The viscous seed mucilage of flax (Linum usitatissimum) is a mixture of rhamnogalacturonan I and arabinoxylan with novel side group substitutions. The rhamnogalacturonan I has numerous single nonreducing terminal residues of the rare sugar L-galactose attached at the O-3 position of the rhamnosyl residues instead of the typical O-4 position. The arabinoxylan is highly branched, primarily with double branches of nonreducing terminal L-arabinosyl units at the O-2 and O-3 positions along the xylan backbone. While a portion of each polysaccharide can be purified by anion-exchange chromatography, the side group structures of both polysaccharides are modified further in about one-third of the mucilage to form composites with enhanced viscosity. Our finding of the unusual side group structures for two well-known cell wall polysaccharides supports a hypothesis that plants make a selected few ubiquitous backbone polymers onto which a broad spectrum of side group substitutions are added to engender many possible functions. To this end, modification of one polymer may be accompanied by complementary modifications of others to impart functions to heterocomposites not present in either polymer alone.
Kingella kingae is an encapsulated gram-negative organism that is a common cause of osteoarticular infections in young children. In earlier work, we identified a glycosyltransferase gene called csaA that is necessary for synthesis of the [3)-β-GalpNAc-(1→5)-β-Kdop-(2→] polysaccharide capsule (type a) in K. kingae strain 269–492. In the current study, we analyzed a large collection of invasive and carrier isolates from Israel and found that csaA was present in only 47% of the isolates. Further examination of this collection using primers based on the sequence that flanks csaA revealed three additional gene clusters (designated the csb, csc, and csd loci), all encoding predicted glycosyltransferases. The csb locus contains the csbA, csbB, and csbC genes and is associated with a capsule that is a polymer of [6)-α-GlcpNAc-(1→5)-β-(8-OAc)Kdop-(2→] (type b). The csc locus contains the cscA, cscB, and cscC genes and is associated with a capsule that is a polymer of [3)-β-Ribf-(1→2)-β-Ribf-(1→2)-β-Ribf-(1→4)-β-Kdop-(2→] (type c). The csd locus contains the csdA, csdB, and csdC genes and is associated with a capsule that is a polymer of [P-(O→3)[β-Galp-(1→4)]-β-GlcpNAc-(1→3)-α-GlcpNAc-1-] (type d). Introduction of the csa, csb, csc, and csd loci into strain KK01Δcsa, a strain 269–492 derivative that lacks the native csaA gene, was sufficient to produce the type a capsule, type b capsule, type c capsule, and type d capsule, respectively, indicating that these loci are solely responsible for determining capsule type in K. kingae. Further analysis demonstrated that 96% of the invasive isolates express either the type a or type b capsule and that a disproportionate percentage of carrier isolates express the type c or type d capsule. These results establish that there are at least four structurally distinct K. kingae capsule types and suggest that capsule type plays an important role in promoting K. kingae invasive disease.
Exopolysaccharides (EPS) produced by some lactic acid bacteria are often used by the dairy industry to improve the rheological and physical properties of yogurt, but the relationship between their structure and functional effect is still unclear. The EPS from different species, or different strains from the same species, may differ in terms of molar mass, repeating unit structure, and EPS yield during fermentation of milk. This study aimed to characterize the detailed properties of EPS produced from 7 strains of Streptococcus thermophilus, which is one of the key cultures used for yogurt manufacture. Milk was fermented with strains DGCC 7698, DGCC 7710, DGCC 7785, ST-10255y, St-143, STCth-9204, and ST4239. These strains were selected because they have been used in previous studies on yogurt texture, but a complete description of their EPS structural properties has not yet been reported. All strains were fermented under a similar acidification rate by adjusting the level of supplementation with peptone or the inoculation level, which allowed for a comparison of EPS yields under similar growth conditions (reconstituted skim milk at 40°C). The EPS from each strain was isolated and the weight-average molar mass and z-average root mean square radius determined using size-exclusion chromatography multiangle laser light scattering. The monosaccharide composition of EPS was determined using gas chromatography-mass spectrometry, and repeating unit structure was determined using nuclear magnetic resonance spectroscopy. The weight-average molar mass values of EPS ranged from 0.14 to 1.61 × 10 g/mol. All 7 EPS samples were uncharged. The strains ST-10255y and ST4239 had EPS with the same repeating unit structure. The monosaccharide compositions of the various EPS were mainly composed of glucose and galactose, with low levels of rhamnose in the EPS isolated from DGCC 7698, and N-acetylgalactosamine in the EPS from DGCC 7785, ST-10255y, and ST4239. The yields of EPS (measured when fermented milks reached pH 4.6) ranged from 8.0 to 76.4 mg of glucose equivalents/kg. In addition to (free) EPS, some strains were also able to produce capsular polysaccharide (associated with the bacterial cells) when observed with negative staining technique. The results of our study will help the dairy industry to better understand the mechanism by which different strains of Streptococcus thermophilus affect yogurt texture.
Dimethylsulfoxide-solubilized polysaccharides from delignified corn stover and aspen were characterized. The biomass was delignified by two different techniques; a standard acid chlorite and a pulp and paper QPD technique comprising chelation (Q), peroxide (P), and acid-chlorite (D). Major polysaccharides in all fractions were diversely substituted xylan. Xylan acetylation was intact after chlorite delignification and, as expected, xylan from QPDdelignified fraction was de-acetylated by the alkaline peroxide step. The study of DMSO-extractable xylans from chlorite-delignified biomass revealed major differences in native acetylation patterns between corn stover and aspen xylan. Xylan from cell walls of corn stover contains 2-O-and 3-O-mono-acetylated xylan and [MeGlcA-a-(1 ? 2)][3-OAc]-xylp units. In addition, aspen xylan also contains 2,3-di-O-acetylated xylose. 1,4-b-D-xylp residues substituted with MeGlcA at O-2 position are absent in chlorite-delignified aspen xylan. Sugar composition in accord with NMRspectroscopic data indicated that corn stover xylan is arabinosylated while aspen xylan is not. We have shown that corn stover xylan has similar structure with xylans from other plants of Poales order. No evidence was found to indicate the presence of 1,4-b-D-[MeGlcA-a-(1 ? 2)][Ara-a-(1 ? 3)]-xylp in corn stover xylan fractions.Keywords Biomass Á Biomass delignification Á Plant cell wall Á Glucuronoxylan Á Hardwood xylan Á Corn stover xylan Á Xylan acetylation Á Endoxylanase Á Homo-and-heteronuclear NMR-spectroscopy Á MALDI-TOF MS Abbreviations MGX 4-O-Methylglucuronoxylans GAX Glucuronoarabinoxylans MLG Mixed-linkage glucan MeGlcA 4-O-Methylglucuronic acid QA QPD-delignified DMSO-solubilized HMW fraction from aspen QS QPD-delignified DMSO-solubilized HMW fraction from corn stover CA Chlorite-delignified DMSO-solubilized HMW fraction from aspen CS Chlorite-delignified DMSO-solubilized HMW fraction from corn stover MW Molecular weight DP Degree of polymerization
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