实用临床医药杂志2013，Vol.17Issue(16)：27-30,4.DOI:10.7619/jcmp.201316010
rhuPAa-melittin发酵条件及大规模纯化方法
Optimization of rhuPAa-melittin fermentation and purification conditions on large scale
摘要
Abstract
Objective To obtain rhuPAa-melittin protein with high-purity,the conditions of rhuPAa-melittin in the fermentation and purification progress were optimized to achieve high productivity.Methods The rhuPAa-melittin to linear was used,and rhuPAa-melittin-pPICZαC was transformed into Pichia pastoris.PCR was used to identify the yeast genome,screening highly effective and stable secretion expression rhuPAa-melittin pichia pastoris strain.The conditions like expression time and pH values in the fermentation progress were optimized within small scale and large scale.Results 28-30 ℃,0.5％ were optimized and used in the fermentation of rhuPAa-melittin.Moreover,the running rate of methanol was controlled at 9 mL/ (h · L).The DO value of the initialmedium was used at 25 ％-30％.The pH value of fermentation was 6.0-7.0 and the whole progress lasted for 12 hrs.All these conditions were applied to obtain the rhuPAa-melittin protein with high production,high purity and yield rate.Conclusion The rhuPAa-melittin protein was produced at 350 mg/L with a purity of 96％ and 65％ yield rate.关键词
rhuPAa-melittin/发酵/纯化Key words
rhuPAa-melittin / fermentation / purification分类
医药卫生引用本文复制引用
金世光,李城,王大新,颜炜群..rhuPAa-melittin发酵条件及大规模纯化方法[J].实用临床医药杂志,2013,17(16):27-30,4.基金项目
江苏省十二五"科教兴卫工程"创新团队与领军人才(LI201159) （LI201159）
江苏省科技支撑计划社会发展项目(BE02010697) （BE02010697）
国家973重点项目(2007CB936104). （2007CB936104）
