Purification and methylation analysis of cell wall material from Solanum tuberosum |
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Authors: | Stephen G. Ring Robert R. Selvendran |
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Affiliation: | Agricultural Research Council, Food Research Institute, Colney Lane, Norwich NR4 7UA, U.K. |
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Abstract: | Cell wall material (CWM) of potatoes was prepared by sequentially extracting the wet ball-milled tissue with 1 % aq. Na deoxycholate, PhOHHOAcH2O and 90 % (v/v) aq. DMSO. The purity of the CWM (e.g. absence of residual starch) was established by carbohydrate analysis using different acid hydrolysis conditions and by methylation studies. The partially methylated alditol acetates from the CHCl3MeOH soluble fraction (S) of the methylated CWM were separated into 15 main peaks by GLC. Fourteen of these peaks were carbohydrate derivatives and the identity of most of these was established by MS. Reduction of the hydrolysate of S with NaBD4 was used to identify the carbohydrate derivatives present in peaks 7 and 11 above. The occurrence of 4-linked galacturonosyl residues in the methylated polymers was established after reduction of S with LiAlH4 and LiAlD4. The main glycosidic linkages present in the non-cellulosic polysaccharides of the wall in descending order of concentration are: 4-linked galactose, 4-linked galacturonic acid, 5-linked arabinose and 4,6-linked glucose. The major branch points are those through 0–6 of glucose and 0–4 of rhamnose. Arabinose, galactose and xylose residues constituted the non-reducing ends. Graded acid hydrolysis of the CWM made it possible to assess the relative strengths of some of the glycosidic linkages. The general structural features of the CWM are discussed in the light of these results. |
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Keywords: | Solanaceae potatoes cell wall material extraction and purification methylation analysis. |
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