Purification and characterization of alkaline phosphatase from Bacillus stearothermophilus

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

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19 Citations (Scopus)


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
Issue number3
Publication statusPublished - 1999

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Molecular Medicine
  • Biomedical Engineering
  • Applied Microbiology and Biotechnology
  • Drug Discovery
  • Process Chemistry and Technology


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