Innovative Enzymatic Approach to Resolve Homogalacturonans Based on their Methylesterification Pattern

• Published in: Biomacromolecules 2012-05, Vol.13 (5), p.1615-1624
• Main Authors: Ralet, Marie-Christine, Williams, Martin A. K., Tanhatan-Nasseri, Abrisham, Ropartz, David, Quéméner, Bernard, Bonnin, Estelle
• Format: Article
• Language: English
• Subjects: Applied sciences; Chemical and Process Engineering; Engineering Sciences; Exact sciences and technology; Food engineering; Life Sciences; Methyl Ethers - chemistry; Methyl Ethers - metabolism; Models, Molecular; Natural polymers; Pectins - chemistry; Pectins - metabolism; Physicochemistry of polymers; Polygalacturonase - chemistry; Polygalacturonase - metabolism; Starch and polysaccharides
• ISSN: 1525-7797; 1526-4602
• DOI: 10.1021/bm300329r

Three series of model homogalacturonans (HGs) covering a large range of degree of methylesterification (DM) were prepared by chemical and/or enzymatic means. Randomly demethylesterified HGs, HGs containing a few long demethylesterified galacturonic acid stretches, and HGs with numerous but short demethylesterified blocks were recovered. The analysis of the degradation products generated by the action of a purified pectin lyase allowed the definition of two new parameters, the degree of blockiness, and the absolute degree of blockiness of the highly methylesterified stretches (DBMe and DBabsMe, respectively). By combining this information with that obtained by the more traditional endopolygalacturonase digestion, the total proportion of degradable zones for a given DM was measured and was shown to permit a clear differentiation of the three types of HG series over a large range of DM. This double enzymatic approach will be of interest to discriminate industrial pectin samples exhibiting different functionalities and to evaluate pectin fine structure dynamics in vivo in the plant cell wall, where pectin plays a key mechanical role.