Jpn. J. Food Microbiol., 26(2), 76 80, 2009 Symposium 1 Control of microorganisms in stress environment Mechanisms of Actions of Hypochlorous Acid as Cleaning and Disinfecting Agents in Relation to Injury to Bacteria Satoshi FJ@JO6@> (Industrial Technology Center of Okayama Prefecture, 5301 Haga, Kita-ku, Okayama-shi 701 1296) 1. (HOCl) (NaOCl) ph pk a 7.5, 25 21) ph (OCl ) (H ) HOCl OCl H (1) 1 (HOCl) ph (ph 8 10) OCl (ph 4 6) HOCl ph HOCl (Cl 2 ) HOCl HOCl, OCl,Cl 2 Cl 2 HOCl OCl ph ph HOCl OCl 14) 2. (HOCl) (HOH) (H) (Cl) HOCl 1 Cl 14) Cl C C, C N, C N NH 2, SH (d ) Cl 2 (2e ) Cl 701 1296 5301 1 ph
77 2 HOCl 3 P. fluorescens (HOCl) 3. (1) (ph 7.5 10) (ph 5 6.5) HOCl 4, 12, 16) HOCl 2 HOCl OCl OCl 2 A OCl HOCl HOCl 2 A 2 B OCl HOCl HOCl H ph DpH ATP H ATP HOCl DpH 14) Chick Watson 6, 25) log(n/n 0 ) kct (2) N 0 N T C k log(n/n 0 ) vs. CT 3 Pseudomonas fluorescens 2.5 ppm, ph 5.7 9.3 log(n/n 0 ) vs. CT 16) (C) HOCl ph 1 ph HOCl ph Bacillus 4, 20) k Arrhenius 10 k 2 15) (2) ( SH) HOCl OCl 1) HOCl SH 1, 10, 23) SH
78 Vol. 26 No. 2 2009 4 BSA P. fluorescens SH HOCl SH HOCl 2) ATP / 3) 5) HOCl DNA 10, 19) HOCl SH DNA (3) HOCl (H 2 O 2 ) ( OH) 9, 10) O 2 1 O 2 O 2 H 2 O 2 OH H 2 O (3) (3) 2 B O 2 (SOD) H 2 O 2 H 2 O 2 H 2 O (3) H 2 O 2 H 2 O OH OH HOCl O 2 H 2 O 2 O 2 H 2 O 2 H 2 O 2 HOCl Fe 2 22) Fe 2 H 2 O 2 OH (4) Fenton OH DNA 2 H 2 O 2 Fe 2 OH OH Fe 3 (4) HOCl Fe 2 OH 13) HOCl Fe 2 OH Cl Fe 3 (5) (4) (5) Fe 3 O 2 Fe 2 Haber Weiss HOCl HOCl OH 14) 4. (ph 12) (OH ) OH (ph 7.5 10) OCl 24) (1) OCl OH ph OCl 4 (BSA) P. fluorescens (Al 2 O 3 ) ph (2 10) (120 2,200 ppm) OCl 14) BSA P. fluorescens OCl OCl AC 1,000 ppm OCl 10 ppm ph BSA OCl 24) P. fluorescens
79 Arrhenius 60 OCl 15) (2) ( NH 3 ) HOCl/OCl 17, 18) ( NH CO ) (d ) ( N CO ), Cl OH OCl 7) (3) (ph 12.5) OH OCl ph 10,000 ppm OH OCl SH 11) (1.5 4 ) (200 ppm) OH OCl 8) 5. HOCl 1) Albrich, J. M., McCarthy, C. A. and Hurst, J.: Biological reactivity of hypochlorous acid: Implications for microbicidal mechanisms of leukocyte myeloperoxidase. Proc. Natl. Acad. Sci. USA, 78, 210 214 (1981). 2) Barrette, W. C. Jr., Albrich, J. M. and Hurst, J. K.: Hypochlorous acid-promoted loss of metabolic energy in Escherichia coli. Infect. Immun., 55, 2518 2525 (1987). 3) Barrette, W. C. Jr., Hannum, D. M., Wheeler, W. D. and Hurst, J. K.: General mechanism for the bacterial toxicity of hypochlorous acid: Abolition of ATP production. Biochemistry, 28, 9172 9178 (1989). 4) Brazis, A. R., Leslie, J. E., Kabler, P. W. and Woodward, R. L.: The inactivation of spores of Bacillus globigii and Bacillus anthracis by free available chlorine. Appl. Microbiol., 6, 338 342 (1958). 5) Champer, A. K. and McFeters, G. A.: Chlorine injury and the enumeration of waterborne coliform bacteria. Appl. Environ. Microbial., 37, 633 641 (1979). 6) Chick, H.: An investigation of the laws of disinfection. J. Hyg., 8, 92 158 (1908). 7) Clegg, L. F. L.: Cleaning and sterilization of milking equipment on the farm. In Milk Hygiene (FAO/WHO), p. 195 220, World Health Organization, Geneva (1962). 