Abstract
Alternanase, an endoglucanase that hydrolyzes the bacterial exopolysaccharide alternan, will also hydrolyze the trisaccharide, panose, to produce glucose and a disaccharide that can be formed into a novel, cyclic tetrasaccharide. The glucose can then be selectively and quantitatively measured by enzyme-based reaction which forms the basis of a coupled enzyme assay to quantitate alternanase activity. By this method a preparation of alternanase purified by affinity chromatography on immobilized isomaltose had a maximum reaction rate (V max) of 0.75 μmol glucose min−1 and a K m of 34 mM for panose. Two competitive inhibitors of alternanase activity were also evaluated using this coupled enzyme assay: isomaltose had a K i of 94 mM while the cyclic tetrasaccharide had a K i of 66 mM.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Biely P, Côté GL, Burgess-Cassler A (1994) Purification and properties of alternanase, a novel endo-α-1,3-α-1,6-D-glucanase. Eur. J. Biochem. 226: 633–639.
Côté GL, Ahlgren JA (2001) The hydrolytic and transferase action of alternanase on oligosacchararides. Carbohydr. Res. 332: 373–379.
Côté GL, Biely P (1994) Enzymatically produced cyclic α-1,3-linked and α-1,6-linked oligosaccharides of D-glucose. Eur. J. Biochem. 226: 641–648.
Côté GL, Robyt JF (1982) Isolation and partial characterization of an extracellular glucansucrase from Leuconostoc mesenteroides NRRL B-1355 that synthesizes an alternating (1-6), (1-3)-α-Dglucan. Carbohydr. Res. 101: 57–74.
Rinderknecht H, Wilding P, Haverback BJ (1967) A new method for determination of α-amylase. Experientia 23: 805.
Wyckoff HA, Côté GL, Biely P (1996) Isolation and characterization of microorganisms with alternan hydrolytic activity. Curr. Microbiol. 32: 343–348.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Ahlgren, J.A., Ĉoté, G.L. Development of a coupled enzyme assay for the measurement of alternanase activity**, *** . Biotechnology Letters 24, 1277–1280 (2002). https://doi.org/10.1023/A:1016210025413
Issue Date:
DOI: https://doi.org/10.1023/A:1016210025413