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Purification and Characterization of Alginate Lyase from Mucoid Pseudomonas aeruginosa Strain 214 | ||
| Journal of Genetic Resources | ||
| مقاله 1، دوره 1، شماره 2، آذر 2015، صفحه 55-60 اصل مقاله (246.63 K) | ||
| نوع مقاله: Research Article | ||
| شناسه دیجیتال (DOI): 10.22080/jgr.2015.1164 | ||
| نویسندگان | ||
| Hoda Gol Fakhrabadi؛ Parinaz Ghadam* ؛ Parisa Mohammadi؛ Ahya Adbi Ali | ||
| Department of Biology, Faculty of Science, Alzahra University, Tehran, I.R. Iran | ||
| تاریخ دریافت: 28 اسفند 1391، تاریخ بازنگری: 01 تیر 1392، تاریخ پذیرش: 21 مرداد 1392 | ||
| چکیده | ||
| Pseudomonas aeruginosa is an opportunistic pathogen that causes a variety of infections in compromised patients. The ability of Pseudomonas aeruginosa to produce chronic infection is based in part on its ability to biosynthesis of biofilm, and alginate is the major polysaccharide in the synthesized biofilm. So alginate degradation is very essential in the dispersion of Pseudomonas aeruginosa biofilm. Alginate lyase is an important enzyme in alginate degradation. This enzyme is different, especially with respect to molecular weight, pI and substrate specificity in various bacteria and even in various strains of a bacterium. The amount of alginate in mucoid strains is more than in nonmucoid strains. In this study, P. aeruginosa strain 214 was selected because it forms highly mucoidal colonies and thus it is a good candidate for alginate lyase preparation. Alginate lyase was extracted from the periplasmic space of P. aeruginosa by the use of heat shock method. Thiobarbitoric acid assay was used for measuring the activity of alginate lyase. This enzyme showed the most activity in Tryptic Soy Broth (TSB) medium. The optimum concentration of sodium alginate was 0.02 mg/ml and the optimum activity of the enzyme was found in 20 min reaction time at 37°C. The enzyme was purified by a simple two-step procedure; ammonium sulfate precipitation and ion exchange column chromatography DEAE-Sepharose Cl-6B. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) suggested a molecular weight of approximately 40 kDa for alginate lyase. | ||
| کلیدواژه‌ها | ||
| Alginate؛ Alg lyase؛ Biofilm؛ Pseudomonas aeruginosa | ||
| مراجع | ||
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Abdi-Ali A, Mohammadi-Mehr M, Agha Alaei Y. 2006. Bactericidal activity of various antibiotics against biofilm-producing Pseudomonas aeruginosa. Int J Antimicrob Agent 27: 196-200.
Bartell PF, Orr TE, Lam Gk. 1966. Polysaccharide depolymerases associated with bacteriophage infection. J Bacteriol 92: 56-62.
Boyd A, Chakrabarty AM. 1994. Role of AlgL cell detachment of Pseudomonas aeruginosa. Appl Environ Microb 60: 2355-2364.
Boyd A, Ghosh M, May TB, Shinabarger D, Keogh R, Chakrabarty AM. 1993. Sequence of the algL gene of Pseudomonas aeruginosa and purification of its AlgL product. Gene 131: 1-8.
Boyd J, Turvey JR. 1977. Isolation of a poly-alpha-L-guluronate lyase from Klebsiella aerogenes. Carbohydr Res 57: 163-171.
Doubet RS, Quatrano RS. 1982. Isolation of marine bacteria capable of producing specific lyases for Alginate degradation. Appl Environ Microb 44: 754-760.
Dunne WM, Buckmire FL. 1985. Partial purification and characterization of a polymannuronic acid depolymerase produced by a mucoid strain of Pseudomonas aeruginosa isolated from a patient with cystic fibrosis. Appl Environ Microbiol 50: 562-569.
Eavns LR, Linker A. 1973. Production and characterization of the slime polysaccharide of Pseudomonas aeruginosa. J Bacteriol 16: 915-921.
Eftekhar F, Schiller NL. 1994. Partial purification and characterization of a mannuronan-specific AlgL from Pseudomonas aeruginosa. Curr Microbiol 29: 37-42.
Garron M, Cygler M. 2010. Structural and mechanistic classification of uronic acid containing polysaccharide lyases. Glycobiology 20: 1547-1573.
Hansen JB, Doubet RS, Ram J. 1984. Alginase enzyme production by Bacillus circulance. Appl Environ Microb 47: 704-713.
Hoshino T, Kageyama M. 1980. Purification and properties of a binding protein for branched-chain amino acids in Pseudomonas aeruginosa. J Bacteriol 141: 1055-1063.
Jain S, Ohman D. 2005. Role of an AlgL for Alginate transport in mucoid Pseudomonas aeruginosa. Infect Immun 73: 6429-6436.
Linhardt R, Galliher P, Cooney C. 1986. Polysaccharide lyases. Appl Biochem Biotechnol 12: 135-145.
Linker A, Evans LR. 1984. Isolation and characterization of an alginase from mucoid strains of Pseudomonas aeruginosa. J Bacteriol 159: 958-964.
May TB, Shinabarger D, Maharaj R, Kato J, Chu L, DeVault JD, Roychoudhury S, Zielinski NA, Berry A, Rothmel RK. 1991. Alginate synthesis by Pseudomonas aeruginosa: A key pathogenic factor in chronic pulmonary infection of cystic fibrosis patients. Clin Microbiol Rev 4:191-206.
Nguyen LK, Schiller NL. 1989. Identification of a slime exopolysaccharide depolymerase in mucoid strains of Pseudomonas aeruginosa. Curr Microbiol 18: 323-332.
Periss J, Ashwell G. 1962. Alginic acid metabolism in bacteria. J Biol Chem 237: 309-316.
Ramsey DM, Wozniak DJ. 2005. Understanding the control of Pseudomonas aeruginosa alginate synthesis and the prospects for management of chronic infection in cystic fibrosis. Mol Microbiol 56: 309-322.
Rehm B. 1998. Alginate lysase from Pseudomonas aeruginosa CF1/M1 prefers the hexameric oligomannuronate as substrate, FEMS. Microbiol Lett 165:175-180.
Rehm B, Valla S. 1997. Bacterial Alginate: Biosynthesis and applications. Appl Microbiol Biot 48: 281-289.
Schiller NL, Monday SR, Boyd CM, Keen NT, Ohman DE. 1993. Characterization of the Pseudomonas aeruginosa AlgL gene (algL): Cloning, sequencing, and expression in Escherichia coli. J Bacteriol 175: 4780-4789.
Sutherland IW, Keen GA. 1981. Alginases from Beneckea pelagia and Pseudomonas species. J Appl Biochem 3: 48-57.
Weissbach A, Hurwitz J. 1959. The formation of 2-keto-3 deoxyheptonic acid in extracts of Escherichia coli B. J Biol Chem 4: 705-714.
Wong TY, Preston LA, Schiller NL. 2000. AlgL: Review of major sources and enzyme characteristics, structure-function analysis, biological roles and applications. Annu Rev Microbiol 54: 289-340.
Xiao L, Han F, Yang Z, Lu X, Yu W. 2006. A novel AlgL with high activity on acetylated Alginate of Pseudomonas aeruginosa FRD1 from Pseudomonas sp. QD03. World J Microbiol Biotechnol 22: 81–89.
Yamasaki M, Moriwaki S, Miyake O, Hashimoto W, Murata K, Mikami B. 2004. Structure and function of hypothentical Pseudomonas aeruginosa protein PA1167 classified into family PL-7. J Biol Chem 279: 31863-31872.
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