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Research Article
Volume 1 Issue 1 - 2015
Isolation, Identification and Oil Resistance of Protease Producing Bacillus Subtilis from Automobile Repair Centre Soil, Nanded (India)
Bhagwan N Rekadwad*, Laxmikant V Gumte and Chandrahasya N Khobragade
School of Life Sciences, Swami Ramanand Teerth Marathwada University, Nanded, India
*Corresponding Author: Bhagwan N Rekadawd, School of Life Sciences, Swami Ramanand Teerth Marathwada University, Maharashtra, India.
Received: December 22, 2015; Published: December 29, 2015
Citation: Bhagwan N Rekadwad., et al. “Isolation, Identification and Oil Resistance of Protease Producing Bacillus Subtilis from Automobile Repair Centre Soil, Nanded (India)”. EC Bacteriology and Virology Research 1.1 (2015): 17-23.
Abstract
The natural phenomenon of microbial bioremediation is an eco-friendly measure by which one can clean up petroleum hydrocarbon pollutants from the environments. We have isolated a total 12 isolates capable to degrade variety of hydrocarbons from automobile repair centres, Nanded (India). Of these 12 species, the protease producing bacterium Bacillus subtilis (HD1) was capable to grow and degrade kerosene, diesel, tar alone and in combinations. It is a potent degrader of hydrocarbon such as Diesel and Kerosene + Diesel + Tar hydrocarbon. The ability of the isolated Bacillus subtilis HD1 to grow in the presence of different hydrocarbons is clear evidence that its genome harbour the relevant resistant gene. The isolated strain HD1 may be useful in remediating oil polluted sites. Further research in this area is needed and will make contribution for marked improvement.
Keywords: Oil pollution; Hydrocarbons; Bioremediation; Oxygenases; Bacillus subtilis
Abbreviations: MSRTC: Maharashtra State Road Transport Corporation; CFU: Colony forming unit; MSB: Mineral salt broth; °C: Degree Celsius
Introduction
Petroleum products are one of the major sources of energy for industries and in daily life. Various petroleum products are directly released into environment. This incidence occurred while exploration, production, refining, transport handling and storage of petroleum products. These components also released into environment weather accidently or due to manmade activities take place at filling station repairing of motor vehicles, servicing etc. These activities are the main cause of soil and water pollution. The hydrocarbon containing soil and water causes extensive damage to the entire ecosystem and damaging human health leading to mutation in microorganisms or may cause death of higher organisms. To avoid such consequence, various technologies have been adapted to minimize its harm to the entire ecosystem such as mechanical burning, exploration, dispersion and washing. These useful methods for soil remediation are expensive and are characterized by incompletely removal of hydrocarbon contaminants, additionally, can also cause air pollution [1].
The process of bioremediation is one of the solutions to detoxify or remove pollutants using microorganisms. It is defined as “the use of microorganism to remove pollutants owing to their diverse metabolic capabilities” it is an evolving method for the removal and degradation of many environmental pollutants including the petroleum products. The success of bioremediation depends on environmental/abiotic factors such as temperature, pH, acidity, alkalinity, salinity etc. and biotic factor such as one’s ability to establish and maintain stable condition (because of the presence of stable biomolecule) [2].
The complex process microbial degradation of petroleum compounds depends on the form (nature) and quantity of the hydrocarbon present in environment. Petroleum hydrocarbon is divided into four types such as saturated, aromatics, asphaltenes (phenols, fatty acids, ketones, esters and porphyrins) and resins (pyridines, quinolones, carbazoles, sulfoxides and amides). Degradation of hydrocarbon depends on their size and susceptibility to microbial attack can be generally ranked as follows: linear alkanes > branched alkanes > small aromatics > cyclic alkanes. Microbial bioremediation can be achieved by the alkanes degrading profile such as bacteria (Arthrobacter, Burkholderia, Mycobacterium, Pseudomonas, Sphingomonas, Rhodococcus, Alcaligenes, Acinetobacter lwoffi, Flavobacterium sp., Micrococcus roseus, Corynebacterium ,Gordonia, Brevibacterium, Aeromicrobium, Dietzia); Fungi (Amorphoteca, Neosartorya, Talaromyces, and Graphium)and yeasts (Candida, Yarrowia, and Pichia). These microorganisms were isolated from petroleum contaminated soil and proved to be the potential organisms for hydrocarbon degradation. Similarly, microalgae also take part in hydrocarbon degradation e.g. Protheca20pfi, Green algae, Red algae, Brown algae and Diatoms. These also have the ability to degrade hydrocarbons [3-7].
