Crystal structure of sorbitol dehydrogenase

Johansson, Kenth; El-Ahmad, Mustafa; Kaiser, Christina; Jörnvall, Hans; Eklund, Hans; Höög, Jan‐Olov; Ramaswamy, S.

doi:10.1016/s0009-2797(00)00260-x

Cited by 42 publications

(36 citation statements)

References 10 publications

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“…In contrast to the LADH structures and the 3.0-Å structure of rat SDH (23), but in agreement with the crystal structures of human SDH (22), the structures presented here indicate that the primary coordination sphere of the catalytic zinc includes a water molecule, Glu 67 , His…”

Section: Discussionsupporting

confidence: 83%

“…On this basis it has been predicted that SsGDH and TaGDH will oxidize their sugar substrates via a mechanism similar to that of other, characterized, MDR family members (17). Previous structural and mechanistic studies of MDR family members have centered on liver alcohol dehydrogenase (LADH) (19 -21) and sorbitol dehydrogenase (SDH) (22,23), whereas the structure of TaGDH has only been described in an apo form (17). Thus there are no currently available structures of an MDR family member in complex with a ring form sugar substrate.…”

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confidence: 99%

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The Structural Basis of Substrate Promiscuity in Glucose Dehydrogenase from the Hyperthermophilic Archaeon Sulfolobus solfataricus

Milburn

Lamble

Theodossis

et al. 2006

Journal of Biological Chemistry

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The hyperthermophilic archaeon Sulfolobus solfataricus grows optimally above 80°C and utilizes an unusual, promiscuous, nonphosphorylative Entner-Doudoroff pathway to metabolize both glucose and galactose. The first enzyme in this pathway, glucose dehydrogenase, catalyzes the oxidation of glucose to gluconate, but has been shown to have activity with a broad range of sugar substrates, including glucose, galactose, xylose, and L-arabinose, with a requirement for the glucose stereo configuration at the C2 and C3 positions. Here we report the crystal structure of the apo form of glucose dehydrogenase to a resolution of 1.8 Å and a complex with its required cofactor, NADP ؉ , to a resolution of 2.3 Å . A T41A mutation was engineered to enable the trapping of substrate in the crystal. Complexes of the enzyme with D-glucose and D-xylose are presented to resolutions of 1.6 and 1.5 Å , respectively, that provide evidence of selectivity for the ␤-anomeric, pyranose form of the substrate, and indicate that this is the productive substrate form. The nature of the promiscuity of glucose dehydrogenase is also elucidated, and a physiological role for this enzyme in xylose metabolism is suggested. Finally, the structure suggests that the mechanism of sugar oxidation by this enzyme may be similar to that described for human sorbitol dehydrogenase.The hyperthermophilic archaeon Sulfolobus solfataricus grows optimally at 80 -85°C and pH 2-4, utilizing a wide range of carbon and energy sources, and has been used as a model organism of archaeal sugar metabolism, being subject to extensive and comprehensive investigations (1, 2). Central metabolism in S. solfataricus involves a variant of the Entner-Doudoroff pathway (3). Typically this pathway has been described as non-phosphorylative (3), proceeding with no net ATP production, and with analogous pathways being described for the thermophilic archaea Sulfolobus acidocaldarius (4), Thermoplasma acidophilum (5), and Thermoproteus tenax (6), as well as certain strains of Aspergillus fungi (7,8). In this pathway, glucose dehydrogenase and gluconate dehydratase catalyze the oxidation of glucose to gluconate and the subsequent dehydration of gluconate to 2-keto-3-deoxygluconate (KDG).2 KDG aldolase then catalyzes the cleavage of KDG to glyceraldehyde and pyruvate. The glyceraldehyde is phosphorylated by glycerate kinase to give 2-phosphoglycerate. A second molecule of pyruvate is then produced from this by the actions of enolase and pyruvate kinase. Glucose dehydrogenase and KDG aldolase from S. solfataricus have been reported to have high activity with galactose and 2-keto-3-deoxygalactonate, respectively (9), with recent reports demonstrating the activity of gluconate dehydratase from this organism with galactonate (10, 11). Consequently, it was proposed that the entire central metabolic pathway in this organism is promiscuous for the metabolism of glucose and galactose. This situation is in contrast with other microorganisms, where separate enzymes and pathways are present for the meta...

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Section: Discussionsupporting

confidence: 83%

mentioning

confidence: 99%

The Structural Basis of Substrate Promiscuity in Glucose Dehydrogenase from the Hyperthermophilic Archaeon Sulfolobus solfataricus

Milburn

Lamble

Theodossis

et al. 2006

Journal of Biological Chemistry

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“…The holoenzyme of SDH is a tetramer with MW of 38000 Da and 348 amino acid proteins expression pattern were investigated during the different stages of embryonic development (Wang, et al, 2013). Similarly in the plasma of Rat SDH is tetramer with four identical monomers (Johansson et al, 2001). Number of other enzymes are present as oligomers of identical monomers forming dimmers or tetramers (Traut,1994).…”

Section: Resultsmentioning

confidence: 99%

Purification of sorbitol dehydrogenase from diapause eggs of Dysdercus cingulatus (fabricus 1775)

Shahjahan

Martin

2017

IJARe

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Sorbitol Dehydrogenase (SDH) is important enzymes responsible for the protection of eggs from harsh condition as it convert fructose to sorbitol which serves as polyol and stabilize the soluble proteins and lipid bilayer. The enzyme SDH (dIdiol NAD oxidoreductase EC 1.1.1.14) is purified from the eggs of red cotton bug, Dysdercus cingulatus exposed to low temperature. The SDH from eggs were purified by using different protein purification technique viz, ion exchange chromatography, gel filtration and SDS-PAGE. The ion exchange chromatography showed 0.185U total activity, with a purification fold of 1.48 and yield 3.36%. Gel filtration chromatography showed an increase in purification of SDH activity by 2.73 with 0.095 U total activity and yield of 1.73%. SDS-PAGE revealed that SDH molecular weight of 38 KDa. The Km value of Fructose: NADH is 0.4:0.01 and the Vmax was 0.6 and 0.03. Thus low temperature increases the activity of SDH which convert fructose to sorbitol which protects the cell during diapause condition.

