B. hydrolyticusの "風化型" Cellulaseに就て : 細菌 Hydrolaseに關する研究・第3報
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The occurence of cellulase of "weathering" type in cultures of an aerobe B. hydrolyticus, a non-typical cellulose-organism, has already been suggested by us.In order to confirm this suggestion, we worked out with crude enzye solution derived from the organism. The organism previously adapted on cellulose, was grown in pepton-broth, furnished sometimes with cellulodextrin.After 2〜3-days indubation, the bacteria were collected from te surface of the media and the paste was suspended in phospharte buffer (pH, 7) to be subjected to autolysis in the presence of tuluol or sodium cyanide at 50℃.Autolysate thus prepared, or cell-free enzyne solution, obtained from the autolysate by precipitation with alcohol, followed by extraction with water, was used to react upon disintegrated filter paper at 50℃ and pH 7,which are the conditions nearly optimum for the reaction.Added substrate is recorved almost completely after 20-hours incubation in the sediment of reaction mixture, denoting the lack of liquefying action, but it is attacked by the enzyme so as to become fragile and acid-labile in agrement with the previous results obtained in cellulose-cultures of this organism. The values of R^* are as a whole much greater than those obtained in cellulose-cultures.The extent of enzymic attack upon substrate is not unique, for K^* of attacked cellulose decreases with the extent of acid-hydrolysis, indicating that the penetration of the enzyme into mycel-band of cellulose proceeds very slowly.Reducing action upon hypoiodite is slightly increased with attacked cellulose as compared with intakt control.The cellulase of weathering type is therefore quite different from ordinary saccharogenic cellulase and is characterized distinctly by the mode of attack described above. The nature of the enzymic action may primarily consist in partial hydrolysis at β-glucosidic linkages situated far distant from terminal groups in the molecule.It is presumed, in conenction with the previous report, that the organism produces, basides this enzyme, a very small amount of saccharogenic cellulase in old cellulose-cultures, bur the young cells retain exclusively the enzyme of weathering type. The distribution of this enzyme is obscure, and it is supposed to be difficult to direct it in typical cellulose-organisms, even if they produce it, may be covered by the predominant action of ordinary cellulase.
- 公益社団法人日本生物工学会の論文
- 1949-09-15
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