Skip to main content
Plant Physiology logoLink to Plant Physiology
. 1982 Apr;69(4):864–868. doi: 10.1104/pp.69.4.864

Studies on the Pectic Substances of Plant Cell Walls

III. DEGRADATION OF CARROT ROOT CELL WALLS BY ENDOPECTATE LYASE PURIFIED FROM ERWINIA AROIDEAE

Haruyoshi Konno 1, Yoshiki Yamasaki 1
PMCID: PMC426319  PMID: 16662310

Abstract

Pectate lyase was isolated from the cell extract of Erwinia aroideae. The enzyme was further purified to a high degree by a procedure involving ammonium sulfate fractionation and chromatography on CM-Sephadex C-50 and on Sephadex G-200. The enzyme attacked its substrate in an endo fashion and was more active on the sodium salt of acid-insoluble polygalacturonate or pectic acid than it was on the methoxylated pectin. The enzyme had an optimum pH at 9.3, was stimulated by calcium ions, and was completely inhibited by ethylenediaminetetraacetic acid. In addition, the reaction products showed an absorption maximum between 230 and 235 nm and reacted with thiobarbituric acid. These results indicate that the purified enzyme is an endopectate lyase. The endopectate lyase also had the ability to solubilize effectively the pectic fraction from the cell walls of carrot (Daucus carota) root tissue. The enzyme released 30.5% of the wall as soluble products and also liberated all of the galacturonic acid present in the walls. The total neutral sugars released by the enzyme were 10.6% of the walls, which corresponded to 71.5% of noncellulosic neutral sugars. The soluble products were separated into five fractions by DEAE-Sephadex A-50 column chromatography. Based on the analysis of sugar composition of each fraction, the pectic fraction of carrot cell wall is presented.

Full text

PDF
864

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. BITTER T., MUIR H. M. A modified uronic acid carbazole reaction. Anal Biochem. 1962 Oct;4:330–334. doi: 10.1016/0003-2697(62)90095-7. [DOI] [PubMed] [Google Scholar]
  2. English P. D., Maglothin A., Keegstra K., Albersheim P. A Cell Wall-degrading Endopolygalacturonase Secreted by Colletotrichum lindemuthianum. Plant Physiol. 1972 Mar;49(3):293–298. doi: 10.1104/pp.49.3.293. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Hsu E. J., Vaughn R. H. Production and catabolite repression of the constitutive polygalacturonic acid trans-eliminase of Aeromonas liquefaciens. J Bacteriol. 1969 Apr;98(1):172–181. doi: 10.1128/jb.98.1.172-181.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Karr A. L., Albersheim P. Polysaccharide-degrading Enzymes are Unable to Attack Plant Cell Walls without Prior Action by a "Wall-modifying Enzyme". Plant Physiol. 1970 Jul;46(1):69–80. doi: 10.1104/pp.46.1.69. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Keegstra K., Talmadge K. W., Bauer W. D., Albersheim P. The Structure of Plant Cell Walls: III. A Model of the Walls of Suspension-cultured Sycamore Cells Based on the Interconnections of the Macromolecular Components. Plant Physiol. 1973 Jan;51(1):188–197. doi: 10.1104/pp.51.1.188. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  7. Loescher W., Nevins D. J. Auxin-induced Changes in Avena Coleoptile Cell Wall Composition. Plant Physiol. 1972 Nov;50(5):556–563. doi: 10.1104/pp.50.5.556. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Moran F., Nasuno S., Starr M. P. Extracellular and intracellular polygllacturonic acid trans-eliminases of Erwinia carotovora. Arch Biochem Biophys. 1968 Feb;123(2):298–306. doi: 10.1016/0003-9861(68)90138-0. [DOI] [PubMed] [Google Scholar]
  9. PREISS J., ASHWELL G. Polygalacturonic acid metabolism in bacteria. I. Enzymatic formation of 4-deoxy-L-threo-5-hexoseulose uronic acid. J Biol Chem. 1963 May;238:1571–1583. [PubMed] [Google Scholar]
  10. Talmadge K. W., Keegstra K., Bauer W. D., Albersheim P. The Structure of Plant Cell Walls: I. The Macromolecular Components of the Walls of Suspension-cultured Sycamore Cells with a Detailed Analysis of the Pectic Polysaccharides. Plant Physiol. 1973 Jan;51(1):158–173. doi: 10.1104/pp.51.1.158. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. WEISSBACH A., HURWITZ J. The formation of 2-keto-3-deoxyheptonic acid in extracts of Escherichia coli B. I. Identification. J Biol Chem. 1959 Apr;234(4):705–709. [PubMed] [Google Scholar]

Articles from Plant Physiology are provided here courtesy of Oxford University Press

RESOURCES