Catalytic buffers enable positive-response inhibition-based sensing of nerve agents.
We report herein an efficient method to control pH in reactions catalyzed by hydrolytic enzymes, such as the degradation of paraoxon by phosphotriesterase (E.G. 3.1.8.1; OPH), using urease-catalyzed (E.G. 3.5.1.5) urea hydrolysis as a buffering agent. Given the distinct pH profiles of urease and OPH activities, urease produces base on demand in response to pH drop during paraoxon detoxification. As pH changes, the enzyme activities fluctuate to finally stabilize at a pH "set-point," where the rates of acid and base generation are equal. By varying the urease to OPH ratio, various pH "set-points" ranging between 6.5 and 8.5 were achieved within minutes and could be predicted theoretically. This dynamic approach for pH control was successfully applied to the development of a positive-response inhibition-based sensor.
Russell
Alan J
AJ
Mc Gowan Institute for Regenerative Medicine, Suite 401 Scaife Hall, University of Pittsburgh, Pittsburgh, Pennsylvania 15261, USA. rusell@engrng.pitt.edu
Erbeldinger
Markus
M
DeFrank
Joseph J
JJ
Kaar
Joel
J
Drevon
Geraldine
G
D043303:Aryldialkylphosphatase; D015374:Biosensing Techniques; D001709:Biotechnology / Q000379:methods*; D002021:Buffers; D002619:Chemical Warfare Agents; D004950:Esterases / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D010261:Paraoxon / Q000378:metabolism*; D014508:Urea / Q000378:metabolism; D014510:Urease / Q000737:chemistry / Q000378:metabolism
D002021:Buffers; D002619:Chemical Warfare Agents; D014508:Urea; D004950:Esterases; D043303:Aryldialkylphosphatase; D014510:Urease; D010261:Paraoxon