BMB Reports

Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
권호 표지
DOI QR코드

DOI QR Code

Purification and characterization of hepatic lipase from Todarodes pacificus

  • Park, Jong-Won (Department of Chemistry, Kangnung National University) ;
  • Cho, Soon-Yeong (Department of Food Science, Kangnung National University) ;
  • Choi, Suk-Jung (Department of Chemistry, Kangnung National University)
  • Received : 2007.08.10
  • Accepted : 2007.10.05
  • Published : 2008.03.31
Download PDF
⟨ Previous Next ⟩

Abstract

Lipase was purified from squid (Todarodes pacificus) liver in an attempt to investigate the possibility of applying the enzyme for biotechnological applications. Crude extract of squid liver was initially fractionated by the batch type ion exchange chromatography. The fraction containing lipase activity was further purified with an octyl-Sepharose column. Finally, lipase was purified by eluting active protein from a non-dissociating polyacrylamide gel after zymographic analysis. Molecular weight of the purified enzyme was determined to be 27 kDa by SDS-polyacrylamide gel electrophoresis. The enzyme showed the highest activity at a temperature range of $35-40^{\circ}C$ and at pH 8.0. The activity was almost completely inhibited at 1 mM concentration of $Hg^{2+}$ or $Cu^{2+}$ ion. Partial amino acid sequence of the enzyme was also determined.

