Molecules 2017, 22(12), 2153; https://doi.org/10.3390/molecules22122153
Frozen Microemulsions for MAPLE Immobilization of Lipase
Valeria Califano 1, Francesco Bloisi 2,*
, Giuseppe Perretta 1, Antonio Aronne 3, Giovanni Ausanio 2, Aniello Costantini 3 and Luciano Vicari 2
, Giuseppe Perretta 1, Antonio Aronne 3, Giovanni Ausanio 2, Aniello Costantini 3 and Luciano Vicari 21
Istituto Motori-CNR, via G. Marconi 8, 80125 Napoli, Italy
2
CNR-SPIN and Department of Physics “Ettore Pancini”, University of Naples Federico II, Piazzale V. Tecchio 80, 80125 Napoli, Italy
3
Department of Chemical Engineering, Materials and Industrial Production, University of Naples Federico II, Piazzale V. Tecchio 80, 80125 Napoli, Italy
*
Author to whom correspondence should be addressed.
Received: 21 September 2017 / Revised: 26 November 2017 / Accepted: 27 November 2017 / Published: 5 December 2017
(This article belongs to the Special Issue Lipases and Lipases Modification)
Abstract
Candida rugosa lipase (CRL) was deposited by matrix assisted pulsed laser evaporation (MAPLE) in order to immobilize the enzyme with a preserved native conformation, which ensures its catalytic functionality. For this purpose, the composition of the MAPLE target was optimized by adding the oil phase pentane to a water solution of the amino acid 3-(3,4-dihydroxyphenyl)-2-methyl-l-alanine (m-DOPA), giving a target formed by a frozen water-lipase-pentane microemulsion. Fourier transform infrared (FTIR) spectroscopy and atomic force microscopy (AFM) were used to investigate the structure of MAPLE deposited lipase films. FTIR deconvolution of amide I band indicated a reduction of unfolding and aggregation, i.e., a better preserved lipase secondary structure in the sample deposited from the frozen microemulsion target. AFM images highlighted the absence of big aggregates on the surface of the sample. The functionality of the immobilized enzyme to promote transesterification was determined by thin layer chromatography, resulting in a modified specificity. View Full-TextKeywords:
MAPLE; microemulsion; lipase; thin film; enzyme immobilization
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).
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Califano, V.; Bloisi, F.; Perretta, G.; Aronne, A.; Ausanio, G.; Costantini, A.; Vicari, L. Frozen Microemulsions for MAPLE Immobilization of Lipase. Molecules 2017, 22, 2153.
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