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Use of thermostable Fusarium heterosporum lipase for production of structured lipid containing oleic and palmitic acids in organic solvent-free system

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Journal of the American Oil Chemists' Society

Abstract

A newly developed 1,3-positionally specific thermostable lipase from Fusarium heterosporum (named R275A lipase) was immobilized on Dowex WBA for the production of structured lipid by acidolysis of tripalmitin (PPP) with oleic acid (OA). The immobilized catalyst was fully activated by pretreatment at 50°C in a PPP/OA mixture containing 2% water. The pretreatment caused concomitant hydrolysis, but the hydrolysis was repressed using a substrate without water in the subsequent reactions. The optimal reaction conditions were determined as follows: A mixture of PPP/OA (1∶2, w/w) and 8% immobilized lipase catalyst was incubated at 50°C for 24 h with shaking at 130 oscillations/min. The acidolysis reached 50% under these conditions, and the contents of triolein, 1,3-dioleoyl-2-palmitoyl-glycerol, 1(3),2-dioleoyl-3(1)-palmitoyl-glycerol, 1(3),2-palmitoyl-3(1)-oleoyl-glycerol, 1,3-dipalmitoyl-2-oleoyl-glycerol, and PPP in the reaction mixture were 8, 36, 4, 28, 1, and 6 mol%, respectively. The stabilities of immobilized R275A lipase catalyst and two immobilized catalysts containing Rhizopus delemar or Rhizomucor miehei lipases were compared under the conditions mentioned above, with the catalysts being transferred to fresh substrate every 24 h. The half-life of the R275A lipase catalyst was 370 d, which was significantly longer than those of Rhizopus and Rhizomucor lipase catalysts.

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References

  1. Breckenridge, W.C., L. Marai, and A. Kuksis, Triglyceride Structure of Human Milk Fat, Can. J. Biochem. 47:761–769 (1969).

    Article  CAS  Google Scholar 

  2. Martin, J.C., P. Bougnoux, J.-M. Antoine, M. Lanson, and C. Couet, Triacylglycerol Structure of Human Colostrum and Mature Milk, Lipids 28:637–643 (1993).

    CAS  Google Scholar 

  3. Jensen, C., N.R. Buist, and T. Wilson, Absorption of Individual Fatty Acids from Long-Chain or Medium-Chain Triglycerides in Very Small Infants, Am. J. Clin. Nutr. 43:745–751 (1986).

    CAS  Google Scholar 

  4. Innis, S.M., R. Dyer, and C.M. Nelson, Evidence That Palmitic Acid Is Absorbed as sn-2 Monoacylglycerol from Human Milk by Breast-Fed Infants, Lipids 29:541–545 (1994).

    CAS  Google Scholar 

  5. Shimada, Y., C. Koga, A. Sugihara, T. Nagao, N. Takada, S. Tsunasawa, and Y. Tominaga, Purification and Characterization of a Novel Solvent-Tolerant Lipase from Fusarium heterosporum, J. Ferment. Bioeng. 75:349–352 (1993).

    Article  CAS  Google Scholar 

  6. Nagao, T., Y. Shimada, A. Sugihara, and Y. Tominaga, Cloning and Nucleotide Sequence of cDNA Encoding a Lipase from Fusarium heterosporum, J. Biochem. 116:536–540 (1994).

    CAS  Google Scholar 

  7. Nagao, T., Y. Shimada, A. Sugihara, and Y. Tominaga, Expression of Lipase cDNA from Fusarium heterosporum by Saccharomyces cerevisiae: High-Level Production and Purification, J. Ferment. Bioeng. 81:488–492 (1996).

    Article  CAS  Google Scholar 

  8. Nagao, T., Y. Shimada, A. Sugihara, and Y. Tominaga, C-Terminal Peptide of Fusarium heterosporum Lipase Is necessary for Its Increasing Thermostability, J. Biochem. 124:1124–1129 (1998).

