Amylopectin isolation

Potter and Hassid have shown that subfractions of tapioca amylopectin and maize amylopectin of different molecular size had the same degree of branching, whilst the /3-amylolysis limits reported in Table V, and in Refs. 3 and 4, are fully in accord vdth Myrback and Sill n s calculations. Furthermore, the constancy of the inner CL of amylopectin isolated from starch granules during growth (see Table V) is also in agreement with the suggested theory of starch biosynthesis. 

Enzymic Hydrolysis of Amylopectin. Isolation of a Crystalline Trisaccharide Hen-decaacetate, M. L. Wolfrom, L. W. Georges, Alva Thompson, and I. L. Miller,/. Amer. Chem. Soc., 71, 2873-2875 (1949). 

Successful methods entail precipitation of the amylose from solution as an insoluble complex, which is removed by high-speed centrifuging the amylopectin is isolated from the supernatant liquor by precipitation with alcohol or, more satisfactorily, by freeze-drying. (Precipitation with alcohol does not always appear to be quantitative, and the physical form of the product obtained by freeze-drying is more satisfactory for subsequent dissolution and esterification.) The amylose can then be further purified by reprecipitation with the same or, preferably, a different complexing agent. 

Table I shows the results of periodate oxidation and methylation applied to amylopectins from various sources. If the percentage of amylose in a whole starch is known, the length of unit chain of the amylopectin component can be calculated from the results of periodate oxidation of the whole starch (see Table II). The length of unit chain appears to depend on the botanical species, but not the variety, from which the starch wras isolated. In the case of tapioca and corn amylopectins, sub-fractionation of these by precipitation with methanol, followed by periodate oxidation, showed the sub-fractions had the same degree of branching as the original amylopectins.71 The action of periodate on whole starches and amylopectins is now so well established and accurate, that it may well completely supersede the methylation technique for pure starches, in view of its many advantages.

Amylo-1 —> 6-glucosidase obtained by Cori and Larner218 from rabbit muscles, and R-enzyme isolated by Hobson, Whelan and Peat219 from potatoes and broad beans, are typical debranching enzymes, which will hydrolyze the 6 — 1-a-D-glucosidic linkage rather than the normal 4 —> 1-a-D linkage. These enzymes will therefore be particularly important in determinations of the fine structure of amylopectin, if they can be sufficiently well purified. 

The isolation of 2,3-dimethyl-D-glucose from the hydrolysis products of certain methylated polysaccharides has been an important factor in assigning structures to these polysaccharides. From trimethyl-starch it has been recovered in about 3% yield, together with 2,3,4,6-tetra-methyl- and 2,3,6-trimethyl-D-glucopyranose, and arises from the points of linkage of the repeating chains of the amylopectin component.67,69,70 From a dimethyl-starch the yield is considerably higher (75%).71 Other sources are the methylated capsular polysaccharide of Rhizobium radici-... 

Structural analysis of isolated amylose and amylopectin components has been carried out by standard methods based on methylation, periodate oxidation, and partial acid hydrolysis studies. Methylation and periodate oxidation studies established the linkage types and frequency of... 

Interest in the influence of lipids on pyrazine formation has recently been generated by the identification of long-chain alkyl-substituted heterocyclic compounds in foods and in model systems. Pyrazines in this category include 2-heptylpyrazine isolated from french fried potato flavor (7), and 2-methyl-3(or 6)-pentylpyrazine and 2,5-dimethyl-3-pentylpyrazine, isolated from extruded zein/corn amylopectin/corn oil systems (8, 9). Only the involvement of lipids or lipid-decomposition products in the formation of these compounds could account for the long-chain alkyl substitution on the pyrazine ring. 

The characteristics of the isolated biopolymers depend on their structure. Cellulose and amylose are linear polymers, whereas amylopectin, pectin and hemicelluloses are branched polymers. Pectin and amylopectin contain carboxylic groups, which make interactions with water molecules very important. Amylose has a helix structure, whereas the cellulose molecule looks like a ribbon. The interactions with water and other neighbouring molecules are therefore different. 

Species and/or cultivar differences are also observed in other starch properties and in the properties of isolated amylose and amylopectin. To illustrate, purified amylose samples have been shown to differ in (3-amylolysis limit and average DP.64,67,124 Purified amylopectin samples have also been shown to differ in (3-amylolysis limit, average length of unit chains and viscosity.64,66 67 124,125 Campbell et al.121 observed a range of amylose content from 22.5% to 28.1% in 26 maize inbreds selected for maturity, kernel characteristics and pedigree. Starches from these non-mutant genotypes also differed in thermal properties (DSC), paste viscosities and gel strengths. 

Starch from ae su kernels in a dent background consists of 51-60% amylose, as determined by the blue value procedure (Table 3.6), with the amylose percentage increasing with increasing kernel age.57 Yeh et al.,90 in contrast, reported that ae su reduced amylose concentration from 65% for su to 28% for ae su, based on SEC separation of starch polysaccharides isolated from kernels in a sweet com background. Three fractions were obtained (Table 3.7). The first two were loosely-branched, similar to the amylopectin fractions in ae. Amylose from the third peak fraction was similar in iodine staining to that from normal however, some short chain length amylose as... 

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