title: Improving the catalytic activity of hyperthermophilic Pyrococcus horikoshii prolidase for detoxification of organophosphorus nerve agents over a broad range of temperatures.
abstract: Prolidases hydrolyze Xaa-Pro dipeptides and can also cleave the P-F and P-O bonds found in organophosphorus (OP) compounds, including the nerve agents soman and sarin. Ph1prol (PH0974) has previously been isolated and characterized from Pyrococcus horikoshii and was shown to have higher catalytic activity over a broader pH range, higher affinity for metal, and increased thermostability compared to P. furiosus prolidase, Pfprol (PF1343). To obtain a better enzyme for OP nerve agent decontamination and to investigate the structural factors that may influence protein thermostability and thermoactivity, randomly mutated Ph1prol enzymes were prepared. Four Ph1prol mutants (A195T/G306S-, Y301C/K342N-, E127G/E252D-, and E36V-Ph1prol) were isolated which had greater thermostability and improved activity over a broader range of temperatures against Xaa-Pro dipeptides and OP nerve agents compared to wild type Pyrococcus prolidases.
authors.affiliation: Division of Infectious Diseases, Department of Internal Medicine, The University of Michigan, Ann Arbor, MI 48109, USA.
authors.forename: Casey M
authors.initials: CM
authors.lastname: Theriot
authors.forename: Rebecca L
authors.initials: RL
authors.lastname: Semcer
authors.forename: Saumil S
authors.initials: SS
authors.lastname: Shah
authors.forename: Amy M
authors.initials: AM
authors.lastname: Grunden
mesh_terms: D001711:Biotransformation; D002619:Chemical Warfare Agents / Q000378:metabolism*; D004150:Dipeptidases / Q000235:genetics / Q000378:metabolism*; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D050505:Mutant Proteins / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D010755:Organophosphates / Q000378:metabolism*; D043902:Pyrococcus horikoshii / Q000737:chemistry / Q000201:enzymology* / Q000378:metabolism; D012524:Sarin / Q000378:metabolism; D012999:Soman / Q000378:metabolism; D013696:Temperature
chemical_list: D002619:Chemical Warfare Agents; D050505:Mutant Proteins; D010755:Organophosphates; D012999:Soman; D012524:Sarin; D004150:Dipeptidases; C022449:proline dipeptidase