重组酵母产人IFN-α2a与LL-37融合蛋白的工艺优化

The Process Optimization of Pichia pastoris GS115LI for Producing Fused Protein of LL-37 and IFN-α2a

通过单因素试验探索重组酵母P.pastoris GS115LI产人LL-37基因与IFN-α2a融合蛋白的最佳条件。结果显示,重组酵母菌产蛋白酶的最佳表达条件是利用BMMY为诱导培养基,以接种量OD600=5.5、温度26℃、p H6.0、诱导剂(甲醇)为每隔24 h添加1.0%、振荡速度为200 r/min条件下连续诱导144 h。在最佳培养条件下,发酵液中的LL-37最高抗菌活性达27.9 mm。与初始的发酵条件相比,人LL-37基因与IFN-α2a融合蛋白的产量提高了32.29%。

With experiments of single factor, the optimal process of recombinantP. pastoris GS115LI for producing fused protein of LL-37 and IFN-α2a were using BMMY as inducing medium, inoculum as OD600=5.5, temperature as 26℃, pH6.0, inducing chemical（methanol） adding amount as 1.0% every 24 h, incubating shaking speed as 200 r/min, and inducing time as 144 h. Under those optimal conditions, the 27.9 mm of LL-37 antimicrobial activity could be gotten. Compared with the original conditions, the productivity of fused protein of LL-37 and IFN-α2a was increased as 32.29%.