Diisopropyl-fluorophosphatase as a Catalytic Bioscavenger

  • Hossein Allahyari Applied Biotechnology Research Center, Baqiyatallah University of Medical Sciences, Tehran Iran
  • Ali Mohammad Latifi Applied Biotechnology Research Center, Baqiyatallah University of Medical Sciences, Tehran Iran


Organophosphorus chemicals, used as pesticides and warfare nerve agent, are highly toxic compounds that inhibit acetylcholine esterase enzyme rapidly. A novel effective treatment for nerve gas poising is using of hydrolytic enzymes to degradation of these agents. DFPase from loligo vulgaris is highly stable and robust biocatalyst for the hydrolysis of various chemical warfare agents such as sarin, soman, tabun. Unfortunately, wild-type DFPase prefers less toxic isomers of these agents leading to slower detoxification. Also, due to non-human origin of the enzyme, immunological reactions occur when it injects into body. Engineering of the enzyme for creating of new efficient variants is an interesting research field which leads to occurrence of novel and prominent bioscavengers.


1. Raushel, F.M., Chemical biology: Catalytic detoxification. Nature, 2011. 469(7330): p. 310-311.

2. Wymore, T., et al., Hydrolysis of DFP and the nerve agent (S)-sarin by DFPase proceeds along two different reaction pathways: implications for engineering bioscavengers. The Journal of Physical Chemistry B, 2014. 118(17): p. 4479-4489.

3. Wales, M.E. and T.E. Reeves, Organophosphorus hydrolase as an in vivo catalytic nerve agent bioscavenger. Drug testing and analysis, 2012. 4(3-4): p. 271-281.

4. Nachon, F., et al., Progress in the development of enzyme-based nerve agent bioscavengers. Chemico-biological interactions, 2013. 206(3): p. 536-544.

5. Chandrasekaran, L., T. Belinskaya, and A. Saxena, In search of a catalytic bioscavenger for the prophylaxis of nerve agent toxicity. Chemico-biological interactions, 2010. 187(1): p. 349-354.

6. Masson, P., Evolution of and perspectives on therapeutic approaches to nerve agent poisoning. Toxicology letters, 2011. 206(1): p. 5-13.

7. Melzer, M., et al., Reversed enantioselectivity of diisopropyl fluorophosphatase against organophosphorus nerve agents by rational design. Journal of the American Chemical Society, 2009. 131(47): p. 17226-17232.

8. Chemnitius, J.-M., et al., Organophosphate detoxicating hydrolases in different vertebrate species. Comparative Biochemistry and Physiology Part C: Comparative Pharmacology, 1983. 76(1): p. 85-93.

9. Hoskin, F.C., M.A. Kirkish, and K.E. Steinmann, Two enzymes for the detoxication of organophosphorus compounds—sources, similarities, and significance. Fundamental and Applied Toxicology, 1984. 4(2): p. S165-S172.

10. Hoskin, F.C. and R.J. Long, Purification of a DFP-hydrolyzing enzyme from squid head ganglion. Archives of biochemistry and biophysics, 1972. 150(2): p. 548-555.

11. Little, J.S., et al., Partial characterization of an enzyme that hydrolyzes sarin, soman, tabun, and diisopropyl phosphorofluoridate (DFP). Biochemical pharmacology, 1989. 38(1): p. 23-29.

12. Mazur, A., An enzyme in animal tissues capable of hydrolyzing the phosphorus-fluorine bond of alkyl fluorophosphates. Journal of Biological Chemistry, 1946. 164(1): p. 271-289.

13. Wang, F., M. Xiao, and S. Mu, Purification and properties of a diisopropyl‐fluorophosphatase from squid Todarodes pacificus steenstrup. Journal of biochemical toxicology, 1993. 8(3): p. 161-166.

14. Mirzaei, M., A.M. Latifi, and R. Jafari, Improvement of Thermal Stability of DFPase by In silico Methods. Journal of Applied Biotechnology Reports, 2015. 1(4): p. pp. 155-159.

