Partial characterization of Bacillus pumilus catalase partitioned in poly(ethylene glycol)/sodium sulfate aqueous two-phase systems

Karakus Y., Isik S.

PREPARATIVE BIOCHEMISTRY & BIOTECHNOLOGY, vol.49, no.4, pp.391-399, 2019 (SCI-Expanded) identifier identifier identifier

  • Publication Type: Article / Article
  • Volume: 49 Issue: 4
  • Publication Date: 2019
  • Doi Number: 10.1080/10826068.2019.1573197
  • Journal Name: PREPARATIVE BIOCHEMISTRY & BIOTECHNOLOGY
  • Journal Indexes: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Page Numbers: pp.391-399
  • Keywords: Activity, aqueous two-phase system, Bacillus pumilus, catalase, characterization, purification, BIOCHEMICAL-CHARACTERIZATION, AMSONIA-ORIENTALIS, PURIFICATION, PEROXIDASE, EXTRACTION, PROTEINS, OXIDASE, INVERTASE, EXPRESSION, SEPARATION
  • Kocaeli University Affiliated: Yes

Abstract

Aqueous two-phase partitioning system (ATPS) was used to extract and purify catalase from Bacillus pumilus. The system parameters for effective purification of catalase were optimized. The best catalase recovery (123%) with a 4.6-fold purification was obtained in the bottom phase of ATPS including the mixture of 15% (w/w) PEG4000, 10% (w/w) Na2SO4 and 3% (w/w) NaCl at pH 5.0. The purified enzyme was characterized regarding its activity and stability. The highest enzyme activity was observed at pH 7.0 and 37 degrees C on hydrogen peroxide. The enzyme was quite stable at temperatures between 30 and 55 degrees C and a pH range of 7.0-9.0. The K-m and V-max values were determined from Lineweaver-Burk plot as 11 mM and 1667 mu mole ml(-1) min(-1), respectively. Overall, it can be said that ATPS is a rapid, reasonable, straightforward and cost-effective process for catalase purification in comparison to the chromatographic methods.