![International Journal of Trend in
International Open Access
ISSN No: 2456
@ IJTSRD | Available Online @ www.ijtsrd.com
Effect of Glucose
Bacterial Isolates
Lecturer
Shri Shivaji Science College Nagpur
ABSTRACT
The demand for biosurfactants is gradually increasing
and are thus substituting their chemically synthesized
counterparts [14]. The production of biosurfactants
commercially requires high expenses. For the
production of biosurfactant proper optimization of
physico-chemical parameters is very important. Hence
the research was conducted to study the effect of
glucose as a carbon source for production of
biosurfactant using bacterial isolates from oil
contaminated sites in MSM medium.
KEY WORDS: Biosurfactants, Bacter
Glucose medium, Minimal Salt Medium.
INTRODUCTION
Biosurfactants are amhiphatic compounds
mostly produced by bacterial species
Biosurfactants are reported to be toxic in nature
They are biodegradable in nature as compared to other
commercially available surfactants [1]. Biosurfactants
have varied environmental applications
hydrocarbon remediation from soil, dispersion
spills and enhancement of oil recovery processes
5]. Biosurfactants have potential use in
industry, agriculture, pharmaceutics, petro
paper and pulp industry etc. and therefore research in
the area of biosurfactants has expanded quite a lot in
recent years. The development of this line of research
is of great importance, mainly in view of the present
concern with protection of the environment
International Journal of Trend in Scientific Research and Development (IJTSRD)
International Open Access Journal | www.ijtsrd.com
ISSN No: 2456 - 6470 | Volume - 3 | Issue – 1 | Nov
www.ijtsrd.com | Volume – 3 | Issue – 1 | Nov-Dec 2018
f Glucose on Biosurfactant Production
Isolates from Oil Contaminated Sites
Anjali Sharma
ecturer, Department of Microbiology,
Shri Shivaji Science College Nagpur, Maharashtra, India
The demand for biosurfactants is gradually increasing
and are thus substituting their chemically synthesized
counterparts [14]. The production of biosurfactants
commercially requires high expenses. For the
production of biosurfactant proper optimization of the
chemical parameters is very important. Hence
the research was conducted to study the effect of
glucose as a carbon source for production of
biosurfactant using bacterial isolates from oil
Biosurfactants, Bacterial isolates,
Glucose medium, Minimal Salt Medium.
hiphatic compounds that are
bacterial species[1,4,17].
toxic in nature [7].
biodegradable in nature as compared to other
surfactants [1]. Biosurfactants
environmental applications such as
hydrocarbon remediation from soil, dispersion of oil
enhancement of oil recovery processes [2,
potential use in various
petro chemistry,
therefore research in
expanded quite a lot in
of this line of research
view of the present
concern with protection of the environment. The most
significant advantage of a biosurfactant over
surfactants is its ecological acceptanc
the other advantages of biosurfactants include
selectivity, specific activity at extreme temperatures,
pH and salinity and the possibility of their production
through fermentation [9, 10].
biosurfactants have been report
biosurfactant is influenced by the
enhance the growth of the microbe.
to stimulate biosurfactant production
optimization of ecological, physiological
nutritional conditions [15].
MATERIALS AND METHODS:
In order to study effect of glucose as a carbon source
on production of biosurfactant, M
was used as the basal medium.
