Taka-amylaseの分離法に就て

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The method presented here has been derived from the hypothesis, that, when an enzyme in aqueous solution is dehydrated to some extent, insufficient for the precipitation of the enzyme, by means of electrolytes or solvents, it may be adsorbed efficiently by some weak adsorbents, on which little adsorption will take place if the activity of water is high enough. Aqueous extracts of koji, prepared from wheatbran, on which Asp. Oryzae has been grown, are used as starting materials in our experiments. It merits consideration, that Taka-amylase is hardly salted out by sodium sulfate in 100% saturation at 30℃, while the amylases in malt extract are precipitated almost quantitatively under these conditions. The difference in behaviors is analyzed, and it is found that malt extract contains a substance, which is salted out easily and the precipitate plays the role of an adsorbent in the sense of the hypothesis mentioned above. The responsible substance is thermostable and can be applied for the precipitation of Taka-amylase with good results in the above conditions.The quantitative separation of Taka-amylase from the extracts is made possible by the application of hydrocellulose (e. g. in the amt. of 0.2% in dry wt.), agar-sol (e. g. 0.15〜0.5%) or similar substances, followed by the saturation of sodium sulfate at 32℃, but starch granules and native cellulose are unavailable, Protease is also fairly adsorbed, but maltase sparingly in this procedure.In these conditions, casein and egg-albumin provide less efficient adsorbent for amylase as compared with substances of carbohydrate nature, but better for protease.In practice, after the saturation of the salt at 38℃, the filtration is carried out isothermally at 30℃. Elution is brought about without difficulty by the treatment of the filter-cake with small amount of water.In most cases, the adsorbents may be recovered and used repeatedly.
社団法人日本生物工学会の論文
1950-01-15

Taka-amylaseの分離法に就て

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