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Synthesis of Wax Esters from Crude Fish Fat by Lipase of Burkholderia sp. EQ3 and Commercial Lipases

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Journal of the American Oil Chemists' Society

Abstract

The lipase from Burkholderia sp. EQ3 was used to synthesize wax esters in comparison with commercial lipases. The supernatant of Burkholderia sp. EQ3 was collected from a liquid basal medium with 1 % fish oil after 12 h cultivation (1.90 U/ml of lipase activity). The crude lipase was prepared by acetone precipitation of the culture supernatant (4.70 U/mg and 9.40 purification folds). The crude fish fat obtained by hexane extraction of waste fat from the wastewater pond of a canned tuna factory and cetyl alcohol were used for wax esters synthesis. Five commercial lipases were screened in comparison with crude lipase from Burkholderia sp. EQ3 in wax esters synthesis. The optimum conditions for wax esters synthesis from crude fish fat using Novozyme 435 were enzyme 1 U, substrate molar ratio of crude fish fat to cetyl alcohol 1:2 (115.30 mg of crude fish fat and 66.67 mg of cetyl alcohol) in hexane at 37 °C and 200 rpm with 90.81 % (TLC–FID peak area) after one h of reaction. The optimum conditions for the synthesis by crude lipase from Burkholderia sp. EQ3 were crude lipase 40 U, substrate molar ratio of crude fish fat and cetyl alcohol 1:2 in isooctane at 30 °C and 200 rpm with 95.07 % (TLC–FID peak area) after 6 h of reaction. The synthesized wax esters were mainly composed of cetyl palmitate and cetyl oleate.

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References

  1. Uttamangkabovorn M, Prasertsan P, H-Kittikun A (2005) Water conservation in canned tuna (pet food) plant in Thailand. J Cleaner Prod 13:547–555

    Article  Google Scholar 

  2. Peter TR, Robert B (2001) Beeswax through the ages. Pers Care 10:27–31

    Google Scholar 

  3. Savolainen M, Herder J, Khoo C, Lovqvist K, Dahlqvist C, Glad H, Juppo AM (2003) Evaluation of polar lipid–hydrophilic polymer microparticles. Int J Pharm 262:47–62

    Article  CAS  Google Scholar 

  4. Chen JP, Wang JB (1997) Wax ester synthesis by lipase-catalyzed esterification with fungal cells immobilized on cellulose biomass support particles. Enzyme Microb Technol 18:615–622

    Article  CAS  Google Scholar 

  5. Radzi SM, Basri M, Salleh AB, Ariff A, Mohammad R, Rahman MBA, Rahman RNZRA (2005) Large scale production of liquid wax ester by immobilized lipase. J Oleo Sci 54:203–209

    Article  CAS  Google Scholar 

  6. 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  Google Scholar 

  7. Trani M, Ergan F, Andre G (1991) Lipase-catalyzed production of wax esters. J Am Oil Chem Soc 68:20–22

    Article  CAS  Google Scholar 

  8. Mukherjee KD, Kiewitt I (1988) Preparation of esters resembling natural waxes by lipase-catalyzed reactions. J Agric Food Chem 36:1333–1339

    Article  CAS  Google Scholar 

  9. Poisson L, Jan S, Vuillemard JC, Sarazin C, Seguin JP, Barbotin JN, Ergan F (1999) Lipase-catalyzed synthesis of waxes from milk fat and oleyl alcohol. J Am Oil Chem Soc 76:1017–1021

    CAS  Google Scholar 

  10. Salis A, Solinas V, Monduzzi M (2003) Wax esters synthesis from heavy fraction of sheep milk fat and cetyl alcohol by immobilized lipases. J Mol Catal B-Enzym 21:167–174

    Article  CAS  Google Scholar 

  11. Gunawan ER, Basri M, Rahman MBA, Salleh AB, Rahman RNA (2004) Lipase-catalyzed synthesis of palm oil based wax esters. J Oleo Sci 10:471–477

    Article  Google Scholar 

  12. Rungsilp S (1998) Production of fish oil containing omega 3 PUFA from tuna condensate. MS Thesis, Prince of Songkla University, Thailand

  13. Pawongrat R, Xu X, H-Kittikun A (2007) Synthesis of monoacylglycerol rich in polyunsaturated fatty acids from tuna oil with immobilized lipase AK. Food Chem 104:251–258

    Article  CAS  Google Scholar 

  14. Kaewsuwan S, Cahoon EB, Perroud PF, Wiwat C, Panvisavas N, Quatrano RS, Cove DJ, Bunyapraphatsara N (2006) Cloning and functional characterization of the moss Physcomitrella patens (HEDW) B.S.G. ∆5-desaturase gene involved in arachidonic acid and eicosapentaenoic acid biosynthesis. J Biol Chem 281:21988–21997

    Article  CAS  Google Scholar 

  15. Dandavate V, Jinjala J, Keharia H, Madamwar D (2009) Production, partial purification and characterization of organic solvent tolerant lipase from Burkholderia multivorans V2 and its application for ester synthesis. Bioresour Technol 100:3374–3381

    Article  CAS  Google Scholar 

  16. Lee SY, Rhee JS (1993) Production and partial purification of a lipase from Pseudomonas putida 3SK. Enzyme Microb Technol 15:617–623

    Article  CAS  Google Scholar 

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

    CAS  Google Scholar 

  18. IUPAC (1979) Standard methods for the analysis of oils, fats and derivative. Part I, 6th edn. Pergamon Press, France

    Google Scholar 

  19. Isono Y, Nabetani H, Nakajima M (1995) Lipase-surfactant complex as catalyst of interesterification and esterification in organic media. J Ferment Bioeng 80:170–175

