Separation and Characterization of the endo-<i>β</i>-(1→3)-<scp>d</scp>-Glucanase from<i>Rizoctonia solani</i>

Totsuka, Atsushi; Usui, Taichi

doi:10.1080/00021369.1986.10867433

Cited by 9 publications

(12 citation statements)

References 6 publications

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“…The optimal pH and temperature for enzyme activity as well as the pH and temperature stability of the endo-P-1,3-glucanase from the monokaryon protoclone B No. 5 were within the range of other endo-P-1,3-glucanases from filamentous fungi (Clark et al 1978;Iwamuro et al 1985;Totsuka and Usui 1986;Tangarone et al 1989). The complete inhibition of the endo-P-1,3-glucanase of the monokaryon protoclone B No.…”

Section: Discussionmentioning

confidence: 87%

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

Prokop¹,

Rapp²,

Wagner³

1994

Can. J. Microbiol.

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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 ...

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Section: Discussionmentioning

confidence: 87%

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

Prokop¹,

Rapp²,

Wagner³

1994

Can. J. Microbiol.

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“…25) The molecular mass of the enzyme was estimated under denaturing conditions to be about 94 kDa. Of fungal fJ-l,3-g1ucanases, the molecular mass of fJ-l ,3-g1ucanase of S. elegans is higher than that of Penicillium italicum (68 kDa),21) Rhizopus arrhizus (32.1 kDa),26) R. solani (29 kDa),27) and Trichoderma longibrachiatum (70 kDa). 22) In a previous study, we detected two isoforms of fJ-l,3-glucanase when crude culture filtrate of S. elegans was electrophoresed on a native gel.…”

Section: Characterization Of P-j3-glucanasementioning

confidence: 96%

Purification and Partial Characterization of aβ-1,3-Glucanase Secreted by the MycoparasiteStachybotrys elegans

Tweddell¹,

Jabaji²,

Goetghebeur³

et al. 1995

Bioscience, Biotechnology, and Biochemistry

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A /I-l,3-glucanase secreted by Stachybotrys elegans, when grown on minimal synthetic medium containing Rhizoctonia solani cell wall fragments, was purified to homogeneity. The purification method involved ammonium sulfate precipitation and ion-exchange and size-exclusion chromatographies. The molecular mass of the enzyme was estimated under denaturing conditions to be about 94 kDa. The enzyme had an optimum pH of 5.0 and was most active between 40 and 50°C. Except for Mn 2 +, the enzyme activity was not sensitive to the metal ions tested and K m of 0.18 mg/ml was estimated for laminarin as a substrate. Cell wall lytic activity of purified /I-l,3-glucanase was tested on actively growing R. solani hyphae. /I-l,3-Glucanase induced morphological changes such as hyphal tip swelling, bursting, leakage of cytoplasm, and formation of numerous septae.Lytic enzymes, particularly chitinases and f3-1,3-glucanases, are essential in the life cycles of higher fungi. They have been implicated in apical growth, 1) wall softening during hyphal branching, germination, hyphal fusion,2,3) and conidia formation. 4 ) Recent studies indicate that during mycoparasitism of plant pathogenic fungi, several of these hydrolytic enzymes are produced by the mycoparasites and are involved in the degradation of fungal host cell walls. 5) Among these enzymes, chitinases and f3-1,3-g1ucanases are considered key cell-wall degrading enzymes of higher fungi. 6, 7) In addition, these enzymes have been shown to be produced by several potential biocontrol agents, and may be an important factor in biological control. 8 -11) Recently Stachybotrys elegans (Pidopl.) W. Gams, a soil fungus, was shown to have a strong mycoparasitic activity against the soilborne pathogen Rhizoctonia solani Kuhn in in vitro studies. Ultrastructural studies of the interaction between the mycoparasite and R. solani,12) as well as biochemical studies on the production of hydrolytic enzymes by S. elegans 13 ) strongly suggest the implication of these enzymes, particularly f3-1,3-g1ucanases, in the degradation of R. solani cell walls. As far as we can tell, nothing was reported about the f3-1 ,3-glucanases of this newly discovered mycoparasite. The objectives of this study were to purify and to partially characterize a f3-1,3-g1ucanase secreted by S. elegans, as well as to study its biological effects on the morphology of R. so/ani hyphae. Materials and MethodsOrganism and culture conditions. The mycoparasite S. elegans was isolated from soil in Izmir, Turkey. S. elegans was grown on sterile oat seeds for 10 days as described by Escande and Echandi. 14) The seeds were stored at 4°C and served as stock cultures. Inoculum for the production of fi-I ,3-glucanases in batch cultures consisted of potato dextrose agar t Corresponding author.(PDA; Difco Laboratories, Detroit, Michigan) disks of IO-day old mycelium of S. elegans grown from colonized oat kernels. Large-scale production of fi-I ,3-glucanases was done in 2-liter and 3~liter Erlenmeyer flasks containing respectively 400 and 60...

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“…While low molecular weight β-1,3-glucanases were also present in phytopathogenic fungi like Rhizoctonia solani (Totsuka and Usui, 1986), a high molecular mass extracellular endo-β-1,3-glucanase was produced by the mycoparasitic fungus T. harzianum (CECT 2413). The gene encoding this enzyme was cloned and found to encode a protein of a molecular mass of 78 kDa (De La Cruz et al, 1995).…”

Section: Purification Of the β β β β-13-glucanasementioning

confidence: 99%

Purification and characterization of a beta-Glucanase produced by Trichoderma harzianum showing biocontrol potential

Marco

Félix

2007

Braz. arch. biol. technol.

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Separation and Characterization of the endo-β-(1→3)-d-Glucanase fromRizoctonia solani

Cited by 9 publications

References 6 publications

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

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

Purification and Partial Characterization of aβ-1,3-Glucanase Secreted by the MycoparasiteStachybotrys elegans

Purification and characterization of a beta-Glucanase produced by Trichoderma harzianum showing biocontrol potential

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