Properties of the NAD(P)H-dependent xylose reductase from the xylose-fermenting yeast <i>Pichia stipitis</i>

Verduyn, Cornelis; Kleef, R. van; Frank, Joachim; Schreuder, Henk; Dijken, Johannes P. van; Scheffers, W. A.

doi:10.1042/bj2260669

Cited by 282 publications

(121 citation statements)

References 14 publications

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“…An expected problem in the fungal pathway is the imbalance of redox cofactors. The two reduction steps are NADPH-linked: the aldose reductase is NADPH-specific, or in the case of P. stipitis, has a preference for NADPH (9). The L-xylulose reductase, which was purified from A. niger, was also shown to be NADPH-specific (29).…”

Section: Cloning Of L-arabinitol 4-dehydrogenasementioning

confidence: 99%

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Cloning and Expression of a Fungal l-Arabinitol 4-Dehydrogenase Gene

Richard¹,

Londesborough²,

Putkonen³

et al. 2001

Journal of Biological Chemistry

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Section: Cloning Of L-arabinitol 4-dehydrogenasementioning

confidence: 99%

“…The first enzyme in both pathways is an aldose reductase (EC 1.1.1.21). The corresponding enzymes in Saccharomyces cerevisiae (8) and P. stipitis (9) have been characterized. They are unspecific and can use either L-arabinose or D-xylose with approximately the same rate to produce L-arabinitol or xylitol, respectively.…”

mentioning

confidence: 99%

Cloning and Expression of a Fungal l-Arabinitol 4-Dehydrogenase Gene

Richard¹,

Londesborough²,

Putkonen³

et al. 2001

Journal of Biological Chemistry

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“…However, there is only limited knowledge about the enzymes of this pathway in yeast. Aldose reductases, which are active with L-arabinose and D-xylose, were described for the yeasts S. cerevisiae (8) and P. stipitis, for example (9). The enzymes have similar affinity toward D-xylose and L-arabinose and convert both sugars with a similar rate.…”

mentioning

confidence: 99%

A Novel NADH-linked L-Xylulose Reductase in the L-Arabinose Catabolic Pathway of Yeast

Verho

Putkonen

Londesborough

et al. 2004

Journal of Biological Chemistry

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An NADH-dependent L-xylulose reductase and the corresponding gene were identified from the yeast Ambrosiozyma monospora. The enzyme is part of the yeast pathway for L-arabinose catabolism. A fungal pathway for L-arabinose utilization has been described previously for molds. In this pathway L-arabinose is sequentially converted to L-arabinitol, L-xylulose, xylitol, and D-xylulose and enters the pentose phosphate pathway as D-xylulose 5-phosphate. In molds the reductions are NADPH-linked, and the oxidations are NAD ؉ -linked. Here we show that in A. monospora the pathway is similar, i.e. it has the same two reduction and two oxidation reactions, but the reduction by L-xylulose reductase is not performed by a strictly NADPH-dependent enzyme as in molds but by a strictly NADH-dependent enzyme. The ALX1 gene encoding the NADH-dependent Lxylulose reductase is strongly expressed during growth on L-arabinose as shown by Northern analysis. The gene was functionally overexpressed in Saccharomyces cerevisiae and the purified His-tagged protein characterized. The reversible enzyme converts Lxylulose to xylitol. It also converts D-ribulose to Darabinitol but has no activity with L-arabinitol or adonitol, i.e. it is specific for sugar alcohols where, in a Fischer projection, the hydroxyl group of the C-2 is in the L-configuration and the hydroxyl group of C-3 is in the D-configuration. It also has no activity with C-6 sugars or sugar alcohols. The K m values for L-xylulose and D-ribulose are 9.6 and 4.7 mM, respectively. To our knowledge this is the first report of an NADH-linked L-xylulose reductase.

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“…In this organism, xylose is converted into xylulose by two oxidoreductases. First, xylose is reduced to xylitol by an NADPH/NADH-linked xylose reducatase (XR) [6], and then xylitol is oxidized to xylulose by an NAD-linked xylitol dehydrogenase (XDH) [7]. Finally, xylulokinase (XK) phosphorylates xylulose into xylulose-5-phosphate, which is metabolized further via the pentose phosphate pathway (PPP) (Fig.…”

Section: Introductionmentioning

confidence: 99%

Conversion of Waste Agriculture Biomass to Bioethanol by Recombinant <i>Saccharomyces cerevisiae</i>

et al. 2010

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Agricultural waste biomass has already been transferred to bioethanol and used as energy related products, although many issues such as efficiency and productivity still to be overcome. In this study, the protein engineering was applied to generate enzymes with completely reversed coenzyme specificity and developed recombinant yeasts containing those engineered enzymes for construction of an efficient biomass-ethanol conversion system. Recombinant yeasts were constructed with the genes encoding a wild type xylose reductase (XR) and the protein engineered xylitol dehydrogenase (XDH) (with NADP) of Pichia stipitis. These recombinant yeasts were characterized based on the enzyme activity and fermentation ability of xylose to ethanol. The protein engineered enzymes were expressed significantly in Saccharomyces cerevisiae as judged by the enzyme activity in vitro. Ethanol fermentation was measured in batch culture under anaerobic conditions. The significant enhancement was found in Y-ARS strain, in which NADP + -dependent XDH was expressed; 85% decrease of unfavorable xylitol excretion with 26% increased ethanol production, when compared with the reference strain expressing the wild-type XDH.

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Properties of the NAD(P)H-dependent xylose reductase from the xylose-fermenting yeast Pichia stipitis

Cited by 282 publications

References 14 publications

Cloning and Expression of a Fungal l-Arabinitol 4-Dehydrogenase Gene

Cloning and Expression of a Fungal l-Arabinitol 4-Dehydrogenase Gene

A Novel NADH-linked L-Xylulose Reductase in the L-Arabinose Catabolic Pathway of Yeast

Conversion of Waste Agriculture Biomass to Bioethanol by Recombinant <i>Saccharomyces cerevisiae</i>

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