Changes In The Stability And Kinetic Parameters Up On Glycation Of Thermostable A-Amylase From Bacillus Subtilis

Abstract

Glycation of the thermostable α-amylase, KLE, from Bacillus subtilis occurred during incubation with maltodextrin at 95C. This was revealed by the release of 5-hydroxymethyl-2-furfuraldehyde from the acid hydrolysis of glycated KLE (gKLE), the differences in the protein band patterns on SDS and Native-PAGE, and the shifting of the pI value from the range of 5.6-6.5 to that of 5.2-6.5. After glycation, the activity of gKLE was still retained. Furthermore, gKLE was more resistant to heat and pH compared with the nonglycated enzyme. The K m, reaction rate and efficiency to convert gelatinized cornstarch into maltodextrin of KLE were remained unchanged after glycation. This was different from the result obtained for BAN, another thermostable α-amylase produced by B. amyloliquefaciens. Glycation in BAN decreased the activity in converting gelatinized cornstarch into maltodextrin. Moreover, the stability and kinetic parameters of BAN were found to be negatively affected by glycation. © 2010, Wiley Periodicals, Inc.