    CAS  Google Scholar 

  9. Nagao, T., Y. Shimada, A. Sugihara, and Y. Tominaga, Amino Acid Residues Contributing Stabilization of Fusarium heterosporum Lipase, J. Biosci. Bioeng. 89:446–450 (2000).

    Article  CAS  Google Scholar 

  10. Shimada, Y., T. Nagao, A. Kawashima, M. Yamamoto, A. Sugihara, S. Komemushi, and Y. Tominaga, Immobilization of Rhizopus delemar Lipase, Kagaku to Kogyo 74:117–121 (2000).

    CAS  Google Scholar 

  11. Sugihara, A., Y. Shimada, and Y. Tominaga, Separation and Characterization of Two Molecular Forms of Geotrichum candidum Lipase, J. Biochem. 107:426–430 (1990).

    CAS  Google Scholar 

  12. Shimada, Y., K. Maruyama, S. Okazaki, M. Nakamura, A. Sugihara, and Y. Tominaga, Enrichment of Polyunsaturated Fatty Acids with Geotrichum candidum Lipase, J. Am. Oil Chem. Soc. 71:951–954 (1994).

    CAS  Google Scholar 

  13. Sugihara, A., M. Ueshima, Y. Shimada, S. Tsunasawa, and Y. Tominaga, Purification and Characterization of a Novel Thermostable Lipase from Pseudomonas cepacia, J. Biochem. 112:598–603 (1992).

    CAS  Google Scholar 

  14. Official Methods and Recommended Practices of the American Oil Chemists’ Society, 5th edn., AOCS Press, Champaign, 1997, AOCS Official Method Ch 3-91.

  15. Shimada, Y., A. Sugihara, K. Maruyama, T. Nagao, S. Nakayama, H. Nakano, and Y. Tominaga, Production of Structured Lipid Containing Docosahexaenoic and Caprylic Acids Using Immobilized Rhizopus delemar Lipase, J. Ferment. Bioeng. 81:299–303 (1996).

    Article  CAS  Google Scholar 

  16. Kosugi, Y., K. Takahashi, and C. Lopez, Large-Scale Immobilization of Lipase from Pseudomonas fluorescens Biotype I and an Application for Sardine Oil Hydrolysis, J. Am. Oil Chem. Soc. 72:1281–1285 (1995).

    CAS  Google Scholar 

  17. Shimada, Y., M. Suenaga, A. Sugihara, S. Nakai, and Y. Tominaga, Continuous Production of Structured Lipid Containing γ-Linolenic and Caprylic Acids by Immobilized Rhizopus delemar Lipase, Ibid. 76:189–193 (1999).

    CAS  Google Scholar 

  18. Shimada, Y., A. Sugihara, H. Nakano, T. Yokota, T. Nagao, S. Komemushi, and Y. Tominaga, Production of Structured Lipids Containing Essential Fatty Acids by Immobilized Rhizopus delemar Lipase, Ibid. 73:1415–1420 (1996).

    Article  CAS  Google Scholar 

  19. Shimada, Y., T. Nagao, Y. Hamasaki, K. Akimoto, A. Sugihara, S. Fujikawa, S. Komemushi, and Y. Tominaga, Enzymatic Synthesis of Structured Lipid Containing Arachidonic and Palmitic Acids, Ibid. 77:89–93 (2000).

    CAS  Google Scholar 

  20. Xu, X., A.R.H. Skands, C.-E. Høy, H. Mu, S. Balchen, and J. Adler-Nissen, Production of Specific-Structured Lipids by Enzymatic Interesterification: Elucidation of Acyl Migration by Response Surface Design, Ibid. 75:1179–1186 (1998).

    CAS  Google Scholar 

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Correspondence to Yuji Shimada.

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Nagao, T., Shimada, Y., Sugihara, A. et al. Use of thermostable Fusarium heterosporum lipase for production of structured lipid containing oleic and palmitic acids in organic solvent-free system. J Amer Oil Chem Soc 78, 167–172 (2001). https://doi.org/10.1007/s11746-001-0238-7

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  • DOI: https://doi.org/10.1007/s11746-001-0238-7

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