title: Strategies for the one-step immobilization-purification of enzymes as industrial biocatalysts.
abstract: 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 adsorbent 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.
authors.affiliation: Departamento de Química, Facultad de Ciencias de la Universidad del Tolima, Ibagué, Colombia.
authors.forename: Oveimar
authors.initials: O
authors.lastname: Barbosa
authors.affiliation: Escuela de Bacteriología y Laboratorio Clínico, Universidad Industrial de Santander, Bucaramanga, Colombia.
authors.forename: Claudia
authors.initials: C
authors.lastname: Ortiz
authors.affiliation: Instituto Universitario de Materiales, Departamento de Química Inorgánica, Universidad de Alicante, Campus de San Vicente del Raspeig, Ap. 99-03080, Alicante, Spain.
authors.forename: Ángel
authors.initials: Á
authors.lastname: Berenguer-Murcia
authors.affiliation: Laboratorio de Biotecnología, Instituto Colombiano del Petróleo-Ecopetrol, Piedecuesta, Bucaramanga, Colombia.
authors.forename: Rodrigo
authors.initials: R
authors.lastname: Torres
authors.affiliation: 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. Electronic address: rafaelcrodrigues@ufrgs.br.
authors.forename: Rafael C
authors.initials: RC
authors.lastname: Rodrigues
authors.affiliation: Departamento de Biocatalisis, ICP-CSIC, Campus UAM-CSIC, Cantoblanco, 28049 Madrid, Spain. Electronic address: rfl@icp.csic.es.
authors.forename: Roberto
authors.initials: R
authors.lastname: Fernandez-Lafuente
mesh_terms: D001709:Biotechnology; D002852:Chromatography, Ion Exchange; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D004800:Enzymes, Immobilized; D011506:Proteins
keywords: Chimeric proteins; Controlled immobilization; Covalent attachment; Enzyme stabilization; IMAC; Ionic exchange; Multimeric enzymes