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Activity enhancement of a Bacillus circulans xylanase by introducing ion-pair interactions into an α-helix

Paper ID Volume ID Publish Year Pages File Format Full-Text
34545 45033 2013 7 PDF Available
Title
Activity enhancement of a Bacillus circulans xylanase by introducing ion-pair interactions into an α-helix
Abstract

•We enhanced the activity of Bacillus circulans xylanase (1.66–3.58 times).•Ion-pair was introduced at the α-helix region.•Effect was observed at distantly located active site cleft (23.85–25.15 Å).•Active-site flexibility was increased while the helix flexibility was decreased.•There is increased mobility of the active-site cleft lining residues in strand B8.

IntroductionIt is widely accepted that ion-pair increases rigidity and thermostability. There are numerous studies on ion-pairs and thermostability, but none are available about the effect of ion-pair on the activity of enzymes. This paper studies whether an ion-pair allows flexible movement in an enzyme molecule and affects its activity.Materials and methodsIon-pairs are designed at the α-helix region of a Bacillus circulans xylanase, and they are far from the active-sites (23.85–25.15 Å). Two ion-pairing mutations are situated at the C-terminus (D151/E151-K154 ion-pairs) of the helix. One mutation is double-site (F48R-N151D), which introduces both the tertiary (R48-D151) and intra-helical (D151-K154) ion-pairs.Results and discussionAll of the mutants enhanced the catalytic efficiency against xylan (1.66–3.58 times). The double-site mutation showed a synergistic effect on the activity. Overall, the ion-pairs decreased the flexibility (increased rigidity) of the α-helix region and increased the active-site flexibility. The ion-pairs were destabilizing and surface-located; this means that the weaker destabilizing ion-pair still allows flexible movement in the active-site. There is higher mobility of the strand B4 where the active site residue E172 is located. Moreover, the residues lining the active-site cleft (strand B8) showed increased flexibility upon substrate binding.ConclusionIncrease in the activity was due to the increase in active-site flexibility and increased mobility of the residues lining the active-site cleft (strand B8).

Keywords
Bacillus circulans xylanase; Activity; Ion-pair interaction; α-Helix; Flexibility; Molecular dynamics simulation
First Page Preview
Activity enhancement of a Bacillus circulans xylanase by introducing ion-pair interactions into an α-helix
Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 48, Issue 10, October 2013, Pages 1495–1501
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering