Abstract
A procedure was developed for the separation and purification of amylose and amylopectin isolated from cotton leaves. Cotton leaves were homogenized in 0.02 molar phosphate buffer at pH 7.0 containing HgCl2 plus toluene. Crude starch granules were collected by centrifugation and partially purified by treating with acetone and toluene. The starch granules were then dispersed in dimethylsulfoxide and precipitated with ethyl alcohol. The precipitate was suspended in boiling water. Amylose was separated from amylopectin and cell wall particles on a Sepharose 2B column and further purified with thymol and butanol. Amylopectin was then separated from the colloidal cell wall contaminants by its specific interaction with concanavalin A. Purities of starch components were verified by specific biochemical and enzymic tests in addition to their iodine-binding capacity. This procedure should also be suitable for purification of starch components from other plant sources.
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- BLIGH E. G., DYER W. J. A rapid method of total lipid extraction and purification. Can J Biochem Physiol. 1959 Aug;37(8):911–917. doi: 10.1139/o59-099. [DOI] [PubMed] [Google Scholar]
- Chang C. W. Manometric method for determining bicarbonate content in plant leaves. Anal Biochem. 1975 May 26;66(1):185–193. doi: 10.1016/0003-2697(75)90736-8. [DOI] [PubMed] [Google Scholar]
- GREENWOOD C. T., THOMSON J. Studies on the biosynthesis of starch granules. 2. The properties of the components of starches from smooth- and wrinkled-seeded peas during growth. Biochem J. 1962 Jan;82:156–164. doi: 10.1042/bj0820156. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hopkins R. H., Jelinek B. The fractionation of potato starch. Biochem J. 1948;43(1):28–32. doi: 10.1042/bj0430028. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Juliano B. O., Varner J. E. Enzymic degradiation of starch granules in the cotyledons of germinating peas. Plant Physiol. 1969 Jun;44(6):886–892. doi: 10.1104/pp.44.6.886. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kennedy J. F., Rosevear A. An assessment of the fractionation of carbohydrates on concanavalin A-sepharose 4B by affinity chromatography. J Chem Soc Perkin 1. 1973;19:2041–2046. doi: 10.1039/p19730002041. [DOI] [PubMed] [Google Scholar]
- Lloyd K. O. The preparation of two insoluble forms of the phytohemagglutinin, concanavalin A, and their interactions with polysaccharides and glycoproteins. Arch Biochem Biophys. 1970 Apr;137(2):460–468. doi: 10.1016/0003-9861(70)90463-7. [DOI] [PubMed] [Google Scholar]