Producing human ceramide-NS by metabolic engineering using yeast Saccharomyces cerevisiae

• Published in: Scientific reports Vol. 5; no. 1; p. 16319
• Main Authors: Murakami, Suguru, Shimamoto, Toshi, Nagano, Hideaki, Tsuruno, Masahiro, Okuhara, Hiroaki, Hatanaka, Haruyo, Tojo, Hiromasa, Kodama, Yukiko, Funato, Kouichi
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 17.11.2015; Nature Publishing Group
• Subjects: 101/58; 45/29; 631/326/2522; 631/61/318; 82/16; Amidohydrolases - genetics; Amidohydrolases - metabolism; Amino Acid Sequence; Antifungal Agents - pharmacology; C gene; Ceramidase; Ceramide; Ceramides - analysis; Ceramides - metabolism; Chromatography, High Pressure Liquid; Depsipeptides - pharmacology; Desaturase; Endoplasmic reticulum; Endoplasmic Reticulum - metabolism; Humanities and Social Sciences; Humans; Industrial Microbiology - methods; Inositol; Localization; Metabolic Engineering; Microscopy, Fluorescence; Mixed Function Oxygenases - deficiency; Mixed Function Oxygenases - genetics; multidisciplinary; Oxidoreductases - genetics; Oxidoreductases - metabolism; Phospholipase C; Saccharomyces cerevisiae - drug effects; Saccharomyces cerevisiae - metabolism; Saccharomyces cerevisiae Proteins - genetics; Saccharomyces cerevisiae Proteins - metabolism; Science; Skin; Sphingolipids; Sphingosine - analysis; Sphingosine - metabolism; Tandem Mass Spectrometry; Type C Phospholipases - genetics; Type C Phospholipases - metabolism; Yeast
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/srep16319

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.