Masayasu Nomura

Enzyme formation in lysozyme lysate of Bacillus subtilis

An active lysate of B. subtilis was obtained by treatment with lysozyme in the presence of polyvinyl sulfate and protease inhibitor from potato extract. Such preparations retain the ability to form amylase, incorporate glycine into protein and nucleic acid, and oxidize glucose at rates comparable to those of intact cells. Such activities can be observed in the absence of osmotic regulation. The effectiveness of the addition of polyvinyl sulfate was interpreted to be due to an inhibition of the destructive action of intracellular RNAase.

Stimulation of amylase formation by a ribonucleic acid preparation from Bacillus subtilis

Abstract Further studies were performed on the nature of active factor (ribonucleic acid [RNA] preparation) which can stimulate the formation of amylase in Bacillus subtilis . Ribonuclease (RNAase) treatment of the acid-precipitated active factor apparently does not destroy the stimulating effect. However, the active factor becomes acid-soluble after RNAase treatment. This indicates that active factor is RNA or substance(s) attached to RNA. In contrast to ribonuclease treatment, alkaline hydrolysis destroys activity. The activity of RNA and its RNAase dugest from various sources were examined and compared with the active preparation obtained from an amylase-producing culture. Yeast RNA and its RNAase digest cannot replace the active factor. Preparations derived from a culture which is very poor in amylase production are also inactive. Specificity of action of this active factor preparation was examined. The preparations stimulates the incorporation of 15 C-glycine into protein to some extent. However, the same degree of stimulation is obserced with RNAase digest of yeast RNA. Therefore, this stimulation can ve interpreted as being due to non-specific action of general RNA in the preparation. In contrast to incorporation, protease formation is stimulated far better by factor preparation than by RNAase digest of yeast RNA, as in the case of amylase formation. In contrast to polynucleotide preparation reported by other workers, the active factor cannot induce amylase formation in cells which have been poor in amylase formation in a growing medium.