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 language | English |
|---|---|
| Pages (from-to) | 235-239 |
| Number of pages | 5 |
| Journal | Biotechnology and Applied Biochemistry |
| Volume | 29 |
| Issue number | 3 |
| Publication status | Published - 1999 |
ASJC Scopus subject areas
- Biotechnology
- Bioengineering
- Molecular Medicine
- Biomedical Engineering
- Applied Microbiology and Biotechnology
- Drug Discovery
- Process Chemistry and Technology