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Combined Application of Targeted and Untargeted Proteomics Identifies Distinct Metabolic Alterations in the Tetraacetylphytosphingosine (TAPS) Producing Yeast Wickerhamomyces Ciferrii

Combined Application of Targeted and Untargeted Proteomics Identifies Distinct Metabolic Alterations in the Tetraacetylphytosphingosine (TAPS) Producing Yeast Wickerhamomyces Ciferrii

Daniel Wolff, Frank ter Veld, Tim Köhler, Ansgar Poetsch

J Proteomics. 2013 Apr 26;82:274-87.

PMID: 23500128

Abstract:

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 (15)N-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.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP13018489	Tetraacetylphytosphingosine		13018-48-9	Price

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