Effect of Culture Temperature and Dissolved Oxygen Concentration on Expression of α-Amylase Gene in Batch Culture of Spore-Forming Host, Bacillus subtilis 1A289

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Spore-forming recombinant Bacillus subtilis 1A289 containing plasmid pC194 carrying an α-amylase encoding gene was used for the production of the cloned gene product in batch culture. In flask culture, the α-amylase activity at a culture temperature of 45℃ was 643 U/ml which was 2-fold higher than that at 37℃. Nospore formation was detected int he culture. When the cells were cultured in a jar fermentor at 45℃ with a dissolved oxygen (DO) level of 90%, the enzyme activity was only 184 U/ml which was only 30% of that in the flask culture. However, when the DO level was controlled at 5%, the activity was 846 U/ml. In the case of cultures at 45℃ with a DO level higher than 50%, spore concentration and spore fraction were more than 3.3×10^8 spores/ml and 85-100%, respectively. These values were 10 times higher with respect to spore concentartion and 6 times higher with respect to the fraction of cells forming spores than those at a DO level of 5%. There were three types of cells in the fermentor : cells resistant to chloramphenical and producing α-amylase; cells having lost plasmid pC194; cells forming spores. The specific enzyme activity of vegetative cells was 1.9kU/mg dry cell weight, while that of spores was 0.02 kU/mg dry cell wieight. The α-amylase gene expression of the spore-forming recombinant B. subtilis was affected strongyl by culture temperature, DO level, and fraction of cells forming spores.
社団法人日本生物工学会の論文
1997-07-25

Effect of Culture Temperature and Dissolved Oxygen Concentration on Expression of α-Amylase Gene in Batch Culture of Spore-Forming Host, Bacillus subtilis 1A289

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