Enhancement of production of cloned glucoamylase under conditions of low aeration from recombinant yeast using a SUC2 promoter

Abstract

The effect of aeration rate on the production of cloned glucoamylase in a recombinant yeast was investigated. This system consisted of Saccharomyces cerevisiae transformed with the 2 μ-based plasmid YEpSUCSTA which contains the SUC2 promoter, the STA signal sequence, and the STA structural gene. In contrast to typical yeast expression reports, high production of cloned glucoamylase was achieved at low aeration level (0·3 vvm). The recombinant yeast grown at 0·3 vvm aeration produced more glucoamylase (0·94 units/ml) than when grown at 0·0 vvm, 0·6 vvm, or 0·9 vvm (9·4, 1·4, and 3·1 times more, respectively). A high dissolved oxygen level early in the cultivation was important for cell growth and a low dissolved oxygen level during the production stage was important for glucoamylase production. In large scale processes for the production of recombinant proteins, the maintenance of aeration and dissolved oxygen at high levels is difficult and expensive. In this work, we have evaluated the coordination of oxygen level with growth and protein production and developed optimal conditions. Since a low aeration rate was optimal, our results demonstrate that the method described at the laboratory scale should be successfully applied at an industrial scale.