PARTIAL PURIFICATION AND CHARACTERIZATION OF AN EXTRACELLULAR METALLOPEPTIDASE PRODUCED BY <i>Bacillus amyloliquefaciens</i> FE-K1
Fundagul Erem, Mehmet Inan, Barcin Karakas Budak, Muharrem Certel
article (Uluslararası, Hakemli) | Yazar(lar): Fundagul Erem, Mehmet Inan, Barcin Karakas Budak, Muharrem Certel | Yıl: 2020 | Dergi/Kurum: TRAKYA UNIVERSITY JOURNAL OF NATURAL SCIENCES | İndeks: WOS.ESCI | DOI: 10.23902/trkjnat.647525 — Özet: The aim of this study was to purify and characterize the peptidase of Bacillu
The aim of this study was to purify and characterize the peptidase of Bacillus amyloliquefaciens (Fukumoto) (strain FE-Kt) isolated from ropey bread. Peptidases were purified from crude enzyme solution by affinity chromatography with an efficiency of 25 % and a purification coefficient of 1.53. The optimum pH of partially purified peptidase (PPPase) solution was determined as 7.5 and the peptidases retained approximately 90 % of their initial activity in the pH range 7.0-8.5 following incubation at 37 degrees C for 2 h. The optimum temperature for the PPPase was 60 degrees C. The approximate molecular weight of the PPPase was determined as 36 kDa. Inactivation of the PPPase in the presence of O-FEN and EDTA showed them to be metallopeptidases and 5 mM of K-+(1) and 5 mM of Mn+2 ions increased the enzyme activity by 4 % and 6.15 %, respectively. The presence of Hg+2, Fe+3 and SDS (0.1-1.0 % w/v) caused inactivation whereas the enzyme retained most of its activity in the presence of 0.1-1.0 % (v/v) Triton X-100, Tween 20 and Tween 80 and 1-20 % (v/v) xylene, ethanol, acetone and acetonitrile. Characterization of the PPPase revealed the enzyme as a neutral serine metallopeptidase compatible with some organic solvents and surfactants.