摘要
研究毕赤酵母工程菌GS-LM-18产高温碱性脂肪酶的发酵培养基。通过单因素及正交试验对发酵培养基种类、初始pH及甲醇诱导浓度进行优化,并进行10L发酵罐放大培养研究。结果表明,在诱导温度为30℃,甲醇诱导浓度为1.0%时,最优发酵培养基配方为葡萄糖2%,蛋白胨2%,酵母浸粉1.5%,硫酸镁0.05%,磷酸二氢钾0.11%,磷酸氢二钾0.044%;发酵初始pH为8。按优化条件进行10L发酵罐放大培养,获得发酵液酶活高达2 749U/mL,是摇瓶发酵水平的3.9倍,蛋白含量为1.8mg/mL,OD600为290。该毕赤酵母工程菌发酵生产高温碱性脂肪酶,具有工业化生产的潜力。
To research the fermentation medium of high-temperature alka-line lipase produced by Pichia pastoris GS-LM-18. The medium, the pH and the methanol concentration for induction were optimized by single factor experiments and orthogonal experimental design, and the results of optimization were verified by enlarged fermentation in 10 L fermentor. The results showed that the optimum fermentation medium for the lipase production from Pichia pastoris GS-LM-18 were as follows： glucose 2 %, peptone 2%, yeast extract 1.5%, Magnesium sulfate 0. 05%, potassium dihydrogen phosphate 0.11%, potassium hydrogen phosphate 0. 044%, the optimum pH 8, and the inducing temperature was 30 ℃ and the optimum inducing concentration of methanol was 1%. According to the optimum conditions, the final enzyme activity was as high as 2 749 U/ mL in 10 L fermentor, which was 3.9 times of the fermentation level in shake flask. And the final cell density（OD600） was 290, 1.8 mg protein was obtained in lmL fermentation broth. The high-temperature alkaline lipase produced by Pichia pastoris GS-LM-18 will be of industrial production potential.
出处
《食品与机械》
CSCD
北大核心
2014年第4期196-199,210,共5页
Food and Machinery
基金
山东省技术创新项目(编号:201340516010)
关键词
毕赤酵母
高温碱性脂肪酶
培养基
优化
Pichia pastoris
high-temperature alkaline lipase medium
optimization