the glucose was kept at 1% (w/v). 1% inoculum
culture of isolated bacterial
IHD19, IHD21, IHD36, IHD44, IHD58, IHD80,
IHD89, IHD96, IHD112, IHD 148, IHD152, IHD157,
IHD 176, IHD178, and IHD188) were
100 ml of Minimal Salt Medium containing glucose
as carbon source and were incubated in rotar
at 120rpm at 37 ⁰C for 7 days. Growth of cells was
monitored by measuring the absorbance (Optical
Density), A540. The biosurfactant production was
determined by measuring the EI% as described under
[2, 12, 13, 16, 18]
Research and Development (IJTSRD)
www.ijtsrd.com
1 | Nov – Dec 2018
Dec 2018 Page: 730
n Biosurfactant Production using
rom Oil Contaminated Sites
significant advantage of a biosurfactant over chemical
is its ecological acceptance [6,8]. Some of
the other advantages of biosurfactants include
selectivity, specific activity at extreme temperatures,
pH and salinity and the possibility of their production
through fermentation [9, 10]. A large variety of
biosurfactants have been reported [11]. Production of
biosurfactant is influenced by the factors which also
enhance the growth of the microbe. One strategy used
to stimulate biosurfactant production has been the
optimization of ecological, physiological and
MATERIALS AND METHODS:
glucose as a carbon source
on production of biosurfactant, Minimal Salt Medium
as the basal medium. The concentration of
was kept at 1% (w/v). 1% inoculum
strains (IHD3, IHD13,
IHD19, IHD21, IHD36, IHD44, IHD58, IHD80,
IHD89, IHD96, IHD112, IHD 148, IHD152, IHD157,
IHD 176, IHD178, and IHD188) were inoculated into
edium containing glucose
were incubated in rotary shaker
C for 7 days. Growth of cells was
monitored by measuring the absorbance (Optical
. The biosurfactant production was
determined by measuring the EI% as described under](https://image.slidesharecdn.com/89effectofglucoseonbiosurfactantproductionusingbacterialisolatesfromoilcontaminatedsites-190123113503/85/Effect-of-Glucose-on-Biosurfactant-Production-using-Bacterial-Isolates-from-Oil-Contaminated-Sites-1-320.jpg)
![International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456
@ IJTSRD | Available Online @ www.ijtsrd.com
RESULTS AND DISCUSSION:
Table.1. Effect of Glucose as Carbon Sources
pH = 7.00, Temperature: 37 ⁰C. Inoculum = 1 vol%, engine oil = 2 vol%, Incubation time = 7days,
Bacteri
al
isolates
IH
D 3
IH
D
13
IH
D
19
IH
D
21
IH
D
36
EI % 64 78 88 69 76
O.D
600 nm
0.7 1.6
1.8
2
0.8
1.7
6
A number of carbon sources have been used by many
researchers for biosurfactant production. The quality
and quantity of produced biosurfactant are affected
and influenced by the nature of the carbon substrate.
Out of the various carbon sources screened for
production of biosurfactant in the current study,
glucose was found to be the best carbon source for
growth and biosurfactant production. The highest
Emulsification activity was obtained using glucose at
concentration of (1% w/v) as source of carbon along
with engine oil, resulted in higher emulsifier activity
(EI% = 88 % and EA = 1.82). The results were in
disagreement with the founding of A. Khopade
et.al.,[11] where they obtained maximum activity
(EI%= 80%) when using sucrose as source of carbon
along with engine oil by marine isolate
sp. B3.
Maximum growth of bacterial cell and biosurfactant
production by bacterial species isolated from
petroleum contaminated soil were observed by using
glucose as the source of carbon along with engine oil
where the maximum value of emulsification index
64
78
88
69
76
0
10
20
30
40
50
60
70
80
90
100
International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456
www.ijtsrd.com | Volume – 3 | Issue – 1 | Nov-Dec 2018
Table.1. Effect of Glucose as Carbon Sources
Inoculum = 1 vol%, engine oil = 2 vol%, Incubation time = 7days,
shaking speed = 125 r.p.m.
IH
D
44
IH
D
58
IH
D
80
IH
D
89
IH
D
96
IH
D
112
IH
D
148
IH
D
152
68 75 67 54 66 76 62 73
1.2
8
1.5
1.0
7
0.6
2
0.9 0.6 0.4 0.7
A number of carbon sources have been used by many
researchers for biosurfactant production. The quality
and quantity of produced biosurfactant are affected
and influenced by the nature of the carbon substrate.
Out of the various carbon sources screened for
production of biosurfactant in the current study,
glucose was found to be the best carbon source for
growth and biosurfactant production. The highest
Emulsification activity was obtained using glucose at
concentration of (1% w/v) as source of carbon along
with engine oil, resulted in higher emulsifier activity
(EI% = 88 % and EA = 1.82). The results were in
disagreement with the founding of A. Khopade
where they obtained maximum activity
(EI%= 80%) when using sucrose as source of carbon
along with engine oil by marine isolate Streptomyces
Maximum growth of bacterial cell and biosurfactant
bacterial species isolated from
were observed by using
glucose as the source of carbon along with engine oil
where the maximum value of emulsification index
was found to be EI%= 88%. which was given by IHD
19, Intermediate range of Emulsification Index were
recorded between 79% to 64% given by the bacterial
isolates viz.,. IHD 3, IHD 13, IHD 21, IHD 36, IHD
44, IHD 58, IHD 80, IHD 96, IHD 112, IHD 148,
IHD 152, IHD 157, IHD 176. IHD 178 and IHD188.
