Effect of Glucose on Biosurfactant Production using Bacterial Isolates from Oil Contaminated Sites

1. International Journal of Trend in International Open Access International Open Access Journal | www.ijtsrd.com International Journal of Trend in Scientific Research and Development (IJTSRD) Research and Development (IJTSRD) www.ijtsrd.com ISSN No: 2456 ISSN No: 2456 - 6470 | Volume - 3 | Issue – 1 | Nov 1 | Nov – Dec 2018 Effect of Glucose Bacterial Isolates f Glucose on Biosurfactant Production Isolates from Oil Contaminated Sites Anjali Sharma n Biosurfactant Production using rom Oil Contaminated Sites Lecturer ecturer, Department of Microbiology, Shri Shivaji Science College Nagpur, Maharashtra, India 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 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 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, 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] 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 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 physiological and Biosurfactants, Bacterial isolates, Glucose medium, Minimal Salt Medium. MATERIALS AND METHODS: glucose as a carbon source hiphatic compounds that are bacterial on production of biosurfactant, Minimal Salt Medium as the basal medium. The concentration of was kept at 1% (w/v). 1% inoculum species 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 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 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 @ IJTSRD | Available Online @ www.ijtsrd.com www.ijtsrd.com | Volume – 3 | Issue – 1 | Nov-Dec 2018 Dec 2018 Page: 730

2. International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456 International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456 International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456-6470 RESULTS AND DISCUSSION: Table.1. Effect of Glucose as Carbon Sources 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 58 80 89 96 68 75 67 54 66 1.2 8 7 2 pH = 7.00, Temperature: 37 ⁰C. Inoculum = 1 vol%, engine oil = 2 vol%, Incubation time = 7days, Inoculum = 1 vol%, engine oil = 2 vol%, Incubation time = 7days, Bacteri al isolates EI % O.D 600 nm IH D 13 78 IH D 19 88 1.8 2 IH D 21 69 IH D 36 76 1.7 6 IH D IH D IH D IH D IH D 112 76 IH D 148 62 IH D 152 73 IH D 157 79 IH D 176 77 IH D 178 69 IH D 188 71 IH D 3 64 1.0 0.6 0.7 1.6 0.8 1.5 0.9 0.6 0.4 0.7 0.8 1 0.6 0.9 Effect of Glucose Effect of Glucose 100 90 88 80 79 78 77 76 76 70 75 73 71 69 69 68 67 66 60 64 62 50 54 40 30 20 10 0 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 soilwere observed by using glucose as the source of carbon along with engine oil where the maximum value of emulsification index where the maximum value of emulsification index 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 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 Malaysiana, 42 (5): 615 - 623. 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 Maximum growth of bacterial cell and biosurfactant bacterial species isolated from inon, H., Noramiza, S. and Shahidan, R. (2013). Screening of biosurfactants produced by the hydrocarbon degrading were observed by using bacteria. bacteria. Sains glucose as the source of carbon along with engine oil 623. @ IJTSRD | Available Online @ www.ijtsrd.com www.ijtsrd.com | Volume – 3 | Issue – 1 | Nov-Dec 2018 Dec 2018 Page: 731

3. International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456 International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456 International Journal of Trend in Scientific Research and Development (IJTSRD) ISSN: 2456-6470 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 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 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 cometabolism. Applied 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. 10.Hall-Stoodley L., et al. Bacterial Biofilms: From the Natural Environmental to Infectious Disease Nature Review Microbiology. 2004. 2; 95 11.Khopade, A. N, R. Biao X. Liu,, K Mahadik, L Zhang, K. R. Kokare (2012). Production and stability studies of the biosurfactant isolat marine B4 species, Elsevier, 198 12.Mullignn, C.N. Applications for Biosurfactants. Pollution. 133,183-198 13.Oliveira R, Melo L, Oliveira A, Salgueiro R (1994). Polysaccharide production and biofilm formation by Pseudomonas fluorescens pH and surface material. Colloids Surf. 2:41 14.Soberon-Chavez G, Aguirre R(December2005). “The Pseudomonas aeruginosa RhIA enzyme is involved in rhamnolipid and polyhydroxyalkanoate Microbiol. Biotechol.32 (11 15.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. 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: 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. Anjali Sharma, Dr. A. V. Gomashe, ‘Biosurfactant production by bacterial species isolated from Bacterial Biofilms: From the Natural Environmental to Infectious Disease. Nature Review Microbiology. 2004. 2; 95-108 (2018) IJCRT | Volume 6, Issue 2 April 2018 | ISSN: 2320-2882. 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. Anjali Sharma, Dr. A. V. Gomashe, Mayur Rohi, Isolation and Screening Producing Bacterial Species from Petroleum Oil International Journal of Scientific Progress and Research (IJSPR) ISSN: 4689 Issue 120, Volume 42, Number 03, of of Biosurfactant Biosurfactant Mullignn, Applications for Biosurfactants. Environmental C.N. (2005) (2005) Environmental 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. Benincasa, M., Marqués, A., Pinazo, A. and Manresa, A. (2010). Rhamnolipids surfactants: gies. In. Sen,. R. - 184. New and Springer Science Chavez G, Aguirre-Ramirez M, Sanchez R(December2005). “The Pseudomonas aeruginosa RhIA enzyme is involved in rhamnolipid and polyhydroxyalkanoate Microbiol. Biotechol.32 (11-12):675-7. Banat IM, Franzetti A, Gandolfi I, Bestetti G, Martinotti MG, Fracchia L, Smyth TJ, Marchant R. Microbial biosurfactants production”.J. production”.J. In Ind. produc production, Appl Microbiol 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. Bailey, R.W. (1958). The reaction of pentoses Biochemical Journal, 68: 669 - W. R. Finnerty, “Microbial conversion of hydrocarbons products: commercial perspectives,” Bioconversion of WasteMaterials to Industrial , A. M. Amsterdam, The Netherlands, 1991. Bouchez, M., Blanchet, D. and Vandacasteele, J. P. (1995). Degradation of polycyclic aromatic hydrocarbons by pure strains and defined strain associations: inhibition Applied Martin, Martin, Ed., Ed., Elsevier, Elsevier, phenomena Microbiology Microbiology phenomena and and P., Yu, L. and Yu, D-F. (2011). Comparative study of biosurfactant produced by microorganisms isolated from formation water of petroleum reservoir. Colloids Bordoloi, N. K. and Konwar, B. K. (2009). iosurfactant in enhancing solubility and metabolism of petroleum hydrocarbons. Journal Physicochemical Physicochemical , 392: 124 - 130. and and 505. Y. and Miller, R. M. (1992). Enhancement of octadecane dispersion and biodegradation by a rhamnolipid biosurfactant. Applied and Environmental Microbiology, 58: 3276 - Biofilms: Microbial Life on Surfaces J. Infectious Diseases. 2002. 8 (3) @ IJTSRD | Available Online @ www.ijtsrd.com www.ijtsrd.com | Volume – 3 | Issue – 1 | Nov-Dec 2018 Dec 2018 Page: 732