Microbial biotransformation has been performed by these microorganisms with the help of various enzymes produced during metabolism. Various enzyme produced by microorganisms such as dioxygenase, monooxygenase, hydroxylase, bacterial P450 oxygenase system, eukaryotic P450. In addition to these enzymes, amylase, protease, gelatinase, catalase, oxidase etc., are also produced by these microorganisms (Table 1) [8-11].
Enzyme Substrate Microorganisms References
Soluble methane
monooxygenases
C1-C8 alkanes alkenes and cycloalkanes Methylococcus,Methylosinus, Methylocysts Methylomonas, Methylocella McDonald., et al. 2006
Particulate methane
monooxygenases
C1-C5 (halogenated) alkanes and cycloalkanes Methylobacter, Methylococcus, Methylocyctis  
AlkB related
alkane
hydroxylases
C5-C16 Alkanes, fatty acids, alkyl benzenes, cycloalkanes and so forth Pseudomonas , Burkholderia, Rhodococuus, Mycobacterium van Beilen., et al. 2002
Eukaryotic P450 C10-C16 alkanes, fatty acid Candida maltose, Candida tropicalis, Yarrowialipolytica Iida., et al. 2000
Bacterial P450
oxygenase system
C5-C16 alkanes, cycloalkanes Acinetobacter, Caulobacter, Mycobacterium van Beilen., et al. 2006
Dioxygenases C10-C30 alkanes Acinetobacter sp. Maeng., et al.1996
Table 1: Enzyme involved in biodegradation of petroleum hydrocarbon.
The complex structure and high molecular weight hydrocarbon, degradation of petroleum occurred very slowly and hence creates serious problem to the living things. Some living organisms are resistant to hydrocarbon pollutant metabolize these compounds by producing hydrocarbon degrading enzyme. This process is called microbial transformations. The use of microbial catalyst (enzymes) in biodegradation of organic compounds has advanced significantly during the past three decades [12].
In the present research, we have isolated and identified enzyme producing eubacteria and showed their biodegrading ability.
Materials and Methods
All chemicals used were purchased from Hi Media laboratories Ltd., Mumbai (India).
Collection of sample
The composite soil samples were collected in sterile polythene bags from long seasoned automobile repair centres in Nanded city (India) such as Bafna, MSRTC workshop.
Isolation of petroleum degrading bacteria
Step I: The under mentioned petroleum products (at 1% concentration) were combined with enrichment medium such as
a. Kerosene
b. Diesel
c. Kerosene + Diesel (1:1)
d. Kerosene +Diesel + Tar (1:1:1)
Step II: The collected soil sample (1g) was suspended in 100 mL sterile Zobell broth and Bushnell Haas medium containing 1% of the above mentioned petroleum hydrocarbons separately and incubated in a shaker (at 120 rpm) at 30°C.
Step III: 100 μL of enriched broth were spread onto the Zobell Marine agar plate and then incubated for 24h at 30°C. After incubation period, the colony forming units (CFU) was recorded. The organism showing diverse characteristics has been selected and obtained as pure culture for further study.
Morphological, biochemical characterization and identification of isolated species
All selected isolates were examined for their colony, cell morphology, motility, gram’s staining and biochemical characteristics as per standard methods (Aneja, 2003). Isolated species were identified using Bergey’s Manual of Systematic Bacteriology [13].
