(ed.), Thermophilic bacteria. CRC Press, Boca Raton, Fla. pp.105–184,1992.
The research paper published by IJSER journal is about Novel Thermophilic Anaerobes Phylogenetically Related to Genera Thermoanaerobacter and Clostridium Isolated from Automobile Radiators 1
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Novel Thermophilic Anaerobes Phylogenetically Related to Genera Thermoanaerobacter and Clostridium Isolated from Automobile Radiators
Makanjuola O.R1, Adetunji C.O2, Adetunji J.B3, Koledowo A.K4
1 INTRODUCTION
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The realization that extreme environments harbor different kinds of prokaryote lineage has resulted in a complete reassessment of our concept of microbial evolution and has given considerable impetus to extremophile research [9], [3]. Extremophlic microorganisms especially thermophilic bacteria have developed a variety of molecular
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• 1Ladoke Akintola University of Technology, Ogbomoso, Oyo
State, Nigeria. Email: markosuolale@yahoo.com
• 2Nigerian Stored Products Research Institute, Km 3 Asa dam road, P.M.B 1489, Ilorin, Kwara State. Nigeria. Email:
charliguitar@yahoo.com
• 4Obafemi Awolowo University, Ile-ife, Osun State, Nigeria.Email: onikky1982@yahoo.com
3University of Ilorin, Department of Biochemistry, P.M.B. 1515,
Ilorin, Kwara State:callbumbun@yaho.com
strategies in order to survive in harsh conditions. Thermophiles are microorganisms that thrive at relatively high temperatures, between 45 °C and 80°C. It is generally believed that at high temperature, biomolecules such as enzymes denature thereby losing their function and hence, stopping the metabolism. Also, the fluidity of membranes increases significantly thereby disrupting the cell. The molecular basis for adaptations of thermophilic organisms to extreme environments is to prevent denaturation and degradation. Their membrane lipids contain more saturated and straight chain fatty acids than do mesophiles, which grow typically between 15 °C and
40 °C. This allows thermophiles to grow at higher temperatures by providing the right degree of fluidity needed for membrane function. The presence of
chaperones which refold denatured proteins increase
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The research paper published by IJSER journal is about Novel Thermophilic Anaerobes Phylogenetically Related to Genera Thermoanaerobacter and Clostridium Isolated from Automobile Radiators 2
ISSN 2229-5518
the stability of thermophilic proteins [4]. Also, thermophilic proteins appear to be smaller and in some cases more basic, which may also result in increased stability [8].
A number of thermophiles, belonging to different taxonomic groups, produce thermostable enzymes. This class of enzyme is important not only for the study of biocatalysis and protein stability at extreme conditions but also for the many biotechnological opportunities they offer. This group of organism facilitate enzymatic degradation of polymeric substrates such as starch, cellulose, xylan, pectin and chitin
Hence, the current study was carried out
with the objective to isolate and identify thermophilic organisms from automobile radiators.
Water sample from three different automobile radiators; Honda, Jetta and Passat were collected using sterile syringe, one for each and labeled as sample A, B and C respectively. The samples were immediately taken to the laboratory for subsequent sampling to reduce the proliferation of mesophilic organisms.
0.2ml of each sample recovered by sterile syringe was transferred to each of the bottles, consisting
of 10 ml sterilized nutrient broth (pH 7.65) in a McCantney bottles which were labeled accordingly. The samples were incubated in a water bath at 70°C for 7 days. A control medium consisted of sterilized nutrient broth only was incubated in the water bath alongside with the samples. Growth was followed by measuring the turbidity at 600nm at the 7th day of incubation.
After the 7th day of incubation in water bath, the cultures were introduced into a prepared plate containing nutrient agar using streak method and incubated for 2 days. Isolation of pure culture was done using spread plate method and streak plate method recommended by [7].
The thermophilic isolates were identified
by the use of conventional methods for the presumptive identification by biochemical tests.
Out of 3 samples cultured, all samples grew on nutrient broth and nutrient agar and they showed a clear growth distinction compared to the control. The growth rate (Turbidity) of the thermophilic organisms in the nutrient broth at the 7th day of culturing were determined spectrophotometrically and the results of the optical density (OD) were 0.743nm, 0.427nm,
0.130nm and 0.050nm for sample A, B,C and Control
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The research paper published by IJSER journal is about Novel Thermophilic Anaerobes Phylogenetically Related to Genera Thermoanaerobacter and Clostridium Isolated from Automobile Radiators 3
ISSN 2229-5518
respectively as shown in Fig.1 below.
The result of the optical density (OD) showed that sample A has the highest growth rate follow by sample B and C. The increasing order of the growth rate is; Control < Sample C < Sample B < Sample A .
Clostridium spp.
