Protease and urease production during utilization of diesel by fluorescent Pseudomonas species isolated from local soil
Abstract
Background and objectives: Bacteria, most prevalently the Pseudomonas species possess high capacity to utilize and de- grade petroleum hydrocarbons and are classified as the hydrocarbonoclastic microorganisms. Many species of the genus Pseudomonas are notorious for their aerobic degradation capacity, extracellular enzyme production and are metabolically versatile organisms capable of utilizing a wide range of hydrocarbons and other compounds. In this study, the ability of diesel utilization by some locally isolated Pseudomonas species was tested.
Materials and Methods: From a local red laterite soil, four different Pseudomonas species were isolated on King’s B me- dium, characterized, identified and tested their potential in utilizing diesel, a petroleum hydrocarbon. At the same time, pro- duction of protease and urease enzymes during the utilization of diesel was also assayed following the standard procedures.
Results: The isolates were grown well on diesel and subsequently produced the extracellular enzymes protease and urease at significant levels when compared to their production in the absence of diesel. Optimum temperature and pH for increased growth by four isolates was found to be 37oC and pH 8.0 indicating the maximum utilization of diesel. All the isolates showed maximum growth in medium with 100% diesel than 100% glycerol as carbon source, when tested with different proportions of diesel and glycerol as carbon sources. Plasmid profile of the isolates revealed that, all four Pseudomonas isolates harbored two low molecular weight plasmids; one with 3 Kb size and the other with 10 kb to 12 Kb size.
Conclusion: The four Pseudomonas isolates of the present study were found to have potential in diesel degradation and can be recommended for bioremediation of sites that are contaminated with diesel.
Leahy JG, Colwell RR. Microbial degradation of hy- drocarbons in the environment. Microbiol Mol Biol Rev 1990; 54: 305-315.
Holloway BW, Escuadra MD, Morgan AF, Saffery R, Krishnapillai V. The new approaches to whole ge- nome analysis of bacteria. FEMS Microbiol Lett 1992; 100:101-106.
Wongsa P, Tanaka M, Ucno A, Hasanuzzaman M, Yumoto I, Okuyama H. Isolation and characterization of novel strains of Pseudomonas aeruginosa and Ser- ratia marcescens possessing high efficiency to degrade gasoline, kerosene, diesel oil and lubricating oil. Curr Microbiol 2004;49: 415-422.
Song H, Wang X, Bartha R. Bioremediation Potential of Terrestrial Fuel Spills. Appl Environ Microbiol 1990;56: 652-656.
Raaijmakers JM, Sluis L, Bakker PAHM, Schippers B, Koster M, Weisbeek PJ. Utilization of heterolo- gous siderophores and rhizosphere competence of fluorescent Pseudomonas spp. Can J Microbiol 1995;41:126-135.
Godfrey T, Reichelt J (1983). Industrial enzymology.Nature press, New York. pp 1-4.
Moses V, Cape RE (1991). Biotechnology, the science and business UK. Harwood Academic publishers. pp 322-326.
King EO, Ward MK, Raney DE. Two simple media for the demonstration of pyocyanin and fluorescein. J Lab Clin Med 1954; 44:301-307.
Nannipieri S, Cerevelli, Matarese E. Analysis of urease, protease, catalase from soil. Soil Sci Soc Am J 1980;44:1021-1028.tion in crude oil-containing medium. World J Microbiol Biotechnol 2006; 22:1-8.
Caballero AR, Morean JM, Engel LS, Marquart ME, Hill JM, Callaghan RJ. Pseudomonas aeruginosa pro- tease IV enzyme assays and comparison to other pseu- domonal proteases. Analytical Biochem 2001; 290:330-337.
Himelbloom BH, Hassan HM. Effects of Cysteine on Growth, Protease Production, and Catalase Activity of Pseudomonas fluorescens. Appl Environ Microbiol 1986; 51: 418-421.
Ishikawa H, Ishimi K, Sugiura M, Sowa A, Fujiwara N. Kinetics and mechanism of enzymatic hydrolysis of gelatin layers of X-ray film and release of silver parti- cles. J Ferment Bioeng 1993; 76: 300-305.
Yagci A, Tuc Y, Soyletir G. Elastase and alkaline pro- tease production by Pseudomonas aeruginosa strains: comparison of two procedures. New Microbiol 2002;25:223-229.
Jensen SE, Fecycz IT, Campbell JN. Nutritional factors controlling exocellular protease production by Pseu- domonas aeruginosa. J Bacteriol 1980; 144: 844-847.
Hastie AT, Hingley ST, Kueppers F, Higgins ML, Tan- nenbaum CS, Weinbaum G. Protease Production by Pseudomonas aeruginosa Isolates from Patients with Cystic Fibrosis. Infect Immun 1983; 40: 506-513.
Dutta JR, Banerjee R. Isolation and characterization of a newly isolated Pseudomonas mutant for protease pro- duction. Braz Arch of Biol Technol 2006; 49:37- 47.
Margesin R, Schinner F. Bioremediation of diesel-oil- contaminated alpine soil at low temperatures. Appl Microbiol Biotech 1997; 47: 462-468.