Keywords

References

  1. Houde, A., Kademi, A. and Leblanc, D. (2004) Lipases and their industrial applications. Appl. Biochem. Biotechnol. 118, 155-170 https://doi.org/10.1385/ABAB:118:1-3:155
  2. Jaeger, K.-E., Dijkstra, B. W. and Reetz, M. T. (1999) Bacterial biocatalysts: molecular biology, three-dimensional structures, and biotechnological applications of lipases. Annu. Rev. Microbiol. 53, 315-351 https://doi.org/10.1146/annurev.micro.53.1.315
  3. Schmid, R. D. and Verger, R. (1998) Lipases: interfacial enzymes with attractive applications. Angew. Chem. Int. Ed. 37, 1608-1633 https://doi.org/10.1002/(SICI)1521-3773(19980703)37:12<1608::AID-ANIE1608>3.0.CO;2-V
  4. Modi, M. K., Reddy, J. R., Rao, B. V. and Prasad, R. B. (2007) Lipase-mediated conversion of vegetable oils into biodiesel using ethyl acetate as acyl acceptor. Bioresour. Technol. 98, 1260-1264 https://doi.org/10.1016/j.biortech.2006.05.006
  5. Noureddini, H., Gao, X. and Philkana, R. S. (2005) Immobilized Pseudomonas cepacia lipase for biodiesel fuel production from soybean oil. Bioresour. Technol. 96, 769-777 https://doi.org/10.1016/j.biortech.2004.05.029
  6. Reddy, R. R., Chadha, A. and Bhattacharya, E. (2001) Porous silicon based potentiometric triglyceride biosensor. Biosens. Bioelectron. 16, 313-317 https://doi.org/10.1016/S0956-5663(01)00129-4
  7. Wu, L.-C. and Cheng, C.-M. (2005) Flow-injection enzymatic analysis for glycerol and triacylglycerol. Anal. Biochem. 346, 234-240 https://doi.org/10.1016/j.ab.2005.08.031
  8. Ferrone, M., Raimondo, M. and Scolapio, J. S. (2007) Pancreatic enzyme pharmacotherapy. Pharmacotherapy 27, 910-920 https://doi.org/10.1592/phco.27.6.910
  9. Liese, A., Seelbach, K. and Wandrey, C. (2000) Industrial Biotransformations. Wiley-VCH, Weinheim, Germany
  10. Gottsche, J. R., Nielsen, N. S., Nielsen, H. H. and Mu, H. (2005) Lipolysis of different oils using crude enzyme isolate from the intestinal tract of rainbow trout, Oncorhynchus mykiss. Lipids 40, 1273-1279 https://doi.org/10.1007/s11745-005-1495-9
  11. Gjellesvik, D. R., Lombardo, D. and Walther, B. T. (1992) Pancreatic bile salt dependent lipase from cod (Gadus morhua): purification and properties. Biochim. Biophys. Acta 1124, 123-134 https://doi.org/10.1016/0005-2760(92)90088-D
  12. Hazel, J. R. and Sidell, B. D. (2004) The substrate specificity of hormone-sensitive lipase from adipose tissue of the Antarctic fish Trematomus newnesi. J. Exp. Biol. 207, 897-903 https://doi.org/10.1242/jeb.00823
  13. Leger, C., Bauchart, D. and Flanzy, J. (1977) Some properties of pancreatic lipase in Salmo gairdnerii Rich.:$K_m$, effects of bile salts and $Ca^{2+}$, gel filtrations. Comp. Biochem. Physiol. B 57, 359-363 https://doi.org/10.1016/0305-0491(77)90067-0
  14. Patton, J. S. (1975) High levels of pancreatic nonspecific lipase in rattlesnake and leopard shark. Lipids 10, 562-564 https://doi.org/10.1007/BF02532361
  15. Skinner, E. R. and Youssef, A. M. (1982) The characterization of lipoprotein lipase isolated from the post-heparin plasma of the rainbow trout, Salmo gairdneri Richardson. Biochem. J. 203, 727-734 https://doi.org/10.1042/bj2030727
  16. Sukarno, Takahashi, K., Hatano, M. and Sakurai, Y. (1996) Lipase from neon flying squid hepatopancreas: purification and properties. Food Chem. 57, 515-521 https://doi.org/10.1016/S0308-8146(96)00044-1
  17. Kurooka, S. Okamoto, S. and Hashimoto, M. (1977) A novel and simple colorimetric assay for human serum lipase. J. Biochem. 81, 361-369 https://doi.org/10.1093/oxfordjournals.jbchem.a131467
  18. Choi, S.-J, Hwang, J. M. and Kim, S. I. (2003) A colorimetric microplate assay method for high throughput analysis of lipase activity. J. Biochem. Mol. Biol. 36, 417-420 https://doi.org/10.5483/BMBRep.2003.36.4.417
  19. Bastida, A., Sabuquillo, P., Armisen, P., Fernandez- Lafuente, R., Huguet, J. and Guisan, J. M. (1998) A single step purification, immobilization, and hyperactivation of lipases via interfacial adsorption on strongly hydrophobic supports. Biotechnol. Bioeng. 58, 486-493 https://doi.org/10.1002/(SICI)1097-0290(19980605)58:5<486::AID-BIT4>3.0.CO;2-9
  20. Bompensieri, S., Gonzalez, R., Kok, R., Miranda, M. V., Nutgeren-Roodzant, I., Hellingwerf, K. J., Cascone, O. and Nudel, B. C. (1996) Purification of a lipase from Acinetobacter calcoaceticus AAC323-1 by hydrophobic-interaction methods. Biotechnol. Appl. Biochem. 23, 77-81
  21. Diogo, M. M., Silva, S., Cabral, J. M. and Queiroz, J. A. (1999) Hydrophobic interaction chromatography of Chromobacterium viscosum lipase on polypropylene glycol immobilised on Sepharose. J. Chromatogr. A 849, 413-419 https://doi.org/10.1016/S0021-9673(99)00619-6
  22. Fernandez-Lorente, G., Ortiz, C., Segura, R. L., Fernandez-Lafuente, R., Guisan, J. M and Palomo, J. M. (2005) Purification of different lipases from Aspergillus niger by using a highly selective adsorption on hydrophobic supports. Biotechnol. Bioeng. 92, 773-779 https://doi.org/10.1002/bit.20656
  23. Khyami-Horani, H. (1996) Thermotolerant strain of Bacillus licheniformis producing lipase. World J. Microbiol. Biotechnol. 12, 399-401 https://doi.org/10.1007/BF00340219
  24. Patkar, S. A. and Bjorkling, F. (1994) Lipase inhibitors; in Lipases-their structure, biochemistry and application. Woolley, P. and Petersen, S. B. (eds.) pp. 207-224, Cambridge University Press, Cambridge, UK
  25. Snellman, E. A., Sullivan, E. R. and Colwell, R. R. (2002) Purification and properties of the extracellular lipase, Lip A, of Acinetobacter sp. RAG-1. Eur. J. Biochem. 269, 5771-5779 https://doi.org/10.1046/j.1432-1033.2002.03235.x
  26. Sugihara, A., Tani, T. and Tominaga, Y. (1991) Purification and characterization of a novel thermostable lipase from Bacillus sp. J. Biochem. 109, 211-216
  27. Gabriel, O. (1971) Locating enzymes on gels; in Methods in Enzymology, Jakoby, W. B. (ed.), pp. 343-352, Academic Press Inc., New York, USA
  28. Farias, R. N., Torres, M. and Canela, R. (1997) Spectrophotometric determination of the positional specificity of nonspecific and 1,3-specific lipases. Anal. Biochem. 252, 186-189 https://doi.org/10.1006/abio.1997.2240

Cited by

  1. Purification and characterization of an intracellular lipase from pleopods of whiteleg shrimp (Litopenaeus vannamei) vol.158, pp.1, 2011, https://doi.org/10.1016/j.cbpb.2010.10.004
  2. Effectivity of Immunoglobulin Y Anti Lipase as a Pancreatic Lipase Inhibitor for Prevention of Obesity* vol.15, pp.8, 2016, https://doi.org/10.3923/pjn.2016.752.759
  3. Compositions and yield of lipids extracted from hepatopancreas of Pacific white shrimp (Litopenaeus vannamei) as affected by prior autolysis vol.134, pp.2, 2012, https://doi.org/10.1016/j.foodchem.2012.02.188
  4. Identification of enzyme genes in the liver of the Bleeker’s squid Loligo bleekeri by expressed sequence tag analysis vol.76, pp.1, 2010, https://doi.org/10.1007/s12562-009-0182-1
  5. Purification and Biochemical Characterization of Digestive Lipase in Whiteleg Shrimp vol.13, pp.2, 2011, https://doi.org/10.1007/s10126-010-9298-7
  6. Biochemical Quality Changes During Iced Storage of Indian Octopus (Cistopus indicus) vol.39, pp.5, 2016, https://doi.org/10.1111/jfq.12215