Hyperthermophilic aldolases as biocatalyst for C-C bond formation: rhamnulose 1-phosphate aldolase from Thermotoga maritima

Appl Microbiol Biotechnol. 2015 Apr;99(7):3057-68. doi: 10.1007/s00253-014-6123-7. Epub 2014 Oct 18.

Abstract

The TM1072 gene from Thermotoga maritima codifies for a putative form of a rhamnulose-1-phosphate aldolase (Rha-1PA Tm). To investigate this enzyme further, its gene was cloned and expressed in Escherichia coli. The purified enzyme was activated by Co(2+) as a divalent metal ion cofactor, instead of Zn(2+) as its E. coli homologue, and exhibited a maximum of activity at 95 °C. Furthermore, the enzyme displayed a high stability against extreme reaction conditions, retaining 90 % of its activity in the presence of 40 % of acetonitrile and showing a half-life greater than 3 h at 115 °C. The kinetic parameters at room temperature (R/T) were also studied; the K M was calculated to be 3.6 ± 0.33 mM, while k cat/K M was found to be 0.7 × 10(3) s(-1) M(-1). Given these characteristics, Rha-1PA Tm is an attractive enzyme for use as a biocatalyst for industrial applications, offering intriguing possibilities for practical biocatalysis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aldehyde-Lyases / chemistry
  • Aldehyde-Lyases / genetics*
  • Aldehyde-Lyases / metabolism*
  • Amino Acid Sequence
  • Catalysis
  • Cloning, Molecular
  • Enzyme Stability
  • Escherichia coli / genetics
  • Half-Life
  • Hydrogen-Ion Concentration
  • Kinetics
  • Molecular Sequence Data
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Temperature
  • Thermotoga maritima / enzymology*
  • Thermotoga maritima / genetics

Substances

  • Recombinant Proteins
  • Aldehyde-Lyases
  • rhamnulose-1-phosphate adolase