Lowest EI% was recorded to be 54% which was given
by IHD 89.
CONCLUSION:
Growth and production medium for biosurfactant
production was optimized by using glucose as the
carbon source in the Minimal Salt Medium.
Maximum production of biosurfactant was observed
in MSM medium containing glucose and 2% engine
oil.
REFERENCES:
1. Ainon, H., Noramiza, S. and Shahidan, R. (2013).
Screening of biosurfactants produced by the
hydrocarbon degrading bacteria.
Malaysiana, 42 (5): 615 - 623.
68
75
67
54
66
76
62
73
79
Effect of Glucose
International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456-6470
Dec 2018 Page: 731
Inoculum = 1 vol%, engine oil = 2 vol%, Incubation time = 7days,
IH
D
157
IH
D
176
IH
D
178
IH
D
188
79 77 69 71
0.8 1 0.6 0.9
which was given by IHD
19, Intermediate range of Emulsification Index were
tween 79% to 64% given by the bacterial
isolates viz.,. IHD 3, IHD 13, IHD 21, IHD 36, IHD
44, IHD 58, IHD 80, IHD 96, IHD 112, IHD 148,
IHD 152, IHD 157, IHD 176. IHD 178 and IHD188.
Lowest EI% was recorded to be 54% which was given
Growth and production medium for biosurfactant
production was optimized by using glucose as the
carbon source in the Minimal Salt Medium.
Maximum production of biosurfactant was observed
in MSM medium containing glucose and 2% engine
inon, H., Noramiza, S. and Shahidan, R. (2013).
Screening of biosurfactants produced by the
hydrocarbon degrading bacteria. Sains
623.
77
69 71
Effect of Glucose](https://image.slidesharecdn.com/89effectofglucoseonbiosurfactantproductionusingbacterialisolatesfromoilcontaminatedsites-190123113503/85/Effect-of-Glucose-on-Biosurfactant-Production-using-Bacterial-Isolates-from-Oil-Contaminated-Sites-2-320.jpg)
Effect of Glucose on Biosurfactant Production using Bacterial Isolates from Oil Contaminated Sites
- 1. International Journal of Trend in International Open Access ISSN No: 2456 @ IJTSRD | Available Online @ www.ijtsrd.com Effect of Glucose Bacterial Isolates Lecturer Shri Shivaji Science College Nagpur ABSTRACT The demand for biosurfactants is gradually increasing and are thus substituting their chemically synthesized counterparts [14]. The production of biosurfactants commercially requires high expenses. For the production of biosurfactant proper optimization of physico-chemical parameters is very important. Hence the research was conducted to study the effect of glucose as a carbon source for production of biosurfactant using bacterial isolates from oil contaminated sites in MSM medium. KEY WORDS: Biosurfactants, Bacter Glucose medium, Minimal Salt Medium. INTRODUCTION Biosurfactants are amhiphatic compounds mostly produced by bacterial species Biosurfactants are reported to be toxic in nature They are biodegradable in nature as compared to other commercially available surfactants [1]. Biosurfactants have varied environmental applications hydrocarbon remediation from soil, dispersion spills and enhancement of oil recovery processes 5]. Biosurfactants have potential use in industry, agriculture, pharmaceutics, petro paper and pulp industry etc. and therefore research in the area of biosurfactants has expanded quite a lot in recent years. The development of this line of research is of great importance, mainly in view of the present concern with protection of the environment International Journal of Trend in Scientific Research and Development (IJTSRD) International Open Access Journal | www.ijtsrd.com ISSN No: 2456 - 6470 | Volume - 3 | Issue – 1 | Nov www.ijtsrd.com | Volume – 3 | Issue – 1 | Nov-Dec 2018 f Glucose on Biosurfactant Production Isolates from Oil Contaminated Sites Anjali Sharma ecturer, Department of Microbiology, Shri Shivaji Science College Nagpur, Maharashtra, India The demand for biosurfactants is gradually increasing and are thus substituting their chemically synthesized counterparts [14]. The production of biosurfactants commercially requires high expenses. For the production of biosurfactant proper optimization of the chemical parameters is very important. Hence the research was conducted to study the effect of glucose as a carbon source for production of biosurfactant using bacterial isolates from oil Biosurfactants, Bacterial isolates, Glucose medium, Minimal Salt