression of chitinase was repressed when pH was lower than 5.5 and the optimum condition was pH 5.5～6.0.The optimum concentration of methanol was 1%.The protein expression can be improved greatly when added 0.05% oleic acid.On the basis of these,the recipe of fermentation medium was optimized through orthogonal experimental design.The optimal conditions were found and the amount of protein expression reached 171.99mg/L,chitinase activity 49.58U/mL under the conditions.