High Lipase Production from Geotrichum candidum in Reduced Time using Cottonseed Oil: Optimization, Easy Purification and Specificity Characterization
PDF

Keywords

Fermentation
optimization
lipases
bioproducts
bioprocessing

How to Cite

1.
Patrícia F. de Castro, Natália C. Moreira, Mateus N. Esperanca, Liliane M. de Oliveira, Alberto C. Badino, Olga L. Tavano, Adriano A. Mendes, Rodrigo C. Basso, Roberto Fernández-Lafuente, Daniela B. Hirata. High Lipase Production from Geotrichum candidum in Reduced Time using Cottonseed Oil: Optimization, Easy Purification and Specificity Characterization. J. Chem. Eng. Res. Updates. [Internet]. 2017 Jun. 8 [cited 2024 Nov. 21];3(2):60-9. Available from: https://avantipublisher.com/index.php/jceru/article/view/886

Abstract

Despite the wide variety of potential applications of lipases within industrial processes, the high cost of production and purification is still their main limiting factor. The aim of this work is to optimize the production of Geotrichum candidum lipase (GCL) using submerged fermentation with a combination of statistical experimental design and surface methodology analysis, in order to give a higher production within a shorter time at the lowest possible cost and easy purification. Cottonseed oil, a low-cost by-product of cotton processing, was used as both an inducer and a carbon source. A maximum lipase activity of 27.17 IUmL-1 was achieved after 30h fermentation in a 5L stirred tank bioreactor under optimal conditions: 2.3% (m/v) of casein peptone, 0.8% (v/v) of cottonseed oil and 0.05% (m/v) of MgSO4 and NaNO3. The lipase purification in a single step by immobilization on PHB particles was verified. The combination of these two steps allowed a significant decrease in this lipase cost of production. Moreover, the produced lipase showed high specificity to hydrolyze long-chain fatty acids with cis-9 double bonds, such as oleic and linoleic acids, having an excellent potential for modifying oils in order to produce different bio-products in industrial applications.

https://doi.org/10.15377/2409-983X.2016.03.02.1
PDF

References

Hasan F, Shah AA and Hameed A. Industrial applications of microbial lipases. Enzyme Microb Technol 2006; 39: 235-251. https://doi.org/10.1016/j.enzmictec.2005.10.016

Houde A, Kademi A and Leblanc D. Lipases and their industrial applications—an overview. Appl Biochem Biotech 2004; 118: 155-170. https://doi.org/10.1385/ABAB:118:1-3:155

Salihu A, Alam MdZ, AbdulKarim MI and Salleh HM. Optimization of lipase production by Candida cylindracea in palm oil mill effluent based medium using statistical experimental design. J Mol Catal B Enzym 2011; 69: 66-73. https://doi.org/10.1016/j.molcatb.2010.12.012

Silveira EA, Tardioli PW and Farinas CS. Valorization of palm oil industrial waste as feedstock for lipase production. Appl Biochem Biotech 2016; 179: 558-71. https://doi.org/10.1007/s12010-016-2013-z

Júnior WGM, Kamimura ES, Ribeiro EJ, Pessela BC, Cardoso VL and Resende MM. Optimization of the production and characterization of lipase from Candida rugosa and Geotrichum candidum in soybean molasses by submerged fermentation. Protein Expres Purif 2016; 123: 26-34. https://doi.org/10.1016/j.pep.2016.04.001

Brabcová J, Deminanová Z, Vondrásek J, Jágr M, Zarevúcka M and Palomo JM. Highly selective purification of three lipases from Geotrichum candidum 4013 and their characterization and biotechnological applications. J Mol Catal B Enzym 2013; 98: 62-72. https://doi.org/10.1016/j.molcatb.2013.09.012

Sidebottom CM, Charton E, Dunn PPJ, et al. Geotrichum candidum produces several lipases with markedly different substrate specificities. Eur J Biochem. 1991; 202: 485-491. https://doi.org/10.1111/j.1432-1033.1991.tb16398.x

Jacobsen T, Olsen J and Allerman K. Substrate specificity of Geotrichum candidum lipase preparations. Biotechnol Lett 1990; 12: 121-126. https://doi.org/10.1007/BF01022427

Levy LM, Dehli JR and Gotor V. Lipase-catalyzed resolution of cyclic cis- and trans-beta-hydroxy esters. Tetrahedron- Asymmetr 2003; 14: 2053-8. https://doi.org/10.1016/S0957-4166(03)00366-5