Medium. hiphatic compounds that are bacterial species[1,4,17]. toxic in nature [7]. biodegradable in nature as compared to other surfactants [1]. Biosurfactants environmental applications such as hydrocarbon remediation from soil, dispersion of oil enhancement of oil recovery processes [2, potential use in various petro chemistry, therefore research in expanded quite a lot in of this line of research view of the present concern with protection of the environment. The most significant advantage of a biosurfactant over surfactants is its ecological acceptanc the other advantages of biosurfactants include selectivity, specific activity at extreme temperatures, pH and salinity and the possibility of their production through fermentation [9, 10]. biosurfactants have been report biosurfactant is influenced by the enhance the growth of the microbe. to stimulate biosurfactant production optimization of ecological, physiological nutritional conditions [15]. MATERIALS AND METHODS: In order to study effect of glucose as a carbon source on production of biosurfactant, M was used as the basal medium. the glucose was kept at 1% (w/v). 1% inoculum culture of isolated bacterial IHD19, IHD21, IHD36, IHD44, IHD58, IHD80, IHD89, IHD96, IHD112, IHD 148, IHD152, IHD157, IHD 176, IHD178, and IHD188) were 100 ml of Minimal Salt Medium containing glucose as carbon source and were incubated in rotar at 120rpm at 37 ⁰C for 7 days. Growth of cells was monitored by measuring the absorbance (Optical Density), A540. The biosurfactant production was determined by measuring the EI% as described under [2, 12, 13, 16, 18] Research and Development (IJTSRD) www.ijtsrd.com 1 | Nov – Dec 2018 Dec 2018 Page: 730 n Biosurfactant Production using rom Oil Contaminated Sites significant advantage of a biosurfactant over chemical is its ecological acceptance [6,8]. Some of the other advantages of biosurfactants include selectivity, specific activity at extreme temperatures, pH and salinity and the possibility of their production through fermentation [9, 10]. A large variety of biosurfactants have been reported [11]. Production of biosurfactant is influenced by the factors which also enhance the growth of the microbe. One strategy used to stimulate biosurfactant production has been the optimization of ecological, physiological and MATERIALS AND METHODS: glucose as a carbon source on production of biosurfactant, Minimal Salt Medium as the basal medium. The concentration of was kept at 1% (w/v). 1% inoculum strains (IHD3, IHD13, IHD19, IHD21, IHD36, IHD44, IHD58, IHD80, IHD89, IHD96, IHD112, IHD 148, IHD152, IHD157, IHD 176, IHD178, and IHD188) were inoculated into edium containing glucose were incubated in rotary shaker C for 7 days. Growth of cells was monitored by measuring the absorbance (Optical . The biosurfactant production was determined by measuring the EI% as described under
- 2. International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456 @ IJTSRD | Available Online @ www.ijtsrd.com RESULTS AND DISCUSSION: Table.1. Effect of Glucose as Carbon Sources pH = 7.00, Temperature: 37 ⁰C. Inoculum = 1 vol%, engine oil = 2 vol%, Incubation time = 7days, Bacteri al isolates IH D 3 IH D 13 IH D 19 IH D 21 IH D 36 EI % 64 78 88 69 76 O.D 600 nm 0.7 1.6 1.8 2 0.8 1.7 6 A number of carbon sources have been used by many researchers for biosurfactant production. The quality and quantity of produced biosurfactant are affected and influenced by the nature of the carbon substrate. Out of the various carbon sources screened for production of biosurfactant in the current study, glucose was found to be the best carbon source for growth and biosurfactant production. The highest Emulsification activity was obtained using glucose at concentration of (1% w/v) as source of carbon along with engine oil, resulted in higher emulsifier activity (EI% = 88 % and EA = 1.82). The results were in disagreement with the founding of A. Khopade et.al.,[11] where they obtained maximum activity (EI%= 80%) when using sucrose as source of carbon along with engine oil by marine isolate sp. B3. Maximum growth of bacterial cell and biosurfactant production by bacterial species isolated from petroleum contaminated soil were observed by using