Optimization of Feeding Strategy for Overproduction of Human Lipocortin-I in Saccharomyces cerevisiae Controlled by the GAL10 Promoter

概要

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Fed-batch fermentation was conduced to overproduce human lipocorin-I (LC1) in recombinant Saccharomyces cerevisiae controlled by the GAL10 promoter. To optimize the feeding strategy, various fed-batch culture modes were performed with concentrated feed media containing carbon mixtures of galactose (Gal) and glucose (Glu) at three different concentration ratios (Gal : Glu ratios) : 9 : 1,1 : 1,1 : 9. The cell growth, expression level of LC1,and the plasmid stability were investigaged under these fed-batch culture modes. While both glucose and galactose were being fed to the fermentor, the clugose concentration in the culture broth was kept below 1g/l for efficient gene expression. High cell concentrations of greater than 100 g dry cell weight/l were achieved with these fed-batch culture modese. A significant amount of intact LC1 was found to be secreted into the culture medium, but proteolytically cleaved products (des1-26-LC1) were also observed in the culture medium. The fed-batch fermentation with the feed medium at a Gal : Glu ratio of 1 : 1 resulted in the highest LC1 total (intact LC1+des1-26-LC1) concentration of 500 mg/l, which correspondded to 1.73- and 1.83-fold increases over that produced with the media at Gal : Glu ratios of 1 : 9 and 9 : 1,respectively.
社団法人日本生物工学会の論文
1997-11-25