Purification and properties of extracellular phytase from Bacillus sp. KHU-10

Yang Mun Choi, Hyung Joo Suh, Jin Man Kim

Research output: Contribution to journalArticle

108 Citations (Scopus)

Abstract

Bacillus species producing a thermostable phytase was isolated from soil, boiled rice, and mezu (Korean traditinal koji). The activity of phytase increased markedly at the late stationary phase. An extracellular phytase from Bacillus sp. KHU-10 was purified to homogeneity by acetone precipitation and DEAE-Sepharose and phenyl-Sepharose column chromatographies. Its molecular weight was estimated to be 46 kDa on gel filtration and 44 kDa on SDS-polyacrylamide gel elctrophoresis. Its optimum pH and temperature for phytase activity were pH 6.5-8.5 and 40°C without 10 mM CaCl2 and pH 6.0-9.5 and 60°C with 10 mM CaCl2. About 50% of its original activity remained after incubation at 80°C or 10 min in the presence of 10 mM CaCl2. The enzyme activity was fairly stable from pH 6.5 to 10.0. The enzyme had an isoelectric point of 6.8. As for substrate specificity, it was very specific for sodium phytate and showed no activity on other phosphate esters. The Km value for sodium phytate was 50 μM. Its activity was inhibited by EDTA and metal ions such as Ba2+, Cd2+, Co2+, Cr3+, Cu2+, Hg2+, and Mn2+ ions.

Original languageEnglish
Pages (from-to)287-292
Number of pages6
JournalJournal of Protein Chemistry
Volume20
Issue number4
DOIs
Publication statusPublished - 2001 Oct 16

Fingerprint

6-Phytase
Bacilli
Bacillus
Purification
Phytic Acid
Ions
Agarose Chromatography
Column chromatography
Isoelectric Point
Enzyme activity
Enzymes
Substrate Specificity
Acetone
Edetic Acid
Sepharose
Gel Chromatography
Metal ions
Esters
Soil
Molecular Weight

Keywords

  • Bacillus sp.
  • Calcium
  • Phytase

ASJC Scopus subject areas

  • Biochemistry

Cite this

Purification and properties of extracellular phytase from Bacillus sp. KHU-10. / Choi, Yang Mun; Suh, Hyung Joo; Kim, Jin Man.

In: Journal of Protein Chemistry, Vol. 20, No. 4, 16.10.2001, p. 287-292.

Research output: Contribution to journalArticle

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