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Construction of cellulase hyperproducing strains derived from polyploids of Trichoderma reesei
1Department of Food Science and Technology, Faculty of Horticulture, Minamikyushu University, Miyazaki, Japan.
Abstract:
The mycelial mat of Trichoderma reesei strain QM 6a was treated with 0.1% (w/v) colchicine solution for 14 days and designated M14. The cellulase productivity of strain M14 was not much higher than that of the original strain. When conidia of M14 were treated with ethylmethane sulphonate (EMS) solution, the cellulase hyperproducers, M14-1 and M14-2, were isolated using a selection medium containing Avicel. The DNA content of M14-1 and M14-2 was higher than that of the original strain. Cellulase productivity per mycelium of these strains increased and was higher than that of the original strain. The cellulase productivity did not change through ten generations when these strains were cultivated successively on a medium containing Avicel. It was concluded that cellulase hyperproducers, whose cellulase productivity per mycelium increased, could be obtained when the conidia of strain M14 were treated with EMS.
Insights
Ethylmethane sulphonate (EMS) treatment of Trichoderma reesei strain M14 conidia yielded hyper-cellulase producing strains M14-1 and M14-2. These strains exhibit increased DNA content and stable, enhanced cellulase productivity.
Area of Science:
- Microbiology
- Biotechnology
- Enzyme Engineering
Background:
- Trichoderma reesei is a key fungus for industrial cellulase production.
- Strain improvement is crucial for enhancing cellulase yield and efficiency.
Purpose of the Study:
- To develop hyper-cellulase producing strains of Trichoderma reesei.
- To investigate the effect of ethylmethane sulphonate (EMS) on cellulase productivity.
Main Methods:
- Treatment of Trichoderma reesei strain QM 6a mycelia with colchicine to create strain M14.
- Mutagenesis of M14 conidia using ethylmethane sulphonate (EMS).
- Isolation and selection of hyper-cellulase producers on Avicel medium.
Main Results:
- Colchicine treatment showed limited improvement in cellulase productivity.
- EMS treatment successfully generated hyper-cellulase producing strains M14-1 and M14-2.
- Isolated strains M14-1 and M14-2 exhibited higher DNA content and significantly increased cellulase productivity per mycelium.
- Enhanced cellulase productivity remained stable over ten generations.
Conclusions:
- Ethylmethane sulphonate (EMS) is an effective mutagen for developing hyper-cellulase producing Trichoderma reesei strains.
- Increased DNA content correlates with enhanced cellulase productivity.
- The developed strains offer potential for improved industrial enzyme applications.