Combined application of targeted and untargeted proteomics identifies distinct metabolic alterations in the tetraacetylphytosphingosine (TAPS) producing yeast Wickerhamomyces ciferrii
| Autor: | Tim Köhler, Daniel Wolff, Ansgar Poetsch, Frank ter Veld |
|---|---|
| Rok vydání: | 2013 |
| Předmět: |
Proteomics
Proteome Biophysics Biology Ceramides Tandem mass spectrometry Biochemistry Fungal Proteins Acetyl Coenzyme A Gene Expression Regulation Fungal chemistry.chemical_classification Sphingolipids Nitrogen Isotopes Selected reaction monitoring Acetylation Sphingolipid Yeast carbohydrates (lipids) Metabolic pathway Enzyme chemistry Isotope Labeling Saccharomycetales |
| Zdroj: | Journal of Proteomics. 82:274-287 |
| ISSN: | 1874-3919 |
| DOI: | 10.1016/j.jprot.2013.03.002 |
| Popis: | The Wickerhamomyces ciferrii strain NRRL Y-1031 F-60-10A is a well-known producer of tetraacetylphytosphingosine (TAPS) and used for the biotechnological production of sphingolipids and ceramides. It was our aim to gain new biological insights into the sphingolipid metabolism by employing a dual platform mass spectrometry strategy. The first step comprised metabolic 15N-labeling in combination with label-free proteomics using high resolution LTQ Orbitrap mass spectrometry. Then selected reaction monitoring tandem mass spectrometry served for the targeted quantification of sphingoid base biosynthesis enzymes. The non-producer strain NRRL Y-1031-27 served as a reference strain. In total, 1697 proteins were identified, and 123 enzymes of main metabolic pathways were observed as differentially expressed. Major findings were: 1) the likely presence of higher carbon flux in NRRL Y-1031 F-60-10A, resulting in higher availability of pyruvate and acetyl-CoA; 2) indications of oleaginous yeast-like behavior in NRRL Y-1031 F-60-10A; and 3) approx. 2-fold higher abundance of eight sphingolipid biosynthesis enzymes in NRRL Y-1031 F-60-10A. Taken together, in NRRL Y-1031 F-60-10A, the levels of glycolytic enzymes apparently gear carbon flux towards higher acetyl-CoA synthesis rates, while simultaneously reducing protein biosynthesis in the process. Thereby, C2 building blocks for acyl-moieties and downstream sphingoid base acetylation are provided to maintain TAPS production. Biological significance First quantitative proteome study for a phytosphingosine-producing yeast. |
| Databáze: | OpenAIRE |
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