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Optimization of Lipase Production Using Bacillus subtilis by Response Surface Methodology

Authors: A. Shyamala Devi, K. Chitra Devi, R. Rajendiran


A total of 6 isolates of Bacillus subtilis were isolated from oil mill waste collected in Namakkal district, Tamilnadu, India. The isolated bacteria were screened using lipase screening medium containing Tween 80. BS-3 isolate exhibited a greater clear zone than the others, indicating higher lipase activity. Therefore, this isolate was selected for media optimization studies. Ten process variables were screened using Plackett–Burman design and were further optimized by central composite design of response surface methodology for lipase production in submerged fermentation. Maximum lipase production of 16.627 U/min/ml were predicted in medium containing yeast extract (9.3636g), CaCl2 (0.8986g) and incubation periods (1.813 days). A mean value of 16.98 ± 0.2286 U/min/ml of lipase was acquired from real experiments.

Keywords: Bacillus subtilis, extracellular lipase, Plackett–Burman design, response surface methodology.

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[1] N. Kulkarni, and R.V. Gadre, "Production and properties of an alkaline, thermophilic lipase from Pseudomonas fluorescens NS2W”, Journal of Industrial Microbiology and Biotechnology, vol. 28, pp.344–348, 2002.
[2] S. Fodiloglu, and O. Erkmen, O, "Lipase production by Rhizopus oryzae growing on different carbon and nitrogen sources”. Journal of the Science of Food and Agriculture, vol. 79, pp.1936–1938, 1999.
[3] G. Corzo, and S. Revah, "Production and characteristics of the lipase from Yarrowia lipolytica 681”, Bioresource Technology, vol. 70, pp.173–180, 1999.
[4] P. Sommer, C. Bormann, and F. Gotz, "Genetic and biochemical characterization of a new extracellular lipase from Streptomyces cinnamomeus”, Applied and Environmental Microbiology, vol. 63, pp.3553–3560, 1997.
[5] M.B. Nthangeni, H.G. Patterton, A. van Tander, W.P. Vergeer, and D. Litthauer, "Over-expression and properties of a purified recombinant Bacillus licheniformis lipase: A comparative report on Bacillus lipases”. Enzyme and Microbial Technology, vol. 28, pp.705-712, 2001.
[6] R.N. Rahman, J.H. Chin, A.B. Salleh, and M. Basri, "Cloning and expression of a novel lipase gene from Bacillus sphaericus 205y”. Molecular Genetics and Genomics, vol. 269, pp.252-260, 2003.
[7] C. Ruiz, F.I. Javier Pastor, and P. Diaz, "Isolation and characterization of Bacillus sp. BP-6 LipA, a ubiquitous lipase among mesophilic Bacillus species”. Letters in Applied Microbiology, vol. 37, pp.354-359, 2003.
[8] M.L.M. Fernandes, E.B. Saad, J.A. Meira, L.P. Ramos, D.A. Mitchell, and N. Krieger, "Esterification and transesterification reactions catalysed by addition of fermented solids to organic reaction media, Journal of Molecular Catalysis B: Enzymatic, vol. 44, pp.8-13, 2007.
[9] E. Jurdao, F. Camacho, G. Luzon, and J.M. Viceria, "Kinetics models of activity of β-galactosidase: Influence of pH, ionic concentration and temperature”, Enzyme and Microbial Technology, vol. 34, pp.33-40, 2004.
[10] C. Tari, N. Gogus, and F. Tokatli, "Optimization of biomass, pellet size and polygalacturonase production by Aspergillus sojae ATCC 20235 using response surface methodology”, Enzyme and Microbial Technology, vol. 40, pp.1108-1116, 2007.
[11] F.P.Duta, F.P. de França, and L.M. de Almeida Lopes, "Optimization of culture conditions for exopolysaccharide production in Rhizobium sp. using the response surface method”, Electronic Journal of Biotechnology, vol. 9, pp.391-399, 2006.
[12] C. Liu, Y.Liu, W. Liao, Z. Wen, and S. Chen, "Application of statistically based experimental designs for the optimization of nisin production from whey”. Biotechnology Letters, vol. 25, pp.877-882, 2003.
[13] V.V. Dasu, and T. Panda, "Optimization of microbiological parameters for enhanced griseofulvin production using Response Surface Methodology”. Bioprocess Engineering, vol. 22, pp. 45 – 49, 2002.
[14] E. Haba, O. Bresco, C. Ferrer, A. Marques, M. Busquets, and A. Manresa, "Isolation of lipase screening bacteria by developing used frying oil as selective substrate”, Enzyme and Microbial Technology , vol. 26, pp.40-44, 2000.
[15] F. Pedreschi, K. Kaack, and K. Granby, "The effect of asparaginase on acrylamide formation in French fries,” Food Chemistry, vol.109,pp.386– 392,2008.
[16] S. Sadasivam, and A. Manickam, 1996. Biochemical methods, 2nd Eds. New age international (P) limited, New Delhi, 1996.
[17] H. Treichel, D. de Oliveira, M. A. Mazutti, M. D. Luccio, and J. Vladimir Oliveira, "A review on microbial lipases production food,” Bioprocess Technol., vol.3, pp.182–196, 2010.
[18] Y. Wang, K.C. Srivastava, G.J. Shen, and H.Y. Wang, "Thermostable alkaline lipase from a newly isolated thermophilic Bacillus strain, A30– 1 (ATCC 53841)”, Journal of Fermentation and Bioengineering, vol. 79, pp. 433–438, 1995.
[19] F. Hasan, A. A. Shah, and A. Hameed, "Industrial applications of microbial lipases”, Enzyme and Microbial Technology, vol. 39, pp. 235- 251, 2006.
[20] T. Selva Mohan, A. Palavesam, and G. Immanvel. "Isolation and characterization of lipase-producing Bacillus strains from oil mill waste, African Journal of Biotechnology, vol. 7 (15), pp. 2728-2735, 2008.
[21] D. Maccio, A. Fabra, and S. Castro, "Acidity and calcium interaction affect the growth of Bradyrhizobium sp. and attachment to peanut roots,” Soil Biology and Biochemistry, vol. 34, pp.201-208, 2002.
[22] R. Sharma, Y. Chisti, and U.C. Banerjee. "Production, purification, characterization, and applications of lipases”, Biotechnology Advances, vol. 19, pp.627-662, 2001.
[23] M. Hymavathi, T. Sathish, P. Brahmaiah, and R. S. Prakasham, "Impact of carbon and nitrogen sources on L-asparaginase production by isolated Bacillus circulans (MTCC 8574): Application of saturated Plackett- Burman design,” Chemical and Biochemical Engineering Quarterly, vol. 24. (4), pp. 473–480, 2010.
[24] J. H. Panwal, T. Viruthagiri and G. Baskar, "Statistical modeling and optimization of enzymatic milk fat splitting by soybean lecithin using response surface methodology”, International Journal of Nutrition and Metabolism, vol. 3(5), pp. 50–57, 2011.
[25] S. Akhnazarova, and V. Kefarov, Experiment optimization in chemistry and chemical engineering, Moscow, Mir Publisher, 1982.