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United States Department of Agriculture

Agricultural Research Service

Title: Purification of Alternanase by Affinity Chromatography

Authors
item Ahlgren, Jeffrey
item Cote, Gregory

Submitted to: International Symposium on Exopolysaccharides from Lactic Acid Bacteria
Publication Type: Abstract Only
Publication Acceptance Date: May 19, 2001
Publication Date: N/A

Technical Abstract: The enzyme alternanase, produced by Bacillus sp. NRRL B-21195, hydrolyzes alternan, a polysaccharide produced by certain strains of Leuconostoc mesenteroides that consists of glucose linked by alternating alpha(1-6), alpha(1-3) linkages. The main product of enzymatic hydrolysis by alternanase is a novel cyclic tetrasaccharide of glucose that also has alternating linkages between the glucose moieties. An improved purification scheme for alternanase has been recently developed that incorporates the use of isomaltosyl units linked to agarose for selectively binding the alternanase enzyme. Bound enzyme was eluted with 0.5 M sodium chloride and was nearly pure after this procedure. When followed by preparative isoelectric focusing, a single band of 116 kDa was observed when the product was analyzed by SDS-PAGE. The purification procedure is relatively simple and rapid and can be scaled to permit large quantities of fenzyme to be purified in high yield. The process produces enzyme of excellent purity which can be used to hydrolyze alternan to its various products for further investigation.

Last Modified: 8/27/2014
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