Purification and characterization of alkaline phosphatase from Bacillus stearothermophilus

Shuji Mori, Motoi Okamoto, Masahiro Nishibori, Mitsuko Ichimura, Junko Sakiyama, Hiroshi Endo

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Soluble alkaline phosphatase from the thermophilic bacterium Bacillus stearothermophilus was purified by a combination of chromatographlc methods, and its properties were examined. The purified enzyme had specific activity of 4.43 μmol p-nitrophenol/min per mg of protein and seemed to be a single band on SDS/PAGE with a molecular mass of 32 kDa. Its apparent K(m) for p-nitrophenyl phosphate was 1.114 mM. The enzyme exhibited an optimal pH of approx. 9.0 and exhibited its highest activity at 60-70°C. It also showed a bivalent cation requirement for activity, with maximal enhancement in the presence of Mg2+. In addition, significant thermal stability was observed in comparison with counterparts from mesophiles. Its partial N-terminal sequence was T1FSIVAFDPATGELGIAVQ19 as estimated by automated Edman degradation method. A search on the SwissProt database did not reveal any similar protein sequences from other sources.

Original languageEnglish
Pages (from-to)235-239
Number of pages5
JournalBiotechnology and Applied Biochemistry
Volume29
Issue number3
Publication statusPublished - 1999

Fingerprint

Geobacillus stearothermophilus
Phosphatases
Bacilli
Purification
Alkaline Phosphatase
Enzymes
Proteins
Protein Databases
Molecular mass
Cations
Polyacrylamide Gel Electrophoresis
Bacteria
Phosphates
Thermodynamic stability
Hot Temperature
Positive ions
Databases
Degradation
Nitrophenols
nitrophenylphosphate

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Biotechnology

Cite this

Purification and characterization of alkaline phosphatase from Bacillus stearothermophilus. / Mori, Shuji; Okamoto, Motoi; Nishibori, Masahiro; Ichimura, Mitsuko; Sakiyama, Junko; Endo, Hiroshi.

In: Biotechnology and Applied Biochemistry, Vol. 29, No. 3, 1999, p. 235-239.

Research output: Contribution to journalArticle

Mori, S, Okamoto, M, Nishibori, M, Ichimura, M, Sakiyama, J & Endo, H 1999, 'Purification and characterization of alkaline phosphatase from Bacillus stearothermophilus', Biotechnology and Applied Biochemistry, vol. 29, no. 3, pp. 235-239.
Mori, Shuji ; Okamoto, Motoi ; Nishibori, Masahiro ; Ichimura, Mitsuko ; Sakiyama, Junko ; Endo, Hiroshi. / Purification and characterization of alkaline phosphatase from Bacillus stearothermophilus. In: Biotechnology and Applied Biochemistry. 1999 ; Vol. 29, No. 3. pp. 235-239.
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