Glucans structure, enzymic analysis

In an earlier article1 on the application of enzymic techniques to the analysis of the structure of polysaccharides, the -d- and /3-D-glucans were discussed, as well as more-general aspects of the preparation and use of catabolic enzymes in such analyses. The present article describes enzymic contributions to knowledge of the structures of other polysaccharides, but an account of subsequent research on a- and jS-D-glucans is also included. 

The observation47 that the stability of the secondary-ordered structure of xanthan (and as a consequence, its susceptibility to hydrolysis by cellulase) is a function of temperature, pH, and ionic strength has allowed the application of enzymic techniques to structural analysis. Xanthan is hydrolyzed in salt-free, aqueous solutions at elevated temperatures that is, hydrolysis occurs if the chain is unordered. It was proposed that, at lower temperatures and in the presence of salt, the small side-chains organize around the (1 ->4)-/3-D-glucan backbone and protect it from hydrolysis. 

A pentasaccharide and a trisaccharide have been isolated from an enzymic hydrolyzate of hemicellulosic material from oat coleoptiles.224 Methylation analysis of their structures indicated that these compounds may derive from j8-D-(l — 4)-linked D-glucans having terminal D-xylopyranosyl groups on some 0-6 atoms. Xylo-glucans having these structural features are present in the primary cell-walls of dicotyledons.22411... 

Figure 8-4. Methylation analysis of in vitro P-D-glucans. (a) Gas chromatography of the permethyl-ated alditol acetate obtained from methylation analysis of the cellulose synthesized in vitro by the enzyme from blackberry. Peak 1, derivative characteristic of (1 4) linked glucosyl units. Peak 2, internal standard (mj o-inositol). The derivative characteristic of (1 3) linked glucosyl units usually elutes 1 min before the major derivative visible in the chromatogram (not shown see Bulone et al. 1995). (b) Structural characterization by electron impact mass spectrometry of the 1,4,5-tri-0-acetyl-2,3,6-tri-0-methyl-D-glucitol derivative corresponding to peak 1 in A and characteristic of (1 4) linked glucosyl units.

The importance to achieve detailed structural characterization of in vitro products is illustrated by the application of the methods listed in Table 8-2 to the analysis of in vitro (1 3)-P-D-glucans synthesized under various conditions. For instance, it has been shown that for a given plant species the morphology and the stucture of the in vitro products are affected by the nature of the detergent used to extract the membrane-bound synthases (Lai Kee Him et al. 2001). Also, for a given detergent, enzymes from different plant species do not necessarily synthesize products that have the same morphology and structure (Lai Kee Him et al. 2001 Colombani et al. 2004). From these observations, it seems important... 

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