Purification and Characterization of Phytase from Bacillus subtilis

Bacillus subtilis가 생산하는 Phytase의 정제 및 특성

  • Koh Hyun-Jung (Department of Biological Resources and Technology, Yonsei University) ;
  • Chu In-Ho (Department of Biological Resources and Technology, Yonsei University) ;
  • Chung Kun-Sub (Department of Biological Resources and Technology, Yonsei University)
  • 고현정 (연세대학교 생물자원공학과) ;
  • 추인호 (연세대학교 생물자원공학과) ;
  • 정건섭 (연세대학교 생물자원공학과)
  • Published : 2006.03.01

Abstract

A bacterial strain producing high level of a phytase was isolated from cattle feces and identified as Bacillus subtilis, and designated as Bacillus sp. CF 5-26. The production of the phytase from Bacillus sp. CF 5-26 reached the highest level after 72 hours at $37^{\circ}C$. The optimum condition of the media for the production of phytase was 10% rice bran extract, 0.1% whey protein powder, $0.01%\;CaCl_{2},\;0.01%\;KH_{2}PO_4$. The phytase was purified 20.3 folds with ethanol precipitation, Sephadex G-100, CM Sepharose CL-6B and Sephacryl S-100-HR column chromatography. The molecular weight of the purified enzyme was estimated to be 66 kDa on SDS-polyacrylamide gel electrophoresis. The purified phytase activity was stable up pH 5.0, 7.0, 11.0 and the remaining activity was 50% when it was treated at $100^{\circ}C$ for 1 hour. The substrate specificity of phytase was most active against sodium phytate and inositol polyphosphate compound. And the phytase hydrolysed tripolyphosphate and pyrophosphate a little. The Km value for the sodium phytate was 0.64 mM and the Vmax value was $4.41\;{\mu}mol/min$.

Keywords

Phytate;Bacillus subtilis;phytase

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