Determination of Extracellular Hydrolytic Enzyme Production Capabilities of Acidithiobacillus Bacteria Isolated from Awasppi River in Sulaymaniyah, Iraq and 16S rDNA Analysis

Year 2018, Volume: 2 Issue: 1, 17 - 20, 30.06.2018

Kerem Özdemir , Ghalib Abdalla Sharef Metin Ertaş

https://doi.org/10.31594/commagene.392847

Cited By: 1

Abstract

In this study, the 16S rDNA gene regions’ phylogenetic positions based on the sequence analysis were determined by examining some hydrolytic extracellular enzyme activities of the Acidithiobacillus bacteria isolated from the water and mud samples obtained from the Awasspi River in Sulaymaniyah province, northern Iraq and from the Darzilais sulphur cave in Sangaw district. While the catalase activity of the isolates was positive; lipase, protease, xylanase, and amylase activities were found to be negative. Genomic DNAs of the isolates were isolated and 16S rDNA gene region was realized with PCR amplification. To determine the distance between the species, Maximum Likelihood algorithm was used to form a phylogenetic tree. It was observed that gene strands belonging to Acidithiobacillus species acquired from NCBI and 2 isolates we used in our study clustered with a strong homology and their genetic positions were revealed. 

Keywords

Acidithiobacillus, Sulaimani, Enzyme, 16S rDNA

Awa Sppi Nehri’nden (Süleymaniye-Irak) İzole Edilen Acidithiobacillus Cinsi Bakterilerin Ektraselüler Hidrolitik Enzim Üretme Kabiliyetlerinin Belirlenmesi ve 16S rDNA Analizi

Abstract

Bu araştırmada, Kuzey Irak'ın Süleymaniye şehrinde bulunan Awasppi nehrinden ve Sangaw'daki Darzilais sülfür
kaynaklı mağaradan alınan su ve çamur örneklerinden izole edilen Acidithiobacillus bakterilerinin bazı hidrolitik ektraselüler
enzim aktiviteleri incelenerek, 16S rDNA gen bölgelerinin sekans analizi temeline dayalı filogenetik pozisyonları belirlenmiştir.
İzolatların katalaz aktivitesi pozitif iken, lipaz, proteaz, ksilanaz ve amilaz aktivitesi negatif olarak belirlenmiştir.
İzolatların Genomik DNA’ları izole edilerek 16S rDNA gen bölgesi PZR amplifikasyonu ile gerçekleştirilmiştir. Türler
arasındaki uzaklığı belirlemek için Maximum Likelihood algoritması kullanılarak oluşturulan filogenetik ağaç NCBI’dan elde
edilmiş Acidithiobacillus türlerine ait gen dizileri ve çalışmamızda kullandığımız 2 izolat güçlü bir homoloji ile kümelendiği
gözlemlenerek genetik pozisyonları ortaya konmuştur. Karşılaştırmalı 16S rDNA gen dizisi analizi sonucunda, G1 izolatının
Acidithiobacillus thiooxidans’a % 99 benzerlik gösterdiği ve G2 izolatının Acidithiobacillus ferrooxidans’a % 99 benzerlik gösterdiği
belirlenmiştir

Keywords

Acidithiobacillus, Süleymaniye, Enzim, 16S rDNA

References

• Bellenberg, S., Barthen, R., Boretska, M., Zhang, R., Sand, W., & Vera, M. (2015). Manipulation of pyrite colonization and leaching by ironoxidizing Acidithiobacillus species. Applied and Environmental Microbiology, 99, 1435-1449.
• Busse, H.J., Denner, E.B.M., & Lubitz, W. (1996). Classification and identification of bacteria: Current approaches to an old problem. Overview of methods used in bacterial systematics. Journal of Biotechnology, 47, 3-38.
• Cavalier-Smith, T. (2002). The neomuran origin of archaebacteria, the negibacterial root of the universal tree and bacterial megaclassification. International Journal of Systematic and Evolutionary Microbiology, 52, 7-76
• Gonzalez-Toril, E., Llobet-Brossa, E., Casamayor, E.O., Amann, R., & Amils, R. (2003). Microbial Ecology of An Extreme Acidic Environment, The Tinto River. Applied and Environmental Microbiology, 69(8), 4853-4865.
• Hedrich, S., & Johnson, D.B. (2013). Acidithiobacillus ferridurans, sp. nov.; an Acidophilic Iron, Sulfur and Hydrogen-metabolizing Chemolithotrophic Gammaproteobacterium. International Journal of Systematic and Evolutionary Microbiology, 63, 4018-4025.
• Holt, J.G., Krieg, N.R., Sneath, P.H.A., Staley, J.T., & Williams, S.T. (1994). Bergey’s Manual of Determinative Bacteriology, 9th ed. Baltimore: Williams & Wilkins.
• Janeta Starosvetsky A., Udy Zukerman B., Robert H., & Armon A. (2012). A simple medium modification for isolation, growth and enumeration of Acidithiobacillus thiooxidans (syn. Thiobacillus thiooxidans) from water samples. Journal of Microbiological Methods, 92, 178–182.
• Kamimura, K., Higashino, E., Moriya, S., & Sugio, T. (2003). Marine acidophilic sulfur- oxidizing bacterium requiring salts for the oxidation of reduced inorganic sulfur compounds. Extremophiles, 95- 99.
• Karavaiko, G.I., Turova, T.P., Kondrateva, T.F., Lysenko, A.M., Kolganova, T.V., Ageeva, S. N., Muntyan, L.N., & Pivovarova, T.A. (2003). Phylogenetic heterogeneity of the species Acidithiobacillus ferrooxidians. International Journal of Systematic and Evolutionary Microbiology, 53, 113–119.
• Khan, S., Haq, F., Hasan, F., Saeed, K., & Ullah, R. (2012). Growth and biochemical activities of Acidithiobacillus thiooxidans collected from black shale. Journal of Microbiology Research, 2(4), 78-83.
• Lane, D.J., Stahl, D.A., Olsen, G.J., Heller, D.J., & Pace, N.R. (1985). Phylogenetic analysis of the genera Thiobacillus and Thiomicrospira by 5S rRNA sequences. Journal Bacteriology, 163, 75–81.
• Rawlings, D. E. (1997). Biomining: Theory, Microbes and Industrial Processes, Berlin: Springer-Verlag.
• Rawlings, D.E. (2005). Characteristics and adaptability of iron and sulfur‐ oxidizing microorganisms used for the recovery of metals from minerals and their concentrates. Microbial Cell Factories, 4, 13.
• Starosvetsky, J., Zukerman, U., & Armon, R.H. (2013). A simple medium modification for isolation, growth and enumeration of Acidithiobacillus thiooxidans (syn. Thiobacillus thiooxidans) from water samples. Journal of Microbiological Methods, 92(2), 178-182.
• Valenzuela L., Chi A., Beard S., Orell A., Guiliani N., & Shabanowitz J. (2006). Genomics, metagenomics and proteomics in biomining microorganisms. Biotechnology Advances, 24, 197–211.
• Van den Burg, B. (2003). Extromophiles as a source for novel enzyme. Current Opinion in Microbiology, 6, 213–218.
• Vandamme, P., Pot, B., Gillis, M., de Vos, P., Kersters, K., & Swings, J. (1996). Polyphasic taxonomy, a consensus approach to bacterial systematics. Microbiological Reviews, 60(2), 407-438.