High production, purification, biochemical characterization and gene analysis of a novel catalase from the thermophilic bacterium Ureibacillus thermosphaericus FZSF03

Int J Biol Macromol. 2017 Oct:103:89-98. doi: 10.1016/j.ijbiomac.2017.05.034. Epub 2017 May 10.

Abstract

A catalase-producing thermophilic bacterium, Ureibacillus thermosphaericus FZSF03, was isolated from high-temperature compost. Catalase production in this strain increased 31 times and reached 57,630U/mL after optimization in a shake flask, which might represent the highest catalase activity level among reported wild strains. This catalase was further purified and identified. The purified enzyme showed a specific activity of 219,360U/mg, higher than many other catalases. The molecular weight of this enzyme is 52kDa according to sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and the enzyme was identified as a monofunctional haeme catalase of Ureibacillus thermosphaericus by liquid chromatography-mass spectrometry (LC-MS)/MS. The optimal reaction temperature for this catalase was found to be 60°C. Stability was observed at 60°C and at a pH of 10.0, indicating the superiority of this enzyme at a high temperature and under alkaline conditions. Therefore, this catalase is a prospective candidate for industrial production and applications. The gene encoding this catalase is 1503bp. As the amino acid sequence shows low similarity with other catalases, we suggest that this is a novel monofunctional haeme catalase.

Keywords: Catalase; High production; Ureibacillus thermosphaericus.

MeSH terms

  • Amino Acid Sequence
  • Bacillales / drug effects
  • Bacillales / enzymology*
  • Bacillales / genetics
  • Bacillales / physiology
  • Carbon / pharmacology
  • Catalase / biosynthesis*
  • Catalase / genetics*
  • Catalase / isolation & purification
  • Catalase / metabolism
  • Cloning, Molecular
  • Dose-Response Relationship, Drug
  • Fermentation / drug effects
  • Hydrogen-Ion Concentration
  • Kinetics
  • Nitrogen / pharmacology
  • Temperature*

Substances

  • Carbon
  • Catalase
  • Nitrogen