Purification and Partial Characterization of a Developmentally Regulated 1,3- -Glucanase from Penicillium italicum

Sánchez, Miguel; Nombela, César; Villanueva, Julio R.; Santos, T

doi:10.1099/00221287-128-9-2047

Cited by 15 publications

(17 citation statements)

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“…The optimum pH for the purified enzyme was 4.0 to 5.0. Similar results were reported for the laminarinase produced by P. oxalicum (pH 4.0) (Copa-Patiño et al 1989), whereas the laminarinase from P. italicum had an optimum pH of 6.0 (Sánchez et al 1982). Laminarinases with an optimum of pH 5.0 are produced by a wide variety of organisms (Bara et al 2003;Chipoulet and Chararas 1984;Kulminskaya et al 2001;Sena et al 2011).…”

Section: Resultssupporting

confidence: 82%

“…According to Copa-Patiño et al (1989), the denatured molecular weight of P. oxalicum was found to be 79 kDa, which is similar to the molecular weight of the laminarinase from P. rolfsii c3-2(1) IBRL. A laminarinase with a molecular weight of 65 kDa was also detected in P. italicum by SDS-PAGE (Sánchez et al 1982). The effects of pH and temperature on the activity and stability of the laminarinase from P. rolfsii c3-2(1) IBRL are shown in Figs.…”

Section: Resultsmentioning

confidence: 95%

“…However, it was higher than the K m (0.04 mg/mL) of the laminarin produced by P. italicum (Sánchez et al 1982). There are few research reports on the purification of the laminarinases produced by Penicillium fungal strains.…”

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confidence: 85%

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Purification and Characterization of a Thermostable Laminarinase from Penicillium rolfsii c3-2(1) IBRL

Lee¹,

Arai²,

Demirtaş³

et al. 2014

BioResources

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A laminarinase (endo-β-1,3-glucanase) was purified to homogeneity from Penicillium rolfsii c3-2(1) IBRL, which was originally produced in liquid culture containing 1% xylan from birchwood, via anion-exchange chromatography, gel filtration on Sephacryl S-100, and hydrophobic interaction chromatography. A single protein band with a molecular weight of 75 kDa was detected by sodium dodecyl sulfatepolyacrylamide gel electrophoresis, which had an optimum catalytic activity at pH 4.0 to 5.0 and 70 °C. This purified enzyme was most stable in the pH range 4 to 7, while it was thermostable up to 55 °C and retained up to 90% of its activity after 4 h pre-incubation. A substrate laminarin kinetic study yielded estimated K m and V max values of 0.0817 mg/mL and 372.2 µmol/min/mg, respectively. Laminari-oligosaccharide degradation, which was analyzed by thin layer chromatography, yielded the major hydrolysis products laminaribiose and glucose.

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Section: Resultssupporting

confidence: 82%

Section: Resultsmentioning

confidence: 95%

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Purification and Characterization of a Thermostable Laminarinase from Penicillium rolfsii c3-2(1) IBRL

Lee¹,

Arai²,

Demirtaş³

et al. 2014

BioResources

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“…min-' . mg protein -') (Reichelt and Fleet, 1981;Sanchez et al, 1982;Notario, 1982;Usui et al, 1985;Tangarone et al, 1989;de la Cruz et al, 1995). The increase of V,,,, with the size of the substrate, indicates that the number of 1,3-/?-glucosidic bonds/molecule of substrate influence directly the activity of the 74-kDa endo-p-glucanase, suggesting a putative repetitive attack where the enzyme cleaves several times the glucan chain before dissociation (Robyt and French, 1970;Thoma, 1976).…”

Section: Discussionmentioning

confidence: 99%

Purification and Characterization of an Endo‐1,3‐β‐Glucanase from Aspergillus fumigatus