15. Bigley, A.N. and F.M. Raushel, Catalytic mechanisms for phosphotriesterases. Biochimica et Biophysica Acta (BBA)-Proteins and Proteomics, 2013. 1834(1): p. 443-453.

16. Katsemi, V., et al., Mutational and structural studies of the diisopropylfluorophosphatase from Loligo vulgaris shed new light on the catalytic mechanism of the enzyme. Biochemistry, 2005. 44(25): p. 9022-9033.

17. Yeung, D.T., et al., Structure/function analyses of human serum paraoxonase (HuPON1) mutants designed from a DFPase-like homology model. Biochimica et Biophysica Acta (BBA)-Proteins and Proteomics, 2004. 1702(1): p. 67-77.

18. Bird, S.B., A. Dawson, and D. Ollis, Enzymes and bioscavengers for prophylaxis and treatment of organophosphate poisoning. Frontiers in bioscience (Scholar edition), 2009. 2: p. 209-220.

19. Draganov, D.I., Paraoxonase 1 (PON1) as a Potential Catalytic Sca-venger in the Prophylaxis and Treatment of Organo-phosphate Poisoning. Medical Aspects of Chemical and Biological Terrorism, 2005: p. 227.

20. Kirby, S.D., et al., Human paraoxonase double mutants hydrolyze V and G class organophosphorus nerve agents. Chemico-biological interactions, 2013. 203(1): p. 181-185.

21. Tsai, P.-C., et al., Enzymes for the homeland defense: optimizing phosphotriesterase for the hydrolysis of organophosphate nerve agents. Biochemistry, 2012. 51(32): p. 6463-6475.

22. Yeung, D.T., D.E. Lenz, and D.M. Cerasoli, Human paraoxonase I: A potential bioscavenger of organophosphorus nerve agents, in The Paraoxonases: Their Role in Disease Development and Xenobiotic Metabolism. 2008, Springer. p. 151-170.

23. Hoskin, F.C., Diisopropylphosphorofluoridate and tabun: enzymatic hydrolysis and nerve function. Science, 1971. 172(3989): p. 1243-1245.

24. Schulz, W., et al., Enzymatishe hydrolyse von hochtoxischen phosphoroganishchen verbindugen. Auftragsnummer T, 1987.

25. Blum, M.-M., et al., Inhibitory potency against human acetylcholinesterase and enzymatic hydrolysis of fluorogenic nerve agent mimics by human paraoxonase 1 and squid diisopropyl fluorophosphatase. Biochemistry, 2008. 47(18): p. 5216-5224.

26. Hartleib, J. and H. Rüterjans, Insights into the reaction mechanism of the diisopropyl fluorophosphatase from Loligo vulgaris by means of kinetic studies, chemical modification and site-directed mutagenesis. Biochimica et Biophysica Acta (BBA)-Protein Structure and Molecular Enzymology, 2001. 1546(2): p. 312-324.

27. Cheng, T.-c. and J.J. Calomiris, A cloned bacterial enzyme for nerve agent decontamination. Enzyme and microbial technology, 1996. 18(8): p. 597-601.

28. Drevon, G.F., Enzyme immobilization into polymers and coatings. 2002, University of Pittsburgh.

29. Lenz, D.E., et al., Stoichiometric and catalytic scavengers as protection against nerve agent toxicity: a mini review. Toxicology, 2007. 233(1): p. 31-39.

30. Masson, P. and D. Rochu, Catalytic bioscavengers against toxic esters, an alternative approach for prophylaxis and treatments of poisonings. Acta naturae, 2009. 1(1): p. 68.

31. Trovaslet-Leroy, M., et al., Organophosphate hydrolases as catalytic bioscavengers of organophosphorus nerve agents. Toxicology letters, 2011. 206(1): p. 14-23.

32. Aharoni, A., et al., Directed evolution of mammalian paraoxonases PON1 and PON3 for bacterial expression and catalytic specialization. Proceedings of the National Academy of Sciences of the United States of America, 2004. 101(2): p. 482-487.

33. Amitai, G., et al., Enhanced stereoselective hydrolysis of toxic organophosphates by directly evolved variants of mammalian serum paraoxonase. FEBS Journal, 2006. 273(9): p. 1906-1919.

