title: Producing human ceramide-NS by metabolic engineering using yeast Saccharomyces cerevisiae.
abstract: Ceramide is one of the most important intercellular components responsible for the barrier and moisture retention functions of the skin. Because of the risks involved with using products of animal origin and the low productivity of plants, the availability of ceramides is currently limited. In this study, we successfully developed a system that produces sphingosine-containing human ceramide-NS in the yeast Saccharomyces cerevisiae by eliminating the genes for yeast sphingolipid hydroxylases (encoded by SUR2 and SCS7) and introducing the gene for a human sphingolipid desaturase (encoded by DES1). The inactivation of the ceramidase gene YDC1, overexpression of the inositol phosphosphingolipid phospholipase C gene ISC1, and endoplasmic reticulum localization of the DES1 gene product resulted in enhanced production of ceramide-NS. The engineered yeast strains can serve as hosts not only for providing a sustainable source of ceramide-NS but also for developing further systems to produce sphingosine-containing sphingolipids.
authors.affiliation: Department of Biofunctional Science and Technology, Graduate School of Biosphere Science, Hiroshima University, Hiroshima, 739-8528, Japan.
authors.forename: Suguru
authors.initials: S
authors.lastname: Murakami
authors.affiliation: Department of Biofunctional Science and Technology, Graduate School of Biosphere Science, Hiroshima University, Hiroshima, 739-8528, Japan.
authors.forename: Toshi
authors.initials: T
authors.lastname: Shimamoto
authors.affiliation: Suntory World Research Center, Kyoto 619-0284, Japan.
authors.forename: Hideaki
authors.initials: H
authors.lastname: Nagano
authors.affiliation: Department of Biofunctional Science and Technology, Graduate School of Biosphere Science, Hiroshima University, Hiroshima, 739-8528, Japan.
authors.forename: Masahiro
authors.initials: M
authors.lastname: Tsuruno
authors.affiliation: Suntory World Research Center, Kyoto 619-0284, Japan.
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Okuhara
authors.affiliation: Suntory World Research Center, Kyoto 619-0284, Japan.
authors.forename: Haruyo
authors.initials: H
authors.lastname: Hatanaka
authors.affiliation: Department of Biophysics and Biochemistry, Osaka University Graduate School of Medicine, Suita 565-0871, Japan.
authors.forename: Hiromasa
authors.initials: H
authors.lastname: Tojo
authors.affiliation: Suntory World Research Center, Kyoto 619-0284, Japan.
authors.forename: Yukiko
authors.initials: Y
authors.lastname: Kodama
authors.affiliation: Department of Biofunctional Science and Technology, Graduate School of Biosphere Science, Hiroshima University, Hiroshima, 739-8528, Japan.
authors.forename: Kouichi
authors.initials: K
authors.lastname: Funato
mesh_terms: D000581:Amidohydrolases / Q000235:genetics / Q000378:metabolism; D000595:Amino Acid Sequence; D000935:Antifungal Agents / Q000494:pharmacology; D002518:Ceramides / Q000032:analysis / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D047630:Depsipeptides / Q000494:pharmacology; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006801:Humans; D007218:Industrial Microbiology / Q000379:methods*; D060847:Metabolic Engineering; D008856:Microscopy, Fluorescence; D006899:Mixed Function Oxygenases / Q000172:deficiency / Q000235:genetics; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D013110:Sphingosine / Q000032:analysis / Q000378:metabolism; D053719:Tandem Mass Spectrometry; D010738:Type C Phospholipases / Q000235:genetics / Q000378:metabolism
chemical_list: D000935:Antifungal Agents; D002518:Ceramides; D047630:Depsipeptides; D029701:Saccharomyces cerevisiae Proteins; C071398:aureobasidin A; D006899:Mixed Function Oxygenases; D010088:Oxidoreductases; C109313:SUR2 protein, S cerevisiae; C109314:Scs7 protein, S cerevisiae; C108138:dihydroceramide desaturase; C419391:ISC1 protein, S cerevisiae; D010738:Type C Phospholipases; D000581:Amidohydrolases; C000588734:alkaline dihydroceramidase, S cerevisiae; D013110:Sphingosine