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“…2A), suggesting that the stability of the C4 mutant was also increased in vivo. The removal of Zn 2ϩ and concomitant enzyme inactivation has been observed in several zinc-containing ADHs and SDHs (47)(48)(49). The enzyme activity of WT and the C4 mutant decreased to 50 and 90%, respectively, by treatment with 1 mM EDTA (pH 7.5) (Fig.…”

Section: Introduction Of Additional Structural Zinc Atom-becausementioning

confidence: 80%

“…There are some substitution patterns of these four cysteines in the PDHs (Table I). Among them, enzymes with the pattern of DSMD, RDCS, RDCT have been shown to have no structural zinc by enzymatic or crystallographic analyses (15,46,47). These analyses suggested that four fully conserved cysteine residues are necessary for binding to the structural zinc atom and that PsXDH having a pattern of SSYC has no structural zinc atom.…”

Section: Introduction Of Additional Structural Zinc Atom-becausementioning

confidence: 99%

Complete Reversal of Coenzyme Specificity of Xylitol Dehydrogenase and Increase of Thermostability by the Introduction of Structural Zinc

Watanabe

Kodaki

Makino

2005

Journal of Biological Chemistry

172

106

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Pichia stipitis NAD؉ -dependent xylitol dehydrogenase (XDH), a medium-chain dehydrogenase/reductase, is one of the key enzymes in ethanol fermentation from xylose. For the construction of an efficient biomass-ethanol conversion system, we focused on the two areas of XDH, 1) change of coenzyme specificity from NAD ؉ to NADP ؉ and 2) thermostabilization by introducing an additional zinc atom. Site-directed mutagenesis was used to examine the roles of Asp 207 ؉ -dependent XDH mutants constructed in this study decreased the thermostability compared with the wild-type enzyme, we attempted to improve the thermostability of XDH mutants by the introduction of an additional zinc atom. The introduction of three cysteine residues in wild-type XDH gave an additional zinc-binding site and improved the thermostability. The introduction of this mutation in D207A/I208R/F209S and D207A/I208R/F209S/N211R mutants increased the thermostability and further increased the catalytic activity with NADP ؉ .Xylose is one of the major components of hemicellulose, the second most abundant carbohydrate polymer in nature. Efficient utilization of xylose is required to develop economically viable processes for producing biofuels such as ethanol from biomass (for recent reviews, see Refs. 1-4). Yeasts have long been used for the production of alcoholic beverages such as wine, beer, and Japanese sake. In particular, Saccharomyces cerevisiae has been used widely because of the ability to produce high concentrations of ethanol and high inherent ethanol tolerance. The native strains, however, cannot ferment xylose as a carbon source. The major strategy for the generation of xylose-fermenting S. cerevisiae is to introduce genes involved in xylose metabolism from other organisms (Scheme 1). In xylosefermenting fungi such as Pichia stipitis, xylose is converted into xylulose by the sequential action of two oxidoreductases. First, xylose reductase (alditol:NADP ϩ 1-oxidoreductase, EC 1.1.1.21) catalyzes reduction of the C1 carbonyl group of xylose, yielding xylitol as the product. Xylitol is then oxidized by xylitol dehydrogenase (XDH 1 ; EC 1.1.1.9) to give xylulose. S. cerevisiae transformed with these two genes from P. stipitis could ferment xylose to ethanol. There is another problem; the excretion of xylitol occurs unless a co-metabolizable carbon source such as glucose is added. This is probably caused by several combined factors. In particular, intercellular redox imbalance due to a different coenzyme specificity of xylose reductase (with NADPH) and XDH (with NAD ϩ ) has been thought to be one of the main factors (2). The generation of an NADP ϩ -dependent XDH by protein engineering would avoid this problem.XDH is a medium-chain dehydrogenase/reductase (MDR), which constitutes a large enzyme superfamily consisting of about 1000 members (for reviews, see Refs. 5 and 6). The superfamily is classified into eight subfamilies based on amino acid sequence alignment and the structural similarity of substrates. XDH belongs to the polyol dehydrogenase ...

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Crystal structure of sorbitol dehydrogenase

Cited by 42 publications

References 10 publications

The Structural Basis of Substrate Promiscuity in Glucose Dehydrogenase from the Hyperthermophilic Archaeon Sulfolobus solfataricus

The Structural Basis of Substrate Promiscuity in Glucose Dehydrogenase from the Hyperthermophilic Archaeon Sulfolobus solfataricus

Purification of sorbitol dehydrogenase from diapause eggs of Dysdercus cingulatus (fabricus 1775)

Complete Reversal of Coenzyme Specificity of Xylitol Dehydrogenase and Increase of Thermostability by the Introduction of Structural Zinc

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