glucose as the source of carbon along with engine oil where the maximum value of emulsification index 64 78 88 69 76 0 10 20 30 40 50 60 70 80 90 100 International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456 www.ijtsrd.com | Volume – 3 | Issue – 1 | Nov-Dec 2018 Table.1. Effect of Glucose as Carbon Sources Inoculum = 1 vol%, engine oil = 2 vol%, Incubation time = 7days, shaking speed = 125 r.p.m. IH D 44 IH D 58 IH D 80 IH D 89 IH D 96 IH D 112 IH D 148 IH D 152 68 75 67 54 66 76 62 73 1.2 8 1.5 1.0 7 0.6 2 0.9 0.6 0.4 0.7 A number of carbon sources have been used by many researchers for biosurfactant production. The quality and quantity of produced biosurfactant are affected and influenced by the nature of the carbon substrate. Out of the various carbon sources screened for production of biosurfactant in the current study, glucose was found to be the best carbon source for growth and biosurfactant production. The highest Emulsification activity was obtained using glucose at concentration of (1% w/v) as source of carbon along with engine oil, resulted in higher emulsifier activity (EI% = 88 % and EA = 1.82). The results were in disagreement with the founding of A. Khopade where they obtained maximum activity (EI%= 80%) when using sucrose as source of carbon along with engine oil by marine isolate Streptomyces Maximum growth of bacterial cell and biosurfactant bacterial species isolated from were observed by using glucose as the source of carbon along with engine oil where the maximum value of emulsification index was found to be EI%= 88%. which was given by IHD 19, Intermediate range of Emulsification Index were recorded between 79% to 64% given by the bacterial isolates viz.,. IHD 3, IHD 13, IHD 21, IHD 36, IHD 44, IHD 58, IHD 80, IHD 96, IHD 112, IHD 148, IHD 152, IHD 157, IHD 176. IHD 178 and IHD188. Lowest EI% was recorded to be 54% which was given by IHD 89. CONCLUSION: Growth and production medium for biosurfactant production was optimized by using glucose as the carbon source in the Minimal Salt Medium. Maximum production of biosurfactant was observed in MSM medium containing glucose and 2% engine oil. REFERENCES: 1. Ainon, H., Noramiza, S. and Shahidan, R. (2013). Screening of biosurfactants produced by the hydrocarbon degrading bacteria. Malaysiana, 42 (5): 615 - 623. 68 75 67 54 66 76 62 73 79 Effect of Glucose International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456-6470 Dec 2018 Page: 731 Inoculum = 1 vol%, engine oil = 2 vol%, Incubation time = 7days, IH D 157 IH D 176 IH D 178 IH D 188 79 77 69 71 0.8 1 0.6 0.9 which was given by IHD 19, Intermediate range of Emulsification Index were tween 79% to 64% given by the bacterial isolates viz.,. IHD 3, IHD 13, IHD 21, IHD 36, IHD 44, IHD 58, IHD 80, IHD 96, IHD 112, IHD 148, IHD 152, IHD 157, IHD 176. IHD 178 and IHD188. Lowest EI% was recorded to be 54% which was given Growth and production medium for biosurfactant production was optimized by using glucose as the carbon source in the Minimal Salt Medium. Maximum production of biosurfactant was observed in MSM medium containing glucose and 2% engine inon, H., Noramiza, S. and Shahidan, R. (2013). Screening of biosurfactants produced by the hydrocarbon degrading bacteria. Sains 623. 77 69 71 Effect of Glucose
- 3. International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456 @ IJTSRD | Available Online @ www.ijtsrd.com 2. Anjali Sharma, Dr. A. V. Gomashe, ‘ production by bacterial species isolated from petroleum oil contaminated soil’. (2018) IJCRT | Volume 6, Issue 2 April 2018 | ISSN: 2320 3. Anjali Sharma, Dr. A. V. Gomashe “Isolation and Screening of Biosurfactant Producing Bacterial Species from Petroleum Oil Contaminated Sites”. International Journal of Scientific Progress and Research (IJSPR) ISSN: 2349-4689 Issue 120, Volume 42, Number 03, December (2017). 4. Benincasa, M., Marqués, A., Pinazo, A. and Manresa, A. (2010). Rhamnolipids surfactants: alternatives substrates, new strategies. In. Sen,. R. (Ed.). Biosurfactants, pages. 170 York: Landes Bioscience and Springer Science and Business Media. 5. Banat IM, Franzetti A, Gandolfi I, Bestetti G, Martinotti MG, Fracchia L, Smyth TJ, Marchant R. Microbial biosurfactants produc applications and future potential. Appl Microbiol Biotechnol. 