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, 3(1): 828-831
JANUARY- 2017
Address for Correspondence: Mr. A. Ravi, Lecturer, Department of Biotechnology, Vivekananda Degree & PG
College, Karimnagar, Telangana, India
Received: 12 November 2016/Revised: 05 December 2016/Accepted: 27 December 2016
ABSTRACT- Mechanical workshops release huge levels of used engine oil into the surroundings. Many mechanical
workshops do not have proper disposal mechanisms and eventually used engine oil reaches the surrounding soil. The
hydrocarbons present in used engine oil damage the environment and human health. There are microbes in nature which
have the ability to degrade the used engine oil. Such microbes can be isolated and used for the biodegradation of
hazardous and recalcitrant hydrocarbons occurring in used engine oil. The present study deals with the four used engine
oil degrading bacterial species isolated from oil contaminated sites of mechanical workshops. The bacterial species were
Bacillus, Acinetobacter, Pseudomonas and Micrococcus. The optimum pH and temperature on the oil degradation activity
of bacterial species were determined. In addition, effect of various amino acids on the oil degradation ability of these
bacterial species was studied and best amino acid for oil degrading activity was determined. All the four bacterial species
exhibited highest oil degrading activity at pH 7.0. The optimal temperature required for maximum oil degradation activity
by Bacillus and Acinetobacter species was 330C and 350C respectively. At 310C Pseudomonas and Micrococcus species
had shown maximum oil degradation activity. All the bacterial species had shown enhanced oil degradation activity in
methionine supplemented medium. Bacillus species was found to be efficient among all the four species.
Key-words- Mechanical workshops, Used engine oil, Oil degrading bacteria, Optimum, pH, Temperature, Amino acids
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INTRODUCTION
Used engine oil causes potential damage to the
environment. Engine oil is filled in the motor vehicles for
the functioning of engine. The engine oil is combusted
during its usage. At intervals used engine oil is replaced
with fresh engine oil. The used engine oil is disposed into
the environment from mechanical workshops. Used engine
oil consists of hazardous aliphatic, polyaromatic,
organosulfur compounds and heavy metals [1]. These
compounds and heavy metals are mutagenic and
carcinogenic to humans [2]. Nearly six million tons of crude
oil is disposed into the environment from various sources
[3]
.
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DOI: 10.21276/ijlssr.2017.3.1.13
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Inoculum preparation
Each bacterial culture was inoculated into 10 ml of MS
medium with 5% used engine oil and incubated overnight
at 300C. After incubation the culture was centrifuged at
3500 rpm to get cell pellet. After washing each cell
cultures pellet was inoculated into MS medium with 5%
used engine oil and grown till the optical density of each
culture broth was 1 at 600 nm. One ml of such each
bacterial culture serves as inoculum source [8].
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Bacterial
Species
5.0
6.0
7.0
8.0
Bacillus
0.90 0.05
1.20 0.09
1.41 0.05
1.10 0.12
Acinetobacter
0.61 0.12
0.64 0.09
0.71 0.08
0.62 0.05
Pseudomonas
0.84 0.05
1.00 0.05
1.26 0.08
0.98 0.09
Micrococcus
0.72 0.08
0.91 0.05
1.10 0.09
0.87 0.09
Bacterial
Species
300C
310C
320C
330C
340C
350C
360C
370C
380C
390C
400C
Bacillus
1.43
0.12
1.49
0.05
1.56
0.05
1.63
0.09
1.59
0.05
1.44
0.09
1.38
0.05
1.30
0.09
1.22
0.08
1.03
0.08
0.92
0.05
Acinetobacter
0.70
0.08
0.77
0.12
0.84
0.05
0.91
0.08
0.98
0.12
1.18
0.05
1.10
0.08
1.06
0.05
1.03
0.05
1.00
0.12
0.97
0.12
Pseudomonas
1.19
0.09
1.28
0.05
1.11
0.09
1.03
0.05
0.96
0.09
0.89
0.12
0.80
0.08
0.72
0.05
0.67
0.09
0.61
0.12
0.55
0.08
Micrococcus
1.06
0.05
1.11
0.08
1.01
0.08
0.97
0.05
0.91
0.08
0.84
0.08
0.77
0.08
0.69
0.08
0.62
0.05
0.57
0.05
0.50
0.08
Table3: Effect of various amino acids on oil biodegradation activity of bacterial species
S.
No.
Bacterial
Species
OD at 600 nm of bacterial broth cultures in various amino acids with concentration 1mg/ml
(bacteria grown at their respective optimal pH and temperatures)
Pro
Ala
Phe
Tyr
Arg
Lys
Cys
Met
Gln
Bacillus
1.73
0.12
1.69
0.09
1.59
0.09
1.66
0.05
1.62
0.05
1.57
0.09
1.80
0.05
1.84
0.12
1.76
0.09
Acinetobacter
1.34
0.05
1.31
0.12
1.22
0.12
1.25
0.09
1.28
0.12
1.20
0.12
1.40
0.14
1.44
0.09
1.37
0.05
Pseudomonas
1.47
0.08
1.44
0.09
1.30
0.08
1.40
0.16
1.37
0.09
1.33
0.16
1.54
0.05
1.59
0.12
1.50
0.05
Micrococcus
1.40
0.09
1.31
0.05
1.24
0.05
1.28
0.05
1.20
0.09
1.15
0.08
1.46
0.12
1.50
0.05
1.36
0.12
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CONCLUSION
Used engine oil is disposed from various mechanical
workshops reach the surrounding environment. Some
hydrocarbons present in used engine oil are recalcitrant,
hazardous and cause deleterious effects to environment and
human health. These hazardous hydrocarbons present in
used engine oils can be degraded by microbes to clean up
the environment. In the present work used engine oil
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ACKNOWLEDGMENT
We are extremely thankful to the Head, Department of
Biotechnology, Vivekananda Degree & PG College,
Karimnagar, Telangana, India for permitting us to use the
laboratory to carry out the work related to this paper. We
are also thankful to Principal, Administrative officer and
staff for their support. Finally we thank Head, Department
of computer science for permitting us to use the computer
lab and internet during the preparation of the paper.
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