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

Suguru Murakami; Toshi Shimamoto; Hideaki Nagano; Masahiro Tsuruno; Hiroaki Okuhara; Haruyo Hatanaka; Hiromasa Tojo; Yukiko Kodama; Kouichi Funato

doi:10.1038/srep16319

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

Sci Rep. 2015 Nov 17:5:16319. doi: 10.1038/srep16319.

Authors

Suguru Murakami¹, Toshi Shimamoto¹, Hideaki Nagano², Masahiro Tsuruno¹, Hiroaki Okuhara², Haruyo Hatanaka², Hiromasa Tojo³, Yukiko Kodama², Kouichi Funato¹

Affiliations

¹ Department of Biofunctional Science and Technology, Graduate School of Biosphere Science, Hiroshima University, Hiroshima, 739-8528, Japan.
² Suntory World Research Center, Kyoto 619-0284, Japan.
³ Department of Biophysics and Biochemistry, Osaka University Graduate School of Medicine, Suita 565-0871, Japan.

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amidohydrolases / genetics
Amidohydrolases / metabolism
Amino Acid Sequence
Antifungal Agents / pharmacology
Ceramides / analysis
Ceramides / metabolism*
Chromatography, High Pressure Liquid
Depsipeptides / pharmacology
Endoplasmic Reticulum / metabolism
Humans
Industrial Microbiology / methods*
Metabolic Engineering*
Microscopy, Fluorescence
Mixed Function Oxygenases / deficiency
Mixed Function Oxygenases / genetics
Oxidoreductases / genetics
Oxidoreductases / metabolism
Saccharomyces cerevisiae / drug effects
Saccharomyces cerevisiae / metabolism*
Saccharomyces cerevisiae Proteins / genetics
Saccharomyces cerevisiae Proteins / metabolism
Sphingosine / analysis
Sphingosine / metabolism
Tandem Mass Spectrometry
Type C Phospholipases / genetics
Type C Phospholipases / metabolism

Substances

Antifungal Agents
Ceramides
Depsipeptides
Saccharomyces cerevisiae Proteins
aureobasidin A
Mixed Function Oxygenases
Oxidoreductases
SUR2 protein, S cerevisiae
Scs7 protein, S cerevisiae
dihydroceramide desaturase
ISC1 protein, S cerevisiae
Type C Phospholipases
Amidohydrolases
alkaline dihydroceramidase, S cerevisiae
Sphingosine