34. Goldsmith, M., et al., Evolved stereoselective hydrolases for broad-spectrum G-type nerve agent detoxification. Chemistry & biology, 2012. 19(4): p. 456-466.

35. Gupta, R.D., et al., Directed evolution of hydrolases for prevention of G-type nerve agent intoxication. Nature Chemical Biology, 2011. 7(2): p. 120-125.

36. Hu, X., et al., In silico analyses of substrate interactions with human serum paraoxonase 1. Proteins: Structure, Function, and Bioinformatics, 2009. 75(2): p. 486-498.

37. A. Aharoni, L.G., O. Khersonsky, Q.G.S. Mc, C. Roodveldt, D.S. Tawfik, The evolvability’ of promiscuous protein functions. Nat. Genet, 2005. 37(1): p. 73-76.

38. Cleland, W.W.H., A. C, Enzymatic mechanisms of phosphate and sulfate transfer. Chem. Rev, 2006. 106: p. 3252-3278.

39. Chirino.A.J, A.M.L., Marshal.S.A, Minimizing the immunogenicity of protein therapeutics. Drug Discov. Today, 2004. 9(82).

40. Besio.R, A.S., Forlino.A, Lupi.A, Meneghini.C,Minicozzi.V, Profumo.A, and T.R. Stellato.F, Morante.S, Identifying the structure of the active sites of human recombinant prolidase. Eur. Biophys. J, 2009.

41. Cheng.T.C, D.J.J., Wide-range Application of Alteromonas prolidase For Decontamination of G-type Chemical Nerve Agents. CBMTS III, Spiez, Switzerland, 2000.

42. Attaway, H., et al., Bacterial detoxification of diisopropyl fluorophosphate. Applied and environmental microbiology, 1987. 53(7): p. 1685-1689.

43. Ghanem, E. and F.M. Raushel, Detoxification of organophosphate nerve agents by bacterial phosphotriesterase. Toxicology and applied pharmacology, 2005. 207(2): p. 459-470.

44. Tsai, P.-C., et al., Stereoselective hydrolysis of organophosphate nerve agents by the bacterial phosphotriesterase. Biochemistry, 2010. 49(37): p. 7978-7987.

45. An.Q, L.Y., Jia.N, , Effect of site-directed PEGylation of trichosanthin on its biological activity, immunogenicity, and pharmacokinetics. Biomol. Eng, 2007. 24(643).

46. Novikov.B.N, G.R., Kern.J.R, Wild.M, Wales.M, Improved pharmacokinetics and immunogenicity profile of organophosphorus hydrolase by chemical modification with polyethylene glycol. J. Control. Release, 2010. 146(318).

47. Webster.R, D.E., Harris.P, Siegel.N, Stadler.J, Tilbury.L, Smith.D, PEGylated proteins: evaluation of their safety in the absence of definitive metabolism studies. Drug Metab. Dispos, 2007. 35(9).

48. Melzer, M., et al., In vitro and in vivo efficacy of PEGylated diisopropyl fluorophosphatase (DFPase). Drug testing and analysis, 2012. 4(3-4): p. 262-270.

49. Blum, M.M. and A. Richardt, Hydrolytic enzymes for chemical warfare agent decontamination. Decontamination of Warfare Agents: Enzymatic Methods for the Removal of B/C Weapons, 2008: p. 135-162.
How to Cite
ALLAHYARI, Hossein; LATIFI, Ali Mohammad. Diisopropyl-fluorophosphatase as a Catalytic Bioscavenger. Journal of Applied Biotechnology Reports, [S.l.], v. 3, n. 4, p. 477-482, june 2017. ISSN 2423-5784. Available at: <https://journals.bmsu.ac.ir/jabr/index.php/jabr/article/view/104>. Date accessed: 20 oct. 2018.
Review Articles


DFPase; Bioscavenger; Organophosphate; Bioremediation

Most read articles by the same author(s)

Obs.: This plugin requires at least one statistics/report plugin to be enabled. If your statistics plugins provide more than one metric then please also select a main metric on the admin's site settings page and/or on the journal manager's settings pages.