Abstract
The mode of action of the endopolygalacturonase from Fusarium moniliforme was studied towards a series of pectins with different amounts and distribution patterns of substituents. In homogalacturonan, the enzyme hydrolysed the linkages between two galacturonic acid residues according to a multi-chain attack mechanism, and the final products were monomer and dimer. The enzyme was able to degrade methylated pectins with a degree of methylation up to 70 and acetylated homogalacturonans with a degree of acetylation up to 38 to a lower extend but not xylogalacturonans.
When methylated pectins or acetylated homogalacturonan were used as substrates, the final percentage of hydrolysis decreased with increasing the degree of substitution. A blockwise distribution of the methyl-groups was more favourable to a higher velocity than a random one. In every case, the enzyme was shown to follow a multi-chain attack. The final products included methyl esterified oligogalacturonates. The enzyme seemed to degrade first the non-methylated area of the blockwise methylated substrate. The detailed structure of the oligomers was analysed by mass spectrometry, and showed that the enzyme was able to accomodate methylated or acetylated galacturonic acid residues in its active site.
Keywords
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsPreview
Unable to display preview. Download preview PDF.
References
Benen J.A.E., Kester H.C.M. and Visser J. (1999) Kinetic characterization of Aspergillus nigerN400 endopolygalacturonases I, II, and C. Eur. J. Biochem., 259, 577–585.
Bonnin E., Le GofiA., Körner R., van Alebeek G.-J.W.M., Christensen T.M.I.E., Voragen A.G.J., Roepstorff P., Caprari C., Thibault J.-F. (2001a) Study of the mode of action of endopolygalacturonase from Fusarium moniliforme, Biophysica, Biochimica Acta, in press.
Bonnin E., Dolo E., Le Goff A., Thibault J.-F. (2001b) Characterization of pectin subunits obtained by enzymic fractionation, submitted
Caprari A., Bergmann C., Migheli Q., Salvi G., Albersheim P., Darvill A., Cervone F. and De Lorenzo G. (1993) Fusarium moniliforme secretes four endopolygalacturonases derived from a single gene product. Physiology Molecular Plant Pathology, 43, 453–462.
Caprari C., Mattei B., Basile M.L., Salvi G., Crescenzi V., De Lorenzo G. and Cervone F. (1996) Mutagenesis of endopolygalacturonase from Fusarium moniliforme: Histidine residue 234 is critical for enzymatic and macerating activities and not for binding to polygalacturonase-inhibiting protein (PGIP). Molecular Plant-Microbe Interactions, 9, 617–624.
Kester, H.C.M., Magaud, D., Roy, C., Anker, D., Doutheau, A., Shevchik, V., Hugouvieux-Cotte-Pattat, N., Benen, J.A.E. and Visser, J. (1999) Journal ofBiological Chemistry, 274, 37053–37059.
Körner R., Limberg G., Mikkelsen, J.D. and Roepstorff, P. (1998) Characterisation of enzymatic pectin digests by matrix-assisted laser desorption/ionization mass spectrometry. Journal of Mass Spectrometry, 33, 836–842.
Le Goff A., Renard C.M.G.C., Bonnin E. and Thibault J.-F. (2001) Extraction, purification and chemical characterisation of xylogalacturonans from pea hulls. Carbohydrate Polymers, 45, 325–334.
Limberg G., Körner R., Buchholt H.C., Christensen, T.M.I.E., Roepstorff P. and Mikkelsen J.D. (2000a) Analysis of different de-esterification mechanisms for pectin by enzymatic fingerprinting using endopectin lyase and endopolygalacturonase II from Aspergillus niger. Carbohydrate Research, 327, 293–307.
Limberg G., Körner R., Buchholt H.C., Christensen, T.M.I.E., Roepstorff P. and Mikkelsen J.D. (2000b) Quantification of the amount of galacturonic acid residues in blocksequences in pectin homogalacturonan by enzymatic fingerprinting with exo- and endo-polygalacturonase II from Aspergillus niger. Carbohydrate Research, 327, 321–332.
Micard, V. and Thibault, J.-F. (1999) Oxidative gelation of sugar-beet pectins : use of laccases and hydration properties of the cross-linked pectins. Carbohydrate Polymers, 39, 265–273.
Nelson, N. (1944). A photometric adaptation of the Somogyi method for the determination of glucose. Journal ofBiology and Biochemistry, 153, 378–380.
Nelson P.E. (1992) Taxonomy and biology of Fusarium moniliforme. Mycopathology, 117, 29–36.
Parenicova L., Benen J.A.E., Kester H.C.M., Visser J. (2000) pgaA and pgaB encode two constitutively expressed endopolygalacturonases of Aspergillus niger. Biochemistry Journal, 345, 637–644.
Pasculli, R., Geraeds, C.,Voragen, A.G.J., Pilnik, W. (1991) Characterization of polygalacturonases from yeasts and fungi. Lebensmittel Wissenchaft und Technologie, 24, 63–70.
Rombouts, F.M. and Thibault, J.-F. (1986) Enzymic and chemical degradation and the fine structure of pectins from sugar beet pulp. Carbohydrate Research, 154, 189–203.
Schols, H.A., Bakx, E.J., Schipper, D., Voragen, A.G.J. (1995) A xylogalacturonan subunit present in the modified hairy regions of apple pectin. Carbohydrate Research, 279, 265–279.
Solms J. and Deuel H. (1951) Untersuchungen an acetylierter Pektinsäure. Helvetica Chimica Acta, 34, 2242–2249.
Sturgeon, R.J. (1990) Monosaccharides. In Methods in Plant Biochemistry (P.M.Dey, J.B. Harborne, eds), 2, pp. 1–37.
Thibault J.-F. (1983) Aspergillus niger endopolygalacturonase. 3- Action pattern on polygalacturonic acid. Carbohydrate Polymers, 3, 259–272.
Thibault J.-F., Renard C. M. G. C., Axelos M. A. V., Roger P. and Crépeau M.-J. (1993) Studies of the length of homogalacturonic regions in pectins by acid hydrolysis. Carbohydrate Research, 238, 271–286.
Vink, H. (1963) Degradation of some polymers in aqueous solutions Makromolekulare Chemie 67, 105–123
Voragen, A. G. J., Thibault, J.-F., Axelos, M.A.V., Renard, C.M.G.C. & Pilnik W. (1995) Pectins, in A.M. Stephen (ed), Food polysaccharides and their applications, Marcel Dekker Inc., New York, pp. 287–339.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2003 Springer Science+Business Media Dordrecht
About this chapter
Cite this chapter
Bonnin, E., Le Goff, A., Thibault, JF. (2003). Degradation of Pectins with Different Nature, Amount and Distribution of the Substituents by Endopolygalacturonase of Fusarium Moniliforme . In: Voragen, F., Schols, H., Visser, R. (eds) Advances in Pectin and Pectinase Research. Springer, Dordrecht. https://doi.org/10.1007/978-94-017-0331-4_28
Download citation
DOI: https://doi.org/10.1007/978-94-017-0331-4_28
Publisher Name: Springer, Dordrecht
Print ISBN: 978-90-481-6229-1
Online ISBN: 978-94-017-0331-4
eBook Packages: Springer Book Archive