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http://dx.doi.org/10.48022/mbl.2004.04007

Isolation of a Lipolytic and Proteolytic Bacillus licheniformis from Refinery Oily Sludge and Optimization of Culture Conditions for Production of the Enzymes  

Devi, Sashi Prava (Microbial Ecology Laboratory, Department of Botany, Gauhati University)
Jha, Dhruva Kumar (Microbial Ecology Laboratory, Department of Botany, Gauhati University)
Publication Information
Microbiology and Biotechnology Letters / v.48, no.4, 2020 , pp. 515-524 More about this Journal
Abstract
With the increasing demand for enzymes in industrial applications there is a growing need to easily produce industrially important microbial enzymes. This study was carried out to screen the indigenous refinery bacterial isolates for their production of two industrially important enzymes i.e. lipase and protease. A total of 15 bacterial strains were isolated using Soil Extract Agar media from the oil-contaminated environment and one was shown to produce high quality lipase and protease enzymes. The culture conditions (culture duration, temperature, source of nitrogen, carbon, and pH) were optimized to produce the optimum amount of both the lipase (37.6 ± 0.2 Uml-1) and the protease (41 ± 0.4 Uml-1) from this isolate. Productivity of both enzymes was shown to be maximized at pH 7.5 in a medium containing yeast extract and peptone as nitrogen sources and sucrose and galactose as carbon sources when incubated at 35 ± 1℃ for 48 h. Bacterial strain SAB06 was identified as Bacillus licheniformis (MT250345) based on biochemical, morphological, and molecular characteristics. Further studies are required to evaluate and optimize the purification and characterization of these enzymes before they can be recommended for industrial or environmental applications.
Keywords
Bacillus licheniformis; refinery sludge; enzymes; optimization; cultural conditions;
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1 Bhattacharyya JK, Shekdar AV. 2003. Treatment and disposal of refinery sludges: Indian scenario. Waste Manag. Res. 21: 249-261.   DOI
2 Patil KJ, Chopda MZ, Mahajan RT. 2011. Lipase biodiversity. Indian J. Sci. Technol. 4: 971-982.   DOI
3 Sahu GK, Martin M. 2011. Optimization of growth conditions for the production of extracellular lipase by bacterial strains from dairy industry effluents. Biotechnol. Bioinf. Bioeng. 1: 305-311.
4 Hasan F, Shah AA, Hameed A. 2006. Industrial applications of microbial lipases. Enzyme Microb. Technol. 39: 235-251.   DOI
5 Bharathi D, Rajalakshmi G, Komathi S. 2018. Optimization and production of lipase enzyme from bacterial strains isolated from petrol spilled soil. J. King Saud. Univ. Sci. 31: 1-4.   DOI
6 Salihu A, Alam MZ, AbdulKarim MI, Salleh HM. 2012. Lipase production: an insight in the utilization of renewable agricultural residues. Resour. Conserv. Recy. 58: 36-44.   DOI
7 Jaeger KE, Ransac S, Dijkstra BW, Colson C, Heuvel M, Misset O. 1994. Bacterial lipases. FEMS Microbiol. Rev. 15: 29-63.   DOI
8 Bajpai P. 1999. Application of enzymes in the pulp and paper industry. Biotechnol. Progr. 15: 147-157.   DOI
9 Gupta R, Gupta N, Rathi P. 2004. Bacterial lipases: an overview of production, purification and biochemical properties. Appl. Microbiol. Biotechnol. 64: 763-781.   DOI
10 Sharma R, Chisti Y, Banerjee UC. 2001. Production, purification, characterization, and applications of lipases. Biotechnol. Adv. 19: 627-662.   DOI
11 Pandey A, Benjamin S, Soccol CRP, Nigam, Krieger N, Soccol VT. 1999. The realm of microbial lipases in biotechnology. Biotechnol. Appl. Biochem. 29: 119-131.
12 Rao MB, Tanksale AM, Ghatge MS, Deshpande VV. 1998. Molecular and biotechnological aspects of microbial proteases. Microbiol. Mol. Biol. Rev. 62: 597-635.   DOI
13 Karigar CS, Rao SS. 2011. Role of microbial enzymes in the bioremediation of pollutants: a review. Enzyme Res. 2011: 805187.
14 Puri S, Beg QK, Gupta R. 2002. Optimization of alkaline protease production from Bacillus sp. by response surface methodology. Curr. Micobiol. 44: 286-290.   DOI
15 Kumar M, Rejitha R, Devika S, Balakumaran MD, Immaculate NR, Kalaichelvan PT. 2012. Production, optimization and purification of lipase from Bacillus sp. MPTK 912 isolated from oil mill effluent. Adv. Appl. Sci. Res. 3: 930-938.
16 Nigam P. 2013. Microbial enzymes with special characteristics for biotechnological applications. Biomolecules 3: 597-611.   DOI
17 Sarker PK, Talukdar SA, Deb P, Sayem SA, Moshina K. 2013. Optimization and partial characterization of culture conditions for the production of alkaline protease from Bacillus licheniformis P003. SpringerPlus. 2: 506.   DOI
18 Hamaki T, Suzuki M, Fudou R, Jojima Y, Kajiura T, Tabuchi A, et al. 2005. Isolation of novel bacteria and actinomycetes using soil-extract agar medium, J. Biosci. Bioeng. 99: 485-492.   DOI
19 Lee LP, Karbul HM, Citartan M, Gopinath SCB, Lakshmipriya, Tang TH. 2015. Lipase-secreting Bacillus species in an oil-contaminated habitat: Promising strains to alleviate oil pollution. BioMed. Res. Int. 2015: 820575.
