Peroxisomal NAD(H) Homeostasis in the Yeast <i>Debaryomyces hansenii</i> Depends on Two Redox Shuttles and the NAD<sup>+</sup> Carrier, Pmp47
Selva Turkolmez,
Serhii Chornyi,
Sondos Alhajouj,
Lodewijk IJlst,
Hans R. Waterham,
Phil J. Mitchell,
Ewald H. Hettema,
Carlo W. T. van Roermund
Affiliations
Selva Turkolmez
School of Bioscience, University of Sheffield, Sheffield S10 2TN, UK
Serhii Chornyi
Laboratory Genetic Metabolic Diseases, Department of Clinical Chemistry, Amsterdam UMC, University of Amsterdam, Meibergdreef 9, 1105 AZ Amsterdam, The Netherlands
Sondos Alhajouj
School of Bioscience, University of Sheffield, Sheffield S10 2TN, UK
Lodewijk IJlst
Laboratory Genetic Metabolic Diseases, Department of Clinical Chemistry, Amsterdam UMC, University of Amsterdam, Meibergdreef 9, 1105 AZ Amsterdam, The Netherlands
Hans R. Waterham
Laboratory Genetic Metabolic Diseases, Department of Clinical Chemistry, Amsterdam UMC, University of Amsterdam, Meibergdreef 9, 1105 AZ Amsterdam, The Netherlands
Phil J. Mitchell
School of Bioscience, University of Sheffield, Sheffield S10 2TN, UK
Ewald H. Hettema
School of Bioscience, University of Sheffield, Sheffield S10 2TN, UK
Carlo W. T. van Roermund
Laboratory Genetic Metabolic Diseases, Department of Clinical Chemistry, Amsterdam UMC, University of Amsterdam, Meibergdreef 9, 1105 AZ Amsterdam, The Netherlands
Debaryomyces hansenii is considered an unconventional yeast with a strong biotechnological potential, which can produce and store high amounts of lipids. However, relatively little is known about its lipid metabolism, and genetic tools for this yeast have been limited. The aim of this study was to explore the fatty acid β-oxidation pathway in D. hansenii. To this end, we employed recently developed methods to generate multiple gene deletions and tag open reading frames with GFP in their chromosomal context in this yeast. We found that, similar as in other yeasts, the β-oxidation of fatty acids in D. hansenii was restricted to peroxisomes. We report a series of experiments in D. hansenii and the well-studied yeast Saccharomyces cerevisiae that show that the homeostasis of NAD+ in D. hansenii peroxisomes is dependent upon the peroxisomal membrane protein Pmp47 and two peroxisomal dehydrogenases, Mdh3 and Gpd1, which both export reducing equivalents produced during β-oxidation to the cytosol. Pmp47 is the first identified NAD+ carrier in yeast peroxisomes.