Fontaine

Hartland

Beauvais

et al. 1997

European Journal of Biochemistry

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An endo-I ,3-p-glucanase was purified from a cell wall autolysate of Aspcvgillus ,fumigatus. This pglucanase activity was associated with a glycosylated 74-kDa protein. Using a sensitive colorimetric assay and a high-performance anion-exchange chromatography with a pulsed electrochemical detector for product analysis, it was shown that the endoglucanase hydrolysed exclusively linear 1,3-P-glucan chains, had an optimum pH of 7.0 and an optimum temperature of 60°C. A substrate kinetic study gave a K,, value of 0.3 mg/ml for soluble (laminarin and laminari-oligosaccharides) and 1 .18 mg/ml for insoluble (curdlan) 1,3-P-glucan. Laminar-oligosaccharide degradation, analysed by HPLC, showed that the endoglucanase bind to the subtrate at several positions and suggested that the active site of the enzyme recognized five glucose units linked by a 1,3-1 bond. The association of the present endo-1,3-P-glucanase with the cell wall of A. fumigatus suggests a putative role for this enzyme during cell-wall morphogenesis.Keywords: endo-I ,3-P-glucanase ; cell wall : Aspergillus furnigutus; filamentous fungus.1 ,3-P-Glucanases are classified as exo-l,3-/$glucanase (1,3-P-glucan glucohydrolase) and endo-1,3-P-glucanase (1,3-1-glucan glucanohydrolase). Endoglucanase activities cleave inside a glucan chain in a more or less random fashion: while the exoglucanase activities release glucose residues from the non-reducing end. Many yeast and filamentous fungi produce both exocellular and cell-wall-associated exo-p-glucanases and endo-8-glucanases (Notario, 1982;Hien and Fleet, 1983 ;Copa-Patino, et al. 1989;Stahmann et al., 1993). 1,3-p-Glucan is a predominant polysaccharide of the fungal cell wall, and is thought to be responsible for the shape and rigidity of the wall (Cabib et al., 1982;Fleet, 1991). It has been suggested that 1,3-p-glucanase activities are essential to the continuous rearrangement of the wall 1,3-P-glucans during fungal growth (Cabib et al., 1988;Wessels, 1988). In the filamentous fungus Coprinus cinereus, endo-p-glucanases seem to play an essential role in stipe elongation (Kamada et al., 1985). In Sacchuronzyces cerevisiae, the production of exo-P-glucanases is growth-associated and cellcycle regulated (del Rey et al., 1979;Larriba et al., 1984;Jiang et al., 1995), suggesting that their activities are required at very specific stages during morphogenesis. For example, bud initiation, cell expansion, cell conjugation and sporulation are characterised by a higher activity of particular 1,3-~-glucanases (del Rey et al., 1982; Cenamor et al., 1987). However, gene disruption of yeast exo-p-glucanase activities does not modify the cellular growth (Larriba et al., 1995).Correspondence to T. Fontaine, lnstitut Pasteur, Laboratoire des Aspergillus, 25 rue du Docteur E. Roux, F-75724 Paris cedex 15, FranceAbbreviafions. Abz(OH)ONHNHZ, pamino-hydroxybenzoic acid hydrazide; Con-A, concanavalin A lcctin; dp, degree of polymerisation; Gn, oligosaccharide containing n 1,3-P-linked glucose residues ; G,,r, reduced oligosacchari...

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“…Penicilliupn oxalicum was found to form four enzymes during the so-called autolytic phase (CopaPatino et al, 1989). In Penicillium italicum (Sancez et al, 1982) three developmentally-regulated 1,3-P-glucanases were identified. Sclerotinia sclerotiorum (Jones et al, 1973) and Sclerotium glucanicum (Rapp, 1989) produce endo-and exo-173-P-glucanases.…”

Section: Introductionmentioning

confidence: 99%

Purification and characterization of four extracellular 1,3- -glucanases of Botrytis cinerea

Stahmann

Schimz

Sahm

1993

Journal of General Microbiology

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~ ~~~The filamentous fungus Botvytis cinerea was found to produce four 1,3+glucanases (Glu I, II, 111, IV), These enzymes were visualized by activity-staining after separation by native polyacrylamide gel electrophoresis (PAGE). During growth on glucose as the single carbon source, only Glu III was detectable in the culture supernatant of B. cinerea. After glucose was exhausted from the medium, the extracellular (1,3)( 1,6)-/%~-glucan (cinerean) capsule of the fungus was degraded. In this phase the other three enzymes became detectable and the amount of all four enzymes increased. The enzyme with the greatest activity was Glu II, which was purified to homogeneity by SDS-PAGE. Its M, was 75000 and its isoelectric point (PI) was 5.2. Glycosylation of Glu I1 was shown by the periodic acid/Schiff reaction after SDS-PAGE. Glu I1 cleaved cinerean and laminarin. Both substrates were degraded in an exo-manner as shown by product characterization, and by studying the viscosity decrease in comparison with the liberation of reducing groups. The concentration of substrate that gave halfmaximal velocity (Se5) for Glu II was 580 pg ml-' for cinerean and 152 pg ml-' for laminarin. For Glu 111, also purified to homogeneity by SDS-PAGE, an M, of 84 000 and a PI of 3-6 were determined. Glu III cleaved laminarin (So.5 119 pg ml-') but not cinerean. Glu I and Glu IV were purified by activity-stained native PAGE. Both enzymes cleaved cinerean and laminarin in an exo-manner. Glu I focused at PI 4.9; its Se5 was 275 pg ml-' with cinerean and 138 pg ml-' with laminarin. The PI of Glu IV was 3.4; its So.5 was 171 pg ml-' for cinerean and 27 pg ml-' for lamina rin.

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Purification and Partial Characterization of a Developmentally Regulated 1,3- -Glucanase from Penicillium italicum

Cited by 15 publications

References 4 publications

Purification and Characterization of a Thermostable Laminarinase from Penicillium rolfsii c3-2(1) IBRL

Purification and Characterization of a Thermostable Laminarinase from Penicillium rolfsii c3-2(1) IBRL

Purification and Characterization of an Endo‐1,3‐β‐Glucanase from Aspergillus fumigatus

Purification and characterization of four extracellular 1,3- -glucanases of Botrytis cinerea

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