Ramos EZ, Miotti Júnior RH, Castro PF, et al. Production and immobilization of Geotrichum candidum lipase via physical adsorption on eco-friendly support: Characterization of the catalytic properties in hydrolysis and esterification reactions. J. Mol Catal B Enzym 2015; 118: 43-51. https://doi.org/10.1016/j.molcatb.2015.05.009

Maldonado RR, Burkert JFM, Mazutti MA, Maugeri F and Rodrigues MI. Evaluation of lipase production by Geotrichum candidum in shaken flasks and bench-scale stirred bioreactor using different impellers. Biocatal Agric Biotechnol 2012; 1: 147-151. https://doi.org/10.1016/j.bcab.2012.01.003

Soares CMF, Castro HF, Zanin GM and Moraes FF. Characterization and utilization of Candida rugosa lipase immobilized on controlled pore silica. Appl Biochem Biotechnol 1999; 77-79: 745-757. https://doi.org/10.1385/ABAB:79:1-3:745

Borgstrom B. Investigation on lipid separation methods: Separation of cholesterol esters, glycerides and free fatty acids. Acta Physiol Scand 1952; 25: 111-119. https://doi.org/10.1111/j.1748-1716.1952.tb00863.x

Belfrage P and Vaughan M. Simple liquid-liquid partition system for isolation of labeled oleic acid from mixtures with glycerides. J Lipid Res 1969; 10: 341-344.

Hartman L and Lago RCA. Rapid preparation of fatty acid methyl esters from lipids. Lab Pract 1973; 22: 475-476.

Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227: 680-685. https://doi.org/10.1038/227680a0

Burkert JMF, Maugeri F and Rodrigues MI. Optimization of extracellular lipase production by Geotrichum sp. using factorial design. Bioresour Technol 2004; 91: 77-84. https://doi.org/10.1016/S0960-8524(03)00152-4

Burkert JFM, Maldonado RR, Maugeri Filho F and Rodrigues MI. Comparison of lipase production by Geotrichum candidum in stirring and airlift fermenters. J Chem Technol Biotechnol 2005; 80: 61-67. https://doi.org/10.1002/jctb.1157

Jacobsen T, Olse J and Allermann K. Production, partial purification, and immunochemical characterization of multiple forms of lipase from Geotrichum candidum. Enzyme Microb Technol 1989; 11: 90-95. https://doi.org/10.1016/0141-0229(89)90066-5

Kumar S, Kikon K, Upadhyay A, Kanwar SS and Gupta R. Production, purification, and characterization of lipase from thermophilic and alkaliphilic Bacillus coagulans BTS-3. Protein Expres Purif 2005; 41: 38-44. https://doi.org/10.1016/j.pep.2004.12.010

Padhiar J, Das A and Bhattacharya S. Optimization of process parameters influencing the submerged fermentation of extracellular lipases from Pseudomonas aeruginosa, Candida albicans and Aspergillus flavus. Pakistan J Biol Sci 2011; 14: 1011-1018. https://doi.org/10.3923/pjbs.2011.1011.1018

Fabiszewska AU, Stolarzewicz IA, Zamojska WM and Biatecka-Florjanczyk E. Carbon source impact on Yarrowia lipolytica KKP 379 lipase production. Appl Biochem Biotechnol 2014; 50: 404-410. https://doi.org/10.1134/s000368381404005x

Hlavsová K, Zarevácka M, Wimmer Z, Macková M and Sovová H. Geotrichum candidum 4013: Extracellular lipase versus cell-bound lipase from the single strain. J. Mol Catal B Enzym 2009; 61: 188-193. https://doi.org/10.1016/j.molcatb.2009.06.012

Baillargeon MW, Bistline RG and Sonnet PE. Evaluation of strains of Geotrichum candidum for lipase production and fatty acid specificity. Appl Microb Biotechnol 1989; 30: 92-96. https://doi.org/10.1007/BF00256003

Shimada Y, Sugihara A, Nagao T and Tominaga Y. Induction of Geotrichum candidum lipase by long-chain fatty acids. J Ferment Bioeng 1992; 2: 77-80. https://doi.org/10.1016/0922-338X(92)80004-3

Creative Commons License

This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

Copyright (c) 2017 Patrícia F. de Castro, Natália C. Moreira; Mateus N. Esperanca; Liliane M. de Oliveira, Alberto C. Badino, Olga L. Tavano, Adriano A. Mendes, Rodrigo C. Basso, Roberto Fernández-Lafuente, Daniela B. Hirata