Cloning, overexpression, and characterization of a thermostable nitrilase from an Antarctic Pyrococcus sp.
Extremophiles
; 21(5): 861-869, 2017 Sep.
Article
in En
| MEDLINE
| ID: mdl-28744780
ABSTRACT
Nitriles are important chemical building blocks for the synthesis of intermediates in fine chemical and pharmaceutical industries. Here, we report a new highly thermostable nitrilase from an Antarctic Pyrococcus sp. MC-FB, a hyperthermophilic archaeon. A gene that encoded a nitrilase was identified and subsequently cloned and overexpressed in Escherichia coli. The recombinant nitrilase, named NitMC-FB, is active as a homodimer (60 kDa) with an optimal temperature and pH of 90 °C and 7.0, respectively. NitMC-FB hydrolyzes preferentially aromatic nitriles, being the first aromatic nitrilase from an archaeon described so far. The K M and V max parameters were determined to be 13.9 mM and 3.7 µmol/min*mg, respectively, with 2-cyanopyridine as the substrate. Additionally, the recombinant nitrilase is highly thermostable with a half-life of 8 h at 90 °C.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Pyrococcus
/
Archaeal Proteins
/
Aminohydrolases
Language:
En
Journal:
Extremophiles
Journal subject:
BIOLOGIA
Year:
2017
Type:
Article
Affiliation country:
Chile