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PublicationBiotechnology Advances

Strategies for the one-step immobilization-purification of enzymes as industrial biocatalysts

Authors:Roberto Fernandez-Lafuente
Groups of research:Optimización de biocatalizadores y bioprocesos enzimáticos
Oveimar Barbosaa, Claudia Ortizb, Ángel Berenguer-Murciac, Rodrigo Torresd
Rafael C. Rodriguese,*, Roberto Fernandez-Lafuentef,*.
a Departamento de Química, Facultad de Ciencias Universidad del Tolima, Ibagué, Colombia
b Escuela de Bacteriología y Laboratorio Clínico, Universidad Industrial de Santander, Bucaramanga, Colombia.
c Instituto Universitario de Materiales, Departamento de Química Inorgánica, Universidad de Alicante, Campus de San Vicente del Raspeig, Ap. 99 - 03080 Alicante, Spain.
d Laboratorio de Biotecnología, Instituto Colombiano del Petróleo-Ecopetrol, Piedecuesta, Bucaramanga, Colombia.
e  Biocatalysis and Enzyme Technology Lab, Institute of Food Science and Technology, Federal University of Rio Grande do Sul, Av. Bento Gonçalves, 9500, P.O. Box 15090, ZC 91501-970, Porto Alegre, RS, Brazil.
f Departamento de Biocatalisis. ICP-CSIC. Campus UAM-CSIC. Cantoblanco. 28049 Madrid. Spain.

In this review, we detail the efforts performed to couple the purification and the immobilization of industrial enzymes in a single step. The use of antibodies, the development of specific domains with affinity for some specific supports will be revised. Moreover, we will discuss the use of domains that increase the affinity for standard matrices (ionic exchangers, silicates).  We will show how the control of the immobilization conditions may convert some unspecific supports in largely specific ones. The development of tailor-made heterofunctional supports as a tool to immobilize-stabilize-purify some proteins will be discussed in deep, using low concentration of adsorbents groups and a dense layer of groups able to give an intense multipoint covalent attachment. The final coupling of mutagenesis and tailor made supports will be the last part of the review.
Keywords:Enzyme stabilization, multimeric enzymes, controlled immobilization, chimeric proteins, covalent attachment, ionic exchange, IMAC
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