Rubber Seed Kernel as Potent Solid Substrate for the Production of Lipase by Pseudomonas aeruginosa Strain BUP2

K. N. Unni, Panichikkal Abdul Faisal, Prakasan Priji, Sreedharan Sajith, Sreedharan Sreedevi, E. S. Hareesh, Trikariyoor Asokan Nidheesh Roy, Sailas Benjamin

Enzyme Technology Laboratory, School of Biosciences, University of Calicut, Kerala, India.

DOI: 10.4236/aer.2015.32004                

Abstract

This study explored the utility of flours of rubber seed, coconut and groundnut kernels, and de-oiled cakes of coconut and groundnut as solid substrate for the production of lipase by Pseudomonas aeruginosa strain BUP2 (MTCC No. 5924), a novel bacterium reported from the rumen of Malabari goat. Various proportions (10%, 20%, 30%, 40% or 50%) of flours or cakes were prepared (w/v) with BUP medium (pH 4, 5, 6, 7 or 8), and incubated at different temperature (25°C, 28°C, 30°C or 32°C) for 24 to 96 h. The samples were assayed for lipase activity at 24 h intervals. The rubber seed flour (20%)-BUP medium supported the maximum lipase production (871 U/gds) at 48h incubation (pH 6, 28°C), followed by ground nut flour (398 U/gds), while ground nut cake supported the least lipase production (244 U/gds). From this, it is evident that the cheaply available rubber seed is an efficient substrate for the production of lipase, irrespective of its known demerit that it contains the limarin, a toxin; in fact, we could not detect limarin in the fermented matter. Thus, the utility of rubber seed for the production of a costly enzyme is reported from a novel rumen bacterium, which would be advantageous for rubber farmers.

Keywords

Pseudomonas aeruginosa BUP2, Lipase, Solid-State Fermentation, BUP Medium, Rubber Seed Flour

