Skip to main content
Log in

Optimization of extracellular psychrophilic alkaline lipase produced by marine Pseudomonas sp. (MSI057)

  • Original Paper
  • Published:
Bioprocess and Biosystems Engineering Aims and scope Submit manuscript

Abstract

An endosymbiotic Pseudomonas sp. (MSI057), which could produce high yields of lipase, was isolated from marine sponge Dendrilla nigra, collected from the peninsular coast of India. Maximum production of enzyme was obtained in minimal medium supplemented with 1% tributyrin. Catabolite repression was observed when the medium was supplemented with readily available carbon sources. The optimum temperature and pH for the enzyme production was 30 °C and 9.0, respectively. The enzyme exhibited maximum activity in pH range of 8–9 with an optimum pH 9.0. The activity of purified enzyme was optimum at 37 °C and showed 80% activity at 20 °C and the enzyme activity decreased dramatically above 50 °C. Based on the present findings, the enzyme was characterized as psychrophilic alkaline lipase, which can be developed for industrial applications.

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

Access this article

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

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9
Fig. 10

Similar content being viewed by others

References

  1. Abbas H, Hiol A, Deyris V, Comeau L (2002) Isolation and characterization of an extracellular lipase from Mucor sp strain isolated from palm fruit. Enzyme Microbial Technol 31:968–975

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  3. Altschul SF, Gise W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment tool. J Mol Biol l215:403–410

    Google Scholar 

  4. Altschul SF, Thomas LM, Alejandro AS, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402

    Article  CAS  Google Scholar 

  5. Benjamin S, Pandey A (1997) Coconut cake—a potent substrate for the production of lipase by Candida rugosa in solid state fermentation. Acta Biotechnol 17:241–251

    Article  CAS  Google Scholar 

  6. Bora L, Kalita MC (2007) Production and optimization of thermostable lipase from a thermophilic Bacillus sp LBN 4. Internet J Microbiol 4

  7. Burkert JFM, Maugeri F, Rodrigues MI (2004) Optimization of extracellular lipase production by Geotrichum sp using factorial design. Bioresour Technol 91:77–84

    Article  CAS  Google Scholar 

  8. Camargo MM, Morais MA, Melo EHM, Lima FJL (1998) Production of extracellular lipase by a Candida rugosa strain isolated in Pernambuco. Bras Rev Microb 29:134–137

    Google Scholar 

  9. Cappucino JG, Sherman N (2004) Microbiology: a laboratory manual. Pearson Education, Singapore, pp 491

    Google Scholar 

  10. Collins CH, Lyne PM, Grange JM (1989) Microbiological methods. Butterworths, London

    Google Scholar 

  11. Corzo G, Revah S (1999) Production and characterization of the lipase from Yarrowia lipolytica 681. Bioresour Technol 70:173–180

    Article  CAS  Google Scholar 

  12. Deive FJ, Costas M, Longo MA (2003) Production of thermostable extracellular lipase by Kluyveromyces marxianus. Biotechnol Lett 25:1403–1406

    Article  CAS  Google Scholar 

  13. Dimitris P, Paul C, Dimitris K, Basil JM (1992) Optimizing production of extracellular lipase from Rhodotorula glutinis. Biotechnol Lett 14:397–402

    Article  Google Scholar 

  14. Enticknap JJ, Kelly M, Peraud O, Hill RT (2006) Characterization of a culturable alphaproteobacterial symbiont common to many marine sponges and evidence for vertical transmission via sponge larvae. Appl Environ Microbiol 72:3724–3732

    Article  CAS  Google Scholar 

  15. Fadiloglu S, Erkmen O (2002) Effects of carbon and nitrogen sources on lipase production by Candida rugosa. Turk J Eng Environ Sci 26:249–254

    CAS  Google Scholar 

  16. Fadnavis NW, Deshpande A (2002) Synthetic applications of enzymes entrapped in reverse micelles and organo-gels. Curr Organic Chem 6:393–410

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  18. Gao XG, Cao SG, Zhang KC (2000) Production, properties and application to non-aqueous enzymatic catalysis of lipase from a newly isolated Pseudomonas strain. Enzyme Microbiol Technol 27:74–82

    Article  Google Scholar 

  19. George V, Diwan AM (1993) Simultaneous staining of proteins during polyacrylamide gel electrophoresis in acidic gels by counter migration of Coomasie brilliant blue R-250. Anal Bacteriol 4:662–666

