重组大肠杆菌生产谷胱甘肽发酵条件的研究

研究了重组Ｅ．Ｃｏｌｉ产ＧＳＨ的发酵条件，重点考察了添加酵母膏、前体氨基酸和ＡＴＰ的影响。结果发现，前体氨基酸和ＡＴＰ均能促进胞内ＧＳＨ的积累，若在发酵０ｈ和１２ｈ分别加入２０ｇ／ＬＡＴＰ和９ｍｍｏｌ／Ｌ前体氨基酸，则细胞干重和胞内ＧＳＨ含量可分别比对照提高２４％和１４倍。应用正交试验得出的针对细胞干重和ＧＳＨ总量的最佳组合，最大细胞干重和ＧＳＨ总量比原试验中的最好结果分别提高了１０％和２６％。在分析了该菌对葡萄糖利用情况的基础上，对该菌进行了指数流加培养，２５ｈ细胞干重与发酵液内ＧＳＨ总量分别达到８０ｇ／Ｌ和８８０ｍｇ／Ｌ，比摇瓶最好结果分别提高了８３和４６倍。

Fermentation conditions for production of glutathione by recombinant Escherichia coli]

The fermentation conditions for production of glutathion e by recombinant E.coli were investigated, the addition of certain materials, such as yeast extract, precusor amino acids and ATP were mainly focused on. The results showed that the addition of precusor amino acids and ATP could promote intracellulat GSH accumulation. DCW and intracellular GSH content would be 24% and 1.4 times higher than that of fermentation without additions when 2.0g/L ATP and 9mmol/L precusor amino acids were added at the begining and 12 h of fermentation, respectively. By using the optimized combination of additives obtained from orthogonal experiments, the maximal DCW and total GSH in broth could be improved 10% and 26% higher than the best results in orthogonal experiments, respectively. Based on the analysis of glucose utilizing ability of this strain, an exponential fed batch culture process was conducted. DCW and total GSH in broth culd be 8.3 and 4.6 times higher than that of shaking flask culture and finally reached 80g/L and 880mg/L, respectively.