Introducing transglycosylation activity in Bacillus licheniformis alpha-amylase by replacement of His235 with Glu

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Title
Introducing transglycosylation activity in Bacillus licheniformis alpha-amylase by replacement of His235 with Glu
Author(s)
P L Tran; H J Cha; J S Lee; S H Park; Eui-jeon Woo; K H Park
Bibliographic Citation
Biochemical and Biophysical Research Communications, vol. 451, no. 4, pp. 541-547
Publication Year
2014
Abstract
To understand the role of His and Glu in the catalytic activity of Bacillus licheniformis α-amylase (BLA), His235 was replaced with Glu. The mutant enzyme, H235E, was characterized in terms of its mode of action using labeled and unlabeled maltooctaose (Glc8). H235E predominantly produced maltotridecaose (Glc13) from Glc8, exhibiting high substrate transglycosylation activity, with Km = 0.38 mM and kcat/Km = 20.58 mM-1 s-1 for hydrolysis, and Km2 = 18.38 mM and kcat2/Km2 = 2.57 mM-1 s-1 for transglycosylation, while the wild-type BLA exhibited high hydrolysis activity exclusively. Glu235 - located on a wide open groove near subsite +1 - is likely involved in transglycosylation via formation of an α-1,4-glycosidic linkage and may recognize and stabilize the non-reducing end glucose of the acceptor molecule.
Keyword
Bacillus licheniformis thermostableBinding-subsite mappingSite-directed mutagenesisSubstrate transglycosylationTransfer productα-amylase
ISSN
0006-291X
Publisher
Elsevier
DOI
http://dx.doi.org/10.1016/j.bbrc.2014.08.019
Type
Article
Appears in Collections:
Division of Biomedical Research > Disease Target Structure Research Center > 1. Journal Articles
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