甲醇毕赤酵母表达木质素过氧化物酶的研究

Expression of Lignin Peroxidase in Pichia methanolica PMAD16

将含有黄孢原毛平革菌(Phanerochaetechrysosprium)木质素过氧化物酶基因的甲醇毕赤酵母工程菌进行了鉴定和筛选,筛选得到木质素过氧化物酶活力高的菌株PMLIP08。确定了一步法发酵的最优葡萄糖浓度,优化其发酵培养条件,结果表明葡萄糖的添加量为10g/L时,发酵条件为pH3.0,诱导温度24℃,培养时间12h,甲醇添加量1.1%,诱导时间72h后发酵液中酶活可达4888U/L。

Engineered strains of Pichia methanolica （PM） containing lignin peroxidase were characterized and screened, and obtained a strain PM-LIP-08 with high lignin peroxidase activity. Then the most optimum glucose concentration for one-step-method fermentation was determined to optimize the fermentation culturing conditions.The results showed that with the adding amount of glucose at 10 g/L in YEPD medium, fermentation condition at pH 3.0, inducing temperature at 24 ℃ , culture for 12 hours,induce for 72 hours enzyme activity of the fermentation liquidthe adding amount of methanol at 1.1% （v/v）, after reached up to 4 888 U/L.