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https://hdl.handle.net/11147/1887
Title: | Structural alteration of cofactor specificity in Corynebacterium 2,5-diketo-D-gluconic acid reductase | Authors: | Şanlı Mohamed, Gülşah Banta, Scott Anderson, Stephen Blaber, Michael |
Keywords: | Corynebacterium Aldo keto reductase Ascorbic acid Enzyme engineering Vitamin C |
Issue Date: | Feb-2004 | Publisher: | John Wiley and Sons Inc. | Source: | Şanlı Mohamed, G., Banta, S., Anderson, S., and Blaber, M. (2004). Structural alteration of cofactor specificity in Corynebacterium 2,5-diketo-D-gluconic acid reductase. Protein Science, 13(2), 504-512. doi:10.1110/ps.03450704 | Abstract: | Carynebacterium 2,5-Diketo-D-gluconic acid reductase (2,5-DKGR) catalyzes the reduction of 2,5-diketo-D-gluconic acid (2,5-DKG) to 2-Keto-L-gulonic acid (2-KLG). 2-KLG is an immediate precursor to L-ascorbic acid (vitamin C), and 2,5-DKGR is, therefore, an important enzyme in a novel industrial method for the production of vitamin C. 2,5-DKGR, as with most other members of the aldo-keto reductase (AKR) superfamily, exhibits a preference for NADPH compared to NADH as a cofactor in the stereo-specific reduction of substrate. The application of 2,5-DKGR in the industrial production of vitamin C would be greatly enhanced if NADH could be efficiently utilized as a cofactor. A mutant form of 2,5-DKGR has previously been identified that exhibits two orders of magnitude higher activity with NADH in comparison to the wild-type enzyme, while retaining a high level of activity with NADPH. We report here an X-ray crystal structure of the holo form of this mutant in complex with NADH cofactor, as well as thermodynamic stability data. By comparing the results to our previously reported X-ray structure of the holo form of wild-type 2,5-DKGR in complex with NADPH, the structural basis of the differential NAD(P)H selectivity of wild-type and mutant 2,5-DKGR enzymes has been identified. | URI: | http://doi.org/10.1110/ps.03450704 http://hdl.handle.net/11147/1887 |
ISSN: | 0961-8368 0961-8368 1469-896X |
Appears in Collections: | Chemistry / Kimya PubMed İndeksli Yayınlar Koleksiyonu / PubMed Indexed Publications Collection Scopus İndeksli Yayınlar Koleksiyonu / Scopus Indexed Publications Collection WoS İndeksli Yayınlar Koleksiyonu / WoS Indexed Publications Collection |
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