Automated determination of microbial peroxidase activity in fermentation samples using hydrogen peroxide as the substrate and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonate) as the electron donor in a flow injection system
- 1 January 1995
- journal article
- research article
- Published by Royal Society of Chemistry (RSC) in The Analyst
- Vol. 120 (8) , 2101-2105
- https://doi.org/10.1039/an9952002101
Abstract
An automated flow injection method has been developed for the determination of microbial peroxidase activity. The substrate used was hydrogen peroxide and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonate)(ABTS) was used as the electron donor. In the presence of hydrogen peroxide, peroxidase catalyses the dehydrogenation of ABTS, resulting in the formation of a resonance-stabilized radical cation of ABTS. The green–blue colour formed, recorded at 418 nm, is taken as a measure of the peroxidase activity. The general technical conditions and the general enzymic kinetics have been optimized. Conditions for activation and stabilization of the enzyme were found, e.g., ammonium sulfate acts as a peroxidase activator. The resulting method has a good precision, sensitivity and speed.Keywords
This publication has 12 references indexed in Scilit:
- Crystal structure of the fungal peroxidase from Arthromyces ramosus at 1·9 Å resolutionJournal of Molecular Biology, 1994
- PeroxidasesCurrent Opinion in Biotechnology, 1993
- Spectroscopic parameters, electrode potentials, acid ionization constants, and electron exchange rates of the 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonate) radicals and ionsThe Journal of Physical Chemistry, 1993
- Amino acid sequence of Coprinus macrorhizus peroxidase and cDNA sequence encoding Coprinus cinereus peroxidaseEuropean Journal of Biochemistry, 1993
- Chemical deglycosylation of Horseradish peroxidase and surglycosylation using a new glycosylating reagent: effects on catalytic activity and stabilityPublished by Elsevier ,1993
- Comparison of structure and activities of peroxidases from Coprinus cinereus, Coprinus macrorhizus and Arthromyces ramosusBiochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology, 1992
- Purification, Crystallization, and Characterization of Peroxidase from Coprinus cinereusThe Journal of Biochemistry, 1988
- Plant peroxidases. Their primary, secondary and tertiary structures, and relation to cytochrome c peroxidaseEuropean Journal of Biochemistry, 1985
- Radical-cations as reference chromogens in kinetic studies of ono-electron transfer reactions: pulse radiolysis studies of 2,2′-azinobis-(3-ethylbenzthiazoline-6-sulphonate)Journal of the Chemical Society, Perkin Transactions 2, 1982
- The steady-state kinetics of peroxidase with 2,2′-azino-di-(3-ethyl-benzthiazoline-6-sulphonic acid) as chromogenBiochemical Journal, 1975