Abstract
Multiple enzyme forms of β-mannanase activity fromPolyporus versicolor were puritied to molecular homogeneity by a sequence involving DEAE Bio-Gel A chromatography, gel filtration on Sephadex G-100 and high-performance liquid chromatography using anion exchange and hydrophobic Interaction media. Overall, 7.6% of input activity was recovered in four β-mannanases, A, B, C and 2A, which were purified 112.6-, 165.5-, 143.7-and 19.9-fold respectivety. The β-mannanases were acidic proteins displaying isoelectric points from 3.75 to 4.6, molecular weights in the range of 33,900 to 57,500 and increasing hydrophobicity in the order of C>B>2A>A. Optimal pH and temperature for the hydrotysis of glucomannan by all activities were pH 5.5 and 65°C, respectively. All preparations exhibited activity after 30 min at 65°C, or after protease digestion. Although the response of individual enzymes to selected ions was variable, all β-mannanases were inhibited in decreasing order of Hg2+>Cu2+>Zn2+>Mn2+. All activities functioned as endomannanases.
Résumé
De multiples formes enzymatiques de l'activité β-mannanasique dePolyporus verslcolor ont été purifiées jusqu'à l'homogénéité moléculaire par une séquence impliquant la chromatographie sur Bio-gel DEAE A, la filtration sur gel de Sephadex G-100, et la chromatographie liquide à haute performance utilisant l'échange anionique et les milieux à interaction hydrophobique. On a récupéré en tout 7.6% de l'activité Initiale dans quatre β-mannanases, A, B, C, et 2A, qui ont été purifiées respectivement 112.6, 165.5, 143.7 et 19.9 fois. Les β-mannanases sont des protéines acidiques exhibant des pointsiso-électriques de 3.75 à 4.6, des poids moléculaires compris entre 33 900 et 57 500, et une hydrophobicité croîssante dans l'ordre C>B>2A>A. Les pH et température optimum pour l'hydrolyse de la glucamannane par toutes les activités sont de 5.5 et 65°C respectivement. Toutes les préparations exhibent encore une activité après 30 minutes et 65°C ou après la digestion protéolytique. Bien que la réponse individuelle des enzymes à quelques ions choisis était variable, toutes les β-mannanases sont inhibées dans l'ordre décroissant: Hg2+>Cu2+>Zn2+>Mn2+. Toutes les activités fonctionnent comme endomannanases.
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References
Biely, P. 1985 Microbial xylanolytic systems.Trends in Biotechnology 3, 239–246.
Bradford, M. 1976 A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.Analytical Biochemistry 72, 248–254.
Dea, C.M., Morris, E.R., Rees, D.A., Welsh, E.J., Barnes, H.A. &Price, J. 1977 Associations of like and unlike polysaccharides: mechanism and specificity in galactomannans, interacting bacterial polysaccharides, and related systems.Carbohydrate Research 57, 249–272.
Derochers, M., Jurasek, L. &Paice, M.C. 1981 Production of cellulase, β-glucosidase, and xylanase bySchizophyllum commune grown on a cellulose-peptone mediumDevelopments in Industrial Microbiology 22, 679–684.
Eriksson, O., Goring, D.A.I. &Lindgren, B.O. 1980. Structural studies on the chemical bonds between lignins and carbohydrates in spruce wood.Wood Science and Technology 14, 267–279.
Gierer, J. & Wannström, S. 1985 Formation of lignin-carbohydrate ether bonds during alkaline pulping processes.Proceedings of the International Symposium on Wood and Pulping Chemistry (Vancouver, Canada, 26–30 August 1985).
Haemmerli, S.D., Leisola, M.S.A. &Fiechter, A. 1986 Polymerization of lignins by ligninases fromPhanerochaete chrysosporium.FEMS Microbiology Letters 35, 33–36.
Iversen, T. &Wannström, S. 1986 Lignin-carbohydrate bonds in a residual lignin isolated from pine kraft pulps.Holzforschung 40, 19–22.
