F. SAMAAN, S., R. ALANI, S., M. AL-SHWAIKH, R. (2012). PURIFICATION AND CHARACTERIZATION OF PROTEASE FROM Pseudomonas aeruginosa ISOLATED FROM SOME WOUND AND BURN INFECTION.. Alustath, 3(3), 37-44. doi: 10.37652/juaps.2009.15593
SAMEER F. SAMAAN; SOUOD R. ALANI; RANA M. AL-SHWAIKH. "PURIFICATION AND CHARACTERIZATION OF PROTEASE FROM Pseudomonas aeruginosa ISOLATED FROM SOME WOUND AND BURN INFECTION.". Alustath, 3, 3, 2012, 37-44. doi: 10.37652/juaps.2009.15593
F. SAMAAN, S., R. ALANI, S., M. AL-SHWAIKH, R. (2012). 'PURIFICATION AND CHARACTERIZATION OF PROTEASE FROM Pseudomonas aeruginosa ISOLATED FROM SOME WOUND AND BURN INFECTION.', Alustath, 3(3), pp. 37-44. doi: 10.37652/juaps.2009.15593
F. SAMAAN, S., R. ALANI, S., M. AL-SHWAIKH, R. PURIFICATION AND CHARACTERIZATION OF PROTEASE FROM Pseudomonas aeruginosa ISOLATED FROM SOME WOUND AND BURN INFECTION.. Alustath, 2012; 3(3): 37-44. doi: 10.37652/juaps.2009.15593

PURIFICATION AND CHARACTERIZATION OF PROTEASE FROM Pseudomonas aeruginosa ISOLATED FROM SOME WOUND AND BURN INFECTION.

Journal of University of Anbar for Pure Science
Article 12, Volume 3, Issue 3, December 2009, Pages 37-44    PDF (613.73 K)
Document Type: Research Paper
DOI: 10.37652/juaps.2009.15593
Authors
SAMEER F. SAMAAN* 1; SOUOD R. ALANI2; RANA M. AL-SHWAIKH* 3
1Al-Mustansiriya University - College of Science
2The Ministry of Science and Technology
3University of Baghdad - College of Education Ibn Al-Haytham
Abstract
Twenty Four isolates of Pseudomonas aeruginosa were identified. The isolates were
8(33%) from wound infections, 16(66%) isolates from burn infections. The sensitivity of Pseudomonas
aeruginosa isolates was been tested against (10) antibiotics showed isolates version resistance with
different percentage against antibiotics. Pseudomonas aeruginosa exhibited (100%) resistance to
Ampicillin. While percentages of resistance to Cefixime and Ceftazidime were (95.8%) and (79%)
respectively. Resistance percentages to Tobramycin, Piperacillin, Norfloxacillin, Ciprofloxacin were
(41.6%), (20.8%), (20.8%) and (4%) respectively. All isolates of Pseudomonas aeruginosa were highly
sensitive (100%) to Aztronam, Imipenem, Cefepime. The optimum conditions for protease production
were in LB medium with a pH (8) after (48) hrs of incubation at (35) Cº. Purification of the protease
was done using ion exchange chromatography DEAE-cellulose and gel filtration with sephadex G-100.
Molecular weight of the purified protease was measured by sephadex G-100 and it was found to be
around (21379) Dalton. The optimum temperature of enzyme activity was (35) Cº. However, the pH (8)
was for activity and stability of this enzyme. Zn++ and Ca++ ions may play a role in the enhancement
and stability of the enzyme. Enzyme activity was not inhibited in the presence of reducing agent such
as Cysteine, but it was inhibited in the presence of EDTA
Keywords
Pseudomonas aeruginosa; Protease
References
1.Pellegrino , F. P. C. ; Teixeira , L. M. ; Carvalho , M. G. S. ; Nouer , S.A. ; Oliveire , M. P. ; Sampaio , J. L. M. ; Freitas , A. D. ; Ferreira , A. L. P. ; Amorim , E.L.T.  ; Riley , L.  W. and Moreira, B. M. (2002). Occurrence of a multidrug resistant Pseudomonas aeruginosa clone in different hospitals in Rio de Janeiro, Brazil. J Clin Microb. 40(7):2420-2424.

2.Song, Z.; Wu, H.; Ciofu, O.; Kong,  K.; Hoiby , N.; Rygaard , J. ; Kharazmi, A. and Mathee,K. (2003) . Pseudomonas aeruginosa alginate is refractory to Th1 immune response and impedes host immune clearance in a mouse model of acute lung infection. J Med Microb. 52: 731-740.

3.Mariencheck, W.I.; Alcorn, J. F.; Palmer, S.M.  and Wright, J. R. (2003). Pseudomonas aeruginosa Elastase Degrades surfactant protein A and D. Am J Res Mol Bio. 28:528–537.

4.Al-Douri, S.S. (2003). Characteristics of alkaline protease produced by Pseudomonas aerations. MSC. Thesis Al –Nahrain University. Baghdad. Iraq.

  1. المحمداوی،خوله جبر خلف.(2006). دراسة کیموحیویة وجزیئیة للبروتین المنتج من العزلةالمحلیة لبکتریا Pseudomonas aeruginosa . أطروحة دکتوراه. کلیة العلوم– الجامعة المستنصریة.
6.Foca, M.; Jakob , K. ; Whittier,  S.; Dello-Latta , P. ; Factor, S.; Rubenstein, D. and Saiman, L.(2005). Endemic Pseudomonas aeruginosa infection in a Neonatal Intensive Care Unit. New Englan J Med. 343 (10): 695-700.

