The fungal L-arabinose catabolic pathway

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

Research output: Contribution to conferenceConference articleScientific

Abstract

L-arabinose is a major constituent of plant material so that L-arabinose catabolism is of relevance for microorganisms living on decaying plant material but also in biotechnology where cheap raw materials are used. The fungal pathway, which is distinctly different to the bacterial pathway consists of two NADPH-linked reductases and two NAD-linked dehydrogenases and xylulokinase. Genes coding for enzymes of this pathway are known except for L-arabinitol 4-dehydrogenase (EC 1.1.1.12) and L-xylulose reductase (EC 1.1.1.10). We recently identified these two unknown genes of the fungal pathway. The L-arabinitol 4-dehydrogenase was purified from the mould Trichoderma reesei. Amino acid sequence of peptide fragments were obtained from the purified protein and the corresponding gene cloned by using PCR. The L-xylulose reductase was identified by screening a cDNA library of T. reesei. A strain of S. cerevisiae was constructed which contained all genes of the pathway except a gene for the L-xylulose reductase. To this strain we then transformed a T. reesei cDNA library in a yeast expression vector and screened for growth on L-arabinose. We could demonstrate that the whole pathway is active in S. cerevisiae, in a strain where all the genes of this pathway are over-expressed, leading to growth on L-arabinose and ethanol production under anaerobic conditions, however at a very low rate. We discuss the limiting steps of L-arabinose catabolism and show ways to improve it.
Original languageEnglish
Publication statusPublished - 2003
EventXXI International Conference on Yeast Genetics and Molecular Biology - Gothenburg, Sweden
Duration: 7 Jul 200312 Jul 2003

Conference

ConferenceXXI International Conference on Yeast Genetics and Molecular Biology
CountrySweden
CityGothenburg
Period7/07/0312/07/03

Fingerprint

arabinose
xylulose
Trichoderma reesei
arabinitol
genes
cDNA libraries
metabolism
NAD (coenzyme)
ethanol production
NADP (coenzyme)
anaerobic conditions
molds (fungi)
biotechnology
raw materials
amino acid sequences
peptides
yeasts
screening
microorganisms
enzymes

Cite this

Richard, P., Verho, R., Putkonen, M., Londesborough, J., & Penttilä, M. (2003). The fungal L-arabinose catabolic pathway. Paper presented at XXI International Conference on Yeast Genetics and Molecular Biology, Gothenburg, Sweden.
Richard, Peter ; Verho, Ritva ; Putkonen, Mikko ; Londesborough, John ; Penttilä, Merja. / The fungal L-arabinose catabolic pathway. Paper presented at XXI International Conference on Yeast Genetics and Molecular Biology, Gothenburg, Sweden.
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Richard, P, Verho, R, Putkonen, M, Londesborough, J & Penttilä, M 2003, 'The fungal L-arabinose catabolic pathway' Paper presented at XXI International Conference on Yeast Genetics and Molecular Biology, Gothenburg, Sweden, 7/07/03 - 12/07/03, .

The fungal L-arabinose catabolic pathway. / Richard, Peter; Verho, Ritva; Putkonen, Mikko; Londesborough, John; Penttilä, Merja.

2003. Paper presented at XXI International Conference on Yeast Genetics and Molecular Biology, Gothenburg, Sweden.

Research output: Contribution to conferenceConference articleScientific

TY - CONF

T1 - The fungal L-arabinose catabolic pathway

AU - Richard, Peter

AU - Verho, Ritva

AU - Putkonen, Mikko

AU - Londesborough, John

AU - Penttilä, Merja

N1 - CA2: BEL2 CA: BEL

PY - 2003

Y1 - 2003

N2 - L-arabinose is a major constituent of plant material so that L-arabinose catabolism is of relevance for microorganisms living on decaying plant material but also in biotechnology where cheap raw materials are used. The fungal pathway, which is distinctly different to the bacterial pathway consists of two NADPH-linked reductases and two NAD-linked dehydrogenases and xylulokinase. Genes coding for enzymes of this pathway are known except for L-arabinitol 4-dehydrogenase (EC 1.1.1.12) and L-xylulose reductase (EC 1.1.1.10). We recently identified these two unknown genes of the fungal pathway. The L-arabinitol 4-dehydrogenase was purified from the mould Trichoderma reesei. Amino acid sequence of peptide fragments were obtained from the purified protein and the corresponding gene cloned by using PCR. The L-xylulose reductase was identified by screening a cDNA library of T. reesei. A strain of S. cerevisiae was constructed which contained all genes of the pathway except a gene for the L-xylulose reductase. To this strain we then transformed a T. reesei cDNA library in a yeast expression vector and screened for growth on L-arabinose. We could demonstrate that the whole pathway is active in S. cerevisiae, in a strain where all the genes of this pathway are over-expressed, leading to growth on L-arabinose and ethanol production under anaerobic conditions, however at a very low rate. We discuss the limiting steps of L-arabinose catabolism and show ways to improve it.

AB - L-arabinose is a major constituent of plant material so that L-arabinose catabolism is of relevance for microorganisms living on decaying plant material but also in biotechnology where cheap raw materials are used. The fungal pathway, which is distinctly different to the bacterial pathway consists of two NADPH-linked reductases and two NAD-linked dehydrogenases and xylulokinase. Genes coding for enzymes of this pathway are known except for L-arabinitol 4-dehydrogenase (EC 1.1.1.12) and L-xylulose reductase (EC 1.1.1.10). We recently identified these two unknown genes of the fungal pathway. The L-arabinitol 4-dehydrogenase was purified from the mould Trichoderma reesei. Amino acid sequence of peptide fragments were obtained from the purified protein and the corresponding gene cloned by using PCR. The L-xylulose reductase was identified by screening a cDNA library of T. reesei. A strain of S. cerevisiae was constructed which contained all genes of the pathway except a gene for the L-xylulose reductase. To this strain we then transformed a T. reesei cDNA library in a yeast expression vector and screened for growth on L-arabinose. We could demonstrate that the whole pathway is active in S. cerevisiae, in a strain where all the genes of this pathway are over-expressed, leading to growth on L-arabinose and ethanol production under anaerobic conditions, however at a very low rate. We discuss the limiting steps of L-arabinose catabolism and show ways to improve it.

M3 - Conference article

ER -

Richard P, Verho R, Putkonen M, Londesborough J, Penttilä M. The fungal L-arabinose catabolic pathway. 2003. Paper presented at XXI International Conference on Yeast Genetics and Molecular Biology, Gothenburg, Sweden.