Ebbe Nordlander
Professor
An investigation of the peroxidase activity of Vitreoscilla hemoglobin
Author
Summary, in English
In order to investigate the ability of the Vitreoscilla hemoglobin (VHb) to act as a peroxidase, the protein was overexpressed in Escerichia coli and purified using a 6xHis-tag. The peroxidase activity of VHb was studied using 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), ferrocene carboxylic acid (FcCOOH) dopamine and L-dopa as substrates. The effects of external agents such as pH, salt concentration/ionic strength, and the thermal stability of VHb on the catalytic activity were assessed. The optimum pH for VHb using ABTS as a substrate was estimated to be 6-7. The VHb protein proved to be stable up to 80 degrees C, as judged by its peroxidase activity. Furthermore, NaCl concentrations up to 100 mM did not exert any significant effect on the activity. The catalytic activity against ABTS and FcCOOH was similar to that measured for horseradish peroxidase, whereas in the case of the phenolic substrates dopamine and L-dopa the activity was several orders of magnitude lower. The Michaelis constants, K-m(H2O2), were in good agreement with the data for human and bovine hemoglobin. No activity could be detected for the negative controls lacking VHb. These results demonstrate that VHb exhibits peroxidase activity, a finding in line with the hypothesis that VHb has cellular functions beyond the role as an oxygen carrier.
Department/s
- Pure and Applied Biochemistry
- Department of Chemistry
- Chemical Physics
Publishing year
2007
Language
English
Pages
324-334
Publication/Series
Journal of Biological Inorganic Chemistry
Volume
12
Issue
3
Document type
Journal article
Publisher
Springer
Topic
- Biological Sciences
- Atom and Molecular Physics and Optics
Keywords
- peptide
- peroxidase
- protopoporphyrin IX
- catalysis
Status
Published
ISBN/ISSN/Other
- ISSN: 1432-1327