Abstract
Aspergillus flavus grown in a liquid medium containing pectin as the sole carbon source produced extracellular enzymes which degraded the 1,4-α-d-glycosidic bonds of pectin. The products of degradation were characteristic of substances produced by transeliminase. Synthesis of this enzyme was repressed by the addition of sucrose, glucose, fructose and maltose. The crude enzyme was partially purified by a combination of ultrafiltration and ammonium sulfate precipitation. The partially purified enzyme was separated by molecular exclusion chromatography into three components A, B and C, with molar masses ranging from 13.2 to 64 kDa. Only fraction B exhibited enzymic activity and further fractionated by ion-exchange chromatography into four components I–IV. Among these components, only fractions I and II possessed transeliminase activity. Both fractions had an optimum activity at pH 8.5 and 35°C, and were stimulated by Ca2+, Mg2+, Na+ and K+ but inhibited by EDTA and DNP. The apparentKm for the degradation of pectin by fractions I and II were 6.2 and 8.0 g/L, respectively.
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Alaňa A., Gabilondo A., Hernando F., Moragues M.D., Domingues J.B., Llama M.J., Serra J.L.: Pectin lyase production by aPenicillium italicum strain.Appl. Environ. Microbiol.55, 1612–1616 (1989).
Alaňa A., Alkorta I., Dominguez J.B., Llama M.J., Serra J.L.: Pectin lyase activity in aPenicillium italicum strain.Appl. Environ. Microbiol.56, 3755–3759 (1990).
Albersheim P., Killias U.: Studies relating to the purification of pectin transeliminase.Arch. Biochem. Biophys.97, 107–115 (1962).
Albersheim P., Jones T.M., English P.D.: Biochemistry of the cell wall in relation to infective processes.Ann. Rev. Phytopathol.7, 171–194 (1969).
Albersheim P., Neukom H.: Splitting reaction of pectin chain molecules in neutral solutions.Arch. Biochem. Biophys.90, 46–51 (1960).
Auzla S.S., Chohan J.S., Dutt S.: Production of polygalacturonase by both producer and non-producer isolates ofAspergillus flavus exFriesin vitro andin vivo.Indian Phytopath.28, 294–295 (1975).
Bateman D.F.: The “macerating enzyme” ofRhizoctonia solani.Phytopathology3, 1178–1186 (1963).
Bateman D.F.: The enzymatic maceration of plant tissue.Neth. J. Plant Pathol.74, 67–80 (1968).
Bateman D.F., Millar R.L.: Pectic enzymes in tissue degradation.Ann. Bot. N.S.1, 59–65 (1966).
Dean M., Wood R.K.S.: Cell wall degradation by pectate transeliminase.Nature214, 408–410 (1967).
Dixon M., Webb E.C.:Enzymes. William Clowes & Sons, Great Britain 1971.
Dube H.C., Mathur S.: Hydrolytic and transeliminative degradation of pectic substances by extracellular enzyme ofVerticilium dahlie.Indian Phytopathol.28, 346–351 (1975).
Durrands P.K., Cooper R.M.: The role of pectinases in vascular wilt disease as determined by defined mutants' ofVerticillium albo-atrum.Physiol. Mol. Plant Pathol.32, 363–371 (1988).
Garibaldi A., Bateman D.F.: Pectic enzymes produced byErwinia chrysanthemi and their effects on plant tissue.Physiol. Plant Pathol.1, 25–40 (1971).
Gomes I., Saha R.K., Mohiuddin G., Hoo M.M.: Isolation and characterization of cellulase-free pectinolytic and hemicellulolytic thermophilic fungus.World J. Microbiol. Biotechnol.8, 589–592 (1992).
Hagar S.S., McIntyre G.A.: Pectic enzymes produced byPseudomonas fluorescens, an organism associated with “pink eye” disease of potato tubers.Can. J. Bot.50, 2479–2488 (1972).
Hancock J.G., Millar R.L., Lobeer J.W.: Role of pectolytic and cellulolytic enzymes inBotrytic leaf blight of onion.Phytopathology54, 932–935 (1964).
Hsu E.J., Vanghn R.H.: Production and catabolic repression of the constitutive polygalacturonic acid transeliminase ofAeromonas liquefaciens.J. Bacteriol.98, 172–181 (1969).
Hunter W.J., Elkan G.H.: Role of pectic and cellulolytic enzymes in the invasion of the soybean byRhizobium japonicum.Can. J. Microbiol.21, 1254–1258 (1975).