8) Cousins, C. M. and Allan, C. D.: Sporicidal properties of some halogens. J. Appl. Bacteriol., 30, 168 174 (1967). 9) Dukan, S., Belkin, S. and Touati, D.: Reactive oxygen species are partially involved in the bacteriocidal action of hypochlorous acid. Arch. Biochem. Biophys., 367, 311 316 (1999). 10) Dukan, S. and Touati, D.: Hypochlorous acid stress in Escherichia coli: Resistance, DNA damage, and comparison with hydrogen peroxide stress. J. Bacteriol., 178, 6145 6150 (1996). 11) Estrela, C., Estrela, C. R. A., Barbin, E. L., Spanó, J. C., Marchesan, M. A. and Pécora, J. D.: Mechanism of action of sodium hypochlorite. Braz. Dent. J., 13, 113 117 (2002). 12) Fair, G. M. and Morris, J. C., Chan, S. L, Weil, I. and Burden, R. P.: The behavior of chlorine as a water disinfectant. J. Am. Water Works Assoc., 40, 1051 1061 (1948) 13) Folkes, L. K., Candeias, L. P. and Wardman, P.: Kinetics and mechanisms of hypochlorous acid reactions. Arch. Biochem. Biophys., 323, 120 126 (1995). 14) Fukuzaki, S.: Mechanisms of actions of sodium hypochlorite in cleaning and disinfecting processes. Biocontrol Sci., 11, 147 157 (2006). 15) Fukuzaki, S., Urano, H., Takahashi, K., Yamada, S. and Takagi, A.: Kinetic study of the e#ect of temperature on the cleaning and disinfecting actions of sodium hypochlorite. Bokin Bobai, 37, 253 262 (2009) (in Japanese). 16) Fukuzaki, S., Urano, H. and Yamada, S.: E#ect of ph
80 Vol. 26 No. 2 2009 on the e$cacy of sodium hypochlorite solution as cleaning and bactericidal agents. J. Surface Finish. Soc. Jpn., 58, 465 469 (2007). 17) Hawkins, C. L. and Davies, M. J.: Hypochloriteinduced damage to proteins: formation of nitrogencentred radicals from lysine residues and their role in protein fragmentation. Biochem. J., 332, 617 625 (1998). 18) Hawkins, C. L. and Davies, M. J.: Hypochloriteinduced oxidation of proteins in plasma: Formation of chloramines and nitrogen-centred radicals and their role in protein fragmentation. Biochem. J., 340, 539 548 (1999). 19) McKenna, S. M. and Davies, K. J. A.: The inhibition of bacterial growth by hypochlorous acid: Possible role in the bactericidal activity of phagocytes. Biochem. J., 254, 685 692 (1988). 20) Marks, H. C., Wyss, O. and Stradskov, F. B.: Studies on the mode of action of compounds containing available chlorine. J. Bacteriol., 49, 299 305 (1945). 21) Morris, J. C.: The acid ionization constant of HOCl from 5 to 35. J.Phys. Chem., 70, 3798 3805 (1966). 22) Rosen, H. and Klebano#, S.: Oxidation of Escherichia coli iron centers by the myeloperoxidase-mediated microbial system. J. Biol. Chem., 257, 13731 13735 (1982). 23) Thomas, E. L.: Myeloperoxidase-hydrogen peroxidechloride antimicrobial system: E#ect of exogenous amines on antibacterial action against Escherichia coli. Infect. Immun., 25, 110 116 (1979). 24) Urano, H. and Fukuzaki, S.: The mode of action of sodium hypochlorite in the cleaning process. Biocontrol Sci., 10, 21 29 (2005). 25) Watson, H. E.: A note on the variation of the rate of disinfection with change in the concentration of the disinfectant. J. Hyg., 8, 536 (1908).