Browse DORAS
Browse Theses
Latest Additions
Creative Commons License
Except where otherwise noted, content on this site is licensed for use under a:

Effects of mutations in the helix G region of horseradish peroxidase

Ryan, Barry J. and Ó Fágáin, Ciarán (2008) Effects of mutations in the helix G region of horseradish peroxidase. Biochimie, 90 (9). pp. 1414-1421. ISSN 0300-9084

Full text available as:

[img]PDF - Requires a PDF viewer such as GSview, Xpdf or Adobe Acrobat Reader


Horseradish peroxidase (HRP) has long attracted intense research interest and is used in many biotechnological fields, including diagnostics, biosensors and biocatalysis. Enhancement of HRP catalytic activity and/or stability would further increase its usefulness. Based on prior art, we substituted solvent-exposed lysine and glutamic acid residues near the proximal helix G (Lys 232, 241; Glu 238, 239) and between helices F and F′ (Lys 174). Three single mutants (K232N, K232F, K241N) demonstrated increased stabilities against heat (up to 2-fold) and solvents (up to 4-fold). Stability gains are likely due to improved hydrogen bonding and space-fill characteristics introduced by the relevant substitution. Two double mutants showed stability gains but most double mutations were non-additive and non-synergistic. Substitutions of Lys 174 or Glu 238 were destabilising. Unexpectedly, notable alterations in steady-state Vm/E values occurred with reducing substrate ABTS (2,2′-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid)), despite the distance of the mutated positions from the active site.

Item Type:Article (Published)
Uncontrolled Keywords:Recombinant; Horseradish peroxidase; Site-specific mutagenesis; Protein stabilisation;
Subjects:Biological Sciences > Biotechnology
Biological Sciences > Enzymology
DCU Faculties and Centres:DCU Faculties and Schools > Faculty of Science and Health > School of Biotechnology
Research Initiatives and Centres > National Centre for Sensor Research (NCSR)
Official URL:
Copyright Information:Copyright © 2008 Elsevier Masson SAS
Use License:This item is licensed under a Creative Commons Attribution-NonCommercial-Share Alike 3.0 License. View License
ID Code:541
Deposited On:02 Sep 2008 17:24 by DORAS Administrator. Last Modified 13 Feb 2017 11:49

Download statistics

Archive Staff Only: edit this record