Journal
CARBOHYDRATE POLYMERS
Volume 301, Issue -, Pages -Publisher
ELSEVIER SCI LTD
DOI: 10.1016/j.carbpol.2022.120340
Keywords
RG-I-AGPs; RG-I; AGPs; Covalent linkage; RG-I size; NMR
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This study confirmed the covalent linkage between RG-I and AGP in purified RG-I material. It also identified RG-I-AGP as the major component of traditionally prepared RG-I.
To characterize a purified rhamnogalacturonan-I (RG-I) containing both RG-I and arabinogalactan-protein (AGP) types of glycosyl residues, an AGP-specific beta-1,3-galactanase that can cleave the AG backbone and release the AG sidechain was applied to this material. Carbohydrate analysis and NMR spectroscopy verified that the galactanase-released carbohydrate consists of RG-I covalently attached to the AG sidechain, proving a covalent linkage between RG-I and AGP. Size exclusion chromatography-multiangle light scattering-refractive index detection revealed that the galactanase-released RG-I has an average molecular weight of 41.6 kDa, which, together with the percentage of pectic sugars suggests an RG-I-AGP comprising one AGP covalently linked to two RG-I glycans. Carbohydrate analysis and NMR results of the RG-I-AGP, the galactanase-released glycans, and the RG lyase-released glycans demonstrated that the attached RG-I glycans are decorated with alpha-1,5-arabinan, beta-1,4-galactan, xylose, and 4-O-Me-xylose sidechains. Our measurement suggests that the covalently linked RG-I-AGP is the major component of the traditionally prepared RG-I.
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