English   español  
Please use this identifier to cite or link to this item: http://hdl.handle.net/10261/169003
Share/Impact:
Statistics
logo share SHARE logo core CORE   Add this article to your Mendeley library MendeleyBASE

Visualizar otros formatos: MARC | Dublin Core | RDF | ORE | MODS | METS | DIDL
Exportar a otros formatos:
Title

Stabilization of Enzymes by Multipoint Covalent Attachment on Aldehyde-Supports: 2-Picoline Borane as an Alternative Reducing Agent

AuthorsOrrego, Alejandro H.; Romero-Fernández, Maria; Millán-Linares, María del Carmen ; Yust, María del Mar ; Guisán, José Manuel ; Rocha-Martín, Javier
KeywordsEnzyme immobilization
2-picoline borane
Biocatalysis
Glyoxyl agarose
Schiff base
Enzyme stabilization
Issue Date15-Aug-2018
PublisherMultidisciplinary Digital Publishing Institute
CitationCatalysts 8(8): 333 (2018)
AbstractEnzyme immobilization by multipoint covalent attachment on supports activated with aliphatic aldehyde groups (e.g., glyoxyl agarose) has proven to be an excellent immobilization technique for enzyme stabilization. Borohydride reduction of immobilized enzymes is necessary to convert enzyme–support linkages into stable secondary amino groups and to convert the remaining aldehyde groups on the support into hydroxy groups. However, the use of borohydride can adversely affect the structure–activity of some immobilized enzymes. For this reason, 2-picoline borane is proposed here as an alternative milder reducing agent, especially, for those enzymes sensitive to borohydride reduction. The immobilization-stabilization parameters of five enzymes from different sources and nature (from monomeric to multimeric enzymes) were compared with those obtained by conventional methodology. The most interesting results were obtained for bacterial (R)-mandelate dehydrogenase (ManDH). Immobilized ManDH reduced with borohydride almost completely lost its catalytic activity (1.5% of expressed activity). In contrast, using 2-picoline borane and blocking the remaining aldehyde groups on the support with glycine allowed for a conjugate with a significant activity of 19.5%. This improved biocatalyst was 357-fold more stable than the soluble enzyme at 50 ◦C and pH 7. The results show that this alternative methodology can lead to more stable and active biocatalysts.
Publisher version (URL)https://doi.org/10.3390/catal8080333
URIhttp://hdl.handle.net/10261/169003
DOI10.3390/catal8080333
ISSN2073-4344
Appears in Collections:(ICP) Artículos
(IG) Artículos
Files in This Item:
File Description SizeFormat 
catalysts-08-00333-v2.pdf1,32 MBAdobe PDFThumbnail
View/Open
Show full item record
Review this work
 


WARNING: Items in Digital.CSIC are protected by copyright, with all rights reserved, unless otherwise indicated.