English   español  
Please use this identifier to cite or link to this item: http://hdl.handle.net/10261/31288
Share/Impact:
Statistics
logo share SHARE   Add this article to your Mendeley library MendeleyBASE

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

Title

Preparation and characterization of a bifunctional aldolase/kinase enzyme. A more efficient biocatalyst for C-C bond formation

AuthorsIturrate Montoya, Laura; Sánchez-Moreno, Israel ; Oroz-Guinea, Isabel ; Pérez-Gil, Jesús; García-Junceda, Eduardo
KeywordsAldolases
Biocatalysis
circular dichroism
Fusion enzymes
Protein engineering
Issue Date6-Apr-2010
PublisherWiley-Blackwell
CitationChemistry - a European Journal 16(13): 4018-4030 (2010)
AbstractA bifunctional aldolase/kinase enzyme named DLF has been constructed by gene fusion through overlap extension. This fusion enzyme consists of monomeric fructose-1,6-bisphosphate aldolase (FBPA) from Staphylococcus carnosus and the homodimeric dihydroxyacetone kinase (DHAK) from Citrobacter freundii CECT 4626 with an intervening five amino acid linker. The fusion protein was expressed soluble and retained both kinase and aldolase activities. The secondary structure of the bifuctional enzyme has been analysed by CD spectroscopy, as well as that of the parental enzymes, in order to study the effect of the covalent coupling of the two parent proteins on the structure of the fused enzyme. Since S. carnosus FBPA is a thermostable protein, the effect of the fusion on the thermal stability of the bifunctional enzyme has also been studied. The proximity of the active centres in the fused enzyme promotes a kinetic advantage as the 20-fold increment in the initial velocity of the overall aldol reaction indicates. Experimental evidence supports that this increase in the reaction rate can be explained in terms of substrate channelling
Description“This is the pre-peer reviewed version of the following article: Iturrate, L., Sánchez-Moreno, I., Oroz-Guinea, I., Pérez-Gil, J., García-Junceda, E. (2010) “Preparation and characterization of a bifunctional aldolase/kinase enzyme. A more efficient biocatalyst for C-C bond formation“ Chem. Eur. J., 16, 4018-4030, which has been published in final form at http://onlinelibrary.wiley.com/doi/10.1002/chem.200903096/abstract”
Publisher version (URL)http://dx.doi.org10.1002/chem.200903096
URIhttp://hdl.handle.net/10261/31288
DOI10.1002/chem.200903096
E-ISSN1521-3765
Appears in Collections:(IQOG) Artículos
Files in This Item:
File Description SizeFormat 
Preparation and characterization of an aldolasekinase enzyme.pdf1,05 MBAdobe PDFThumbnail
View/Open
Show full item record
Review this work
 

Related articles:


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