Immobilization of Lipase and Lipase Producing Yeast, Yarrowia lipolytica NBRC 1658 ​

Abstract

Lipase production was screened out of three Yarrowia strains and a novel Candida tropicalis strain isolated from soil samples and Yarrowia lipolytica NBRC 1658 showed the highest lipolytic activity was selected. Lipase produced by Yarrowia lipolytica NBRC 1658 was partially purified by ammonium sulfate precipitation and dialysis. Crude lipase, partially purified lipase and the yeast Yarrowia lipolytica NBRC 1658 was used for immobilization. Immobilization was carried out on carriers such as Sodium-alginate, k-Carrageenan and Agar- Agar. The highest lipase activity was detected in unimmobilized purified lipase sample as 11.17 U/ml.

Keywords

• Lipase,immobilization,Yarrowia lipolytica

Lipaz ve Lipaz Üreten Yarrowia lipolytica NBRC 1658 Suşunun İmmobilizasyonu

Abstract

Ü ç farklı Yarrowia suşu ve toprak örneklerinden yeni izole edilmiş bir Candida tropicalis suşundan lipaz üretimi araştırılmış ve en yüksek lipolitik aktiviteye sahip suş olan Yarrowia lipolytica NBRC 1658 suşu seçilmiştir. Yarrowia lipolytica NBRC 1658 suşundan üretilen lipaz amonyum sülfat çöktürmesi ve diyaliz ile kısmi olarak saflaştırılmıştır. Kaba enzim, kısmi olarak saflaştırılmış lipaz ve Yarrowia lipolytica NBRC 1658 suşu ayrı ayrı sodyum-aljinat, k-Karragenan ve Agar-Agar’a tutuklanmıştır. En yüksek lipaz aktivitesi ise immobilize edilmeyen kısmi saflaştırılmış suşta 11.17 U/ml olarak saptanmıştır

Keywords

• Lipaz,immobilizasyon,Yarrowia lipolytica

References

1. I.S. Babu and G.H. Rao, Lipase production by Yarrowia lipolytica ncım 3589 in solid state fermentation using mixed substrate, Research J. Microbiol., 2 (2007) 469.
2. G. Corzo, S. Revah, Production and characteristics of the lipase from Yarrowia lipolytica 681, Biores. Technol., 70 (1999) 173.
3. R. Gupta, N. Gupta, P. Rathi, Bacterial lipases: an overview of production, purification and biochemical properties, Appl. Microbiol. Biotechnol., 64 (2004) 763.
4. B. Joseph, P.W. Ramteke, G. Thomas, Cold active microbial lipases: some hot issues andrecent developments, Biotechnol. Adv., 26 (2008) 457.
5. K.E. Jaeger, T. Eggert, Lipases for biotechnology. Curr. Opin. Biotechnol., 13 (2002) 390.
6. M.J. Guisan, Immobilization of enzymes as the 21st century begins. Methods in biotechnology – Immobilization of enzymes and cells - Second Edition, Ed. by J.M. Guisan, Humana Press Inc., 2006. 7. M.J. Eisenmenger, J.I. Reyes-De-Corcuera, High hydrostatic pressure increased stability and activity of immobilized lipase in hexane, Enzym. Microbial Technol., 45 (2009) 118.
7. C. Mateo, J.M. Palomo, G. Fernandez-Lorente, J.M. Guisan, R. Fernandez-Lafuente, Improvement of enzyme activity, stability and selectivity via immobilization Technol., 40 (2007) 1451. Enzym. Microbial
8. G.A. Cunha, G. Fernandez-Lorente, V.J. Bevilaqua, J. Destain, C.M.L. Paiva, G.M. Freire, R. Fernandez- Lafuente, M.J. Guisan, Immobilization of Yarrowia lipolytica Lipase – a comparison of stability of physical adsorption and covalent attachment techniques, Appl. Biochem. Biotechnol., 146 (2008) 49.

9. K.S. Yalcin, Investigation of various parameters affecting citric acid production by Yarrowia lipolytica and assessment of citric acid production capacities of some endogenic yeast species., PhD Thesis, Hacettepe University, The Institute for Graduate Studies in Science, Ankara, 2007.
10. D. Hatzinikolaou, J.B. Macris, P. Christakopoulos, D. Kekos, F.N. Kolisis and G. Fountoukidis, Production and partial characterisation of extracellular lipase from Aspergillus niger, Biotechnol. Lett., 18 (1996) 547.
11. H.M. Rifaat, A.A. El-Mahalawy, H.A. El-Menofy, S.A. Donia, Production, optimization and partial purification of lipase from Fusarium oxysporium, J. Appl. Sci. Env. Sanit., 70 (2010) 84.
12. O.H. Lowry, N.J. Rosebrough, A.L. Farr, R.J. Randall, Protein measurement with the folin phenol reagent, J. Biol. Chem., 193 (1951) 265.
13. A. Sugihara, T. Tani, Y. Tominaga, Purification and characterization of a novel thermostable lipase from Bacillus sp., Jo. Biochem., 109 (1991) 211.
14. S.V. Kamzolova, I.G.Morgunov, A. Aurich, O.A. Perevoznikova, N.V. Shishkanova, U. Stottmeister, T.V. Finogenova, Lipase secretion and citric acid production in Yarrowia lipolytica yeast grown on animal and vegetable fat, Food Technol. Biotechnol., 43 (2005) 113.
15. G. Barth, C. Gaillardin, Physiology and genetics of the dimorphic fungus Yarrowla lipolytica. FEMS Microbiol. Rev., 19 (1997) 219.
16. G. Pignede, H. Wang, F. Fudalej, C. Gaillardin, M. Seman, J.M. Nicaud, Characterization of an extracellular lipase encoded by LIP2 in Yarrowia lipolytica, J. Bacteriol., 182 (2000) 2802.
17. R. Lanciotti, A. Gianotti, D. Baldi, R. Angrisani, G. Suzzi, D. Mastrocola, M.E. Guerzoni, Use of Yarrowia lipolytica strains for the treatment of olive mill wastewater, Biores. Technol., 96 (2005) 317.
18. A.A. Palekar, P.T. Vasudevan, S. Yan, Purification of lipase: A review, Biocatal. Biotransform., 18 (2000) 177.
19. F. Hasan, A.A. Shah, A. Hameed, Methods for detection and characterization of lipases: A comprehensive review, Biotechnol. Adv., 27 (2009) 782.
20. A. Aloulou, A. J. Rodriguez, D. Puccinelli, N. Mouz, J. Leclaire, Y. Leblond, F. Carriere, Purification and biochemical characterization of LIP2 lipase from Yarrowia lipolytica, Biochim. Biophys. Acta, 1771 (2007) 228.
21. M. Yu, S. Qin, T. Tan, Purification and characterization of the extracelular lipase Lip2 from Yarrowia lipolytica. Process Biochem., 42 (2007) 384.
22. P.D. Desai, A.M. Dave, S. Devi, Entrapment of lipase into K-carrageenan beads and its use in hydrolysis of olive oil in biphasic system, J. Mol. Catal. B: Enzym., 31 (2004) 143.
23. N.W. Fadnavis, G. Sheelu, B.M. Mani Kumar, M.U. Bhalerao, A.A. Deshpande, Gelatin blends with alginate: gels for lipase immobilization and purification, Biotechnol. Prog., 19 (2003) 557.