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Horse Liver Alcohol Dehydrogenase: new perspectives for an old enzyme
Quaglia, Daniela; Irwin, Jane A.; Paradisi, Francesca
The EE subunit of horse liver alcohol dehydrogenase (HLADH-EE) has been subcloned in pRSETb vector to generate a fusion His-tag protein. The migration from a multistep purification protocol for this well-known enzyme to a single-step has been successfully achieved. Several adjustments to the traditional purification procedure for Histag proteins have been made to retain protein activity. A full characterization of the fusion enzyme has been carried out and compared with the native one. The Km for EtOH, NAD and NADH in the His-tag version of HLADH are in line with the ones reported in literature for the native enzyme. A shift in optimal pH activity is also observed. The enzyme retains the same stability and quaternary structure as the wild type and can therefore be easily used instead of the native HLADH for biotechnological applications. DG - 30/10/2012 JG 2012-11-12 NB this item should appear in the Chemistry and Chemical Biology collection (not library staff collection) CF DG Names JG 2012-11-23
Keyword(s): DAC; Enzyme characterisation; IMAC; Horse liver alcohol dehydrogenase; Chromatographic analysis; Alcohol dehydrogenase; Enzymes--Analysis
Publication Date:
2012
Type: Journal article
Peer-Reviewed: Unknown
Language(s): English
Institution: University College Dublin
Publisher(s): Springer Science+Business Media
First Indexed: 2012-11-30 05:15:30 Last Updated: 2018-10-11 15:51:12