20 Suganthi C, Mageswari A, Karthikeyan S, Anbalagan M, Sivakumar A, Gothandam KM. 2013. Screening and optimization of protease production from a halotolerant Bacillus licheniformis isolated from saltern sediments. J. Genet. Eng. Biotechnol. 11: 47-52.   DOI
21 Aneja KR. 2003. Experiments in Microbiology, Plant Pathology and Biotechnology. New age international publication, pp. 245-275. New Delhi. Fourth edition.
22 Holt JG. 1994. Bergey's Manual of Determinative Bacteriology. 9th Ed. William & Wilkins, Baltimore,USA
23 Winkler UK, Stuckmann M. 1979. Glucogen, hyaluronate and some other polysaccharides greatly enhance the formation of exolipase by serratia marcescens. J. Bacteriol. 138: 663-670.   DOI
24 Sambrook J, Fritsch EF, Maniatis T. 1989. Molecular Cloning: A Laboratory Manual. 2nd Ed. Cold Spring Harbor Laboratory Press. Cold Spring Harbor, New York.
25 Saitou N, Nei M. 1987. The neighbor-joining method: A new method for reconstructing phylogenetic trees. Mol. Biol. Evol. 4: 406-425.
26 Kumar S, Stecher G, Tamura K. 2016. MEGA7: Molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol. Biol. Evol. 33: 1870-1874.   DOI
27 Kilcawley KN, Wilkinson MG, Fox PF. 2002. Determination of key enzyme activities in commercial peptidase and lipase preparations from microbial or animal sources. Enzym. Microb. Technol. 31: 310-320.   DOI
28 Beg QK, Sahai V, Gupta R. 2003. Statistical media optimization and alkaline protease Production from Bacillus mojavensis in a bioreactor Process. Biochem. 39: 203.   DOI
29 Margesin R, Zimmerbauer A, Schinner F. 1999. Soil lipase activity-A useful indicator of oil biodegradation. Biotechnol. Tech. 13: 859-863.   DOI
30 Riffaldi R, Levi-Minzi R, Cardelli R, Palumbo S, Saviozzi A. 2006. Soil biological activities in monitoring the bioremediation of diesel oil-contaminated soil. Water Air Soil Pollut. 170 : 3-15.   DOI
31 Venugopal M, Saramma AV. 2007. An alkaline protease from Bacillus circulans BM15, newly isolated from a mangrove station: characterization and application in laundry detergent formulations. Indian J. Microbiol. 47: 298-303.   DOI
32 Hasan F, Shah AA, Hameed A. 2009. Methods for detection and characterization of lipases: A comprehensive review. Biotechnol. Adv. 27: 782-798.   DOI
33 Sonune N, Garode A. 2018. Isolation, characterization and identification of extracellular enzyme producer Bacillus licheniformis from municipal wastewater and evaluation of their biodegradability. Biotechnol. Res. Innov. 2: 37-44.   DOI
34 Salleh AB, Musani RM, Basri K, Ampon K, Yunus WMZ, Razak CNA. 1993. Extra- and intra-cellular lipases from a thermophilic Rhizopus oryzae and factors affecting their production. Can. J. Microbiol. 39: 978-981.   DOI
35 Xiong Y, Wang Y, Yu Y, Li Q, Wang H, Chen R, et al. 2010. Production and characterization of a novel bioflocculant from Bacillus licheniformis. Appl. Environ. Microbiol. 76: 2778-2782.   DOI
36 Willerding AL, Oliveira LA, Moreira FW, Germano MG, Chagas Jr. AF. 2011. Lipase activity among bacteria isolated from Amazonian soils. Enzyme Res. 2011: 720194.
37 Abada EAE. 2008. Production and characterization of a mesophilic lipase isolated from Bacillus stearothermophilus AB-1. Pakistan J. Biol. Sci. 11: 1100-1106.   DOI
38 Dong H, Gao S, Han S, Cao S. 1999. Purification and characterization of a Pseudomonas sp. lipase and its properties in non-aqueous media. Biotechnol. Appl. Biochem. 30: 251-256.
39 Dalmau E, Montesinos JL, Lotti M, Casas C. 2000. Effect of different carbon sources on lipase production by Candida rugosa. Enzyme Microb. Technol. 26: 657-663.   DOI
40 Chang RC, Chou SJ, Shaw JF. 1994. Multiple forms and functions of Candida rugosa lipase. Biotechnol. Appl. Biochem. 19: 93-97.
41 Ghanem EH, Al-Sayeed HA, Saleh KM. 2000. An alkalophilic thermostable lipase produced by a new isolate of Bacillus alcalophilus. World. J. Microbiol. Biotechnol. 16: 459-464.   DOI
42 Joo HS, Kumar CG, Park GC, Paik SR, Chang CS. 2003.Oxidant and SDS-stable alkaline protease from Bacillus clausii I-52: production and some properties. J. Appl. Microbiol. 95: 267-272.   DOI
43 Nadeem M, Qazi JI, Baig S, Syed Q. 2006. Effect of medium composition on commercially important alkaline protease production by Bacillus licheniformis N-2. Food Technol. Biotechnol. 46: 388-394.