    Article  CAS  Google Scholar 

  20. Makowski M, Ohlmeyer M, Meier D (2005) Long-term development of VOC emission from OSB after hot-pressing. Holzforschung 59:519–523

    Article  CAS  Google Scholar 

  21. Rathi P, Goswami VK, Sahai V, Gupta R (2002) Statistical medium optimization and production of a hyperthermostable lipase from Burkholderia cepacia in bioreactor. J Appl Microbiol 93:930–936

    Article  CAS  Google Scholar 

  22. Mahler GF, Kok RG, Cordenons A, Hellingwerf KJ, Nudel BC (2000) Effects of carbon sources on extracellular lipase production and lipA transcription in Acinetobacter calcoaceticus. J Ind Microbiol Biotechnol 24:25–30

    Article  CAS  Google Scholar 

  23. Takeda Y, Aono R, Doukyu N (2006) Purification, characterization, and molecular cloning of organic-solvent-tolerant cholesterol esterase from cyclohexane-tolerant Burkholderia cepacia strain ST-200. Extremophiles 10:269–277

    Article  CAS  Google Scholar 

  24. Bloomer S, Adlercreutz P, Mattiasson B (1992) Facile synthesis of fatty acid esters in high yields. Enzyme Microb Technol 14:546–552

    Article  CAS  Google Scholar 

  25. Decagny B, Jan S, Vuillemard JC, Sarazin C, Seguin JP, Gosselin C, Barbotin JN, Ergan F (1998) Synthesis of wax ester through triolein alcoholysis: choice of the lipase and study of the mechanism. Enzyme Microb Technol 22:578–582

    Article  CAS  Google Scholar 

  26. Carta G, Gainer JL, Benton AH (1991) Enzymatic synthesis of esters using an immobilized lipase. Biotechnol Bioeng 37:1004–1009

    Article  CAS  Google Scholar 

  27. McGilvery RW, Goldstein GW (1983) Rates of enzymatic reaction. In: Biochemistry: a functional approach, 3rd edn. W. B. Saunders Company, Philadelphia, London, Toronto Mexico City and Tokyo, pp 296–307

  28. Yankah VV, Akoh CC (2000) Lipase-catalyzed acidolysis of tristearin with oleic or caprylic acids to produce structured lipids. J Am Oil Chem Soc 77:495–500

    Article  CAS  Google Scholar 

  29. Kanasawud P, Phutrakul S, Bloomer S, Adlercreutz P, Mattiasson B (1992) Triglyceride interesterification by lipases 3. Alcoholysis of pure triglycerides. Enzyme Microb Technol 14:959–965

    Article  CAS  Google Scholar 

  30. Claon PA, Akoh CC (1994) Enzymatic synthesis of geranyl acetate in n-hexane with Candida antarctica lipase. J Am Oil Chem Soc 71:1177–1182

    Article  Google Scholar 

  31. Rahman MBA, Yong KC, Basri M, Rahman RNZ (2001) Synthesis of oleyl oleate, a liquid wax ester, using lipozyme. Malays J Chem 3:10046–10050

    Google Scholar 

  32. Laane C, Boeren VK, Veeger C (1987) Rule of optimization of biocatalysis in organic solvent. Biotechnol Bioeng 30:81–87

    Article  CAS  Google Scholar 

  33. Basri M, Ampon K, Yunus WMZ, Razak CNA, Salleh AB (1995) Enzymatic synthesis of fatty ester by hydrophobic lipase derivatives immobilized on organic polymer beads. J Am Oil Chem Soc 72:407–411

    Article  CAS  Google Scholar 

  34. Song J, Kahveci D, Chen M, Guo Z, Xie E, Xu X, Besenbacher F, Dong M (2012) Enhanced catalytic activity of lipase encapsulated in PCL nanofibers. Langmuir 28:6157–6162

    Article  CAS  Google Scholar 

  35. Liu Y, Zhang X, Tan H, Yan Y, Hameed BH (2010) Effect of pretreatment by different organic solvents on esterification activity and conformation of immobilized Pseudomonas cepacia lipase. Process Biochem 45:1176–1180

    Article  CAS  Google Scholar 

  36. Zaks A, Klibanov AM (1985) Enzyme-catalyzed processes in organic solvents. Proc Nat Acad Sci USA 82:3192–3196

    Article  CAS  Google Scholar 

  37. Sulong MR, Rahman RNZRA, Salleh AB, Basri M (2006) Novel organic solvent tolerant lipase from Bacillus sphaericus 205y: extracellular expression of a novel OST-lipase gene. Protein Express Purif 49:190–195

    Article  CAS  Google Scholar 

  38. Huynh L-H, Do Q-D, Kasim NS, Ju Y-H (2011) Isolation and analysis of wax esters from activated sludge. Bioresour Technol 102:9518–9523

    CAS  Google Scholar 

Download references

Acknowledgments

The research was supported by the Agro-Industry Practice School (APS) from the National Center for Genetic Engineering & Biotechnology (BIOTEC), National Science & Technology Development Agency. Financial support was also given by the Graduated School of Prince of Songkla University and the Higher Education Research Promotion and National Research University Project of Thailand, Office of the Higher Education Commission. The authors would like to thank Songkhla Canning Public Company Limited (Hat Yai, Thailand) for providing raw materials in this study.

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Correspondence to A. H-Kittikun.

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Ungcharoenwiwat, P., H-Kittikun, A. Synthesis of Wax Esters from Crude Fish Fat by Lipase of Burkholderia sp. EQ3 and Commercial Lipases. J Am Oil Chem Soc 90, 359–367 (2013). https://doi.org/10.1007/s11746-012-2183-y

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  • DOI: https://doi.org/10.1007/s11746-012-2183-y

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