Studies on the Cellulase. (V) -Fractionation of Cellulolytic Complex produced by Trichoderma $viride(O_2-1)$

섬유소(纖維素) 분해효소(分解酵素)에 관(關)한 연구(硏究) (제5보(第5報)) -Trichoderma $(O_2-1)$가 생성(生成)하는 Cellulolytic Complex의 분별(分別)에 대(對)하여-

  • Published : 1969.09.15

Abstract

The yield of cellulase derived from Trichoderma $(O_2-1)$ was remarkably varied with various concentration of ethanol and acetone in purification of the enzyme. In the purification with ethanol of ${\beta}-glucosidase$, the best result was obtained in the concentration of 60% and, of CMCase and of filter paper disintegrating enzyme 80%. And in the purification with acetone of ${\beta}-glucosidase$, filter paper disintegrating enzyme, and CMCase, in the concentration of 60%, 80%, and 90% respectively, was shown the best yield. The activities of crude Cellulase preparation could be seperated into few of fractions by column chromatography with Silica gel, Cellulose powder, and gauze. Most of CMCase, avicelase, and ${\beta}-glucosidase$ were eluted, but most of filter paper disintegrating enzyme and the rest of enzymes mentioned the above were absorbed, and were eluted with water. Therefore, it was considered that CMCase is different from filter paper disintegrating enzyme in properties. The relative activity of CMCase was different from that of avicelase in the peak of elusion part. And it was considered that filter paper disintegrating enzyme and cellulose powder saccharifying enzyme was seperated respectively as absorption part and non absorption part. The auther came to the conclusion that at least there were more than three sorts of cellulase in Trichoderma $(O_2-1)$ cellulase preparation.

Trichoderma $(O_2-1)$의 조효소(粗酵素)를 Ethanol 및 Acetone 농도별(濃度別)에 따른 회수율(回收率)을 실험(實驗)한 결과(結果) 각(各) cellulase는 현저(顯著)한 차이(差異)가 나타났다. 즉(卽) Ethanol을 사용(使用)하였을 경우 ${\beta}-glucosidase$는 60% C.M.C분해효소(分解酵素), 여지붕괴(濾紙崩壞濾) 효소(酵素)는 80% 일때가 좋았고 Acetone을 사용(使用)하였을 경우 ${\beta}-glucosidase$는 60% 여지붕괴(濾紙崩壞濾) 효소(酵素)는 80%, C.M.C분해효소(分解酵索)는 90% 농도(濃度)일때가 좋은 결과(結果)를 나타내었다. 조정제(粗精製)한 Cellulase를 Silicagel, cellulose powder, Gauze를 사용(使用)한 column chromatography에 의(依)하여 분별(分別)란 결과(結果) 수치(數個)의 구분(區分)으로 분별(分別) 할 수 있었다. C.M.C 분해효소(分解酵索)와 Avicel 분해효소(分解酵索), ${\beta}-glucosidase$의 대부분(大部分)은 흡착(吸着)이 되지않고 유출(流出)되었으며, 대부분(大部分)의 여지붕괴효소(濾紙崩壞濾酵素)와 일부분(一部分)의 Avicel분해효소(分解酵索)는 흡착(吸着)되었다가 증류수(蒸溜水)로서 유출(流出)함으로씨 용출(溶出)되었다. 따라서 C.M.C 분해효소(分解酵索)와 여지붕괴효소(濾紙崩壞濾酵素)는 상이(相異)한 Cellulase 성분(咸分)임을 알았다. 또 C.M.C 분해효소(分解酵索)와 Avicel 분해효소(分解酵索)는 용출구분(溶出區分)에 있어 peak가 상이(相異)한 다른 구분(區分)에서 상대활성(相對活性)이 다르므로 역시 별개(別個)의 종류(種類)로 구분(區分)되었다. 또 여지붕괴효소(濾紙崩壞濾酵素)와 Cellulose powder (여지분말(濾紙粉沫)) 당화효소(糖化酵素)는 흡착구분(吸着區分)과 비흡착(非吸着) 구분(區分)으로 분별(分別)되니 사이(相異)한 Cellulase 성분(成分)이라고 생각된다. 따라서 Trichoderma viride $(O_2-1)$의 Cellulase는 적어도 3종이상(種以上)의 Cellulase 성분(成分)으로 되어 있다고 추찰(推察)된다.

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