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Engineering Candida tenuis Xylose reductase for improved utilization of NADH: antagonistic effects of multiple side chain replacements and performance of site-directed mutants under simulated in vivo conditions.
Appl Environ Microbiol. 2005 Oct; 71(10):6390-3.AE

Abstract

Six single- and multiple-site variants of Candida tenuis xylose reductase that were engineered to have side chain replacements in the coenzyme 2'-phosphate binding pocket were tested for NADPH versus NADH selectivity (R(sel)) in the presence of physiological reactant concentrations. The experimental R(sel) values agreed well with predictions from a kinetic mechanism describing mixed alternative coenzyme utilization. The Lys-274-->Arg and Arg-280-->His substitutions, which individually improved wild-type R(sel) 50- and 20-fold, respectively, had opposing structural effects when they were combined in a double mutant.

Authors+Show Affiliations

Institute of Biotechnology and Biochemical Engineering, Graz University of Technology, Petersgasse 12/I, A-8010 Graz, Austria.No affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

16204564

Citation

Petschacher, Barbara, and Bernd Nidetzky. "Engineering Candida Tenuis Xylose Reductase for Improved Utilization of NADH: Antagonistic Effects of Multiple Side Chain Replacements and Performance of Site-directed Mutants Under Simulated in Vivo Conditions." Applied and Environmental Microbiology, vol. 71, no. 10, 2005, pp. 6390-3.
Petschacher B, Nidetzky B. Engineering Candida tenuis Xylose reductase for improved utilization of NADH: antagonistic effects of multiple side chain replacements and performance of site-directed mutants under simulated in vivo conditions. Appl Environ Microbiol. 2005;71(10):6390-3.
Petschacher, B., & Nidetzky, B. (2005). Engineering Candida tenuis Xylose reductase for improved utilization of NADH: antagonistic effects of multiple side chain replacements and performance of site-directed mutants under simulated in vivo conditions. Applied and Environmental Microbiology, 71(10), 6390-3.
Petschacher B, Nidetzky B. Engineering Candida Tenuis Xylose Reductase for Improved Utilization of NADH: Antagonistic Effects of Multiple Side Chain Replacements and Performance of Site-directed Mutants Under Simulated in Vivo Conditions. Appl Environ Microbiol. 2005;71(10):6390-3. PubMed PMID: 16204564.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Engineering Candida tenuis Xylose reductase for improved utilization of NADH: antagonistic effects of multiple side chain replacements and performance of site-directed mutants under simulated in vivo conditions. AU - Petschacher,Barbara, AU - Nidetzky,Bernd, PY - 2005/10/6/pubmed PY - 2005/12/13/medline PY - 2005/10/6/entrez SP - 6390 EP - 3 JF - Applied and environmental microbiology JO - Appl Environ Microbiol VL - 71 IS - 10 N2 - Six single- and multiple-site variants of Candida tenuis xylose reductase that were engineered to have side chain replacements in the coenzyme 2'-phosphate binding pocket were tested for NADPH versus NADH selectivity (R(sel)) in the presence of physiological reactant concentrations. The experimental R(sel) values agreed well with predictions from a kinetic mechanism describing mixed alternative coenzyme utilization. The Lys-274-->Arg and Arg-280-->His substitutions, which individually improved wild-type R(sel) 50- and 20-fold, respectively, had opposing structural effects when they were combined in a double mutant. SN - 0099-2240 UR - https://www.unboundmedicine.com/medline/citation/16204564/Engineering_Candida_tenuis_Xylose_reductase_for_improved_utilization_of_NADH:_antagonistic_effects_of_multiple_side_chain_replacements_and_performance_of_site_directed_mutants_under_simulated_in_vivo_conditions_ L2 - http://aem.asm.org/cgi/pmidlookup?view=long&pmid=16204564 DB - PRIME DP - Unbound Medicine ER -