重组人生长激素在毕赤酵母中的表达研究

Production and secretion of recombinant human growth hormone by pichia pastoris

通过单因素分析和正交实验法优化高密度发酵培养基和诱导条件，根据优化结果指导发酵罐发酵．得到改良BSM培养基的最佳配比为甘油30g／L、酵母粉14g／L、K2SO4 13g／L、MgSO4·7H2O11g／L、CaSO4 0．3g／L、PTM14．4mL、0．1mol／L磷酸盐缓冲液（pH=6．5）；得到摇瓶培养最佳诱导条件为种龄24h，诱导时间30h，甲醇浓度2％，pH6．3～6．9．瑞士Bioengineering公司L1523型发酵罐发酵结果显示，细胞光密度（OD600）达到294，rHGH表达量最高达到0．54g／L．

The optimum medium ingredients and induction conditions in shake-flask culture were investigated by monofactorial analysis and orthogonal experiment. The results of shake-flask culture were scaled up to 12.8L fermenter under the optimum conditions. The optimum medium ingredients are ： glycerol 30g/L,yeast extract 14g/L,K2SO4 13g/L,MgSO4 · 7H2O11g/L,CaSO4 0. 3g/L,PTM1 4.4mL, 0. lmol/L phosphate buffer saline（pH= 6.5）, the optimum induction conditions include the optimal seed growth time is 24h, induction time is 30h, methanol concentration is 2%, pH is 6.3～6.9. In fermentation, the maximum yield of recombinant human growth hormone reached 0.54g/L, the OD600 reached 294.