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- Title
Cloning, overexpression, and characterization of a thermostable nitrilase from an Antarctic Pyrococcus sp.
- Authors
Cabrera, Ma; Blamey, Jenny
- 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 and V 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.
- Subjects
CLONING; GENETIC overexpression; NITRILASES; ARCHAEBACTERIA; PYROCOCCUS; ESCHERICHIA coli
- Publication
Extremophiles, 2017, Vol 21, Issue 5, p861
- ISSN
1431-0651
- Publication type
Article
- DOI
10.1007/s00792-017-0948-9