English   español  
Please use this identifier to cite or link to this item: http://hdl.handle.net/10261/22452
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:

High-level production of recombinant His-tagged rhamnulose 1-phosphate aldolase in Escherichia coli

AuthorsVidal, Luis; Durany, Olga; Suau, T.; Ferrer, Pau; Benaiges, M. D.; Caminal, Glòria
KeywordsRecombinant aldolase
E coli
Issue Date18-Sep-2003
CitationJournal of Chemical Technology and Biotechnology 78(11): 1171-1179 (2003)
AbstractAn expression system based on Escherichia coli and the T5 promoter allowed the overproduction of a his-tagged rhamnulose-1-phosphate aldolase (RhuA; EC, an enzyme with applications in the production of deoxyazasugars and deoxysugars compounds. Shake flask and bioreactor cultivation with E coli M15 (pQErham) were performed under different media and inducing conditions for RhuA expression. A Defined Medium (DM) with glucose as carbon source gave a high volumetric and enzyme productivity (3460 AU dm-3 and 288 AU dm-3 h-1 respectively) compared with Luria-Bertoni (LB) medium (2292 AU dm- 3 and 255 AU dm-3 h-1). The minimum quantity of (isopropyl--D-thiogalactoside) IPTG for optimal induction was estimated in 18-20 ºmol IPTG gDCW-1. The highest volumetric production of RhuA (8333 AU dm-3) was obtained when IPTG was added in the late log-phase. No significant differences were found in specific RhuA activity for induction temperatures of 30 and 37 ºC. An effective two-step purification process comprising affinity chromatography and gel permeation has been developed (overall recovery 66.5%). These studies provide the basis for the further development of an integrated process for recombinant RhuA production suitable for biotransformation applications.
Description9 pages, 8 figures, 2 tables.-- Printed version published Nov 2003.
Publisher version (URL)http://dx.doi.org/10.1002/jctb.909
Appears in Collections:(IQAC) Artículos
Files in This Item:
There are no files associated with this item.
Show full item record

Related articles:

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