摘要
研究了重组表达人源过氧化氢酶酵母菌株G13的发酵工艺和纯化方法。对接种量、生长期培养基、生长时间、诱导时间等因素进行了优化,并确立了10 L发酵罐的发酵条件。结果表明,菌体最高密度达到了0.15g/mL,比原重组菌株S4提高了1倍,人源过氧化氢酶表达量也由原来的平均600 U/mL提高到了1500U/mL。同时建立了二步法纯化该酶工艺,获得了电泳纯的人源过氧化氢酶。所得到的重组蛋白经SDS-PAGE和Western-blot鉴定,证实产物为人源过氧化氢酶。
The fermentation conditios such as inoculum size, growth medium, growth phase and induced time by a new recombinant yeast strain G13 were investigated and its new purification method was presented. Under the optimized conditions, the fermentation in 10 L fermentor was performed. The results showed that the expression level of rhcatalase was increased from 600 U/mL to 1500 U/mL. The purified rhcCatalase was obtained through an ion-exchange resin and verified by Western-blot. Its specific activity was equal to the levcl of commercial products.
出处
《工业微生物》
CAS
CSCD
2011年第3期71-75,共5页
Industrial Microbiology
基金
国家自然科学基金资助(编号30572276)
关键词
人源过氧化氢酶
毕氏酵母
重组表达
优化
纯化
human catalase
pichia pastoris
fermentation
optimization
purification