Margesin R, Schinner F, Zimmerbauer A. Monitoring of bioremediation by soil biological activities. Chemos- phere 2000; 40: 339.
Vetrova AA, Nechaeva IA, Ignatova AA, Puntus IF, Arinbasarov MU, Filonov AE, et al. Effect of catabolic plasmids on physiological parameters and efficiency of oil destruction by Pseudomonas bacteria. Mikrobiologiia 2007; 76: 354 -360.
Tabatabai MA, Bremner J. Assay of urease, catalase in soil. Soil Biol Biochem 1972; 4:479-482.
Holt JG, Krieg NR, Sneath PHA, Stanley JT, William ST (1994). Bergey’s Manual of Determinative Bacteri- ology. Baltimore, USA: William and Wilkins.
Gould WD, Hagedorn C, Bardinelli TR, Zablotowicz RM. New selective media for enumeration and recov- ery of fluorescent pseudomonads from various habitats. Appl Environ Microbiol 1985; 49: 28-32.
ModiVV, Patel RN. Salicylate formation from Naph- thalene by Pseudomonas aeruginosa. Appl Microbiol 1968; 16: 172-173.
Tang X, Zhu Y, Meng Q. Enhanced crude oil biodegrad- ability of Pseudomonas aeruginosa ZJU after preserva- tion in crude oil-containing medium. World J Microbiol Biotechnol 2006; 22:1-8.
Caballero AR, Morean JM, Engel LS, Marquart ME, Hill JM, Callaghan RJ. Pseudomonas aeruginosa pro- tease IV enzyme assays and comparison to other pseu- domonal proteases. Analytical Biochem 2001; 290:330-337.
Himelbloom BH, Hassan HM. Effects of Cysteine on Growth, Protease Production, and Catalase Activity of Pseudomonas fluorescens. Appl Environ Microbiol 1986; 51: 418-421.
Ishikawa H, Ishimi K, Sugiura M, Sowa A, Fujiwara N. Kinetics and mechanism of enzymatic hydrolysis of gelatin layers of X-ray film and release of silver parti- cles. J Ferment Bioeng 1993; 76: 300-305.
Yagci A, Tuc Y, Soyletir G. Elastase and alkaline pro- tease production by Pseudomonas aeruginosa strains: comparison of two procedures. New Microbiol 2002;25:223-229.
Jensen SE, Fecycz IT, Campbell JN. Nutritional factors controlling exocellular protease production by Pseu- domonas aeruginosa. J Bacteriol 1980; 144: 844-847.
Hastie AT, Hingley ST, Kueppers F, Higgins ML, Tan- nenbaum CS, Weinbaum G. Protease Production by Pseudomonas aeruginosa Isolates from Patients with Cystic Fibrosis. Infect Immun 1983; 40: 506-513.
Dutta JR, Banerjee R. Isolation and characterization of a newly isolated Pseudomonas mutant for protease pro- duction. Braz Arch of Biol Technol 2006; 49:37- 47.
Margesin R, Schinner F. Bioremediation of diesel-oil- contaminated alpine soil at low temperatures. Appl Microbiol Biotech 1997; 47: 462-468.
Margesin R, Schinner F, Zimmerbauer A. Monitoring of bioremediation by soil biological activities. Chemos- phere 2000; 40: 339.
Vetrova AA, Nechaeva IA, Ignatova AA, Puntus IF, Arinbasarov MU, Filonov AE, et al. Effect of catabolic plasmids on physiological parameters and efficiency of oil destruction by Pseudomonas bacteria. Mikrobiologiia 2007; 76: 354 -360.
Izmalkova TY, Sazanova OI, Sokolov SL, Kosheleva IA, Boronin AM. The p-7 Incompatibility Group plas- mids responsible for biodegradation of naphthalene and salicylate in fluorescent pseudomonads. Microbiol 2005;74: 290-295.
Park W, Jeon CO, Hohnstock-Ashe AM, Winans SC, Zylstra GJ, Madsen EL. Identification and characteriza- tion of the conjugal transfer region of the pCgl plasmid from naphthalene-degrading Pseudomonas putida Cgl. Appl Environ Microbiol 2003; 69: 3263-3271.
Thavasi R, Jayalakshmi S, Radhakrishnan R, Balasub- ramanian T. Plasmid incidence in four species of hydro- carbonoclastic bacteria isolated from oil polluted marine environment. Biotechnol 2007; 6: 349-352.
Deshpande NM, Dhakephalkar PK, Kanekar P.Plasmid mediated dimethoate degradation in Pseu- domonas aeruginosa MCMB-427. Lett Appl Micro- biol 2001; 33: 275 - 279.
Devereux R, Sizemore RK. Plasmid incidence in ma- rine bacteria isolated from petroleum polluted sites on different petroleum hydrocarbons. Mar Poll Bull 1982; 13: 198-202.
Files | ||
Issue | Vol 1 No 3 (2009) | |
Section | Articles | |
Keywords | ||
Fluorescent Pseudomonas diesel plasmid protease urease |
Rights and permissions | |
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |