A novel NADH-linked L-xylulose reductase in the L-arabinose catabolic pathway of yeast

R Verho, M Putkonen, J Londesborough, M Penttila, P Richard*

*Corresponding author for this work

Research output: Contribution to journalArticleScientificpeer-review

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

Keywords

  • D-ARABINITOL DEHYDROGENASE
  • SACCHAROMYCES-CEREVISIAE
  • PICHIA-STIPITIS
  • XYLOSE REDUCTASE
  • XYLITOL-DEHYDROGENASE
  • CANDIDA-TROPICALIS
  • MOLECULAR-CLONING
  • ASPERGILLUS-NIGER
  • GENE
  • EXPRESSION

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