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Characterization of two cell-wall polysaccharides from Fusicoccum amygdali.

ABSTRACT: 1. The nature of two polysaccharides (s(0) (20) values 6S and 2S respectively in 1m-sodium hydroxide), comprising a fragment (fraction BB, [alpha](D) +236 degrees in 1m-sodium hydroxide), previously isolated from cell walls of Fusicoccum amygdali, has been investigated. 2. Both the major (2S) and minor (6S) components were affected by incubation with alpha-amylase. The 6S polysaccharide was also attacked by exo-beta-(1-->3)-glucanase, which is evidence that it contained both alpha-(1-->4)- and beta-(1-->3)-glucopyranose linkages. By fractionation of the products of alpha-amylase-treated fraction BB it was possible to obtain a water-insoluble polysaccharide, fraction P ([alpha](D) +290 degrees in 1m-sodium hydroxide, 67% of fraction BB) and a water-soluble polysaccharide, fraction Q ([alpha](D) +16 degrees in 1m-sodium hydroxide, 11% of fraction BB), both of which sedimented as single boundaries with s(0) (20) values (in 1m-sodium hydroxide) of 1.7S and 4.6S respectively. 3. Evidence from periodate oxidation, methylation analysis, i.r. spectroscopy and partial acid hydrolysis showed that fraction P consisted of linear chains of alpha-(1-->3)-glucopyranose units with blocks of one or two alpha-(1-->4)-glucopyranose units interspersed at intervals along the main chain. The 2S polysaccharide, from which fraction P is derived, evidently also contains longer blocks of alpha-(1-->4)-glucopyranose units, that are susceptible to alpha-amylase action. 4. Fraction Q consisted of glucose (88%) with small amounts of galactose, mannose and rhamnose. Evidence from digestion with exo- and endo-beta-(1-->3)-glucanases, periodate oxidation and methylation analysis suggests that fraction Q consists of a branched galactomannorhamnan core, to which is attached a beta-(1-->3)-, beta-(1-->6)-glucan. In the cell wall, chains of alpha-(1-->4)-linked glucopyranose units are linked to fraction Q to form the 6S component of fraction BB.

SUBMITTER: Obaidah MA

PROVIDER: S-EPMC1178081 | biostudies-other | 1971 Nov

REPOSITORIES: biostudies-other

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Project description:Symbiotic bacteria inhabiting the human gut have evolved under intense pressure to utilize complex carbohydrates, primarily plant cell wall glycans in our diets. These polysaccharides are not digested by human enzymes, but are processed to absorbable short chain fatty acids by gut bacteria. The Bacteroidetes, one of two dominant bacterial phyla in the adult gut, possess broad glycan-degrading abilities. These species use a series of membrane protein complexes, termed Sus-like systems, for catabolism of many complex carbohydrates. However, the role of these systems in degrading the chemically diverse repertoire of plant cell wall glycans remains unknown. Here we show that two closely related human gut Bacteroides, B. thetaiotaomicron and B. ovatus, are capable of utilizing nearly all of the major plant and host glycans, including rhamnogalacturonan II, a highly complex polymer thought to be recalcitrant to microbial degradation. Transcriptional profiling and gene inactivation experiments revealed the identity and specificity of the polysaccharide utilization loci (PULs) that encode individual Sus-like systems that target various plant polysaccharides. Comparative genomic analysis indicated that B. ovatus possesses several unique PULs that enable degradation of hemicellulosic polysaccharides, a phenotype absent from B. thetaiotaomicron. In contrast, the B. thetaiotaomicron genome has been shaped by increased numbers of PULs involved in metabolism of host mucin O-glycans, a phenotype that is undetectable in B. ovatus. Binding studies of the purified sensor domains of PUL-associated hybrid two-component systems in conjunction with transcriptional analyses demonstrate that complex oligosaccharides provide the regulatory cues that induce PUL activation and that each PUL is highly specific for a defined cell wall polymer. These results provide a view of how these species have diverged into different carbohydrate niches by evolving genes that target unique suites of available polysaccharides, a theme that likely applies to disparate bacteria from the gut and other habitats.

Project description:BackgroundCollenchyma cells occur widely in eudicotyledons and provide mechanical support for growing organs. At maturity, the cells are elongated and have thick, non-lignified walls, which in celery contain cellulose and pectic polysaccharides, together with xyloglucans and heteroxylans and heteromannans. A previous study suggested that at least some of the collenchyma cell wall in celery is laid down after expansion has stopped and is thus secondary. In the present study, we re-examined this. We used chemical analysis and immunomicroscopy to determine changes in the polysaccharide compositions of these walls during development. Additionally, solid-state NMR spectroscopy was used to examine changes in polysaccharide mobilities during development.ResultsWe showed the collenchyma walls are deposited only during cell expansion, i.e. they are primary walls. During cell-wall development, analytical and immunomicroscopy studies showed that within the pectic polysaccharides there were no overall changes in the proportions of homogalacturonans, but there was a decrease in their methyl esterification. There was also a decrease in the proportions of the (1 → 5)-α-L-arabinan and (1 → 4)-β-D-galactan side chains of rhamnogalacturonan I. The proportions of cellulose increased, and to a lesser extent those of xyloglucans and heteroxylans. Immunomicroscopy showed the homogalacturonans occurred throughout the walls and were most abundant in the middle lamellae and middle lamella junctions. Although the (1 → 4)-β-D-galactans occurred only in the rest of the walls, some of the (1 → 5)-α-L-arabinans also occurred in the middle lamellae and middle lamella junctions. During development, the location of the xyloglucans changed, being confined to the middle lamellae and middle lamella junctions early on, but later occurred throughout the walls. The location of the heteroxylans also changed, occurring mostly in the outer walls in young cells, but were more widely distributed in mature cells. Solid-state NMR spectroscopy showed that particularly cellulose, but also homogalacturonans, decreased in mobility during development.ConclusionsOur studies showed that celery collenchyma cell walls are primary and that during their development the polysaccharides undergo dynamic changes. Changes in the mobilities of cellulose and homogalacturonans were consistent with the cell walls becoming stiffer as expansion ceases.

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Characterization of two cell-wall polysaccharides from Fusicoccum amygdali.

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