Liao C.H., Wells J.M.: Association of pectolytic strains ofXanthomonas campestris with soft rots of fruits and vegetables at retail markets.Phytopathology77, 418–422 (1987).
Lisker N., Katan J., Henis Y.: Sequential production of polygalacturonase, cellulase, and pectin lyase byRhizotonia solani.Can. J. Microbiol.21, 1298–1304 (1975).
Lund B.M., Brocklehurst T.F.: Pectic enzyme of pigmented strains ofClostridium.J. Gen. Microbiol.104, 59–66 (1978).
Mount M.S., Bateman D.F., Basham H.G.: Induction of electrolyte loss, tissue maceration, and cellular death of potato tissue by an endopolygalacturonate transeliminase.Phytopathology60, 924–931 (1970).
Nagel C.W., Anderson M.M.: Action of a bacterial transeliminase on normal and unsaturated oligogalacturonic acids.Arch. Biochem. Biophys.112, 332–330 (1965).
Nasuno S., Starr M.P.: Polygalacturonic acid transeliminase ofXanthomonas campestris.Biochem. J.104, 178–185 (1967).
Olutiola P.O.: Some environmental and nutritional factors affecting growth and sporulation ofAspergillus flavus.Trans. Brit. Mycol. Soc.66, 131–136 (1976).
Olutiola P.O., Akintunde O.A.: Pectin lyase and pectin methylesterase production byPenicillium citrinum.Trans. Brit. Mycol. Soc.72, 49–55 (1979).
Oyediran J.O.: Internal mouldiness of commercial cocoa in Ibadan, Western State, Nigeria.Rep. Niger. Stored Prod. Res. Tech. Inst. Rep.2, 51–53 (1970).
Perez-Artes E., Tena M.: Pectic enzyme from two races ofFusarium oxysporum f. sp.ciceri. Enzyme production in culture and enzymatic activity on isolated chickpea cell walls.J. Phytopathol.124, 39–51 (1989).
Pope C.R.: Pectolytic enzyme production in culture of two pathogens ofRumex species.Trans. Brit. Mycol. Soc.58, 523–526 (1972).
Reddy M.N., Stuteville D.L., Sorenson E.L.: Pectolytic activity of byXanthomonas alfalfae in vitro and in infected alfalfa plants.Phytopathology59, 887–888 (1969).
Sherwood R.T.: Pectin lyase and polygalacturonase production byRhizoctonia solani and other fungi.Phytopathology56, 279–286 (1966).
Smith W.K.: Chromatographic examination of the products of digestion of pectic materials by culture solutions of plant pathogenic and other bacteria.J. Gen. Microbiol.18, 42–47 (1958).
Talboys P.W., Busch L.V.: Pectic enzymes produced byVerticillium species.Trans. Brit. Mycol. Soc.55, 367–381 (1970).
Trevelyan W.E., Procter D.R., Harrison J.S.: Detection of sugars on paper chromatograms.Nature166, 444–445 (1950).
Tribe H.T.: Studies in the physiology of parasitism. XIX. On the killing of plant cells by enzymes fromBotrytis cinerea andBacterium aroidae.Ann. Bot.19, 351–368 (1955).
Wang S.C., Pinckard J.A.: Pectic enzymes produced byDiplodia gossipina in vitro and in infected cotton bolls.Phytopathology61, 1118–1124 (1971).
Warren L.: Thiobarbituric acid spray reagent for deoxy sugars and sialic acids.Nature186, 237 (1960).
Whitaker D.R., Hamson K.R., Datta P.K.: Improved procedure for preparation and characterisation ofMyrothecium cellulase. 2. Purification procedures.Can. J. Biochem. Physiol.41, 671–696 (1963).
Wood R.K.S.: Pectic and cellulolytic enzymes in plant diseases.Ann. Rev. Plant Physiol.11, 299–322 (1960).
Wood R.K.S.:Physiological Plant Pathology. Blackwell, Oxford 1967.
Zucker M., Hankin L.: Regulation of pectate lyase synthesis inPseudomonas fluorescens andErwinia carotovora.J. Bacteriol.104, 13–18 (1970).
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Famurewa, O., Oyede, M.A. & Olutiola, P.O. Pectin transeliminase complex fromAspergillus flavus . Folia Microbiol 38, 459–466 (1993). https://doi.org/10.1007/BF02814396
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DOI: https://doi.org/10.1007/BF02814396