Purification and characterization of a glycoside hydrolase family 5 endoglucanase from Tricholoma matsutake grown on barley based solid-state medium
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Strain and culture conditions
T. matsutake strain NBRC30605 was obtained from the NITE Bioresource Research Center (Chiba, Japan). T. matsutake mycelia were grown in SY medium (2% soluble starch from corn, 0.3% yeast extract and 2% agar, w/v, pH 5.1) for 40 days at 24°C with constant conditions. Standard solid-state medium contained 30 g of rolled barley and 15 g of vermiculite with a 60% moisture content in 100 mL Erlenmeyer flask. Liquid contents of the media were prepared with 0.3% yeast extract, although the water
Purification and identification of the endoglucanase from T. matsutake grown in solid-state culture
To determine the productivity of the endoglucanase activity in T. matsutake, we examined cultures in the presence of several compositions of barley based solid-state medium (40 days incubation) and GYL medium (30 and 60 days incubation), as shown in Fig. 1. Of the media tested, rolled barley gave the fastest vegetative mycelial growth, followed by a mixed weight ratio of 2:1 of rolled barley and vermiculite (Fig. 1A). Moreover, this condition had the highest activities with barley β-glucan and
Discussion
Complete hydrolysis of cellulose requires the cooperative actions of three types of cellulases: endoglucanase (EC 3.2.1.4) that randomly cleaves the internal β-1,4-glycosidic bonds; two types of cellobiohydrolases, CBHI and CBHII (EC 3.2.1.176 and EC 3.2.1.91) that processivity act on the chain termini to release cellobiose; and β-glucosidase (EC 3.2.1.21) that hydrolyzes cellobiose to glucose (28). Endoglucanase also hydrolyzes the β-1,3-1,4-branched β-glucans converting into oligosaccharides.
Acknowledgments
This study was supported in part by a grant by the Strategic Research Foundation Grant-aided Project for Private Universities from the Ministry of Education, Culture, Sports, Science, and Technology (MEXT), 2015-2017 (S1512004).
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