The attractive recombinant phytase from Bacillus licheniformis: biochemical and molecular characterization

Appl Microbiol Biotechnol. 2014 Jul;98(13):5937-47. doi: 10.1007/s00253-013-5421-9. Epub 2013 Dec 13.

Abstract

The phyL gene encoding phytase from the industrial strain Bacillus licheniformis ATCC 14580 (PhyL) was cloned, sequenced, and overexpressed in Escherichia coli. Biochemical characterization demonstrated that the recombinant enzyme has an apparent molecular weight of nearly 42 kDa. Interestingly, this enzyme was optimally active at 70-75 °C and pH 6.5-7.0. This enzyme is distinguishable by the fact that it preserved more than 40 % of its activity at wide range of temperatures from 4 to 85 °C. This new phytase displayed also a high specific activity of 316 U/mg. For its maximal activity and thermostability, this biocatalyst required only 0.6 mM of Ca(2+) ion and exhibited high catalytic efficiency of 8.3 s(-1) μM(-1) towards phytic acid.

MeSH terms

  • 6-Phytase / chemistry
  • 6-Phytase / genetics*
  • 6-Phytase / metabolism*
  • Amino Acid Sequence
  • Bacillus / enzymology*
  • Bacillus / genetics*
  • Calcium / metabolism
  • Cations, Divalent / metabolism
  • Cloning, Molecular
  • Coenzymes / metabolism
  • Enzyme Stability
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Gene Expression
  • Hydrogen-Ion Concentration
  • Kinetics
  • Molecular Sequence Data
  • Molecular Weight
  • Phytic Acid / metabolism*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Sequence Alignment
  • Temperature

Substances

  • Cations, Divalent
  • Coenzymes
  • Recombinant Proteins
  • Phytic Acid
  • 6-Phytase
  • Calcium

Associated data

  • GENBANK/AAU22048