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Murtadha Nabeel, Abdul-Gani
- Purification and Characterization of Chondroitinase ABC from Proteus vulgaris, an Iraqi Clinically Isolate
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Authors
Affiliations
1 Department of Biology, University of Madenat Al-Elem, Al-Kadhmiya, Baghdad, IQ
2 Department of Biology, College of Science, Baghdad University, Baghdad, IQ
1 Department of Biology, University of Madenat Al-Elem, Al-Kadhmiya, Baghdad, IQ
2 Department of Biology, College of Science, Baghdad University, Baghdad, IQ
Source
Current Science, Vol 113, No 11 (2017), Pagination: 2134-2140Abstract
Forty-eight isolates were found belonging to Proteus sp. with isolation percentage of 38 out of 125 urine samples which were collected from urinary tract infected patients. Proteus vulgaris and Proteus mirabilis represent 18.7% (9 isolates), and 81.25% (39 isolates) respectively. The production of chondroitinase enzyme was studied in a medium which contained chondroitin sulphate as substrate. One hundred per cent of P. vulgaris showed the ability to produce chondroitinase. P.v 8 isolate produced the highest enzymatic activity which reached about 150 units/ml. Chondroitinase was extracted and purified by precipitation with 60% saturation of ammonium sulphate, dialysis, and followed by column chromatography on Sephadex 6B. The specific activity increased to 5000 (units/mg), with 35.2% yield and 17.4 fold purification. Optimum pH for activity and stability was 8.0; the optimal temperature for activity was 37°C, but the stability of chondroitinase was maintained 100% at 20–40°C for 30 min. Chondroitinase activity increased to 120% and 113% when the enzyme was incubated with Mg+2 and Ca+2 respectively, while NH4Cl and KCl reduced the activity to 95% and 86% respectively. The human cartilage was degradable by the purified enzyme after incubation for 14 days at 37°C. Direct injection of chondroitinase at 0.1 mg/ml in knee cartilage of mice, showed changes in tissues. It may be concluded that chondroitinase enzyme may work as a virulence factor by catalysing the hydrolysis of chondroitin sulphate of cartilage and increasing the tissue permeability to invade the cartilage tissue by P. vulgaris which causes destruction of cartilage and inflammation in humans.Keywords
Chondroitinase ABC, Chondroitinase Extraction, Proteus vulgaris, Purification of Chondroitinase.Full Text
References
- Mobley, H. L. T. and Belas, R., Swarming and pathogenicity of Proteus mirabilis in the urinary tract. Trends Microbiol., 1995, 3, 280–284.
- Longo, D. L., Kasper, D. L., Jameson, J. L., Fauci, A. S., Hauser, S. L. and Loscalzo, J. (eds), Harrison’s Principles of Internal Medicine, McGraw Hill, New York, 2012, 18th edn.
- Collier, M., Understanding wound inflammation. Nurs. Times, 2003, 99(25), 63–64.
- Jacobsen, S. M., Stickler, D. J., Mobley, H. L. T. and Shirtliff, M. E., Complicated catheter-associated urinary tract infections due to Escherichia coli and Proteus mirabilis. Clin. Microbiol. Rev., 2008, 21(1), 26–59.
- Mordi, R. M. and Momoh, M. I., Incidence of Proteus species in wound infections and their sensitivity pattern in the University of Benin Teaching Hospital. Afr. J. Biotechnol., 2009, 8(5), 725–730.
- Smith, R. F. and Willett, N. P., Rapid plate method for screening hyaluronidase and chondroitin sulfatase-production microorganisms. Appl. Microbiol., 1968, 16(9), 1434–1436.
- Kato, F., Mimatsu, K., Iwata, H. and Miura, T., Comparison of tissue reaction with chondroitinase ABC and chymopapain in rabbits as the basis of clinical application in chemonucleolysis. Clin. Orthop. Relat. Res., 1993, 288, 294–302.
