Production, purification, and characterization of an extracellular endo-β-1, 3-glucanase from a monokaryon of Schizophyllum commune ATCC 38548 defective in exo-β-1, 3-glucanase formation

Abstract

1994. Production, purification, and characterization of an extracellular endo-a-l,3-glucanase from a monokaryon of Schizophyllum commune ATCC 38548 defective in exo-P-1,3-glucanase formation. Can. J. Microbiol. 40: 18-23. Production of extracellular P-1,3-glucanase activity by a monokaryotic Schizophyllum commrtne strain was monitored and results indicated that the P-glucanase activity consisted of an endo-P-l,3-glucanase activity, besides a negligible amount of P-1,6-glucanase and P-glucosidase activity. Unlike the P-I ,3-glucanase production of the dikaryotic parent strain S. commune ATCC 38548, the P-1.3-glucanase formation of the monokaryon was not regulated by catabolite repression. The endo-P-I ,3-glucanase of the monokaryon was purified from the culture filtrate by lyophilization, anion exchange chromatography on Mono Q, and gel filtration on Sephacryl S-100. It appeared homogeneous on SDS-PAGE with a molecular mass of 35.5 kDa and the isoelectric point was 3.95. The enzyme was only active toward glucans containing P-1,3-linkages, including lichenan, a P-1 ,3-1,4-D-glucan. It attacked laminarin in an endo-like fashion to form laminaribiose, laminaritriose, and high oligosaccharides. While the extracellular P-glucanases from the dikaryotic S. commune ATCC 38548 degraded significant amounts of schizophyllan, the endo-P-l,3-glucanase from the monokaryon showed greatly reduced activity toward this high molecular mass fi-1,3-/P-1,6-glucan. The K, of the endoglucanase, using laminarin as substrate, was 0.28 mg/mL. Optimal pH and temperature were 5.5 and 50°C, respectively. The enzyme was stable between pH 5.5 and 7.0 and at temperatures below 50°C. The enzyme was completely inhibited by 1 mM H~~+ .Growth of the monokaryotic S. commune strain was not affected by its constitutive endo-P-l,3-glucanase formation. . 1994. Production, purification, and characterization of an extracellular endo-P-l,3-glucanase from a monokaryon of Schizophyllutn commune ATCC 38548 defective in exo-P-1,3-glucanase formation. Can. J. Microbiol. 40 : 18-23. Les auteurs ont CtudiC la production d'une activitC P-1,3-glucanase extracellulaire par une souche monocaryote de Schizophyllum cotnmune; les rCsultats ont indiquC que l'activitt P-glucanase est composCe d'une endo-P-l,3-glucanase, en plus d'une quantitC negligeable d'activitC P-1,6-glucanase et P-glucosidase. Contrairement i la production de P-1,3-glucanase de la souche parentale dicaryote de S. commutle ATCC 38548, la synthbse de P-1,3-glucanase du monocaryon n'a pas Ct C dCterminCe par la repression d'un catabolite. L'endo-P-1,3-glucanase du monocaryon a Ct C purifiCe du filtrat de culture par lyophilisation, chromatographie Cchangeuse d'anion sur Mono Q et filtration sur gel de Sephacryl S-100. L'Clectrophorbse en gel dCnaturant de polyacrylamide (SDS-PAGE) a rCvClt une protCine homogbne de masse molCculaire de 3 5 3 kDa et le point isoClectrique a CtC trouvC i 3,95. L'enzyme fut active seulement envers les glucanes contenant des liaisons P-1,3, incluant la lichenane, une p -1 3 1 ...

Characterization of an endo-1,3-β-D-glucanase produced during the interaction between the mycoparasite Stachybotrys elegans and its host Rhizoctonia solani

“…The affinity of the 75-kDa glucanase for laminarin is among the highest reported in the literature. The K m value of 20 µg·mL -1 is similar to that found for T. longibrachiatum (16 µg·mL -1 ) (Tangarone et al 1989) and lower than those observed for the 94-kDa glucanase from Stachybotrys elegans (180 µg·mL -1 ) (Tweddell et al 1995) and for glucanases obtained from Penicillium italicum (40 µg·mL -1 ) (Sanchez et al 1982), Schizophyllum commune ATCC 38548 (280 µg·mL -1 ) (Prokop et al 1994), and T. harzianum (1180 µg·mL -1 ) (Noronha and Ulhoa 1996).…”

Characterization of a 29-kDa β-1,3-glucanase fromTrichoderma harzianum

“…β‐1,3‐Glucanases of low molecular mass have also been reported previously in different fungi [1]. Values of 29.0, 32.1 and 35.5 kDa have been obtained for Rhizoctonia solani, Rhizopus arrhizus QM 1032 and Schizophyllum commune ATCC 38548 [14–16], respectively.…”

Characterization of a novel endoglucanase from Ganoderma lucidum

“…Future gene (mRNA) isolation and sequencing as well as genome analyses have to enlighten this issue further. Thus far, molecular weights of endoglucanases from various white‐rot fungi range from 15.5 to 65 kDa, as summarized in Table .…”