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	Pandey, A., Benjamin, S., Soccol, C.R., Nigam, P., Krieger, N. and Soccol, V.T. (1999) The Realm of Microbial Lipases in Biotechnology. Biotechnology and Applied Biochemistry, 29, 119-131.
[2]	Benjamin, S. and Pandey, A. (1997) Coconut Cake—A Potent Substrate for the Production of Lipase by Candida rugosa in Solid-State Fermentation. Acta Biotechnologica, 17, 241-251. http://dx.doi.org/10.1002/abio.370170308
[3]	Alkan, H., Baysal, Z., Uyar, F. and Dogru, M. (2007) Production of Lipase by a Newly Isolated Bacillus coagulans under Solid-State Fermentation Using Melon Waste. Applied Biochemistry and Biotechnology, 136, 183-192. http://dx.doi.org/10.1007/BF02686016
[4]	Kempka, A.P., Lipke, N.L., Pinheiro, T.D.L.F., Menoncin, S., Treichel, H., Freire, D.M.G, Luccio, M.D. and de Oliveira, D. (2008) Response Surface Method to Optimize the Production and Characterization of Lipase from Penicillium verrucosum in Solid-State Fermentation. Bioprocess and Biosystems Engineering, 31, 119-125. http://dx.doi.org/10.1007/s00449-007-0154-8
[5]	Aravindan, R., Anbumathi, P. and Viruthagiri, T. (2007) Lipase Applications in Food Industry. Indian Journal of Biotechnology, 6, 141-158.
[6]	Unni, K.N., Priji, P., Geoffroy, V.A., Doble, M. and Benjamin, S. (2014) Pseudomonas aeruginosa BUP2—A Novel Strain Isolated from Malabari Goat Produces Type 2 Pyoverdine. Advances in Bioscience and Biotechnology, 5, 874- 855.
[7]	Kilcawley, K.N., Wilkinson, M.G. and Fox, P.F. (2002) Determination of Key Enzyme Activities in Commercial Peptidase and Lipase Preparations from Microbial or Animal Sources. Enzyme and Microbial Technology, 31, 310-320. http://dx.doi.org/10.1016/S0141-0229(02)00136-9
[8]	Ramachandran, S.S., Singh, S.K., Larroche, C., Soccol, C.R. and Pandey, A. (2007) Oil Cakes and Their Biotechnological Applications—A Review. Bioresource Technology, 98, 2000-2009. http://dx.doi.org/10.1016/j.biortech.2006.08.002
[9]	Chakraborty, R. and Srinivasan, M. (1993) Production of a Thermostable Alkaline Protease by a New Pseudomonas sp. by Solid Substrate Fermentation. Journal of Microbiology and Biotechnoogy, 8, 7-16.
[10]	Benjamin, S. and Pandey, A. (1998) Mixed-Solid Substrate Fermentation—A Novel Process for Enhanced Lipase Production by Candida rugosa. Acta Biotechnologica, 18, 315-324. http://dx.doi.org/10.1002/abio.370180405
[11]	Kaur, S., Vohra, R.M., Kapoor, M., Beg, Q.K. and Hoondal, G.S. (2001) Enhanced Production and Characterization of Highly Thermostable Alkaline Protease from Bacillus sp. P-2. World Journal of Microbiology and Biotechnology, 17, 125-129. http://dx.doi.org/10.1023/A:1016637528648
[12]	Smitha, R.B., Jisha, V.N., Pradeep, S., Josh, M.S. and Benjamin, S. (2013) Potato Flour Mediated Solid-State Fermentation for the Enhanced Production of Bacillus thuringiensis-Toxin. Journal of Bioscience and Bioengineering, 116, 595-601. http://dx.doi.org/10.1016/j.jbiosc.2013.05.008
[13]	Jisha, V.N., Smitha, R.B., Priji, P., Sajith, S. and Benjamin, S. (2014) Biphasic Fermentation Is an Efficient Strategy for the Overproduction of δ-Endotoxin from Bacillus thuringiensis. Applied Biochemistry and Biotechnology, 175, 1519-1535.
[14]	Selvakumar, P. and Pandey, A. (1999) Solid-State Fermentation for the Synthesis of Inulinase from the Strains of Staphylococcus sp. and Kluyveromyces marxianus. Process Biochemistry, 34, 851-855. http://dx.doi.org/10.1016/S0032-9592(99)00008-4
[15]	Prakasham, R.S., Rao, C.S. and Sarma, P.N. (2006) Green Gram Husk—An Inexpensive Substrate for Alkaline Protease Production by Bacillus sp. in Solid-State Fermentation. Bioresource Technology, 97, 1449-1454. http://dx.doi.org/10.1016/j.biortech.2005.07.015
[16]	Benjamin, S. and Pandey, A. (1996) Optimization of Liquid Media for Lipase Production by Candida rugosa. Bioresource Technology, 55, 167-170. http://dx.doi.org/10.1016/0960-8524(95)00194-8
[17]	Singhania, R.R., Soccol, C.R. and Pandey, A. (2008) Application of Tropical Agro-Industrial Residues as Substrate for Solid-State Fermentation Processes. In: Pandey, A., Soccol, C.R. and Larroche, C., Eds., Current Development in Solid-State Fermentation, Springer, New York, 412-442. http://dx.doi.org/10.1007/978-0-387-75213-6_18
[18]	Faisal, P.A., Hareesh, E.S., Priji, P., Unni, K.N., Sajith, S., Sreedevi, S., Josh, M.S. and Benjamin, S. (2014) Optimization of Parameters for the Production of Lipase from Pseudomonas sp. BUP6 by Solid State Fermentation. Advances in Enzyme Research, 2, 125-133. http://dx.doi.org/10.4236/aer.2014.24013
[19]	Mahanta, N., Gupta, A. and Khare, S.K. (2008) Production of Protease and Lipase by Solvent Tolerant Pseudomonas aeruginosa PseA in Solid-State Fermentation Using Jatropha curcas Seed Cake as Substrate. Bioresource Technology, 99, 1729-1735. http://dx.doi.org/10.1016/j.biortech.2007.03.046