The missing link in the fungal L-arabinose catabolic pathway, identification of the L-xylulose reductase gene

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

Research output: Contribution to journalArticleScientificpeer-review

78 Citations (Scopus)

Abstract

The fungal L-arabinose pathway consists of five enzymes, aldose reductase, L-arabinitol 4-dehydrogenase, L-xylulose reductase, xylitol dehydrogenase, and xylulokinase. All the genes encoding the enzymes of this pathway are known except for that of L-xylulose reductase (EC 1.1.1.10). We identified a gene encoding this enzyme from the filamentous fungus Trichoderma reesei (Hypocreajecorina). The gene was named lxf1. It was overexpressed in the yeast Saccharomyces cerevisiae, and the enzyme activity was confirmed in a yeast cell extract. Overexpression of all enzymes of the L-arabinose pathway in S. cerevisiae led to growth of S. cerevisiae on L-arabinose; i.e., we could show that the pathway is active in a heterologous host. The lxr1 gene encoded a protein with 266 amino acids and a calculated molecular mass of 28 428 Da. The LXRI protein is an NADPH-specific reductase. It has activity with L-xylulose, D-xylulose, D-fructose, and L-sorbose. The highest affinity is toward L-xylulose (Km = 16 mM). In the reverse direction, we found activity with xylitol, D-arabinitol, D-mannitol, and D-sorbitol. It requires a bivalent cation for activity. It belongs to the protein family of short chain dehydrogenases. The enzyme is catalytically similar and homologous in sequence to a D-mannitol:NADP 2-dehydrogenase (EC 1.1.1.138).

Original languageEnglish
Pages (from-to)6432-6437
Number of pages6
JournalBiochemistry
Volume41
Issue number20
DOIs
Publication statusPublished - 21 May 2002
MoE publication typeA1 Journal article-refereed

Fingerprint

Arabinose
Yeast
Xylulose
Genes
Enzymes
Oxidoreductases
Gene encoding
L-arabinitol 4-dehydrogenase
Saccharomyces cerevisiae
Mannitol
NADP
mannitol 2-dehydrogenase (NADP)
D-Xylulose Reductase
Sorbose
Mannitol Dehydrogenases
Xylitol
Yeasts
Aldehyde Reductase
Proteins
Sorbitol

Keywords

  • genes
  • gene identification
  • L-Arabinose
  • bioethanol
  • bioenergy

Cite this

Richard, Peter ; Putkonen, Mikko ; Väänänen, Ritva ; Londesborough, John ; Penttilä, Merja. / The missing link in the fungal L-arabinose catabolic pathway, identification of the L-xylulose reductase gene. In: Biochemistry. 2002 ; Vol. 41, No. 20. pp. 6432-6437.
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The missing link in the fungal L-arabinose catabolic pathway, identification of the L-xylulose reductase gene. / Richard, Peter; Putkonen, Mikko; Väänänen, Ritva; Londesborough, John; Penttilä, Merja.

In: Biochemistry, Vol. 41, No. 20, 21.05.2002, p. 6432-6437.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - The missing link in the fungal L-arabinose catabolic pathway, identification of the L-xylulose reductase gene

AU - Richard, Peter

AU - Putkonen, Mikko

AU - Väänänen, Ritva

AU - Londesborough, John

AU - Penttilä, Merja

PY - 2002/5/21

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N2 - The fungal L-arabinose pathway consists of five enzymes, aldose reductase, L-arabinitol 4-dehydrogenase, L-xylulose reductase, xylitol dehydrogenase, and xylulokinase. All the genes encoding the enzymes of this pathway are known except for that of L-xylulose reductase (EC 1.1.1.10). We identified a gene encoding this enzyme from the filamentous fungus Trichoderma reesei (Hypocreajecorina). The gene was named lxf1. It was overexpressed in the yeast Saccharomyces cerevisiae, and the enzyme activity was confirmed in a yeast cell extract. Overexpression of all enzymes of the L-arabinose pathway in S. cerevisiae led to growth of S. cerevisiae on L-arabinose; i.e., we could show that the pathway is active in a heterologous host. The lxr1 gene encoded a protein with 266 amino acids and a calculated molecular mass of 28 428 Da. The LXRI protein is an NADPH-specific reductase. It has activity with L-xylulose, D-xylulose, D-fructose, and L-sorbose. The highest affinity is toward L-xylulose (Km = 16 mM). In the reverse direction, we found activity with xylitol, D-arabinitol, D-mannitol, and D-sorbitol. It requires a bivalent cation for activity. It belongs to the protein family of short chain dehydrogenases. The enzyme is catalytically similar and homologous in sequence to a D-mannitol:NADP 2-dehydrogenase (EC 1.1.1.138).

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KW - genes

KW - gene identification

KW - L-Arabinose

KW - bioethanol

KW - bioenergy

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DO - 10.1021/bi025529i

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JF - Biochemistry

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