Bacillus subtilisのα-Amylase生産系の特質について : α-Amylase生産のmRANの安定性

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Employing a strain of Bacillus subtilis highoy active in α-amylase production and a few strains derived therefrom, physiological studies were undertaken in order to elucidate the behavior of α-amylase forming system which underlies our thoery of process kinetics for α-amylase production. The results indicate : (a) that the measured increase in α-amylase activity represents the corresponding de novo synthesis of α-amylase protein; (b) that the mRNA specific for α-amylase is much more stable than the ordinary mRNA for the great majority of cell protein including alkaline phosphatase. The experimental results are summarized as follows. (1) In replacement cultures of a leucine-less mutuant, both growth and α-amylase formation were arrested quickly when cells were brought in the condition of leucine starvation. The above result and the results of inhibition tests with p-fluorophenylalanine, 5-methyltryptophan, chloramphenicol and puromycin indicate that the measured increase in α-amylase activity in cultures represents the de novo synthesis of the enzyme protein. (2) The addition of actinomycin D (0.6μg/ml) to a growing culture inhibited completely both growth and alkaline phosphatase formation without lowering the rate of α-amylase formation. The latter rate observed at the time of drug addition was maintained for not less than 1 hr. In the same condition, the incorporation of ^<14>C-adenine into the 105,000×g pellet fraction of cells was stopped by the drug. (3) In cultures of an uracyl-less mutant, RNA synthesis, growth and alkaline phosphatase formation were interrupted when the cells were subjected to uracyl starvation. In the same condition, the initial rate of α-amylase formation was consistently maintained; no sign of rate-lowering was perceived for not less than 150 min. (4) The specific radioactivity of the purified preparation of α-amylase synthesized in the presence of ^<14>C-leucine by a leucine-less mut ant was not affected by actinomycine D added to the culture. (5) In a synchronized culture, alkaline phosphatase activity was increased stepwise in tune with growth, while no sign of slow-down was observed in the rate of amylase production.
社団法人日本生物工学会の論文
1968-06-25

Bacillus subtilisのα-Amylase生産系の特質について : α-Amylase生産のmRANの安定性

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