- Garrity, G. M., Brenner, D. J., Krieg, N. R. and Staley, J. T., Bergey’s Manual of Systematic Bacteriology, Springer, 2005, pp. 745–751.
- Yamagata, T., Saito, H., Habuchi, O. and Suzuki, S., Purification and properties of bacterial chondroitinases and chondrosulfatases. J. Biol. Chem., 1968, 243, 1523–1535.
- Abdul-Gani, M. N., Investigation chondroitinase from P. vulgaris and studying its effect on animal tissue. M. Sc. thesis. Biology Department, Collage of Science, Baghdad University, Iraq, 2015.
- Lohse, D. L. and Linhardt, R. J., Purification and characterization of heparin lyases from Flavobacterium heparinum. J. Biol. Chem., 1992, 267, 24347–24355.
- Ke, T., Zhangfu, L., Qing, G., Yong, T., Hong, J., Hongyan, R., Kun, L. and Shigui, L., Isolation of Serratia marcescens as a chondroitinase-producing bacterium and purification of a novel chondroitinase AC. Biotechnol. Lett., 2005, 7, 489–493.
- Whitaker, J. R. and Granum, P. E., An absolute method for protein determination based on difference in absorbance at 235 and 280 nm. Anal. Biochem., 1980, 109, 156–159.
- Laemmli, U. K., Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 1970, 227, 680–685.
- Humason, C. L., Animal Tissue Techniques, W.H. Freeman Company, 1972, 3rd edn, p. 641.
- Al-Bassam, W. W. and Al-Kazaz, A., The isolation and characterization of Proteus mirabilis from different clinical samples. J. Biotech. Res. Center, 2013, 7, 24-30.
- Uwaezuoke, J. C. and Ogbulie, J. N., Antibiotic sensitivity pattern of urinary tract pathogens in Port–Harcourt, Nigeria. J. Appl. Sci., 2006, 10(3), 103–107.
- Malik, S. N. A., Extraction and purification of lipopolysaccharide of locally isolated Proteus mirabilis and study of its effect to prevent urinary tract infection in an animal model. M.Sc. Thesis. College of Science, Biology Department, Baghdad University, Iraq, 2006.
- Orret, F. A., Prevalence of Proteus species in urinary tract infections in a regional hospital in Trinidad. Zhonghua Yi Xue Za Zhi (Tapei), 1999, 62, 438–442.
- Hamai, A., Hashimoto, N., Mochizuki, H., Kato, F., Makiguchi, Y., Horie, K. and Suzuki, S., Two distinct chondroitin sulfate ABC lyases: an endoeliminase yielding tetrasaccharides and an exoeliminase acting on oligosaccharides. J. Biol. Chem., 1997, 272, 9123–9130.
- Hashimoto, N., Mochizuki, H. and Harnai, A., Producing pure chondroitinase ABC from, US Pat. 5,763,205, 9 June 1998.
- Prabhakar, V., Capila, I., Bosques, C. J., Pojasek, K. and Sasisekharan, R., Chondroitinase ABC I from Proteus vulgaris: cloning, recombinant expression and active site identification. Biochem. J., 2005, 386, 103–112.
- Michelacci, Y. M., Horton, D. S. and Población, C. A., Isolation and characterization of an induced chondroitinase ABC from Flavobacterium heparium. Biochim. Biophys. Acta, 1987, 923, 291–301.
- Sawheney, S. K., Introductory Practical Biochemistry, Narosa Publishing House, Mumbai, 2008, p. 362.
- Berry, A. M., Lock, R. A., Thomas, S. M., Rajan, D. P., Hansman, D. and Paton, J. C., Cloning and nucleotide sequence of the Streptococcus pneumoniae hyaluronidase gene and purification of the enzyme from recombinant Escherichia coli. Infect. Immun., 1994, 62, 1101–1108.
- Howell, D. S., Sapolsky, A. I. and Pita, J. C., The Pathogenesis of Degenerative Joint Disease, UpJohn Company, Kalamazoo, MI, 1976, p. 28.