摘要
采用5 L发酵罐对重组大肠杆菌E.coli BL21(DE3)/pET30-PFDH外源表达NADP^+型甲酸脱氢酶进行高密度发酵研究。通过对培养基、溶氧控制、诱导温度、诱导pH、诱导种类和时机的系统优化,在DO-star补料方式下,甲酸脱氢酶发酵液的酶活达到56.9 U/mL,相对于优化前提高了47.8%。在此基础上,利用200 L中试发酵罐对甲酸脱氢酶重组菌进行放大培养及工艺验证,重复6个批次,平均菌体质量浓度和平均酶活达到201.1 g/L和52.3 U/mL。优化后的高密度发酵条件放大重复性好,具备产业化前景。
The high-density fermentation study of NADP+-dependent formate dehydrogenase in recombinant E.coli BL21(DE3)/pET30-PFDH was conducted in 5 L fermentation tank.Through the systematic optimization of the medium,dissolved oxygen control,induction temperature,induction pH,induction species and timing,the enzyme activity of NADP+-dependent formate dehydrogenase fermentation broth reached 56.9 U/mL under the DO-star feeding method,which was 47.8%higher than before the optimization.On this basis,therecombinant Escherichia coli producing formate dehydrogenasewere used for scale-up culture and the process was verified by using a 200 L fermentation tank.Six batches were repeated,and the average cell mass concentration and enzyme activity reached 201.1 g/L and 52.3 U/mL.The optimized high-density fermentation conditions have good repeatability and industrialization prospect.
作者
赵翔
钟浩
孙超
汤利文
许岗
ZHAO Xiang;ZHONG Hao;SUN Chao;TANG Liwen;XU Gang(Hunan Flag Bio-technology Co.,Ltd.,Changsha 410100,China)
出处
《发酵科技通讯》
CAS
2020年第2期72-75,共4页
Bulletin of Fermentation Science and Technology
关键词
重组大肠杆菌
NADP^+型甲酸脱氢酶
高密度发酵
诱导
recombinant Escherichia coli
NADP+-dependent formate dehydrogenase
high density fermentation
induction
