Use of immobilized lipases for lipase purification via specific lipase-lipase interactions

Journal of Chromatography. a
Jose M PalomoRoberto Fernández-Lafuente

Abstract

Lipase from Pseudomonas fluorescens (PFL), an enzyme with a great tendency to yield bimolecular aggregates, was immobilized via multipoint covalent attachment on glyoxyl-agarose in the presence of Triton X-100. This strategy permitted to obtain the enzyme with the active center oriented towards the reaction medium. This immobilized enzyme presents the capacity of specifically adsorbing PFL molecules, that can be easily desorbed by the use of detergents. More interesting, the enzyme was also able to adsorb other lipases. That is, the lipase from Bacillus thermocatenulatus (BTL2) cloned in Escherichia coli was selectively adsorbed on this immobilized enzyme, enabling a very simple purification strategy. Similar results were achieved with some other lipases (those from Rhizomucor miehei (RML), Rhizopus oryzae (ROL), and Humicola Lanuginosa (HLL)). In all cases, the enzyme could be easily desorbed by incubation with Triton X-100. The matrix could be used several cycles without any detrimental effect on the adsorption capacity.

References

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Aug 28, 1998·Chemistry and Physics of Lipids·R Fernandez-LafuenteJ M Guisán
Feb 14, 2003·Biotechnology and Bioengineering·Gloria Fernández-LorenteRoberto Fernández-Lafuente
Mar 11, 2003·Biomacromolecules·José M PalomoJosé M Guisán
Mar 13, 2003·Biotechnology and Applied Biochemistry·Chang Woo SuhEun Kyu Lee
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Dec 1, 1958·Biochimica Et Biophysica Acta·L SARDA, P DESNUELLE

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Citations

Oct 24, 2012·Applied Biochemistry and Biotechnology·Alberta N A Aryee, Benjamin K Simpson
Jun 5, 2007·Nature Protocols·Cesar MateoJose M Guisan
Nov 11, 2011·International Journal of Molecular Sciences·Dongming LanYonghua Wang
Jun 12, 2014·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Cristina Garcia-GalanRoberto Fernandez-Lafuente
Jun 26, 2014·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Rubens A SilvaDenise F S Petri
Nov 25, 2011·Enzyme and Microbial Technology·Don A Cowan, Roberto Fernandez-Lafuente
Nov 25, 2011·Enzyme and Microbial Technology·Karel HernandezRoberto Fernandez-Lafuente
Aug 30, 2008·Bio Systems·Claudio NicoliniEugenia Pechkova
May 17, 2011·Biotechnology Progress·Marco FiliceGloria Fernandez-Lorente
Apr 21, 2006·Biochimica Et Biophysica Acta·Kalyani MondalMunishwar N Gupta
Jul 4, 2015·Enzyme and Microbial Technology·Jose C S dos SantosRoberto Fernandez-Lafuente
Mar 15, 2015·Enzyme and Microbial Technology·Evelin A ManoelRoberto Fernandez-Lafuente
May 20, 2016·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Nazzoly RuedaRoberto Fernandez-Lafuente
Jun 15, 2016·Scientific Reports·Jiandong CuiShiru Jia
Mar 28, 2018·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Lionete Nunes de LimaRaquel de Lima Camargo Giordano
Dec 23, 2008·Medical Mycology·Valeriano Antonio CorbelliniMarco Flores Ferrão
Jan 10, 2017·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Jose J Virgen-OrtízRoberto Fernandez-Lafuente
Mar 21, 2018·Journal of Materials Chemistry. B, Materials for Biology and Medicine·Zhixian LeiZian Lin
Dec 6, 2008·The Journal of Biological Chemistry·César Carrasco-LópezJuan A Hermoso
Sep 7, 2019·International Journal of Biological Macromolecules·Nathalia Saraiva RiosLuciana Rocha Barros Gonçalves

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