A Novel Enzyme Activity Involving the Demethylation of Specific Partially Methylated Oligogalacturonides

Overview

Journal Biochem J

Specialty Biochemistry

Date 2002 Jul 5

PMID 12097140

Citations 1

Authors

Martin A K Williams

Jacques A E Benen

Affiliations

Soon will be listed here.

Abstract

Studies of the enzymic digestion of pectic substrates using different polygalacturonase (PG) preparations have revealed evidence for a previously unreported enzyme activity carried out by a contaminating enzyme in one of the preparations. This observed activity involves the demethylation of specific oligogalacturonides, namely 2-methyltrigalacturonic acid and 2,3-dimethyltetragalacturonic acid. However, no large-scale demethylation of highly methylated polymeric substrates is found, demonstrating that the enzyme responsible is not a conventional pectin methylesterase (PME). Furthermore, it has been shown that a commercial sample of fungal PME from Aspergillus niger demethylates all of the oligogalacturonides present as primary products of endo-PG digestion, in contrast with the activity observed here. On the basis of the known methyl ester distribution of the endo-PG-generated fragments and knowledge of which of these oligogalacturonides are demethylated, it is concluded that the observed activity can be explained by the existence of an exo-acting methylesterase that attacks the non-reducing end of the oligogalacturonide molecules.

Citing Articles

Study of the mode of action of a polygalacturonase from the phytopathogen Burkholderia cepacia.

Massa C, Clausen M, Stojan J, Lamba D, Campa C Biochem J. 2007; 407(2):207-17.

PMID: 17627609 PMC: 2049012. DOI: 10.1042/BJ20061833.

References

Benen J, Kester H, Visser J . Kinetic characterization of Aspergillus niger N400 endopolygalacturonases I, II and C. Eur J Biochem. 1999; 259(3):577-85. DOI: 10.1046/j.1432-1327.1999.00080.x. View

Korner R, Limberg G, Christensen T, Mikkelsen J, Roepstorff P . Sequencing of partially methyl-esterified oligogalacturonates by tandem mass spectrometry and its use to determine pectinase specificities. Anal Chem. 1999; 71(7):1421-7. DOI: 10.1021/ac981240o. View

Kester H, Magaud D, Roy C, Anker D, Doutheau A, Shevchik V . Performance of selected microbial pectinases on synthetic monomethyl-esterified di- and trigalacturonates. J Biol Chem. 1999; 274(52):37053-9. DOI: 10.1074/jbc.274.52.37053. View

Kester H, Benen J, Visser J, Warren M, Orlando R, Bergmann C . Tandem mass spectrometric analysis of aspergillus niger pectin methylesterase: mode of action on fully methyl-esterified oligogalacturonates. Biochem J. 2000; 346 Pt 2:469-74. PMC: 1220875. View

Limberg G, Korner R, Buchholt H, Christensen T, Roepstorff P, Mikkelsen J . 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. Carbohydr Res. 2000; 327(3):321-32. DOI: 10.1016/s0008-6215(00)00068-9. View

Ridley B, ONeill M, Mohnen D . Pectins: structure, biosynthesis, and oligogalacturonide-related signaling. Phytochemistry. 2001; 57(6):929-67. DOI: 10.1016/s0031-9422(01)00113-3. View

Catoire L, Pierron M, Morvan C, du Penhoat C, Goldberg R . Investigation of the action patterns of pectinmethylesterase isoforms through kinetic analyses and NMR spectroscopy. Implications In cell wall expansion. J Biol Chem. 1998; 273(50):33150-6. DOI: 10.1074/jbc.273.50.33150. View

de Vries R, Visser J . Aspergillus enzymes involved in degradation of plant cell wall polysaccharides. Microbiol Mol Biol Rev. 2001; 65(4):497-522, table of contents. PMC: 99039. DOI: 10.1128/MMBR.65.4.497-522.2001. View

Williams M, Buffet G, Foster T . Analysis of partially methyl-esterified galacturonic acid oligomers by capillary electrophoresis. Anal Biochem. 2002; 301(1):117-22. DOI: 10.1006/abio.2001.5481. View

10.

Kester H, Visser J . Purification and characterization of polygalacturonases produced by the hyphal fungus Aspergillus niger. Biotechnol Appl Biochem. 1990; 12(2):150-60. View

11.

Kester H, Muller Y, Visser J . Primary structure and characterization of an exopolygalacturonase from Aspergillus tubingensis. Eur J Biochem. 1996; 240(3):738-46. DOI: 10.1111/j.1432-1033.1996.0738h.x. View

12.

Daas P, Arisz P, Schols H, De Ruiter G, Voragen A . Analysis of partially methyl-esterified galacturonic acid oligomers by high-performance anion-exchange chromatography and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Anal Biochem. 1998; 257(2):195-202. DOI: 10.1006/abio.1997.2554. View

13.

Henrissat B, Coutinho P, Davies G . A census of carbohydrate-active enzymes in the genome of Arabidopsis thaliana. Plant Mol Biol. 2001; 47(1-2):55-72. View