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Study of the methyl ester distribution in pectin with endo ‐polygalacturonase and high‐performance size‐exclusion chromatography
Author(s) -
Daas Piet J. H.,
Voragen Alphons G. J.,
Schols Henk A.
Publication year - 2001
Publication title -
biopolymers
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.556
H-Index - 125
eISSN - 1097-0282
pISSN - 0006-3525
DOI - 10.1002/1097-0282(200102)58:2<195::aid-bip80>3.0.co;2-c
Subject(s) - pectin , chemistry , pectinase , molar mass distribution , polymer , polymerization , size exclusion chromatography , molecular mass , degree of polymerization , silylation , chromatography , organic chemistry , enzyme , biochemistry , catalysis
A method was developed that enables the study of the methyl ester distribution in the polymers of pectin on a molecular level. Endo ‐polygalacturonase was used to extensively degrade three 70% methyl esterified pectins. The molecular weight distribution of the non‐ and enzymatically degraded pectins was determined with high‐performance size‐exclusion chromatography. Next, the molecular weight distribution was converted into a degree of polymerization distribution of galacturonan fragments. Monte Carlo methods were employed for the reconstruction of the parental polymers from their enzymatic degradation products. The results for the random methyl esterified pectin revealed that the enzyme‐degradable sites were indeed randomly distributed, which confirmed the correctness of the procedure developed. The two other pectins studied differed greatly in the amount of non‐, low‐, and high‐esterified regions present in the reconstructed pectic molecules of a given molecular mass. That the approach developed is able to reveal such detailed information makes it unique. The information on the fragmental composition of pectic polymers obtained is an important addition to the study of the methyl ester distribution and the functional properties of pectin. © 2001 John Wiley & Sons, Inc. Biopolymers 58: 195–203, 2001