title: Production of tetraacetyl phytosphingosine (TAPS) in Wickerhamomyces ciferrii is catalyzed by acetyltransferases Sli1p and Atf2p.
abstract: Wickerhamomyces ciferrii secretes tetraacetyl phytosphingosine (TAPS), and in this study, the catalyzing acetyltransferases were identified using mass spectrometry-based proteomics. The proteome of wild-type strain NRRL Y-1031 served as control and was compared to the tetraacetyl phytosphingosine defective mating type NRRL Y-1031-27. Acetylation of phytosphingosine in W. ciferrii is catalyzed by acetyltransferases Sli1p and Atf2p, encoded by genes similar to Saccharomyces cerevisiae YGR212W and YGR177C, respectively. Ablation of SLI1 resulted in an almost complete loss of tri- and tetraacetyl phytosphingosines, whereas the loss ATF2 resulted in an 15-fold increase in triacetyl phytosphingosine. Most likely, it is the concerted action of these two acetyltransferases that yields tetraacetyl phytosphingosine, in which Sli1p catalyzes initial O- and N-acetylation, producing triacetyl phytosphingosine. Finally, Atf2p catalyzes final O-acetylation to yield tetraacetyl phytosphingosine. The current study demonstrates that mass spectrometry-based proteomics can be employed to identify key steps in ill-explored metabolite biosynthesis pathways of nonconventional microorganisms. Furthermore, the identification of phytosphingosine as substrate for alcohol acetyltransferase Atf2p broadens the known substrate range of this enzyme. This interesting property of Atf2p may be exploited to enhance the secretion of heterologous compounds.
authors: Department of Plant Biochemistry, Ruhr University Bochum, Bldg. ND, Fl. 3, Rm. 130, Universitätsstraße 150, 44801, Bochum, Germany, Frank.terVeld@rub.de.
authors: Frank
authors: F
authors: Ter Veld
authors: Daniel
authors: D
authors: Wolff
authors: Christoph
authors: C
authors: Schorsch
authors: Tim
authors: T
authors: Köhler
authors: Eckhard
authors: E
authors: Boles
authors: Ansgar
authors: A
authors: Poetsch
mesh_terms: D000123:Acetyltransferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017353:Gene Deletion; D013058:Mass Spectrometry; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics; D004718:Saccharomycetales / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D013110:Sphingosine / Q000031:analogs & derivatives* / Q000378:metabolism
chemical_list: D029701:Saccharomyces cerevisiae Proteins; C520613:tetraacetyl-phytosphingosine; D000123:Acetyltransferases; C492826:SLI1 protein, S cerevisiae; C116217:ATF2 protein, S cerevisiae; D013110:Sphingosine