Skip to main content
SpringerLink
Log in

Purification and Characterization of Extracellular Lipase from Serratia marcescens VITSD2

  • Research Article
  • Published:
Proceedings of the National Academy of Sciences, India Section B: Biological Sciences Aims and scope Submit manuscript

Abstract

Lipases represent a group of extracellular enzymes and hold a prominent place among the enzymes. In the present research work Serratia marcescens isolated from soil was selected for lipase production. Effects of various parameters such as substrate, pH, temperature, incubation time and inoculum size were analysed for the enhanced lipase production. The optimum substrate was found to be olive oil with the concentration of 0.7 mL in 100 mL of production medium. Serratia marcescens produced maximum lipase after 48 h of incubation at 30 °C with the optimum pH as 7.0. The inoculum concentration of 4 % was more favourable for the production of lipase. The extracted lipase was purified by two steps: ultra-filtration and Carboxymethyl(CM)-Cellulose chromatography. The specific activity of the purified enzyme was determined to be 65.0 U/mg with 2.55 purification fold and yield of 67.5 %. Polyacrylamide gel electrophoresis (SDS-PAGE) analysis revealed the presence of a single band with a molecular weight of around 35 kDa. Based on Fourier transformation-infrared spectroscopy analysis the functional groups of lipase were determined and high performance liquid chromatography analysis with the retention time of 1.995 min reveals the presence of lipase in the purified extract.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  1. Kirk O, Borchert TV, Fuglsang CC (2002) Industrial enzyme applications. Curr Opin Biotechnol 13:345–351

    Article  CAS  PubMed  Google Scholar 

  2. Jaeger KE, Ransac S, Dijkstra BW, Colson C, Heuvel MV, Misset O (1994) Bacterial lipases. FEMS Microbiol Rev 15:29–63

    Article  CAS  PubMed  Google Scholar 

  3. Jaeger KE, Dijkstra BW, Reetz MT (1999) Bacterial biocatalysts: molecular biology, three-dimensional structures and biotechnological applications of lipases. Ann Rev Microbiol 53:315–351

    Article  CAS  Google Scholar 

  4. Pandey A, Benjamin S, Nigam CP, Krieger N, Soccol V (1999) The realm of microbial lipases in biotechnology. Biotechnol Appl Biochem 29:119–131

    CAS  PubMed  Google Scholar 

  5. Gupta R, Gupta N, Rathi P (2004) Bacterial lipases: an overview of production, purification and biochemical properties. Appl Microbiol Biotechnol 64:763–781

    Article  CAS  PubMed  Google Scholar 

  6. Immanuel G, Jwbadhas A, Palaresam A (2008) Investigation of lipase production by milk isolate Serratia rubidaea. Food Technol Biotechnol 46:60–65

    CAS  Google Scholar 

  7. Wiseman A (1995) Introduction to principles. In: Wiseman A (ed) Handbook of enzyme biotechnology. T.J. Press Ltd, Ellis Horwood Ltd, Padstow, Cornwall, UK, pp 3–8

    Google Scholar 

  8. Abdou AM (2003) Purification and partial characterization of psychotropic Serratia marcescens lipase. J Dairy Sci 86:127–132

    Article  CAS  PubMed  Google Scholar 

  9. Hines DA, Saurugger PN, Ihler GM, Benedik MJ (1998) Genetic analysis of extracellular proteins of Serratia marcescens. J Bacteriol 170:4141–4146

    Article  Google Scholar 

  10. Alquati C, Gioia LD, Lotti M (2002) The cold-active lipase of Pseudomonas fragi. Eur J Biochem 269:3321–3328

    Article  CAS  PubMed  Google Scholar 

  11. Lima V, Mitchell D, Fontana JD (2003) Effect of nitrogen and carbon sources on lipase production by Penicillium aurantiogriseum. Food Technol Biotechnol 41:105–110

    CAS  Google Scholar 

  12. Jeuniaux CC (1996) Complex carbohydrates. In: Neufeld EF, Ginsburg V (eds) Methods in enzymology, vol 8. Academic Press Inc, New York, pp 644–650

    Google Scholar 

  13. Arima K, Liu WH, Beppu T (1972) Isolation and identification of lipolytic and thermophilic fungus. Agric Biol Chem 36:1913–1917

    Article  Google Scholar 

  14. Fox PF, Stepaniak L (1983) Isolation and some properties of extracellular heat-stable lipases from Pseudomonas fluorescens strain AFT-36. Dairy Res 50:77–89

    Article  CAS  Google Scholar 

  15. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Article  CAS  PubMed  Google Scholar 