The bacteria showed some morphological and biochemical characteristics. Pure culture isolates were observed and identified from each of the sample cultured on nutrient agar while no growth was observed in the control. Thermoanaerobacter spp was isolated from sample A while Clostridium spp was isolated from sample B and C. Clostridium is a genus of Gram-positive bacteria, belonging to the Firmicutes. They are obligate anaerobes capable of producing endospores. Individual cells are rod-shaped with positive motility, negative lactose fermentation, lipase activity as opposed to positive indole production.
The genus Thermoanaerobacter falls into
Clusters V in the phylogenetic interrelationship of Clostridium [2]. Cells are curved rods, 3–7 X
0.5µm, obligately anaerobe and gram positive.
Growth substrates are glucose, fructose, lactose, rhamnose, arabinose, xylose, sorbitol, inositol,
mannitol, sucrose, lactose, maltose, starch, pectin, pyruvate, and lactate. No growth on cellulose, succinate, citrate, formate, acetate. The major metabolic products are H2, CO2, and acetate; minor amounts of ethanol are produced along with trace amounts of lactate.
The thermophilic, anaerobic, endospore-forming bacteria are currently placed in the genera Clostridium, Desulfotomaculum, Thermoanaerobacter, Thermoanaerobacterium, Caloramator, and Moorella [2], [5], [10], [11]. Ecological studies have showed that both aerobic and anaerobic species and many morphological and physiological types of microorganisms can exist in thermophilic environments [1], [6]. This report is in accordance with the thermophilic organisms isolated during this experiment.
From our experiment, we were able to isolate rod shaped, obligately anaerobic and facultatively thermophilic bacteria belonging to the genera Thermoanaerobacter and Clostridium from automobile radiators. These organisms can be used in the production of various enzymes for industrial and biotechnological importance.
[1] Brock, D. T..The Origins of Research on
Thermophiles. in Thermophiles; Biodiversity,
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Ecology and Evolution, edited by A.L.Reysenbach (Kluwer Academic/Plenum Publishers,New York, pp.1-22,2001.
[2] Collins, M. D., P. A. Lawson, A. Willems, J. J.
Cordoba, J. Fernandez- Garayzabal, P. Garcia, J. Cai, H. Hippe, and J. A. E. Farrow. The phylogeny of the genus Clostridium: proposal of five new genera and eleven new species combinations. Int. J. Syst. Bacteriol. Vol.44,pp.812–826.1994.
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their possible potential in biotechnology. J Biotechnol,vol.64,pp.39-52, 1998.
[4] Jaenicke, R.Structure and
stability of hyperstable proteins. Adv. Protein Chem. Vol.48, pp.181–269,1996
[5] Lee, Y. E., M. K. Jain, C. Lee, S. E. Lowe, and
J. G. Zeikus. Taxonomic distinction
of saccharolytic thermophilic anaerobes: description of Thermoanaerobacterium xylanolyticum gen. nov., sp. nov., and Thermoanaerobacterium saccharolyticum gen. nov., sp. nov.; reclassification of Thermoanaerobium brockii, Clostridium thermosulfurogenes, and Clostridium thermohydrosulfuricum E100-69 as Thermoanaerobacter brockii comb. nov., Thermoanaerobacterium thermosulfurigenes
comb. nov., and Thermoanaerobacter
thermohydrosulfuricus comb. nov., respectively; and transfer of Clostridium thermohydrosulfuricum 39E to Thermoanaerobacter ethanolicus. Int. J. Syst. Bacteriol.volpp. 43:41–51. 1993.
[6] Madigan, M.T., Martinco, J.M., Parker, J..
Ecology And Biotechnological Applications of Thermophiles and Hyperthermophiles. in Brock Biology of Microorganisms, (Prentice-Hall, Inc.,),pp.
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[7] Rath C.C, Subramanyam V.R. Isolation of thermophilic bacteria from hotsprings of Orissa, India. Geobios, vol.25,No.2.pp.113-
119,1998.
[8] S. Kumar, R. Nussinov, How do thermophilic proteins deal with heat? Cell. Mol. Life
Sci. vol.58,pp.1216–1233,2001.
[9] Stetter ,K.O.Hyperthermophiles: isolation, classification and properties. In Extremophiles — Microbial Life in Extreme Environments. Edited by Horikoshi K, Grant WD. New York: Wiley-Liss;pp.1-
24,1998
[10] Wiegel, J. Genus Thermoanaerobacter,
In P. H. A. Sneath, N. S. Mair, M. E. Sharpe, and J. G. Holt (ed.), Bergey’s manual of systematic bacteriology, vol. 2,p.1379–1383,1986. The Williams and Wilkins Co., Baltimore, Md.
[11] Wiegel, J.. The obligate anaerobic
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The research paper published by IJSER journal is about Novel Thermophilic Anaerobes Phylogenetically Related to Genera Thermoanaerobacter and Clostridium Isolated from Automobile Radiators 5
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thermophilic bacteria, In J. K. Kristjannson
(ed.), Thermophilic bacteria. CRC Press, Boca Raton, Fla. pp.105–184,1992.
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