A novel analytical method to evaluate directly catalase activity of microorganisms and mammalian cells by ESR oximetry

Keisuke Nakamura, Taro Kanno, Takayuki Mokudai, Atsuo Iwasawa, Yoshimi Niwano, Masahiro Kohno

Research output: Contribution to journalArticlepeer-review

2 Citations (Scopus)

Abstract

Electron spin resonance (ESR) oximetry technique was applied for analysis of catalase activity in the present study. Catalase activity was evaluated by measuring oxygen from the reaction between hydrogen peroxide (H 2O2) and catalase-positive cells. It was demonstrated that the ESR spectra of spin-label probes, 4-hydroxy-2,2,6,6-tetramethylpiperidine 1-oxyl (TEMPOL), 4-oxo-2,2,6,6-tetramethyl-1-piperidinyloxy (4-oxo-TEMPO) and 4-maleimido-2,2,6,6-tetramethyl-1-piperidinyloxy (4-maleimido-TEMPO) in the presence of H2O2 were broadened with the concentrations of catalase. It was possible to make a calibration curve for catalase activity by peak widths of the spectra of each spin-label probe, which are broadened dependently on catalase concentrations. The broadened ESR spectra were also observed when the catalase-positive micro-organisms or the mammalian cells originally from circulating monocytes/macrophages were mixed with TEMPOL and H2O2. Meanwhile, catalase-negative micro-organisms caused no broadening change of ESR spectra. The present study indicates that it is possible to evaluate directly the catalase activity of various micro-organisms and mammalian cells by using an ESR oximetry technique.

Original languageEnglish
Pages (from-to)1036-1043
Number of pages8
JournalFree Radical Research
Volume44
Issue number9
DOIs
Publication statusPublished - 2010 Sep 1

Keywords

  • Catalase activity
  • ESR
  • analytical method
  • bacteria
  • mammalian cells
  • oximetry

ASJC Scopus subject areas

  • Biochemistry

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