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Review

Enzyme-Coated Micro-Crystals: An Almost Forgotten but Very Simple and Elegant Immobilization Strategy

1
Departamento de Engenharia Química, Universidade Federal do Ceará, Campus do Pici, Bloco 709, Fortaleza CEP 60455760, CE, Brazil
2
Instituto de Engenharias e Desenvolvimento Sustentável, Universidade da Integração Internacional da Lusofonia Afro-Brasileira, Campus das Auroras, Redenção CEP 62790970, CE, Brazil
3
Departamento de Química en Ciencias Farmacéuticas, Facultad de Farmacia, Universidad Complutense de Madrid, Plaza de Ramón y Cajal, s/n., 28040 Madrid, Spain
4
Departamento de Biocatálisis, ICP-CSIC, C/Marir Curie 2, Campus UAM-CSIC, 28049 Madrid, Spain
*
Authors to whom correspondence should be addressed.
Catalysts 2020, 10(8), 891; https://doi.org/10.3390/catal10080891
Received: 23 June 2020 / Revised: 24 July 2020 / Accepted: 5 August 2020 / Published: 6 August 2020
(This article belongs to the Special Issue Multienzymatic Catalysis and/or Enzyme Co-immobilization)
The immobilization of enzymes using protein coated micro-crystals (PCMCs) was reported for the first time in 2001 by Kreiner and coworkers. The strategy is very simple. First, an enzyme solution must be prepared in a concentrated solution of one compound (salt, sugar, amino acid) very soluble in water and poorly soluble in a water-soluble solvent. Then, the enzyme solution is added dropwise to the water soluble solvent under rapid stirring. The components accompanying the enzyme are called the crystal growing agents, the solvent being the dehydrating agent. This strategy permits the rapid dehydration of the enzyme solution drops, resulting in a crystallization of the crystal formation agent, and the enzyme is deposited on this crystal surface. The reaction medium where these biocatalysts can be used is marked by the solubility of the PCMC components, and usually these biocatalysts may be employed in water soluble organic solvents with a maximum of 20% water. The evolution of these PCMC was to chemically crosslink them and further improve their stabilities. Moreover, the PCMC strategy has been used to coimmobilize enzymes or enzymes and cofactors. The immobilization may permit the use of buffers as crystal growth agents, enabling control of the reaction pH in the enzyme environments. Usually, the PCMC biocatalysts are very stable and more active than other biocatalysts of the same enzyme. However, this simple (at least at laboratory scale) immobilization strategy is underutilized even when the publications using it systematically presented a better performance of them in organic solvents than that of many other immobilized biocatalysts. In fact, many possibilities and studies using this technique are lacking. This review tried to outline the possibilities of this useful immobilization strategy. View Full-Text
Keywords: enzyme immobilization; enzymes in organic media; crosslinking of immobilized enzymes; enzyme stabilization; enzyme hyperactivation; solid buffers enzyme immobilization; enzymes in organic media; crosslinking of immobilized enzymes; enzyme stabilization; enzyme hyperactivation; solid buffers
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MDPI and ACS Style

Monteiro, R.R.C.; dos Santos, J.C.S.; Alcántara, A.R.; Fernandez-Lafuente, R. Enzyme-Coated Micro-Crystals: An Almost Forgotten but Very Simple and Elegant Immobilization Strategy. Catalysts 2020, 10, 891. https://doi.org/10.3390/catal10080891

AMA Style

Monteiro RRC, dos Santos JCS, Alcántara AR, Fernandez-Lafuente R. Enzyme-Coated Micro-Crystals: An Almost Forgotten but Very Simple and Elegant Immobilization Strategy. Catalysts. 2020; 10(8):891. https://doi.org/10.3390/catal10080891

Chicago/Turabian Style

Monteiro, Rodolpho R.C., José C.S. dos Santos, Andrés R. Alcántara, and Roberto Fernandez-Lafuente. 2020. "Enzyme-Coated Micro-Crystals: An Almost Forgotten but Very Simple and Elegant Immobilization Strategy" Catalysts 10, no. 8: 891. https://doi.org/10.3390/catal10080891

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