Catalytic mechanism and substrate selectivity of aldo-keto reductases: Insights from structure-function studies of Candida tenuis xylose reductase

Kratzer, Regina; Wilson, D. Keith; Nidetzky, Bernd

doi:10.1080/15216540600818143

Cited by 46 publications

(34 citation statements)

References 38 publications

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“…Several groups have used site-directed mutagenesis to study the structural determinants of cofactor specificity in the AKRs (Bohren et al, 1991;Grimshaw et al, 1995;Kratzer et al, 2006;Kubiseski and Flynn, 1995;Kubiseski et al, 1992;Leitgeb et al, 2005;Ma et al, 2000;Ratnam et al, 1999;Tarle et al, 1993;Yamaoka et al, 1992) and there have been a few reports of the broadening of the cofactor specificity to increase the activity of these enzymes with NAD(H) (Banta and Anderson, 2002;Banta et al, 2002b;Khoury et al, 2009;Liang et al, 2007;Sanli et al, 2004). Through these efforts, several hot spots for mutagenesis have been identified.…”

Section: Introductionmentioning

confidence: 97%

Broadening the cofactor specificity of a thermostable alcohol dehydrogenase using rational protein design introduces novel kinetic transient behavior

Campbell

Wheeldon

Banta

2010

Biotech & Bioengineering

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Cofactor specificity in the aldo-keto reductase (AKR) superfamily has been well studied, and several groups have reported the rational alteration of cofactor specificity in these enzymes. Although most efforts have focused on mesostable AKRs, several putative AKRs have recently been identified from hyperthermophiles. The few that have been characterized exhibit a strong preference for NAD(H) as a cofactor, in contrast to the NADP(H) preference of the mesophilic AKRs. Using the design rules elucidated from mesostable AKRs, we introduced two site-directed mutations in the cofactor binding pocket to investigate cofactor specificity in a thermostable AKR, AdhD, which is an alcohol dehydrogenase from Pyrococcus furiosus. The resulting double mutant exhibited significantly improved activity and broadened cofactor specificity as compared to the wild-type. Results of previous pre-steady-state kinetic experiments suggest that the high affinity of the mesostable AKRs for NADP(H) stems from a conformational change upon cofactor binding which is mediated by interactions between a canonical arginine and the 2'-phosphate of the cofactor. Pre-steady-state kinetics with AdhD and the new mutants show a rich conformational behavior that is independent of the canonical arginine or the 2'-phosphate. Additionally, experiments with the highly active double mutant using NADPH as a cofactor demonstrate an unprecedented transient behavior where the binding mechanism appears to be dependent on cofactor concentration. These results suggest that the structural features involved in cofactor specificity in the AKRs are conserved within the superfamily, but the dynamic interactions of the enzyme with cofactors are unexpectedly complex.

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Section: Introductionmentioning

confidence: 97%

Broadening the cofactor specificity of a thermostable alcohol dehydrogenase using rational protein design introduces novel kinetic transient behavior

Campbell

Wheeldon

Banta

2010

Biotech & Bioengineering

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“…Conformational changes in the loop are necessary for the release of the cofactor. Substrate reduction involves a tyrosine residue, acting as an acid along with other active-site residues arginine, histidine, and glutamate [59]. The phenolic hydroxyl group of tyrosine mediates the catalytic reduction of the substrate carbonyl group.…”

Section: Proteins Involved In Metabolic Regulationmentioning

confidence: 99%

Protein Structure–Function Paradigm in Plant Stress Tolerance

Bhatt

Kumar

Bhavesh

2013

Climate Change and Plant Abiotic Stress Tolerance

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Abiotic stresses lead to major losses of crops and fertile land worldwide. The increased percentage of arable land affected by various kinds of abiotic stresses has intensified the need to understand the different biochemical and physiological changes experienced by plants. There is an urgent need for transgenic plants having a high yield, yet also being stress tolerant, to counter this situation. Various plant and fungal proteins have been identified and characterized at the biochemical, physiological, and molecular levels, imparting stress tolerance in various model plant systems. Understanding the molecular mechanism of these proteins at atomic resolution using structural biology approaches gives a distinct edge over traditional approaches. Various proteins involved in countering abiotic stress conditions in various model organism and their future prospects are discussed with respect to their high-resolution structural information.

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“…9,12 A notable exception is the Candida tenuis xylose reductase (CtXR), which reduces ortho-substituted acetophenones better than the correspondent para-and meta-derivatives, provided the substituent had both low steric impact and electron withdrawing effect (i.e., F, Cl, Br); 13 CtXR has a wide binding pocket, able to accommodate a wide variety of substrates, including ortho-substituted acetophenones. 25 In order to explain the different reactivity for the CvO group of 2′-cyanoacetophenone 1u and 2′-nitroacetophenone 1v, we computed the LUMO frontier molecular orbitals, as shown in Fig. 4, panels b (1u) and c (1v).…”

Section: Paper Organic and Biomolecular Chemistrymentioning

confidence: 99%

Enzymatic reduction of acetophenone derivatives with a benzil reductase from Pichia glucozyma (KRED1-Pglu): electronic and steric effects on activity and enantioselectivity

et al. 2016

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A recombinant ketoreductase from Pichia glucozyma (KRED1-Pglu) was used for the enantioselective reduction of various mono-substituted acetophenones. Reaction rates of meta-and para-derivatives were consistent with the electronic effects described by σ-Hammett coefficients; on the other hand, enantioselectivity was determined by an opposite orientation of the substrate in the binding pocket.Reduction of ortho-derivatives occurred only with substrates bearing substituents with low steric impact (i.e., F and CN). Reactivity was controlled by stereoelectronic features (CvO length and charge, shape of LUMO frontier molecular orbitals), which can be theoretically calculated.

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Catalytic mechanism and substrate selectivity of aldo-keto reductases: Insights from structure-function studies of Candida tenuis xylose reductase

Cited by 46 publications

References 38 publications

Broadening the cofactor specificity of a thermostable alcohol dehydrogenase using rational protein design introduces novel kinetic transient behavior

Broadening the cofactor specificity of a thermostable alcohol dehydrogenase using rational protein design introduces novel kinetic transient behavior

Protein Structure–Function Paradigm in Plant Stress Tolerance

Enzymatic reduction of acetophenone derivatives with a benzil reductase from Pichia glucozyma (KRED1-Pglu): electronic and steric effects on activity and enantioselectivity

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