ISOLATION, CHARACTERIZATION AND DETERMINATION OF PETROLEUM HYDROCARBON DEGRADING METABOLIC CAPABILITIES OF INDIGENOUS MICROORGANISMS IN TURKEY

Öz

The aim of this study, to do develop a method and the material is utilized to eliminate organic pollutants (bioremediation) with oil pollution problems in our country. To do this, 33 bacterial strains isolated from oil contaminated soil obtained from Adana, Batman and Adıyaman in bioremediation studies. Identification of isolated bacteria were performed from the taxonomic classification of MIS (Microbial Identification System), oxidase and 3% KOH tests. All isolates were performed their proliferation capacity in crude oil contaminated environment, and high capability of crude oil degradation in liquid culture medium. 6 bacteria (Pseudomons aeruginosa (2 strain), Pseudomonas putida (2 strain), Citrobacter amalonaticus, Acinetobacter genomospecies were identified according to their improvement capacity in crude oil contaminated environment and high capability of crude oil degradation. These bacterial strains provided that the crude oil degradation by 56% at the end of the 120-day incubation period. This study, can be applied is an important step in terms of getting the bioremediation product in polluted areas in our country

Anahtar Kelimeler

• Soil, Crude oil, Soil pollution, Bacteria, Bioremediation

TÜRKİYE’DE PETROL HİDROKARBONLARINI PARÇALAYAN YERLİ BAKTERİLERİN İZOLASYONU, KARAKTERİZASYONU VE HİDROKARBON PARÇALAMA KABİLİYETLERİNİN BELİRLENMESİ

Öz

Bu çalışmada amaç ülkemizde kirlilik sorunlarından olan petrol ve benzeri organik kirleticilerin ortadan kaldırılmasında faydalanılabilecek bir yöntem (biyoremidasyon) ve materyal geliştirmektir. Bunun için Adana, Batman ve Adıyaman illerinin petrol ile kirlenmiş topraklarından 33 adet bakteri izole edilmiştir. İzole edilen bakterilerin ön tanımlaması, taksonomik sınıflaması, Mikrobiyal Tanılama Sistemi (MIS), oksidaz ve % 3 KOH testleri yapılmıştır. Sıvı kültürde ham petrollü ortamda çoğalma yetenekleri ve petrolü parçalama kabiliyetleri test edilmiştir. Ham petrollü ortamda en iyi gelişmeyi gösteren ve ham petrolü parçalama düzeyleri en yüksek olan 6 bakteri suşu Pseudomons aeruginosa (2 suş) Pseudomonas putida (2 suş), Citrobacter amalonaticus, Acinetobacter genomospecies seçilmiştir. 120 günlük inkübasyon sürecinin sonunda bu 6 bakteri suşu toprakta % 56 oranında petrol ayrışması sağlamıştır. Bu çalışma, ülkemizde kirli alanlarda uygulanabilecek bir biyoremidasyon ürünü elde etme açısından önemli bir basamak oluşturmaktadır

Anahtar Kelimeler

• Toprak, Ham petrol, Toprak kirliliği, Bakteriler, Biyoremidasyon

Kaynakça

1. Alexander M. (1999): “Biodegradation and Bioremediation”, (2nd edition) Academic Press, San Diego.
2. Alvarez H. M. (2003): “Relationship between B-oxidaiton Pathway and the Hydrocarbon- Degrading Profile in Actinomycetes Bacteria”, International Biodeterioration and Biodegradation, Sayı 52, s.35-42.
3. Arthi K., Appalaraju B., Parvathi S. (2003) : “Vancomycin Sensitivity and KOH String Test as An Alternative to Gram Staining of Bacteria”, Int. J. Med. Microbiol., Sayı 21, s.121- 123.
4. Atlas R. M. (1978): “Microorganisms and Petroleum Pollutants”, BioScience, Sayı 28, s.387- 391.
5. Atlas R. M. (1981): “Microbial Degradation of Petroleum Hydrocarbons”, An Environmental Perspective Microbial Rev., Sayı 45, s.180:209.
6. Atlas R. M., Bartha R. (1992): “Hydrocarbon Biodegradation and Oil Spill Bioremediation”, Adv. Microb. Ecol., Sayı 12, s.287-338.
7. Bonnier P. D., Akoun G. L., Cadron E. C., Edwards E. D., Hockness W. (1980): “ A Technique for the Disposal of Oily Refinery Wastes”, Rapor No. 3/10, The Hague: Concawe.
8. Boonchan S., Britz M. L., Stanley G. A. (2000): “Degradation and Mineralization of High- Molekular-Weight Polycyclic Aromatic Hydrocarbons by Defined Fungal-Bacterial Cocultures”, Applied and Environmental Microbiology, Cilt 66; No. 3; s.1007-1019.