    Google Scholar 

  20. Gerday C, Aittaleb M, Bentahir M, Chessa JP, Claverie P, Collins T, D’Amico S, Dumont J, Garsoux G, Georlette D, Hoyoux A, Lonhienne T, Meuwis M, Feller G (2000) Cold adapted enzymes: from fundamentals to biotechnology. Trends Biotechnol 18:103–107

    Article  CAS  Google Scholar 

  21. Ghanem EH, Sayeed HA, Saleh KH (2000) An alkalophilic thermostable lipase produced by new isolate of Bacillus alcalophilus. World J Microbiol Biotechnol 16:459–464

    Article  CAS  Google Scholar 

  22. Gilbert EJ, Drozd JW, Jones CW (1991) Physiological regulation and optimization of lipase activity in Pseudomonas aeruginosa EF2. J Gen Microbiol 137:2215–2221

    CAS  Google Scholar 

  23. Gitlesen T, Bauer M, Adlercreutz P (1997) Adsorption of lipase on polypropylene powder. Biochim Biophys Acta 1345:188–196

    CAS  Google Scholar 

  24. Gombert AK, Pinto AL, Castilho LR, Freirc DMG (1999). Lipase by Penicillium restrictum in solid state fermentation using Babassu oil cake as substrate. Process Biochem 35:85–90

    Article  CAS  Google Scholar 

  25. Gopinath SCB, Hilda A, Lakshmipriya T, Annadurai G, Anbu P (2003) Purification of lipase from Geotrichum candidum: conditions optimized for enzyme production using Box-Behnken design. World J Microbiol Biotechnol 19:681–689

    Article  CAS  Google Scholar 

  26. Gulati R, Isar J, Kumar V, Parsad AK, Parmar VS, Saxena RK (2005) Production of a novel alkaline lipase by Fusarium globulosum using neem oil, and its applications. Pure Appl Chem 77:251–262

    Article  CAS  Google Scholar 

  27. Gunstone FD (1999) Enzymes as biocatalysts in the modification of natural lipids. J Sci Food Agric 79:1535–1549

    Article  CAS  Google Scholar 

  28. Haas MJ, Bailey DG, Baker W, Berka TR, Cichowicz DJ, Derewenda ZS, Genuario RR, Joerger RD, Klein RR, Scott K, Woolf DJ (1999) Biochemical and molecular characterization of a lipase produced by the fungus Rhizopus delemar. J Sci Technol Fats Oils Waxes 101:364–370

    CAS  Google Scholar 

  29. Hall TA (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucl Acids Symp Ser 41:95–98

    CAS  Google Scholar 

  30. He YQ, Wang BW, Tan TW (2004) Fermentation production technology of lipase with Candida sp. Chin J Biotechnol 20:921–926

    Google Scholar 

  31. Hiol A, Jonzo MD, Rugani N, Druet D, Sarda L, Comeau LC (2000) Purification and characterization of an extracellular lipase from thermophilic Rhizopus oryzae strain isolated from palm fruit. Enzyme Microbial Technol 26:421–430

    Article  CAS  Google Scholar 

  32. Jaeger KE, Ransac S, Koch HB, Ferraro F, Dijkstra BW (1993) Topological characterization and modeling of the 3D structure of lipase from Pseudomonas aeruginosa. FEBS Lett 332:143–149

    Article  CAS  Google Scholar 

  33. Jaeger KE, Reetz MT (1998) Microbial lipases form versatile tools for biotechnology. Trends Biotechnol 16:396–403

    Article  CAS  Google Scholar 

  34. Kamini NR, Fuji T, Kurosu T, Iefreji H (2000) Production, purification characterization of an extracellular from the yeast Cryptococcus sp S-2. Process Biochem 36:317–324

    Article  CAS  Google Scholar 

  35. Kanwar L, Gogoi B, Goswami P (2002) Production of Pseudomonas lipase in n-alkane substrate and its isolation using an improved ammonium sulphate precipitation technique. Bioresour Technol 84:207–211