Johnson, K.G. 1990 Exocellular β-mannanases from hemi-cellulolytic fungi.World Journal of Microbiology and Biotechnology 6, 000-000.
Johnson, K.G. &Lanthier, P.H. 1986 β-Lactamases fromActinopolyspora halophila, an extremely halophilic actinomycete.Archives for Microbiology 143, 379–386.
Johnson, K.G., Harrison, B.A., Schneider, H., Mackenzie, C.R. &Fontana, J.D. 1988 Xylanhydrolysing enzymes fromStreptomyces spp.Emzyme and Microbial Technology 10, 402–409.
Johnson, K.G., Silva, M.C., Mackenzie, C.R., Schneider, H. &Fontana, J.D. 1989 Microbial degradation of hemicellulosic materials.Applied Biochemistry and Biotechnology 20/21, 245–258.
Joseleau, J.P. &Kesraoui, R. 1986 Glycosidic bonds between lignin and carbohydrates.Holzforschung 40, 163–168.
Jurasek, L. & Paice, M.G., 1988 Biological bleaching of pulp. InProceedings, 1988 Tappi Bleaching Conference (Orlando, FL), pp. 11–13. Tappi Press.
Kantelinen, A., Rättö, M., Sundquist, J., Ranua, M., Viikari, L. &Lnko, M. 1988 Hemicellulases and their potential role in bleaching. InProceedings, 1988 Tappi Bleaching Conference (Orlando, FL), pp. 1–9. Tappi Press, Finland.
McCleary, B.V. 1988 β-d-Mannanase.Methods in Enzymology 160, 596–610.
MacKenzie, C.R., Bilous, D. &Johnson, K. G. 1984 Purification and characterization of an exoglucanase fromStreptomyces flavogriseus.Canadian Journal of Microbiology 30, 1171–1178.
Minor, J.L. 1986 Chemical linkage of polysaccharides to residual lignin in Loblolly pine kraft pulps.Journal of Wood Chemistry and Technology 6, 185–201.
Mora, F., Comtat, F., Barnoud, F., Pla, F. &Noe, P. 1986 Action of xylanases on chemical pulp fibers. Part I. Investigation on cell-wall modifications.Journal of Wood Chemistry and Technology 6, 147–165.
Noe, P., Chevalier, J., Mora, F. &Comtat, J. 1986 Action of xylanases on chemical pulp fibers. Part II. Enzymatic beating.Journal of Wood Chemistry and Technology 6, 167–184.
Paice, M.B. &Jurasek, L. 1984 Removing hemicellulose from pulps by specific enzymic hydrolysis.Journal of Wood Chemistry and Technology 6, 187–198.
Sjöström, E. 1977 The behaviour of wood polysaccharides during alkaline pulping processes.Tappi 60, 151–154.
Smith, M. M. &Hartley, R. D. 1983 Occurrence and nature of ferulic acid substitution of cell-wall polysaccharides in graminaceous plants.Carbohydrate Research 118, 65–80.
Somogyi, M. 1952 Notes on sugar determination.Journal of Biological Chemistry 195, 19–23.
Takahashi, R., Kusakabe, I., Kobayashi, H., Murakami, K., Maekawa, A. &Suzuki, T. 1984 Purification and some properties of mannanase fromStreptomyces sp.Agricultural and Biological Chemistry 48, 2189–2195.
Viikari, L., Ranua, M., Kantelinen, A., Linko, M. & Sundquist, J. 1987 Applications of enzymes in bleaching. InProceedings of the International Symposium of Wood Pulping Chemistry (Paris, France)1, 151–154. Tappi Press.
Watanabe, T., Kaizu, S. & Koshijima, T. 1986 Binding sites of carbohydrate toward lignin in ‘lignin-carbohydrate complex’ fromPinus densiflora wood.Chemistry Letters, 1871–1874.
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Johnson, K.G., Ross, N.W. & Schneider, H. Purification and some properties of β-mannanase fromPolyporus versicolor . World J Microbiol Biotechnol 6, 245–254 (1990). https://doi.org/10.1007/BF01201292
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DOI: https://doi.org/10.1007/BF01201292