An efficient thermostable organophosphate hydrolase and its application in pesticide decontamination
| Autor: | Luigia Merone, Elena Porzio, Immacolata Del Giudice, Giuseppe Manco, Teresa Maria Carusone, Franz Worek, Luigi Mandrich, Rossella Coppolecchia |
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| Rok vydání: | 2015 |
| Předmět: |
0301 basic medicine
chemistry.chemical_classification biology Paraoxon ved/biology Kinetics Mutant Sulfolobus solfataricus ved/biology.organism_classification_rank.species Bioengineering Applied Microbiology and Biotechnology Combinatorial chemistry 03 medical and health sciences Hydrolysis 030104 developmental biology Enzyme Biochemistry chemistry Lactonase biology.protein medicine Systematic evolution of ligands by exponential enrichment Biotechnology medicine.drug |
| Zdroj: | Biotechnology and Bioengineering. 113:724-734 |
| ISSN: | 0006-3592 |
| DOI: | 10.1002/bit.25843 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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