Mass-Spectrometry-Msms-Identification-Msms-Fragmentation-Methods-Generate-Fragments-Monosaccharides-Connectivity-i

We examined the potential of helium-charge transfer dissociation (He-CTD) as a method of MSMS to decipher the connectivity of β-1,4- and β-1,3-linked oligosaccharides. In contrast to collision-induced dissociation (CID), He-CTD of isolated oligosaccharide precursors produced both glycosidic and cross-ring cleavages of each monosaccharide. The radical-driven dissociation in He-CTD induced single cleavage events, without consecutive fragmentations, which facilitated structural interpretation. He-CTD of various standards up to a degree of polymerization of 7 showed that β-1,4- and β-1,3-linked carbohydrates can be distinguished based on diagnostic 3,5A fragment ions that are characteristic for β-1,4-linkages. Overall, fragment ion spectra from He-CTD contained sufficient information to infer the connectivity specifically for each glycosidic bond. When testing He-CTD to resolve the order of β-1,4- and β-1,3-linkages in mixed-linked oligosaccharide standards, He-CTD spectra sometimes provided less confident assignment of connectivity.

2'-FL -mass spectrometry (IMS-MS) of the standards indicated that ambiguity in the He-CTD spectra was caused by isobaric impurities in the mixed-linked oligosaccharides. Radical-driven dissociation induced by He-CTD can thus expand MSMS to carbohydrate linkage analysis, as demonstrated by the comprehensive fragment ion spectra on native oligosaccharides. The determination of connectivity in true unknowns would benefit from the separation of isobaric Galectin-8-N-domain recognition mechanism for sialylated and sulfated glycans.Galectin-8 has much higher affinity for 3'-O-sulfated or 3'-O-sialylated glycoconjugates and a Lewis X-containing glycan than for oligosaccharides terminating in Galβ1→34GlcNAc, and this specificity is mainly attributed to the N-terminal carbohydrate recognition domain (N-domain, CRD) (Ideo, H., Seko, A., Ishizuka, I., and Yamashita, K.

( 2'-Fucose lactose ) Glycobiology 13, 713-723). In this study, we elucidated the crystal structures of the human galectin-8-N-domain (-8N) in the absence or presence of 4 ligands. The apo molecule forms a dimer, which is different from the canonical 2-fold symmetric dimer observed for galectin-1 and -2. In a galectin-8N-lactose complex, the lactose-recognizing amino acids are highly conserved among the galectins. However, Arg(45), Gln(47), Arg(59), and the long loop region between the S3 and S4 β-strands are unique to galectin-8N. These amino acids directly or indirectly interact with the sulfate or sialic acid moieties of 3'-sialyl- and 3'-sulfolactose complexed with galectin-8N. Furthermore, in the LNF-III-galectin-8N complex, van der Waals interactions occur between the α1-3-branched fucose and galactose and between galactose and Tyr(141), and these interactions increase the affinity toward galectin-8N.

Based on the findings of these x-ray crystallographic analyses, a mutagenesis study using surface plasmon resonance showed that Arg(45), Gln(47), and Arg(59) of galectin-8N are indispensable and coordinately contribute to the strong binding of galectins-8N to sialylated and sulfated oligosaccharides. Arg(59) is the most critical amino acid for binding in the S3-S4 loop region.Antibacterial activities and preservative effect of chitosan oligosaccharide Maillard reaction products on Penaeus vannamei.The purpose of this study was to understand the influence of Maillard reaction activity against Pseudomonas aeruginosa and Vibrio parahaemolyticus and preservative effect of two kinds of COS MR products (MRPs) on Penaeus vannamei were evaluated. The structures of CG (MRPs of COS and glucose) and CM (MRPs of COS and maltose) were characterized by FT-IR and their molecular weights were measured by gel permeation chromatography (GPC). The results showed that CG has enhanced antibacterial activity and preservative effect compared to COS by evaluating total volatile base-nitrogen (TVB-N), trimethylamine (TMA), pH, total bacterial count (TBC) and sensory evaluation. Meanwhile, CM showed decreased antibacterial activity and preservative effect compared to COS by evaluating TMA, pH and TBC.

The results indicated that antibacterial activity and preservative effect of COS increased or decreased after MR. In other words, the type of reducing sugar involved in MR had a great impact on the functional properties of COS-saccharide MRPs and their application to be used as a food [Oligosaccharides from Streptococcus lactis.