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Xylose Utilisation: Cloning and Characterisation of the Xylitol Dehydrogenase from Galactocandida mastotermitis

  • Andreas Habenicht , Hassan Motejadded , Michael Kiess , Angelika Wegerer and Ralf Mattes
Published/Copyright: June 1, 2005
Biological Chemistry
From the journal Volume 380 Issue 12

Abstract

We cloned and successfully expressed the gene for xylitol dehydrogenase from Galactocandida mastotermitis in Escherichia coli. The amino acid sequence revealed that the enzyme belongs to the superfamily of zinc containing, medium-chain alcohol dehydrogenases. The enzyme catalyses the second step in the xylose utilising pathway converting xylose to xylulosephosphate. Xylulose-phosphate is further degraded by the transaldolase and transketolase reactions of the pentose phosphate pathway.

The purified xylitol dehydrogenase from G. mastotermitis was subjected to partial amino acid sequence analysis. The resulting amino acid information was then used to construct oligonucleotide probes for PCR amplification. The PCR product was used to screen a genomic library. The identified xdh gene includes one short intron at its 5′ end. Putative regulatory signals were identified with the help of Saccharomyces cerevisiae regulatory sequence databases. An intronless xdh transcript, cloned by RT-PCR, was actively expressed in pBTac1 at 37°C to approximately 8% of the soluble E. coli protein. Furthermore, the kinetic parameters were determined and conditions were found to stabilise the soluble and active protein.

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Published Online: 2005-06-01
Published in Print: 1999-12-20

Copyright © 1999 by Walter de Gruyter GmbH & Co. KG

Articles in the same Issue

  1. To Our Authors, Readers and Subscribers
  2. Editor's Note
  3. A Chimpanzee Millennium
  4. Molecular Genetics of Dopa-Responsive Dystonia
  5. In Vitro Transcription of a TATA-Less Promoter: Negative Regulation by the Not1 Protein
  6. Fast Control of DNA Replication in Response to Hypoxia and to Inhibited Protein Synthesis in CCRF-CEM and HeLa Cells
  7. The Inhibition of NF-B Activation Pathways and the Induction of Apoptosis by Dithiocarbamates in T Cells Are Blocked by the Glutathione Precursor N-Acetyl-L-Cysteine
  8. Xylose Utilisation: Cloning and Characterisation of the Xylose Reductase from Candida tenuis
  9. Xylose Utilisation: Cloning and Characterisation of the Xylitol Dehydrogenase from Galactocandida mastotermitis
  10. Thermodynamic Properties and DNA Binding of the ParD Protein from the Broad Host-Range Plasmid RK2/RP4 Killing System
  11. Dipeptidyl Peptidase III from Rat Liver Cytosol: Purification, Molecular Cloning and Immunohistochemical Localization
  12. Genomic Expansion Across the Albumin Gene Family on Human Chromosome 4q Is Directional
  13. Comparison of the Tamoxifen Regulated Chimeric Cre Recombinases MerCreMer and CreMer
  14. The Human Cathepsin F Gene a Fusion Product between an Ancestral Cathepsin and Cystatin Gene
  15. Characterization of a New Member of the TNF Family Expressed on Antigen Presenting Cells
  16. Vascular Endothelial Growth Factor (VEGF) and Its Receptors in Tumor-Bearing Dogs
  17. Molecular Cloning and Characterization of a cDNA Encoding a Transferrin Homolog from Bombyx mori
  18. Mitochondria-Derived and Extra-Mitochondrial Human Type-1 Porin Are Identical as Revealed by Amino Acid Sequencing and Electrophysiological Characterisation
  19. Acknowledgement
  20. Content Index
  21. Author Index
  22. Subject Index
https://doi.org/10.1515/BC.1999.180

Articles in the same Issue

  1. To Our Authors, Readers and Subscribers
  2. Editor's Note
  3. A Chimpanzee Millennium
  4. Molecular Genetics of Dopa-Responsive Dystonia
  5. In Vitro Transcription of a TATA-Less Promoter: Negative Regulation by the Not1 Protein
  6. Fast Control of DNA Replication in Response to Hypoxia and to Inhibited Protein Synthesis in CCRF-CEM and HeLa Cells
  7. The Inhibition of NF-B Activation Pathways and the Induction of Apoptosis by Dithiocarbamates in T Cells Are Blocked by the Glutathione Precursor N-Acetyl-L-Cysteine
  8. Xylose Utilisation: Cloning and Characterisation of the Xylose Reductase from Candida tenuis
  9. Xylose Utilisation: Cloning and Characterisation of the Xylitol Dehydrogenase from Galactocandida mastotermitis
  10. Thermodynamic Properties and DNA Binding of the ParD Protein from the Broad Host-Range Plasmid RK2/RP4 Killing System
  11. Dipeptidyl Peptidase III from Rat Liver Cytosol: Purification, Molecular Cloning and Immunohistochemical Localization
  12. Genomic Expansion Across the Albumin Gene Family on Human Chromosome 4q Is Directional
  13. Comparison of the Tamoxifen Regulated Chimeric Cre Recombinases MerCreMer and CreMer
  14. The Human Cathepsin F Gene a Fusion Product between an Ancestral Cathepsin and Cystatin Gene
  15. Characterization of a New Member of the TNF Family Expressed on Antigen Presenting Cells
  16. Vascular Endothelial Growth Factor (VEGF) and Its Receptors in Tumor-Bearing Dogs
  17. Molecular Cloning and Characterization of a cDNA Encoding a Transferrin Homolog from Bombyx mori
  18. Mitochondria-Derived and Extra-Mitochondrial Human Type-1 Porin Are Identical as Revealed by Amino Acid Sequencing and Electrophysiological Characterisation
  19. Acknowledgement
  20. Content Index
  21. Author Index
  22. Subject Index
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