7.Kenneth Todar University of Wisconsin-Madison Department of Bacteriology. (2004).Todar,s online Textbook of Bacteriology.

8.Cotter, C.S.; Avidano, M. A.; Stringer, S.P. and Schultz, G.S. (1996) .Inhibition of proteases in Pseudomonas otitis media in chinchillas. Otolaryngol Head Neck Surg. 115 (4): 342-51.

9.Baron, E. J.; Finegold, S. M. and Peterson, I. L. R. (1994). Bailey and Scott،s diagnostic microbiology 9th ed. Mosby Company. Missouri.

  1. Vandepitte, J.; Verhaegen, J.; Engbaek, K.; Rohner,P.; Piot, P. and Heuck,C.C.(2003). Basic laboratory procedures in clinical Bacteriology .2nded. World Health Organization Geneva. PP. 109-120.
  2. Peters, J. E. and Galloway, D. R. (1990). Purification and Characterization of an active fragment of the Las a protein from Pseudomonas aeruginosa: Enhancement of Elastase activity. J Bacter. 172 (5):2236 2240.
  3. Senior, B.W. (1999). Investigation of the types and characteristics of the proteolytic enzyme formed by diverse strains of proteus species. J. Med. Microbial. 48: 623 -628.
  4. Scopes, R. K. (1987). Protein purification principles and practice.  2nded. Springer Verlag, New York.
  5. Andrews, P. (1964).  Estimation of the Molecular Weights of proteins by sephadex Gel – filtration. Biochem. J. 91: 222-233.
  6. Pirnay, J.P.; DeVos , D.; Cochez, C. ;Bilocq, F.; Pirson, J.; Struelens, M.; Duinslaeger, L.; Cornelis, P.; Zizi, M. and Vanderkelen, A.(2003). Molecular epidemiology of Pseudomonas aeruginosa colonization in a burn unit: persistence of a multidrug resistant clone and a silver sulfadiazine resistant clone. J Clin Microb.41 (3): 1192-1202.
  7. Kriengkauykiat, J.; Porter, E.; Lomovskaya, O. and Wong-Beringer, A. (2005). Use of an efflux pump inhibitor to determine the prevalence of efflux pump mediated fluoroquinolone resistance and multidrug resistance in Pseudomonas aeruginosa. Antimicrob Agents Chemother .49 (2):565-570.
  8. Mansour, A. and Enayat, K. (2004). Bacteriological Monitoring of hospital borne septicemia in burn patients in Ahvaz, Iran. J. Burns and Sury Wound Care. 11: 42-51.
  9. Livermore, D.M. (2002). Multiple Mechanisms of antimicrobial resistance in Pseudomonas aeruginosa: our worst Nightmare. Clin Infect Dis. 34: 634-640.
  10. Flamm, R.K.; Weaver, M. K.; Thornsberry, C.; Jones, M. E.; Karlowsky, J. A. and Sahm, D.F. (2004). Factors associated with relative rates of antibiotic resistance in Pseudomonas aeruginosa Isolates tested in clinical laboratories in the United States from 1999 to 2002. Antimicrobl Agents Chemother.48 (7): 2931-2436.
  11. Neuhauser, M.M.; Weinstein, R.A.; Rydman, R.; Danziger ,L. H.; Karam, G. and Quinn, J.P. (2003).Antibiotic resistance among Gram-negative Bacilli in US Intensive Care Units. JAMA. 289(7): 885-888.
  12. Fluit, A.C.; Verhoef, J.; Schmitz, F. J.; European SEN TRY Participants. (2001). Frequency of isolation and antimicrobial resistance of  gram negative and gram positive bacteria from patients in intensive care unite  of 25 European university  hospitals participating in the European  arm of the SENTRY antimicrobial surveillance program 1997-1998 . Euro. J. Clin. Microbial Infect. Dis. 20 (9): 617-25.
  13. Kapoor, S. and Gathwala, G. (2004). Drug Therapy Aztreonam. Indian Pediatrics. 41 (17): 359-364.
  14. Barequet , I.S. ;  Ben Simon , G.J. ; Safrin , M. ; Ohman, D.E., and Kessler, E. (2004). Pseudomonas aeruginosa Las A protease in treatment of experimental Staphylococcal Keratitis. Antimicro Agents Chemother .48(5): 1681-1687.
  15. Cahan, R.; Axelrad, I.; Safrin, M.; Ohman, D.E. and Kessler, E. (2001). Asecreted amino peptidase of Pseudomonas aeruginosa. J. Biol. Chem. 276 ( Issue 47 )  23 : 43645-43652
  16. Matsumoto, K. (2004). Role of bacterial protease in Pseudomonal and Serratial keratitis. Biol.Chem. 385: 1007-1016.
  17. Anisia, J.; Silva, K.; Jorge, A. and Benitcz, E. (2003). Hemagglutinin/ protease expression and mucin gel penetration in Eltor biotype Vibrio cholerae. Microbiology. 149: 1883-1891.  
  18. Kessler , E. ; Safrin , M. ; Abrams , W.R.; Rosen bloom , J. and Ohman , D.E.(1997) . Inhibitors and Specificity of Pseudomonas aeruginosa   Las A. J.B.C. 272 (15). Issue 11 :9884-9889
  19. Means, G.E. and Feeny, R.E. (1971). Chemical Modification of Proteins. Holden-Day Inc., San Francisco.
Statistics
Article View: 68
PDF Download: 24


Journal Management System. Powered by ejournalplus