    Article  CAS  Google Scholar 

  36. Kunio O, Yonko Y, Junichi S (1994) Lipase production of Aspergillus oryzae [J]. Ferment Bioeng 77:490–495

    Article  Google Scholar 

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

    Article  CAS  Google Scholar 

  38. Landass A, Solberg P (1978) Production and characterization of lipase from a fluorescent Pseudomonad. In: 20th Internatinal Dairy Congress, Paris, pp E:304–E:305

  39. Langen LM, Vroom E, Rantwijk F, Sheldon R (1999) Enzymatic syhthesis of beta-lactamase antibiotics using penicillin-G acrylase in frozen media. FEBS Lett 456:89–92

    Article  Google Scholar 

  40. Lee HK, Ahn MJ, Kwak SH, Song WH, Jeong BC (2003) Purification and characterization of cold active lipase from Psychrotrophic aeromonas sp. LPB 4. J Microbiol 41:22–27

    CAS  Google Scholar 

  41. Lesuisse E, Schanck K, Colson C (1993) Purification and preliminary characterization of extracellular lipase of Bacillus subtilis 168, an extreme basic pH tolerant enzyme. Eur J Biochem 216:155–160

    Article  CAS  Google Scholar 

  42. Macre AR, Hammond AR (1983) Present and future applications of lipases. Biotech Genet Eng Rev 3:193–217

    Google Scholar 

  43. Maliszewka I, Mastalerz P (1992) Production and some properties of lipase from Penicillium citrinum. Enzyme Microb Technol 14:190–193

    Article  Google Scholar 

  44. Margesin R, Shinner F (1994) Properties of cold-adapted microorganisms and their potential role in biotechnology. J Biotechnol 33:1–14

    Article  CAS  Google Scholar 

  45. Matsumae H, Shibatani T (1994) Purification and characterization of the lipase from Serratia marcescens Sr418000 responsible for asymmetric hydrolysis of 3-phenylglycidic acid esters. J Ferment Bioeng 77:152–158

    Article  CAS  Google Scholar 

  46. Muralidhar RV, Marchant R, Nigam P (2001). Lipases in racemic resolutions. J Chem Technol Biotechnol 76:3–8

    Article  CAS  Google Scholar 

  47. Osinga R, Tramper J, Wijffels RH (1998) Cultivation of marine sponges for metabolite production: applications for biotechnology? Trends Biotechnol 16:130–134

    Article  CAS  Google Scholar 

  48. Ota Y, Saroamoto T, Hasuo M (2000) Tributyrin specially induces a lipase with preference for the Sn-2 position of triglyceride in Geotrichum sp. F0401B. Biosci Biotechnol Biochem 64:2497–2499

    Article  CAS  Google Scholar 

  49. Petersen M, Daniel R (2006) Purification and characterization of an extracellular lipase from Clostridium teatnomorphum. World J Microbiol Biotechnol 22:431–435

    Article  CAS  Google Scholar 

  50. Rajmohan S, Dodd CER, Waites WM (2002) Enzymes from isolates of Pseudomonas fluorescens involved in food spoilage. J Appl Microbiol 93:205–213

    Article  CAS  Google Scholar 

  51. Sarkar S, Sreekanth B, Kant S, Banerjee R, Bhattacharyya BC (1998) Production and optimization of microbial lipase. Bioprocess Eng 19:29–32

    Article  CAS  Google Scholar 

  52. Selvin J, Soniya J, Asha KRT, Manjusha WA, Sangeetha VS, Jayaseema DM, Antony MC, Vinitha DAJ (2004) Antibacterial potential of antagonistic Streptomyces sp. isolated from the marine sponge Dendrilla nigra. FEMS Microbiol Ecol 50:117–122

    Article  CAS  Google Scholar 

Download references

Acknowledgments

We wish to thank Dr. A. P. Lipton, Professor and Head, Department of Marine Biotectnology, CMST, Rajakkamangalam, Nagercoil for valuable suggestions throughout this study. J.S. is thankful to DST (SR/FT/L-19/2004), New Delhi and International Foundation for Science (RGA No. F/3776-1), Sweden for financial support.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Joseph Selvin.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kiran, G.S., Shanmughapriya, S., Jayalakshmi, J. et al. Optimization of extracellular psychrophilic alkaline lipase produced by marine Pseudomonas sp. (MSI057). Bioprocess Biosyst Eng 31, 483–492 (2008). https://doi.org/10.1007/s00449-007-0186-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00449-007-0186-0

Keywords

Navigation