9. Butler C. S., Mason J. R. (1997): “Structure–function Analysis of the Bacterial Aromatic Ring–Hydroxylating Dioxygenases”, Adv. Microb. Physiol., Sayı 38, s.47-84.
10. Cerniglia C. E. (1992): “Biodegradation of Polycyclic Aromatic Hydrocarbons”, Biodegradation, Sayı 3, s.351-368.
11. Cooney J. J. (1980): “Microorganisms Capable of Degrading Refractory Hydrocarbons in Ohio Waters”, Ohio Water Resources Center, Columbus, OH for the Office of Water Research and Technology, Washington, D.C., Rapor https://kb.osu.edu/dspace/bitstream/1811/36346/1/OH_WRC_493X.pdf 21.05.2013. No. 493X, Erişim Tarihi:
12. Demir İ., Demirbağ Z. (1999): “Polisiklik Aromatik Hidrokarbonların Biyolojik Olarak Parçalanması”, Tr. J. of Biology, Sayı 23, s.293-302.
13. Desai J. D., Banat J. M. (1997): “Microbial Production of Surfactants and their Commercial Potential”, Microbiol. Mol. Rev., Sayı 61, s.47-64.
14. El-Nawawy A. S., Ghobrial F., Elimam A. (1987): “Feasibility Study of Disposal of Oily Sludge in Kuwait”, Proceeding of the 2. International Conference on New Frontiers for Hazardous Waste Management, Pittsburgh, 27-30 September, s.162-8.
15. Erdoğan E. E. (2010): “Petrol ile Kirlenmiş Toprakların Biyolojik Olarak İyileştirilmesinin Laboratuvar Koşullarında Denenmesi”, Ankara Üniversitesi, Fen Bilimleri Enstitüsü, Toprak Anabilim Dalı, Doktora Tezi.
16. Erdoğan E. E., Şahin F., Karaca A. (2011): “Determination of Petroleum-Degrading Bacteria Isolated from Crude Oil-Contaminated Soil in Turkey”, African Journal of Biotechnology, Cilt 11, No. 21, s. 4853-4859, http://www.academicjournals.org/AJB Erişim Tarihi: 13 Mart 2012.
17. Fall R. R, Brown J. L., Schaeffer T. L. (1979): “Enzyme Recruitment Allows the Biodegradation of Recalcitrant–Branched Hydrocarbons by Pseudomonas Citronellolis”, Appl. Environ. Microbiol., Sayı 38, s.715-722.
18. Hagwell I. S., Delfino L. M., Rao J. J. (1992): “Partitioning of Polycyclic Aromatic Hydrocarbons from oil into water”, Environ. Sci. Technol., Sayı 26, s.2104-2110.
19. Hyina A., Castillo S. M. I., Villarreal S. J. A., Ramirez E. G., Candelas R. J. (2003): “Isolation of Soil Bacteria for Bioremediation of Hydrocarbon Contamination”, Cilt 44, No:1, s.88-91.
20. Holt J. G., Kreig N. R., Sneath P. H. A., Stanely J. T., Williams S. T. (1994): “In: Bergey’s Manual of Systematic Bacteriology”, (Ed: J. G. Holt, N.R. Krieg) Cilt 1, s.141-199.
21. Jack T. R., Lee E., Mueller J. (1985): “Anaerobic Gas Production from Crude Oil”, Int. Bioresources J., 1, s.167-185.
22. Johnson K., Anderson S., Jacobson C. S. (1996): “Phenotypic and Genotypic Characterization of Phenanthrene-Degrading Fluorescent Pseudomonas Biovars”, Appl. Environ. Microbiol., Sayı 62, s.3818-3825.
23. Jùrgensen K. S., Puustinen J. A., Suortti M. (2000): “Bioremediation of Petroleum Hydrocarbon-Contaminated Soil by Composting in Biopiles”, Environmental Pollution, Sayı 107, s.245 - 254.
24. Kapley A., Purohit H. J., Chhatre S., Shanker R., Chakrabati T., Khanna P. (1999): “Osmotolerance and Hydrocarbon Degradation by a Genetically Engineered Microbial Consortium”, Biosource Technology, Sayı 67, s.241-245.
25. Keith L. H., Telliard W. A. (1979): “Priority Pollutants 1-A Perspective View”, Environ. Sci. Technol., Sayı 13, s.416-423.
26. Kim K. H., Ten L. N., Liu Q. M., Im W. T., Lee S .T. (2006): “Sphingobacterium Daejeonense sp. nov., Isolated from a Compost Sample”, Int. J. Syst. Evol. Microbiol., Sayı 56, s.2031–2036.
