The stereospecificity of sequential nicotinamide-adenine dinucleotide-dependent oxidoreductases in relation to the evolution of metabolic sequences

Nascimento, KS; Davies, Daniel

doi:10.1042/bj1490553

Cited by 18 publications

(5 citation statements)

References 16 publications

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“…The stereospecificity of hydride transfer by C. magnoliae MDH is also very similar to that described for many SDRs. The preference of MDH for transfer of the pro-S hydrogen from NAD(P)H is typical of all members of the SDR family that have been studied (3,4,6,9,17). These results suggest that C. magnoliae MDH should be classified as an NAD(P)H-linked SDR.…”

Section: Discussionmentioning

confidence: 69%

Purification and Characterization of a Novel Mannitol Dehydrogenase from a Newly Isolated Strain of Candida magnoliae

Lee¹,

Koo²,

Kim³

et al. 2003

Appl Environ Microbiol

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Mannitol biosynthesis in Candida magnoliae HH-01 (KCCM-10252), a yeast strain that is currently used for the industrial production of mannitol, is catalyzed by mannitol dehydrogenase (MDH) (EC 1.1.1.138). In this study, NAD(P)H-dependent MDH was purified to homogeneity from C. magnoliae HH-01 by ion-exchange chromatography, hydrophobic interaction chromatography, and affinity chromatography. The relative molecular masses of C. magnoliae MDH, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and size-exclusion chromatography, were 35 and 142 kDa, respectively, indicating that the enzyme is a tetramer. This enzyme catalyzed both fructose reduction and mannitol oxidation. The pH and temperature optima for fructose reduction and mannitol oxidation were 7.5 and 37°C and 10.0 and 40°C, respectively. C. magnoliae MDH showed high substrate specificity and high catalytic efficiency (k cat ‫؍‬ 823 s ؊1 , K m ‫؍‬ 28.0 mM, and k cat /K m ‫؍‬ 29.4 mM ؊1 s ؊1 ) for fructose, which may explain the high mannitol production observed in this strain. Initial velocity and product inhibition studies suggest that the reaction proceeds via a sequential ordered Bi Bi mechanism, and C. magnoliae MDH is specific for transferring the 4-pro-S hydrogen of NADPH, which is typical of a short-chain dehydrogenase reductase (SDR). The internal amino acid sequences of C. magnoliae MDH showed a significant homology with SDRs from various sources, indicating that the C. magnoliae MDH is an NAD(P)H-dependent tetrameric SDR. Although MDHs have been purified and characterized from several other sources, C. magnoliae MDH is distinguished from other MDHs by its high substrate specificity and catalytic efficiency for fructose only, which makes C. magnoliae MDH the ideal choice for industrial applications, including enzymatic synthesis of mannitol and salt-tolerant plants.

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Section: Discussionmentioning

confidence: 69%

Purification and Characterization of a Novel Mannitol Dehydrogenase from a Newly Isolated Strain of Candida magnoliae

Lee¹,

Koo²,

Kim³

et al. 2003

Appl Environ Microbiol

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“…The stereochemical course and kinetic mechanism of PTR1 are very similar to those which have been described for many members of the SDR family. The preference of PTR1 for transfer of the pro-S hydrogen from NADPH is observed for all other members of the SDR family which have been studied ( 39 , − . Members of the AKR family () have been observed, like DHFR, to transfer the A-side hydrogen of NADH or NADPH ( − ).…”

Section: Resultsmentioning

confidence: 98%

Leishmania major Pteridine Reductase 1 Belongs to the Short Chain Dehydrogenase Family: Stereochemical and Kinetic Evidence

et al. 1998

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Pteridine reductase 1 (PTR1) is a novel broad spectrum enzyme of pterin and folate metabolism in the protozoan parasite Leishmania. Overexpression of PTR1 confers methotrexate resistance to these protozoa, arising from the enzyme's ability to reduce dihydrofolate and its relative insensitivity to methotrexate. The kinetic mechanism and stereochemical course for the catalyzed reaction confirm PTR1's membership within the short chain dehydrogenase/reductase (SDR) family. With folate as a substrate, PTR1 catalyzes two rounds of reduction, yielding 5,6,7, 8-tetrahydrofolate and oxidizing 2 equiv of NADPH. Dihydrofolate accumulates transiently during folate reduction and is both a substrate and an inhibitor of PTR1. PTR1 transfers the pro-S hydride of NADPH to carbon 6 on the si face of dihydrofolate, producing the same stereoisomer of THF as does dihydrofolate reductase. Product inhibition and isotope partitioning studies support an ordered ternary complex mechanism, with NADPH binding first and NADP+ dissociating after the reduced pteridine. Identical kinetic mechanisms and NAD(P)H hydride chirality preferences are seen with other SDRs. An observed tritium effect upon V/K for reduction of dihydrofolate arising from isotopic substitution of the transferred hydride was suppressed at a high concentration of dihydrofolate, consistent with a steady-state ordered kinetic mechanism. Interestingly, half of the binary enzyme-NADPH complex appears to be incapable of rapid turnover. Fluorescence quenching results also indicate the existence of a nonproductive binary enzyme-dihydrofolate complex. The nonproductive complexes observed between PTR1 and its substrates are unique among members of the SDR family and may provide leads for developing antileishmanial therapeutics.

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“…Therefore, these results imply that the AfM1PDH belongs to the group of pro-S specific dehydrogenases, a functional classification that the fungal enzyme shares with M1PDHs from the bacteria Aerobacter aerogenes and E. coli. 9,18,19 The stereoselectivity of biocatalytic hydrogen transfer to NAD + reflects the relative orientation of substrate and coenzyme upon their binding to the enzyme. It is dictated by the protein structure and therefore expected to be conserved among evolutionary related dehydrogenases.…”

Section: Stereochemical Course Of Hydrogen Transfer Catalyzed By Afm1pdhmentioning

confidence: 99%

Characterization of recombinant Aspergillus fumigatus mannitol-1-phosphate 5-dehydrogenase and its application for the stereoselective synthesis of protio and deuterio forms of d-mannitol 1-phosphate

Krahulec

Armao

Weber

et al. 2008

Carbohydrate Research

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The stereospecificity of sequential nicotinamide-adenine dinucleotide-dependent oxidoreductases in relation to the evolution of metabolic sequences

Cited by 18 publications

References 16 publications

Purification and Characterization of a Novel Mannitol Dehydrogenase from a Newly Isolated Strain of Candida magnoliae

Purification and Characterization of a Novel Mannitol Dehydrogenase from a Newly Isolated Strain of Candida magnoliae

Leishmania major Pteridine Reductase 1 Belongs to the Short Chain Dehydrogenase Family: Stereochemical and Kinetic Evidence

Characterization of recombinant Aspergillus fumigatus mannitol-1-phosphate 5-dehydrogenase and its application for the stereoselective synthesis of protio and deuterio forms of d-mannitol 1-phosphate

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