Engineering of the metabolism of Saccharomyces cerevisiae for anaerobic production of mannitol
| Autor: | Roeland Costenoble, Gunnar Lidén, Lennart Adler, Claes Niklasson |
|---|---|
| Rok vydání: | 2003 |
| Předmět: |
Nitrogen
Mutant Saccharomyces cerevisiae Gene Expression Dehydrogenase Biology medicine.disease_cause Applied Microbiology and Biotechnology Microbiology chemistry.chemical_compound Bioreactors Escherichia coli Glycerol medicine Mannitol Anaerobiosis General Medicine NAD biology.organism_classification chemistry Biochemistry Mutation NAD+ kinase Genetic Engineering Anaerobic exercise medicine.drug |
| Zdroj: | FEMS Yeast Research. 3:17-25 |
| ISSN: | 1567-1364 1567-1356 |
| DOI: | 10.1111/j.1567-1364.2003.tb00134.x |
| Popis: | Under anaerobic conditions, Saccharomyces cerevisiae uses NADH-dependent glycerol-3-phosphate dehydrogenase (Gpd1p and Gpd2p) to re-oxidize excess NADH, yielding substantial amounts of glycerol. In a Deltagpd1 Deltagpd2 double-null mutant, the necessary NAD+ regeneration through glycerol production is no longer possible, and this mutant does not grow under anaerobic conditions. The excess NADH formed can potentially be used to drive other NADH-dependent reactions or pathways. To investigate this possibility, a double-null mutant was transformed with a heterologous gene (mtlD) from Escherichia coli, coding for NADH-dependent mannitol-1-phosphate dehydrogenase. Expression of this gene in S. cerevisiae should result in NADH oxidation by the NADH-requiring formation of mannitol-1-phosphate from fructose-6-phosphate. The strain was characterized using step-change experiments, in which, during the exponential growth phase, the inlet gas was changed from air to nitrogen. It was found that the mutant produced mannitol only under anaerobic conditions. However, anaerobic growth was not regained, which was probably due to the excessive accumulation of mannitol in the cells. |
| Databáze: | OpenAIRE |
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