  16. Senthilmurugan G, Viji S, Sekar L, Suresh K (2013) Enzyme analysis of endophytic new Streptomyces sp. viji10 isolated from velaman roots of orchid plant vandaspathulata (L) SPRENG. Asian J Agri Biol 1(13):149–154

    Google Scholar 

  17. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275

    CAS  PubMed  Google Scholar 

  18. Marzieh MN, Mohsen G, Ali R, Yousefi HA (2010) Fermentative Production of Dextran using food industry wastes. World Acad Sci Eng Technol 44:1241–1243

    Google Scholar 

  19. Sharma CK, Sharma PK, Kanwar SS (2012) Optimization of production conditions of lipase from B. licheniformis MTCC-10498. Res J Recent Sci 1:25–32

    CAS  Google Scholar 

  20. Sirisha EN, Rajasekar M, Lakshmi N (2010) Isolation and optimization of lipase producing bacteria from oil contaminated soils. Adv Biol Res 4:249–252

    CAS  Google Scholar 

  21. Tembhurkar VR, Kulkarni MB, Peshwe SA (2012) Optimization of lipase production by Pseudomonas sp. in submerged batch process in shake flask culture. Sci Res Report 2:2249–2321

    Google Scholar 

  22. Gerritse G, Hommes RWJ, Quax WJ (1998) Development of a lipase fermentation process that uses a recombinant Pseudomonas alcaligenes strain. Appl Environ Microbiol 64:2644–2651

    CAS  PubMed  PubMed Central  Google Scholar 

  23. Eltaweel MA, Rahman RNZRA, Salleh AB, Basri M (2005) An organic solvent-stable lipase from Bacillus sp. strain 42. Ann Microbiol 55:187–192

    CAS  Google Scholar 

  24. Lee DW, Koh YS, Kim KJ, Kim BC, Choi HJ, Kim DS (1999) Isolation and characterization of a thermophilic lipase from Bacillus thermoleovorans ID-1. FEMS Microbiol Lett 179:393–400

    Article  CAS  PubMed  Google Scholar 

  25. Chen J, Ishii T, Shimura S, Kirimura S, Usami S (1992) Lipase production of Tricosporon fermentans WUC12 a newly isolated yeast. J Ferment Bioeng 73:412–414

    Article  CAS  Google Scholar 

  26. Dharmsthiti S, Kuhasuntisuk B (1998) Lipase from Pseudomonas aeruginosa LP602: biochemical properties application for wastewater treatment. J Ind Microbiol Biotechnol 21:75–80

    Article  CAS  Google Scholar 

  27. Kanchan B, Aruna R, Ram R (2001) Identification, purification, and charcterization of a thermally stable lipase from rice bran. A new member of the (phospho) lipase family. Plant Physiol 127:1728–1738

    Article  Google Scholar 

  28. Rakesh K, Arpit S, Arun KDS (2012) Lipase from Bacillus pumilusRK31: production, purification and some properties. World Appl Sci J 16:940–948

    Google Scholar 

  29. Benaki DC, Anggeli A, Chryssikos GD, Yiannopoulos YD, Kamitsos EI, Brumley E, Case ST, Boden N, Hamodrakas SJ (1998) Laser-Raman: FT-IR spectroscopic studies of peptide analogues of silk mothchorion protein segments. Int J Biol Macromol 23:49–59

    Article  CAS  PubMed  Google Scholar 

  30. Raghavan D, Emekalam A (2000) Characterization of starch/polyethylene and starch polyethylene poly (lactic acid) composites. Polym Degrad Stab 72:509–517

    Article  Google Scholar 

Download references

Acknowledgments

Authors are thankful to VIT University for the constant encouragement, help and support for extending necessary facilities.

Author information

Authors and Affiliations

  1. School of Biosciences and Technology, VIT University, Vellore, Tamil Nadu, India

    V. Mohanasrinivasan, C. Subathra Devi, D. Jayasmita, E. Selvarajan & S. Jemimah Naine

Authors
  1. V. Mohanasrinivasan
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. C. Subathra Devi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. D. Jayasmita
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. E. Selvarajan
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. S. Jemimah Naine
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to C. Subathra Devi.

Ethics declarations

Conflict of Interest

Authors have no conflict of interest.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Mohanasrinivasan, V., Devi, C.S., Jayasmita, D. et al. Purification and Characterization of Extracellular Lipase from Serratia marcescens VITSD2. Proc. Natl. Acad. Sci., India, Sect. B Biol. Sci. 88, 373–381 (2018). https://doi.org/10.1007/s40011-016-0763-6

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s40011-016-0763-6

Keywords

Search

Navigation