27. Kishore D., Ashis M. K. (2007): “Crude Petroleum-Oil Biodegradation Effciency of Bacillus Subtilis and Pseudomonas Aeruginosa Strains Isolated from a Petroleum-Oil Contaminated Soil from North-East India”, Bioresource Technology, Sayı 98, s.1339- 1345.
28. Kiyohara H., Takizawa N., Nagao K. (1992): “Natural Distribution of Bacteria Metabolizing Many Kinds of Polyaromatic Hydrocarbons”, J. Ferment. Bioeng., Sayı 74, s.49–51.
29. Klement Z., Rudolph K., Sands D. C. (1990): “Methods in Phytobacteriology”, Akademia Kiado, Budapest.
30. Komukai-Nakamura S., Sugiura K., Yamauchi-Inomata Y., Toki H., Venkateswaran K., Yamamoto S., Tanaka H., Harayama S. (1996): “Construction of Bacterial Consortia that Degrade Arabian Light Crude Oil”, Journal of Fermentation and Bioengineering, Sayı 82, s.570–574.
31. Leahy J. G., Tracy K. D., Eley M. H. (2003): “Degradation of Mixturesof Aromatic and Chloroaliphatic Hydrocarbons by Aromatic Hydrocarbon-Degrading Bacteria”, FEMS Microbiology Ecology, Sayı 43, s.271-276.
32. Matsumoto M., Furuya N., Matsumaya N. (1997): “Characterization of Rhizoctonia Spp., Causal Agents of Shealth Diseases of Rice Plant, by Total Cellular Fatty Acid Analysis”, Ann. Phytopath. Soc. Japon, Sayı 63, s.149-154.
33. Miller I., Berger T. (1985): “Bacteria Identification by Gas Chromatography of Whole Cell Fatty Acids”, Hewlett-Packard Gas Chromatography Application Note, Hewlett-Packard Co., Alto, CA., s.228-238.
34. Odu C. T. I. (1977): “Pollution and the Environment”, Bull. Sci. Assoc. Nig., Cilt 3, No. 2, s.284 -285.
35. Ojumu T. V., Bello O. O., Sonibare J. A., Solomon B. O. (2005): “Evaluation of Microbial Systems for Bioremediation of Petroleum Refinery Effluents in Nigeria”, African Journal of Biotechnology, Cilt 4, No. 1, s.31-35.
36. Olivera N. L., Esteves J. L., Commendatore M. G. (1997): “Alkane Biodegradation by A Microbial Community from Contaminated Sediments in Patagonia, Argentina”, İnt. Biodeterioration and Biodegradation, Cilt 40, No. 1, s.75-79.
37. Ökmen K. G., Algur Ö. F. (2000): “Farklı Karbon Kaynaklarının ve C/N Oranlarının Mikrobiyal Denitrifikasyon Üzerine Etkileri”, Turk J. Biol., Sayı 24, s.533-542.
38. Paisley R. (1995): “MIS Whole Cell Fatty Acid Analysis by Gas Chromatography”, MIDI, Inc., Newark, DE, 5.
39. Peressuttia S. R., Alvarez H. M., Oscar H. P. (2003): “Dynamics Ofhydrocarbon-Degrading Bacteriocenosis Ofan Experimental Oil Pollution in Patagonian Soil”, International Biodeterioration and Biodegradation, Sayı 52, s.21-30.
40. Pokethitiyook P., Sungpetch A., Upathame S., Kruatrachue M. (2002): “Enhancement of Acinetobacter Calcoaceticus in Biodegradation of Tapis Crude Oil”, 17th WCSS, Symposium, No. 42, Thailand.
41. Porta A., Trovato A., McCarty K., Uhler A., Andreotti G. (1998): “Degredation of Saturated and Polycyclic Aromatic Hydrocarbons and Formation of their Metabolites in Bioremediated Crude Oil Containing Sois”, (Ed: B. C. Alleman, A. Leeson), Cilt 1, s.505-510.
42. Rahman K. S. M., Thahina J., Rahman Y., Lakshmanaperumalsamy P., Banat I. M. (2002a): “Towards Efficient Crude Oil Degradation by A Mixed Bacterial Consortium”, Bioresource Technology, Sayı 85, s.257-261.
43. Rahman K. S. M, Banat I. M., Thahira J., Thayumanavan T., Lakshmanaperumalsamy P. (2002b): “Bioremediaton of Gasoline Contaminated Soil by a Bacterial Consortium Amended with Poultry Litter, Coir Pith and Rhamnolipid Biosurfactant”, Bioresource Technology, Sayı 81, s.25-32.