Production extraction and partial purification of protease enzyme
For the production of protease enzyme under submerged state fermentation, Mineral salt medium supplemented with casein (2%) inoculated with 5% 18 hr activated Bacillus subtilis culture and kept in a shaking incubator at 120 rpm for 5 days at 37°C. After completion of incubation period, produced enzyme was harvested by centrifugation of medium at 10,000 rpm for 10 min. at 4°C. The supernatant was collected and the enzyme was precipitated using ammonium sulphate (at 65% saturation and temperature of 4°C). The content was kept in refrigerator at temperature 4°C overnight to precipitate residual protease enzyme. On next day, the precipitated enzyme was separated by centrifugation at 10,000 rpm at 4°C for 20 min. Pellet was collected and kept at 4°C. The collected pellet was dissolved in 5 ml of phosphate buffer (0.1mM, pH 7), filled in dialysis bags (30 KDa) and dialysed against the same buffer (0.05 mM) for 12 hours [14].
Characterization of dialysed protease produced by HD1
Protease assay
Protease was assayed by using casein as a substrate. The protease enzyme activity was determined in sodium carbonate buffer (0.5 M, pH 10) using casein at 1% concentration. The 2 mL reaction mixture containing 0.5 mL casein and 1 mL of appropriately diluted enzyme solution were incubated at 30°C for 10 min, and the reaction was stopped with 1 ml of 10% tri-chloro-acetic acid (TCA). The tubes were centrifuged at 15, 000 rpm for 10 min and the degraded products were measured by modified Lowry method. One unit of alkaline protease activity was defined as the amount of enzyme required to liberate 1 μg of tyrosine/min/ml under the standard assay conditions. The estimations were based on a tyrosine calibration curve [15].
Effect of different temperature on protease activity and stability
Optimum temperature of protease was determined at pH 7 by incubating the enzyme for 10 min. at 10 to 60ºC temperatures. Thermal stability of protease was expressed as percentage of initial activity taken as 100%.
Effect of different pH on Protease activity and stability
Optimum temperature of protease was determined at pH values ranging from 5 to 10 (acetate, pH 5; citrate, pH 6; phosphate, pH 7; Tris-HCl, pH 8; carbonate, pH 9 and 10; sodium phosphate- NaOH). Stability of protease was examined by incubating the enzyme in different buffers at pH values ranging from 3 to 11 for 1 h. Residual activity was estimated as described earlier and expressed as percentage of the initial activity taken as 100% [16-17].
Biodegrading activities of bacteria on petroleum oil
The degrading activities of isolates were obtained by using Mineral salt broth (MSB) containing 1% of each hydrocarbon (petrol, kerosene and diesel) was added and incubated at room temperature for seven days. The optical density at optical density 600 nm was taken using spectrophotometer [18].
Biodegradation studies on minimal medium
94 mL of minimal medium was inoculated with 1 mL hydrocarbon (1%) and 5% inoculum in 250 mL Erlenmeyer flasks for seven days in a shaking incubator (set at 100 rpm) at temperature 30°C. After every 24h, 5 mL medium was taken out. The ability of the isolated strain to degrade hydrocarbon was studied by measuring pH and optical density of the medium [19].
Results and Discussion
Hydrocarbon degrading bacteria were isolated on Zobell Marine agar and Bushnell Haas Medium using petroleum hydrocarbon such as petrol, diesel, petrol and diesel along with tar included in the entire medium. Isolation of bacteria was carried out at 37°C. A total of 12 isolates capable of growth on petroleum products were selected and obtained in pure culture. Of these 12 isolates, nine isolate showing luxuriant growth and diverse characterization has been chosen for further study. These were named as HD1 to HD9. All nine isolate observed were rod shaped endospore forming bacteria. Of these, seven isolate were Gram positive and two isolates were Gram negative. All isolates were capable to grow at 20% NaCl concentration within their optimum temperature and pH. All isolates utilized a variety of sugars and showed diverse enzyme profile including amylase, protease, and urease (Table 2). All the isolate have been identified based on morphological and biochemical characteristics.HD1, HD6 and HD9 were identified as B. subtilis HD2 to HD4 were identified as Bacillus larvae. HD7 and HD8 were identified as Enterobactoraerogenes. Protease produced by Bacillus subtilis (HD1) has optimum temperature of 40oC and optimum pH 7 (Fig. 1-2). Similar type of results has been reported [20].
