Identification of alkane hydroxylase gene in aliphathic-degrading bacteria isolated from waste oil

Subject Areas : Applied Microbiology

Hamid  Tebyanian ¹ (Department of Microbiology, Science and Research Branch, Islamic Azad University, Kerman, Iran)
Mehdi  Hassanshahian ² (Department of Biology, Faculty of Sciences, Shahid Bahonar University of Kerman, Kerman, Iran)
Ashraf  Karimi nik ³ (Department of Microbiology, Science and Research Branch, Islamic Azad university, Kerman, Iran)

Received: 2012-05-24 Accepted : 2012-08-25 Published : 2013-01-01

Keywords: PCR, Biodegradation, Alkane hydrolase,

Abstract :

Background and Objectives: Today hydrocarbon pollution is the most important environmental problem. Biological methods can play an important role in removing these contaminants to the environment. This study was aimed to isolate aliphatic compounds degrading bacteria and identify the best degrading strain. Materials and Methods: Sampling of waste oil depot took in the city of Kerman, Tehran and hydrocarbon-contaminated soils. Alkane degrading bacteria were isolated by using enrichment Bushnel-Hass medium containing hexadecane (as source carbon). In order to identify superior strains, a part of 16SrDNA gene was amplified by PCR and then sequenced. Also, the presence of alkane’s hydroxylase gene in these strains was confirmed by using specific primers. Results: A total of fifteen alakne degrading bacteria were isolated which 8 strains were selected as superior strains. These strains belonged to the genus of Rhodococcus jostii,Stenotrophomonas maltophilia strain M2, Achromobacter piechaudii, Tsukamurell atyrosinosolvens, Pseudomonas fluorescens, Rhodococcus erythropolis,Stenotrophomonas maltophilia strain Q1, Pseudomonas aeruginosa. In this study, the highest growth rate of strians was in concentrations 2.5 percent of hexadecane and the lowest was found in 7 percent. Also, all of the strains have alkane hydroxylase gene. Conclusion: Our results indicated that threre is a high diversity of degradative bacteria in Iran ecosystem and their ability to degrade petroleum wastewater. So, with a proper management of these bacteria can be used to minimize pollution caused by waste oil industries.

References:

1. Fan CY, Krishnamurthy S. Enzymes for enhancing bioremediation of petroleum-contaminated soils: a brief review. J Air Waste Manage Assoc. 1995; 45(6): 453-460.

2. Hua J. Biodegradation of dispersed marine fuel oil in sediment under engineered pre-spill application strategy. Ocean Engineering. 2006; 33(1): 52-67.

3. Gholizadeh H, Mohebatzadeh H. Unique to the study of the effects of gasoline kerosene and gasoline on the growth of several bacteria. The second of the National Congress of Biotechnology, Karaj, Iran. 2001: 78370.

4. Suyama T, Hiroyuki H, Yutaka T. Bacterial isolates degrading aliphatic polycarbonates. FEMS Microbiol Letters. 1998; 161(2): 255-261.

5. Sharma S, Pant A. Biodegradation and conversion of alkanes and crude oil by amarine Rhodococcus sp. Biodegradation. 2000; 11(5): 289-294.

6. Abbasi M, Abduli M, Nasrabadi T, Hoveidi H, Razmkhah N. Solid waste management in tabriz petrochemical complex. J Environ Health Sci Eng. 2006; 3(1): 185-192.

7. Carpenter D, Genovese C. The generation use and disposal of waste crankcase oil in developing countries. Uganda J. 2009; 161(6): 835-841.

8. Muslimsphy V, Sadatnouhi A. The role of bacteria isolated from oil contaminated soil in cleaning the environment. J Biol Sci Res. 2007; 33(2): 154-163.

9. Hassanshahian M, Emtiazi G, Cappello S. Isolation and characterization of crude-oil-degrading bacteria from the Persian Gulf and the Caspian Sea. Marine Pollution Bull. 2012; 64(14): 7-12.

10. Cappello S, Caruso G, Zampino D  Monticelli L, Maimone G, Denaro R, Tripodo B, Troussellier M, Yakimov M, Giuliano L. Microbial community dynamics during assays of harbour oil spill bioremediation, a microscale simulation study. J Appl Microbiol. 2007; 102(1): 184-194.

11. Cappello S, Denaro R, Genovese M, Giuliano L, Yakimov M. Predominant growth of  Alcanivorax during experiments on ‘‘oil spill bioremediation’’ in mesocosms. Microbiol Res. 2007; 162(99): 185-190.

12. Subarna R, Dipak H, Debabrata B, Dipa B, Ranajit K. Survey of petroleum degrading bacteria in coastal waters of Sunderban biosphere reserve. World J Microbiol Biotechnol. 2002; 18(1): 575-581.

13. Pruthi V, Cameotra SS. Rapid identification of biosurfactant-producing bacterial strains using a cell surface hydrophobicity technique. Biotechnol Tech. 1997; 11(3): 671-674.

14. Batista  SB, Mounteer A, Amorim FR, Totola MR. Isolation and characterization of biosurfactant/bioemulsifier-producing bacteria from petroleum contaminated sites. Bioresource Tech. 2006; 97(1): 868-875.

15. Kohno T, Sugimoto Y, Sei K, Mori K. Design of  PCR primers and gene probes for general detection alkane-degrading bacteria. Microbiol Environ. 2002; 17(3): 114-212.

16. Huaa Z, Song R, Dua G, Li H, Chen J. Effects of EDTA and Tween 60 on biodegradation of n-hexadecane with two strains of Pseudomonas aeruginosa. Biochem Eng J. 2007; 36(3): 66-71.

17. Song X, Xu Y, Li G, Zhang Y, Huang T, Hu Z. Isolation, characterization of Rhodococcus sp. P14 capable of degrading high-molecular-weight polycyclic aromatic hydrocarbons and aliphatic hydrocarbons. Marine Pollution Bull. 2011; 62(10): 2122-2128.

18. Das K, Mukherjee AK. Crude petroleum-oil biodegradation efficiency of Bacillus subtilis and Pseudomonas aeruginosa strains isolated from a petroleum-oil contaminated soil from North-East India: Bioresource Tech. 2007; 98(7): 1339-1345.

19. Hassanshahian M, Emtiazi G. Investigation of alkane biodegradation using the microtiter plate method and correlation between biofilm formation, biosurfactant production and crude oil biodegradation. Int Biodeterioration Biodegradation. 2008; 62(1): 170-178.

20. Van Beilen J, Li Z, Duetz W, Smits T,Witholt B. Diversity of alkane hydroxylase systems in the environment. Oil Gas Sci Tech. 2003; 58(4): 427-440.