Skip to main content

IMIS

A new integrated search interface will become available in the next phase of marineinfo.org.
For the time being, please use IMIS to search available data

 

[ report an error in this record ]basket (1): add | show Print this page

one publication added to basket [223230]
A thermostable L-aminoacylase from Thermococcus litoralis: cloning, overexpression, characterization, and applications in biotransformations
Toogood, H.S.; Hollingsworth, E.J.; Brown, R.C.; Taylor, I.N.; Taylor, S.J.C.; McCague, R.; Littlechild, J.A. (2002). A thermostable L-aminoacylase from Thermococcus litoralis: cloning, overexpression, characterization, and applications in biotransformations. Extremophiles 6(2): 111-122. http://dx.doi.org/10.1007/s007920100230
In: Extremophiles. Springer: Tokyo. ISSN 1431-0651; e-ISSN 1433-4909, more
Peer reviewed article  

Available in  Authors 

Keyword
    Marine/Coastal

Authors  Top 
  • Toogood, H.S.
  • Hollingsworth, E.J.
  • Brown, R.C.
  • Taylor, I.N.
  • Taylor, S.J.C.
  • McCague, R.
  • Littlechild, J.A., more

Abstract
    A thermostable L-aminoacylase from Thermococcus litoralis was cloned, sequenced, and overexpressed in Escherichia coli. The enzyme is a homotetramer of 43 kDa monomers and has an 82% sequence identity to an aminoacylase from Pyrococcus horikoshii and 45% sequence identity to a carboxypeptidase from Sulfolobus solfataricus. It contains one cysteine residue that is highly conserved among aminoacylases. Cell-free extracts of the recombinant enzyme were characterized and were found to have optimal activity at 85°C in Tris-HCl at pH 8.0. The recombinant enzyme is thermostable, with a half-life of 25 h at 70°C. Aminoacylase inhibitors, such as mono-tert-butyl malonate, had only a slight effect on activity. The enzyme was partially inhibited by EDTA and p- hydroxymercuribenzoate, suggesting that the cysteine residue and a metal ion are important, but not essential, for activity. Addition of Zn2+ and Co2+ to the apoenzyme increased the enzyme activity, whereas Sn4+ and Cu2+ almost completely abolished enzyme activity. The enzyme was most specific for substrates containing N-benzoyl- or N-chloroacetyl-amino acids, preferring substrates containing hydrophobic, uncharged, or weakly charged amino acids such as phenylalanine, methionine, and cysteine.

All data in the Integrated Marine Information System (IMIS) is subject to the VLIZ privacy policy Top | Authors