44. Rhykerd R. L., Crews B., McInnes K. J., Weaver R. W. (1998): “Impact of Bulking Agents, Forced Aeration, and Tillage on Remediation of Oil-Contaminated Soil”, Bioresource Technology, Sayı 67, s.279-285.
45. Rojas-Avelizapa N. G., Rodriguez-Vazquez R., Enriquez-Villanueva F., Martinez-Cruz J., Poggi-Varaldo H. M. (1999): “Transformer Oil Degradation by An Indigenous Microflora Isolated from A Contaminated Soil”, Resources, Conservation and Recycling, Sayı 27, s.15-26.
46. Sextone A. J., Everett K., Jenkins T., Atlas R. (1978): “Fate of Crude and Refined Oils in North Slope Soils”, Arctic, Sayı 31, s.339-347.
47. Shivaji S., Ray M. K., Rao N. S., Saisree L., Jagannadham M. V., Kumar G. S., Reddy G. S. N., Bhargava P. M. (1992): “Sphingobacterium Antarcticus Sp. Nov., A Psychrotrophic Bacterium from the Soils of Schirmacher Oasis, Antarctica”, Int. J. Syst. Bacteriol., sayı 42, s.102–106.
48. Song H .G., Bartha R. (1990): “Effects of Jet Fuel on The Microbial Community of Soil”, Appl. Environ. Microbiol., Sayı 56, s.646-651.
49. Steffan R., Mccloy J. K., Vainberg S., Condee C. W., Zhang D. (1997): “Biodegradation of the Gasoline Oxygenates Methyle Tert-Butyl Ether, Ethyl Tert- Butyl Ether and tert- Amyl Methyl Ether by Propane-Oxidizing Bacteria”, Appl. Environ. Microbiol., Cilt 63, No. 11, s.4216-4222.
50. Suslow T. V., Schroth M. N., Isaka M. (1982): “Application of A Rapid Method For Gram Differentiation of Plant Pathogenic and Saprophytic Bacteria Without Staining”, Phytopathology, Sayı 72, s.917-918.
51. Şahin F. (1997): “Detection, Identification and Characterization of Strains of Xanthomonas Campestris pv. Vesicatoria by Traditional and Molecular Methods, and Resistance in Capsicum Species to Xanthomonas Campestris pv. Vesicatoria Pepper Race6.”, The Ohio State University, Doktora Tezi.
52. Şahin F., Abbasi P. A., Lewis I. M. L., Zhang J., Miller S. A. (2003a): “Diversity Among Strains of Xanthomonas Campestris pv. Vitians From Lettuce”, Phytopathology, Sayı 93, s.64-70.
53. Şahin F., Kotan R., Abbasi P. A., Miller S.A . (2003b): “Phenotypic and Genotypic Characterization of Xanthomonas Campestris pv. Zinniae Strains”, European J. Plant Pathol., Sayı 109, s.165-172.
54. Takeuchi M., Yokota A. (1992): “Proposals of Sphingobacterium Faecium Sp. Nov., Sphingobacterium Piscium Sp. Nov., Sphingobacterium Heparinum Comb. Nov., Sphingobacterium Thalpophilum Comb. Nov. and Two Genospecies of the Genus Sphingobacterium, and Synonymy of Flavobacterium Yabuuchiae and Sphingobacterium spiritivorum”, J. Gen. Appl. Microbiol., Sayı 38, s.465–482.
55. Walter M. V., Nelson E. C., Firmstone G., Martin D. G., Clayton M. J., Simpson S., Spaulding S. (1997): “Surfactant Enhances Biodegradation of Hydrocarbons: Microcosm and Field Study”, Journal of Soil Contamination, Cilt 6, No. 1, s.61-77.
56. Wilson C. S., Jones K. C. (1993): “Bioremediation of Contaminated With Polynuclear Aromatic Hydrocarbons (PAHs): A Review”, Environmental Pollution, Sayı 81, s.229- 249.
57. Van Hamme J. D., Singh A., Ward O. P. (2003): “Recent Advances in Petroleum Microbiology”, Microbiol. Mol. Biol. Rev., Cilt 67, No. 4, s.503-549.
58. Yabuuchi E., Kaneko T., Yano I., Moss C. W., Miyoshi N. (1983): “Sphingobacterium Gen. Nov., Sphingobacterium Spiritivorum Comb. Nov., Sphingobacterium Multivorum Comb. Nov., Sphingobacterium Mizutae Sp. Nov., and Flavobacterium Indologenes Sp. Nov.: Glucosenonfermenting Gram-Negative Rods in CDC Groups IIK-2 and IIb”, Int J Syst Bacteriol, Sayı 33, s.580–598.