Optimization of the Expression of a Foreign Gene from Galactose-Inducible Promoter in a Recombinant Yeast

概要

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A plasmid which expresses mouse α-amylase under the control of GAL7 promoter was constructed and introduced into Saccharomyces cerevisiae 2230. To optimize the medium for cell growth and gene expression, we cultured the microorganism by shake-flask and fed-batch cultures. In the presence of glucose, gene expression was completely repressed. On the other hand, when glucose was exhausted and the inducer galactose was added to the medium, gene expression and the produciton of mouse α-amylase started immediately, both in shake-flask and fed-batch cultures. In the fed-batch culture, on-off regulation of gene expression could be achieved by changing the sugar substrate during culture. The amount of mouse α-amylase produced was equivalent to 370 mg/l human salivary amylase as evaluated from the specific activity of commercially available human salivary amylase. This value is 60-fold that of the repressive state using glucose as a substrate.
社団法人日本生物工学会の論文
1991-09-25