2010;87:427–444. 6. Bailey, R.W. (1958). The reaction of pentoses with anthrone. Biochemical Journal 672. 7. Bouchez, M., Blanchet, D. and Vandacasteele P. (1995). Degradation of polycyclic aromatic hydrocarbons by pure strains and defined strain associations: inhibition phenomena and cometabolism. Applied Microbiology Biotechnology, 43: 156 - 164. 8. Bordoloi, N. K. and Konwar, B. K. (2009). Bacterial biosurfactant in enhancing solubility and metabolism of petroleum hydrocarbons. of Hazardous Materials, 170: 495 - 505. 9. Donlan R.M. Biofilms: Microbial Life on Surfaces I. Emerging. J. Infectious Diseases. 2002. 8 (3) 881-890. International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456 www.ijtsrd.com | Volume – 3 | Issue – 1 | Nov-Dec 2018 Anjali Sharma, Dr. A. V. Gomashe, ‘Biosurfactant production by bacterial species isolated from (2018) IJCRT | Volume 6, Issue 2 April 2018 | ISSN: 2320-2882. Anjali Sharma, Dr. A. V. Gomashe, Mayur Rohi, Isolation and Screening of Biosurfactant Producing Bacterial Species from Petroleum Oil International Journal of Scientific Progress and Research (IJSPR) ISSN: 4689 Issue 120, Volume 42, Number 03, Benincasa, M., Marqués, A., Pinazo, A. and Manresa, A. (2010). Rhamnolipids surfactants: gies. In. Sen,. R. , pages. 170 - 184. New and Springer Science Banat IM, Franzetti A, Gandolfi I, Bestetti G, Martinotti MG, Fracchia L, Smyth TJ, Marchant R. Microbial biosurfactants production, Appl Microbiol Bailey, R.W. (1958). The reaction of pentoses Biochemical Journal, 68: 669 - Bouchez, M., Blanchet, D. and Vandacasteele, J. P. (1995). Degradation of polycyclic aromatic hydrocarbons by pure strains and defined strain associations: inhibition phenomena and Applied Microbiology Bordoloi, N. K. and Konwar, B. K. 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Shoeb, E., Akhlaq, F., Badar, U., Akhter, J. and Imtiaz, S. (2013). Classification and industrial applications of biosurfactants. Applied Sciences, 4 (3): 243 16. W. R. Finnerty, “Microbial c hydrocarbons products: commercial perspectives,” in Bioconversion of Waste Products, A. M. Martin, Ed., Elsevier, Amsterdam, The Netherlands, 1991. 17. Xia, W-J., Dong, H-P., Yu, L. and Yu, D (2011). Comparative study of biosurfactant produced by microorganisms isolated from formation water of petroleum reservoir. and Surfaces A: Physicochemical and Engineering Aspects, 392: 124 18. Zhang, Y. and Miller, R. M. (1992). Enhancement of octadecane dispersion and biodegradation by a Pseudomonas rhamnolipid biosurfactant. and Environmental Microbiology 3282. International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456-6470 Dec 2018 Page: 732 Bacterial Biofilms: From the Natural Environmental to Infectious Disease. Nature Review Microbiology. 2004. 2; 95-108 Khopade, A. N, R. Biao X. Liu,, K Mahadik, L Zhang, K. R. Kokare (2012). Production and stability studies of the biosurfactant isolated from marine B4 species, Elsevier, 198-204. Mullignn, C.N. (2005) Environmental Applications for Biosurfactants. Environmental Oliveira R, Melo L, Oliveira A, Salgueiro R (1994). Polysaccharide production and biofilm Pseudomonas fluorescens: effects of pH and surface material. Colloids Surf. 2:41-46. Chavez G, Aguirre-Ramirez M, Sanchez R(December2005). “The Pseudomonas aeruginosa RhIA enzyme is involved in rhamnolipid and polyhydroxyalkanoate production”.J. Ind. Microbiol. Biotechol.32 (11-12):675-7. Shoeb, E., Akhlaq, F., Badar, U., Akhter, J. and Imtiaz, S. (2013). Classification and industrial applications of biosurfactants. Part-I: Natural and , 4 (3): 243 - 252. W. R. Finnerty, “Microbial conversion of hydrocarbons products: commercial perspectives,” Bioconversion of Waste Materials to Industrial , A. M. Martin, Ed., Elsevier, Amsterdam, The Netherlands, 1991. P., Yu, L. and Yu, D-F. (2011). Comparative study of biosurfactant produced by microorganisms isolated from formation water of petroleum reservoir. Colloids Physicochemical and , 392: 124 - 130. Y. and Miller, R. M. (1992). Enhancement of octadecane dispersion and biodegradation by a rhamnolipid biosurfactant. Applied and Environmental Microbiology, 58: 3276 -