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섬유소 분해효소 생성을 위한 Trichoderma viride 조절 변이주의 개발
송은경,허태린,이세영 ( Eun Kyong Song,Tae Lin Huh,Se Yong Lee ) 생화학분자생물학회 1982 BMB Reports Vol.15 No.3
An attempt were made to isolate regulatory mutants of Trichoderma viride using a plating technique devised to screen constitutive cellulase mutants and high cellulase producing mutants. With U.V. mutagenesis, 70 mutants which exhibited high cellulase activity in the presence of 2% glucose were initially isolated from Trichoderma viride QM9414, a high celluase producing U.S. Army Natick Laboratory Strain. Upon continuous culture in the liquid medium, however, most of these mutants. were reverted to the glucose sensitive strain. One of the mutant, L40, was stable even after long period of culture in the liquid medium and still showed a high cellulase activity and relative insensitivity to glucose for cellulases production. The characteristics of the mutant L40 and cellulases produced by this mutant were as. follows: 1. When cellulases were induced by Avicel, a crystalline cellulose, the mutant L40 produced approximately 47 % higher cotton activity than the parental strain QM 9414. The induction time for the cotton activity was also shortened appreciably in. the mutant. CMC activity and β-glucosidase activity, however, were not changed in their total activity or induction time. 2. The cellulases induction by Avicel were less sensitive to glucose in mutant L40 than in the parental strain QM9414. 3. When cellulases were induced by carboxy methyl cellulose, a noncrystalline cellulose, there were no appreciable changes in cellulases activities, nor in induction times. 4. When cellulases were induced by cellobiose, the mutant L40 produced approximately 40 % higher cotton activity than the parental QM9414 and the induction time was shortened appreciably in the mutant. The CMC activity was also changed in the mutant but no change in β-glucosidase activity was observed. 5. The enzymic properties of cellulases and the growth characteristics of mutant L40 and the parental QM9414 were the same.
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섬유소 분해효소 생성을 위한 Trichoderma viride 조절 변이주의 개발
송은경,허태린,이세영,Song, Eun-Kyong,Huh, Tae-Lin,Lee, Se-Yong 생화학분자생물학회 1982 한국생화학회지 Vol.15 No.3
구성변이주와 높은 섬유소 분해력을 갖는 돌연변이주를 검색하는 데 이용되는 평판배양법을 써서 Trichoderma νiride의 조절 변이주를 선별하고자 하였다. U.V.를 돌연변이 유발원으로 사용하여, U.S. Army Natick Laboratory균주로써 높은 섬유소 분해력을 갖고 있는 Trichoderma νiride QM9414로부터 2%의 glucose존재하에서도 높은 섬유소 분해력을 나타내는 돌연변이 균주 70개를 선별해서 액침배양한 결과 이들 중 대부분이 glucose에 의해 repression을 받는 것으로 나타났으나 돌연변이 균주의 하나인 L40은 오랜동안의 액침 배양 후에도 높은 섬유소분해력을 보여주었으며, 섬유소 분해효소 생성에 있어서 glucose에 의해 비교적 repession을 덜 받는 것으로 나타났다. 변이주 L40과 변이주 L40에 의해 생성된 섬유소 분해효소의 특성은 다음과 같다. 1. 결정형 섬유소인 Avicel을 유도기질로 사용하여 섬유소 분해효소를 유도하였을 때 결정형 섬유소 분해력인 cotton activity가 약 47 % 증가하였으며 효소 유도시간도 모균주인 QM9414보다 상당히 단축되었다. 비결정형 섬유소 분해력인 CMC activity와 cellobiose를 glucose로 분해하는 ${\beta}$-glucosidase activity에는 큰 변화가 없었다. 2. Avicel을 유도기질로 사용하여 섬유소 분해효소를 유도하였을 경우 돌연변이 균주인 L40이 모균인 QM9414보다 glucose에 의해 repression을 덜 받았다. 3. 비결정 섬유소인 carboxymethyl cellulose로 섬유소 분해효소를 유도했을 때는 섬유소 분해효소의 활성이나 시간이 모균주인 QM9414와 거의 비슷했다. 4. Cellobiose로 섬유소 분해효소를 유도했을 때 돌연변이주 L40의 결정형 섬유소분해력이 모균주 QM9414보다 약 40%증가하였고 효소 유도시간도 약간 단축되었으며 비결정형 섬유소 분해력인 CMC activity도 약간 증가하였으나 유도시간에는 변화가 없었고 ${\beta}$-glucosidase activity는 그 역가에 있어서나 유도시간에 있어서 별다른 변화가 없었다. 5. 변이주인 L40이 모균인 QM9414에 있어서 섬유소 분해효소의 효소적 특성과 성장상의 특정의 차이점은 없었다. An attempt were made to isolate regulatory mutants of Trichoderma viride using a plating technique devised to screen constitutive cellulase mutants and high cellulase producing mutants. With U.V. mutagenesis, 70 mutants which exhibited high cellulase activity in the presence of 2% glucose were initially isolated from Trichoderma viride QM9414, a high celluase producing U.S. Army Natick Laboratory Strain. Upon continuous culture in the liquid medium, however, most of these mutants. were reverted to the glucose sensitive strain. One of the mutant, L40, was stable even after long period of culture in the liquid medium and still showed a high cellulase activity and relative insensitivity to glucose for cellulases production. The characteristics of the mutant L40 and cellulases produced by this mutant were as. follows: 1. When cellulases were induced by Avicel, a crystalline cellulose, the mutant L40 produced approximately 47 % higher cotton activity than the parental strain QM 9414. The induction time for the cotton activity was also shortened appreciably in. the mutant. CMC activity and ${\beta}$-glucosidase activity, however, were not changed in their total activity or induction time. 2. The cellulases induction by Avicel were less sensitive to glucose in mutant L40 than in the parental strain QM9414. 3. When cellulases were induced by carboxy methyl cellulose, a noncrystalline cellulose, there were no appreciable changes in cellulases activities, nor in induction times. 4. When cellulases were induced by cellobiose, the mutant L40 produced approximately 40 % higher cotton activity than the parental QM9414 and the induction time was shortened appreciably in the mutant. The CMC activity was also changed in the mutant but no change in ${\beta}$-glucosidase activity was observed. 5. The enzymic properties of cellulases and the growth characteristics of mutant L40 and the parental QM9414 were the same.
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