An efficient thermostable organophosphate hydrolase and its application in pesticide decontamination.
Biotechnol Bioeng
; 113(4): 724-34, 2016 Apr.
Article
de En
| MEDLINE
| ID: mdl-26416557
ABSTRACT
In vitro evolution of enzymes represents a powerful device to evolve new or to improve weak enzymatic functions. In the present work a semi-rational engineering approach has been used to design an efficient and thermostable organophosphate hydrolase, starting from a lactonase scaffold (SsoPox from Sulfolobus solfataricus). In particular, by in vitro evolution of the SsoPox ancillary promiscuous activity, the triple mutant C258L/I261F/W263A has been obtained which, retaining its inherent stability, showed an enhancement of its hydrolytic activity on paraoxon up to 300-fold, achieving absolute values of catalytic efficiency up to 10(5) M(-1) s(-1). The kinetics and structural determinants of this enhanced activity were thoroughly investigated and, in order to evaluate its potential biotechnological applications, the mutant was tested in formulations of different solvents (methanol or ethanol) or detergents (SDS or a commercial soap) for the cleaning of pesticide-contaminated surfaces.
Mots clés
Texte intégral:
1
Collection:
01-internacional
Base de données:
MEDLINE
Sujet principal:
Pesticides
/
Décontamination
/
Phosphoric monoester hydrolases
/
Sulfolobus solfataricus
Langue:
En
Journal:
Biotechnol Bioeng
Année:
2016
Type de document:
Article
Pays d'affiliation:
Italie