Fine Modulation of the Catalytic Properties of Rhizomucor miehei Lipase Driven by Different Immobilization Strategies for the Selective Hydrolysis of Fish Oil
| Autor: | Jose M. Guisan, Marzia Marciello, Gloria Fernández-Lorente, Marco Filice, Mehdi Mohammadi, Maryam Yousefi |
|---|---|
| Přispěvatelé: | Comunidad de Madrid, Universidad Complutense de Madrid |
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Oriented immobilization
Iminodiacetic acid Pharmaceutical Science Rhizomucor miehei 02 engineering and technology Analytical Chemistry Catalysis Sepharose Rhizomucor miehei lipase lcsh:QD241-441 03 medical and health sciences chemistry.chemical_compound Hydrolysis lcsh:Organic chemistry Drug Discovery Organic chemistry Química farmaceútica Physical and Theoretical Chemistry Lipase 030304 developmental biology omega-3 polyunsaturated fatty acids 0303 health sciences biology Protein chemical modification Organic Chemistry fish oil hydrolysis Lipase immobilization 021001 nanoscience & nanotechnology biology.organism_classification chemistry lipase immobilization Chemistry (miscellaneous) oriented immobilization biology.protein Molecular Medicine Cyanogen bromide protein chemical modification 0210 nano-technology Selectivity |
| Zdroj: | Molecules Volume 25 Issue 3 Molecules, Vol 25, Iss 3, p 545 (2020) E-Prints Complutense. Archivo Institucional de la UCM instname Digital.CSIC. Repositorio Institucional del CSIC |
| ISSN: | 1420-3049 |
| DOI: | 10.3390/molecules25030545 |
| Popis: | This article belongs to the Special Issue Lipases and Lipases Modification 2019. Functional properties of each enzyme strictly depend on immobilization protocol used for linking enzyme and carrier. Different strategies were applied to prepare the immobilized derivatives of Rhizomucor miehei lipase (RML) and chemically aminated RML (NH2-RML). Both RML and NH2-RML forms were covalently immobilized on glyoxyl sepharose (Gx-RML and Gx-NH2-RML), glyoxyl sepharose dithiothreitol (Gx-DTT-RML and Gx-DTT-NH2-RML), activated sepharose with cyanogen bromide (CNBr-RML and CNBr-NH2-RML) and heterofunctional epoxy support partially modified with iminodiacetic acid (epoxy-IDA-RML and epoxy-IDA-NH2-RML). Immobilization varied from 11% up to 88% yields producing specific activities ranging from 0.5 up to 1.9 UI/mg. Great improvement in thermal stability for Gx-DTT-NH2-RML and epoxy-IDA-NH2-RML derivatives was obtained by retaining 49% and 37% of their initial activities at 70 °C, respectively. The regioselectivity of each derivative was also examined in hydrolysis of fish oil at three different conditions. All the derivatives were selective between cis-5,8,11,14,17-eicosapentaenoic acid (EPA) and cis-4,7,10,13,16,19-docosahexaenoic acid (DHA) in favor of EPA. The highest selectivity (32.9 folds) was observed for epoxy-IDA-NH2-RML derivative in the hydrolysis reaction performed at pH 5 and 4 °C. Recyclability study showed good capability of the immobilized biocatalysts to be used repeatedly, retaining 50–91% of their initial activities after five cycles of the reaction. This work was financially supported by the National Institute of Genetic Engineering and Biotechnology (grant number: 587) for which the authors are thankful. M.F. acknowledges the Comunidad Autonoma de Madrid for research project no. 2017-T1/BIO-4992 (“Atracción de Talento” Action) also cofunded by Universidad Complutense de Madrid (UCM). M.M and M.F. are grateful to the Comunidad Autonoma de Madrid and FEDER for the I+D collaborative Program in Biomedicine NIETO-CM (Project reference B2017-BMD3731). |
| Databáze: | OpenAIRE |
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