Parameter Result
Code HD1 HD2 HD3 HD4 HD5 HD6 HD7 HD8 HD9
Cell shape R R R R R R R R R
Gram staining + + + + + + + + +
Endospore P P P P P P P P P
Motility M M M M M M M M M
Colour of colony PY Re W W Y PY Y Y W
Size of colony (mm) 1 1 1 2 2 1 1 1 1
Shape of colony C C C C C C C C C
Margin of colony E E Ir Ir E E E E E
Elevation of colony El F F F EN El El El El
Opacity O O O O O O O O O
Tolerance of NaCl (%) 20 20 20 20 20 20 20 20 20
Optimum temperature (°C) 37 37 37 37 37 37 37 37 37
Optimum pH 7.6 7.6 7.6 7.6 7.6 7.6 7.6 7.6 7.6
Glucose + + + + + + + + +
Lactose + - - + - - + + +
Sucrose + + + + - + + + +
Maltose + + + + + + + + +
Rhamnose + + + + + + + + +
Raffinose + + + + + + + + +
Starch + + + + + + + + +
Casein + + - + - + - + +
Urease - - - - - - - - -
Catalase + + + + + + + + +
Indole production - - - - - - - - -
Methyl red - + + + - + + + +
Voges-Proskauer + - - - + - - - -
Citrate utilization + - - - + + + + +
H2S Production + + + + - + + + +
Note: + = Positive, - = Negative, R= rod, M=motile, NM= non-motile, P, Present, PY= Pale yellow, Re= Reddish, W= White, Y= Yellow, C= Circular, E= Entire, Ir= Irregular, El= Elevated, F= Flat, Cn= Convex, O= Opaque
Table 2: Morphological and biochemical characteristics of isolates.

Figure 1: Effect of different temperatures on stability and activity of HD1 protease.

Figure 2: Effect of different pH on stability and activity of HD1 protease.

The biodegradation studies were performed using B. subtilis (HD1) using minimal medium and mineral salt broth containing 1% hydrocarbon with different combinations. The minimal medium supplemented by 1% kerosene + Diesel + Tar hydrocarbon, Bacillus subtilis showed maximum growth after 48 hours at its optimum temperature and optimum pH. Similarly, we have conducted studies using different combination at 1% concentration on Mineral salt broth (such as kerosene, Diesel, Tar hydrocarbon). HD1 showed noticeable resistant and showed growth in the presence of 1% diesel alone and 1% hydrocarbon mixture (kerosene + Diesel + Tar hydrocarbon) after 72 hrs. (Figure 3). Similar types of studies were performed by different research groups worldwide and reported that Bacillus subtilis species tolerate and grow on higher concentration of diesel, petrol and other hydrocarbon [21-23].

Figure 3: Effect of incubation period on degrading ability of Bacillus subtilis on hydrocarbon.

Conclusion
From the present research, it is clear that the protease producing Bacillus subtilis isolated by us have a high hydrocarbon degrading ability. Bacillus subtilis (HD1) showed as remarkable growth and resistance in the presence of diesel, petrol, and kerosene and tar. Hence, this strain may be used in cleaning oil polluted site.
Acknowledgements
We are thankful to UGC, New Delhi, India for the financial support.
Bibliography
  1. Das N., et al. “Microbial degradation of petroleum hydrocarbon contaminants: an Overview”. Biotechnology Research International (2011): 941810.
  2. Mhatre BA., et al. “Biodegradation of diesel using microbes from a clam (Meretrixmeretrix) shell”. Journal of Geo-Marine Sciences 43 (2014): 877-881.
  3. Bogusławska-Was E., et al. “The seasonal variability of yeasts and yeast-like organisms in water and bottom sediment of the Szczecin Lagoon”. The International Journal of Hygiene and Environmental Health 203.6 (2014): 451-458.
  4. Daugulis AJ., et al. “Microbial degradation of high and low molecular weight poly aromatic hydrocarbons in a two-phase partitioning bioreactor by two strains of Sphingomonas sp”. Biotechnology Letters 25.17 (2003): 1441-1444.
