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

Ritva Verho, Mikko Putkonen, John Londesborough, Merja Penttilä, Peter Richard

Research output: Contribution to journalArticleScientificpeer-review

59 Citations (Scopus)

Abstract

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 L-xylulose 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 L-xylulose to xylitol. It also converts D-ribulose to D-arabinitol 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 Km 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.

Original languageEnglish
Pages (from-to)14746-14751
Number of pages6
JournalJournal of Biological Chemistry
Volume279
Issue number15
DOIs
Publication statusPublished - 9 Apr 2004
MoE publication typeA1 Journal article-refereed

Fingerprint

Arabinose
Xylulose
NAD
Yeast
Yeasts
Molds
Sugar Alcohols
Xylitol
Fungi
Enzymes
NADP
Hydroxyl Radical
Genes
Ribitol
Pentoses
Pentose Phosphate Pathway
Oxidation
Gene encoding
Sugars
Saccharomyces cerevisiae

Keywords

  • reductase
  • L-Xylulose reductase
  • yeasts
  • genes
  • gene identification

Cite this

Verho, Ritva ; Putkonen, Mikko ; Londesborough, John ; Penttilä, Merja ; Richard, Peter. / A Novel NADH-linked L-Xylulose Reductase in the L-Arabinose Catabolic Pathway of Yeast. In: Journal of Biological Chemistry. 2004 ; Vol. 279, No. 15. pp. 14746-14751.
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A Novel NADH-linked L-Xylulose Reductase in the L-Arabinose Catabolic Pathway of Yeast. / Verho, Ritva; Putkonen, Mikko; Londesborough, John; Penttilä, Merja; Richard, Peter.

In: Journal of Biological Chemistry, Vol. 279, No. 15, 09.04.2004, p. 14746-14751.

Research output: Contribution to journalArticleScientificpeer-review

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AB - 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 L-xylulose 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 L-xylulose to xylitol. It also converts D-ribulose to D-arabinitol 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 Km 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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