基因剂量与溶氧对毕赤酵母工程菌发酵产BMP的影响研究

Study on influence of gene dosage and DO on productivity of BMP by engineered P.pastoris

在Pichia pastoris表达系统中,研究了基因拷贝数、甘油补料周期和甲醇流加策略对BMP表达的影响,表明整合有16拷贝BMP基因的工程菌P.pastorisGS115-16B2表达量最高,优化后的发酵条件是：甘油补料周期为4h,采用限制性流加甲醇策略,将溶氧控制在10%~25%之间,诱导72h时,最大菌体密度OD600达到216,此时BMP的表达量为142.42mg/L。

Present work focused on the high expression of Beefy Meaty Peptide（BMP）by Pichia pastoris.The gene copy number,periodicity glycerol feeding and the strategy of methanol feeding for target protein induction were studied.P.pastoris GS115-16B2 was selected for high level of BMP expression.Results showed that the optimized fermentation conditions were 4 h of glycerol feeding period,and the DO should be controlled in about 10%~25% by using limited methanol fed-batch strategy.The maximum cell density and BMP expression levels were 216（OD600）and 142.42 mg/L,respectively,after 72 h of methanol induction.