  5. Chaillan F., et al. “Identification and biodegradation potential of tropical aerobic hydrocarbon degrading microorganisms”. Research in Microbiology 155.7 (2004): 587-595.
  6. Singh H., et al. “Mycoremediation: fungal bioremediation”. Wiley Inderscience, New York, NY, USA (2006):ISBN: 978-0-471-75501-2.
  7. Adebusoye SA., et al. “Microbial degradation of petroleum hydrocarbons in a polluted tropical stream”. World Journal of Microbiology and Biotechnology 23 (2007): 1149-1159.
  8. Maeng JHO., et al. “Isolation and characterization of a novel oxygenase that catalyzes the first step of n-alkane oxidation in Acinetobacter sp. strain M-1”. Journal of Bacteriology 178.13 (1996): 3695-3700.
  9. Iida T., et al. “The cytochrome P450ALK multigene family of an n-alkane-assimilating yeast, Yarrowialipolytica: cloning and characterization of genes coding for new CYP52 family members”. Yeast 16.12 (2000): 1077-1087.
  10. VanBeilen JB., et al. “Rubredoxins involved in alkane Oxidation”. Journal of Bacteriology 184.6 (2002): 1722-1732.
  11. McDonald IR., et al. “Diversity of soluble methane monooxygenase-containing methanotrophs isolated from polluted environments”. FEMS Microbiology Letters 255.2 (2006): 225-232.
  12. VanBeilen JB., et al. “Cytochrome P450 alkane hydroxylases of the CYP153 family are common in alkane-degrading eubacteria lacking integral membrane alkane hydroxylases”. Applied and Environmental Microbiology 72.1 (2006): 59-65.
  13. Bergeys DH., et al. “Bergey’s Manual of Systematic Bacteriology”. Williams and Wilkins, Baltimore (1994).
  14. Sevinc N., et al. “Production of protease by Bacillus sp. N-40 isolated from soil and its enzymatic properties”. Journal of Biodiversity and Environmental Sciences 5.14 (2011): 95-103.
  15. Mala M., et al. “Partial purification and properties of a laundry detergent compatible alkaline protease from a newly isolated Bacillus species Y”. Indian Journal of Microbiology 50.3 (2010): 309-317.
  16. Rekadwad BN., et al. “Characterization of amylase from industrially important thermophilic microorganism: Geobacillusthermo leovorans Strain REKADWADSIS”. International Journal of Life Sciences Biotechnology and Pharma Research 4.1 (2015a): 26-30.
  17. Rekadwad BN., et al. “Enhanced production of thermostable amylase by thermophilic Geobacillusthermoleorans Strain REKADWADSIS isolated from Unkeshwar hot spring sediment”. International Journal of Current Research 7(2015b): 11823-11824.
  18. Boboye B., et al. “Degradative activity of bacteria isolated from Hydrocarbon-polluted site in Ilaje, Ondo State, Nigeria”. African Journal of Microbiology Research 4.23 (2010): 2484-2491.
  19. Darsa KV., et al. “Biodegradation of petroleum compound using the bacterium Bacillus subtilis”. Science International 2.1 (2014): 20-25.
  20. Venugopal M., et al. “An alkaline protease from Bacillus circulansBM15, newly isolated from a mangrove station: characterization and application in laundry detergent formulations”. Indian Journal of Microbiology 47.4 (2007): 298-303.
  21. Meintanis C., et al. “Biodegradation of crude oil by thermophilic bacteria isolated from a volcano island” Biodegradation 17.2 (2006): 105-111.
  22. Nwaogu LA., et al. “Degradation of diesel oil in a polluted soil using Bacillus subtilis”. African Journal of Biotechnology 7.12 (2008): 1939-1943.
  23. Pant G., et al. “Production, optimization and partial purification of protease from Bacillus Subtilis”. Journal of Taibah University for Science 9.1 (2015): 50-55.
Copyright: © 2015 Bhagwan N Rekadwad., et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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