Functional assessment of plant and microalgal lipid pathway genes in yeast to enhance microbial industrial oil production

Biotechnol Appl Biochem. 2018 Mar;65(2):138-144. doi: 10.1002/bab.1573. Epub 2017 Jul 11.

Abstract

As promising alternatives to fossil-derived oils, microbial lipids are important as industrial feedstocks for biofuels and oleochemicals. Our broad aim is to increase lipid content in oleaginous yeast through expression of lipid accumulation genes and use Saccharomyces cerevisiae to functionally assess genes obtained from oil-producing plants and microalgae. Lipid accumulation genes DGAT (diacylglycerol acyltransferase), PDAT (phospholipid: diacylglycerol acyltransferase), and ROD1 (phosphatidylcholine: diacylglycerol choline-phosphotransferase) were separately expressed in yeast and lipid production measured by fluorescence, solvent extraction, thin layer chromatography, and gas chromatography (GC) of fatty acid methyl esters. Expression of DGAT1 from Arabidopsis thaliana effectively increased total fatty acids by 1.81-fold above control, and ROD1 led to increased unsaturated fatty acid content of yeast lipid. The functional assessment approach enabled the fast selection of candidate genes for metabolic engineering of yeast for production of lipid feedstocks.

Keywords: acyltransferase; biofuel; fatty acid accumulation; triacylglycerol.

MeSH terms

  • Arabidopsis / enzymology
  • Arabidopsis / genetics*
  • Arabidopsis / metabolism
  • Biofuels* / analysis
  • Biofuels* / microbiology
  • Diacylglycerol Cholinephosphotransferase / genetics
  • Diacylglycerol Cholinephosphotransferase / metabolism
  • Diacylglycerol O-Acyltransferase / genetics
  • Diacylglycerol O-Acyltransferase / metabolism
  • Fatty Acids / genetics*
  • Fatty Acids / metabolism
  • Industrial Microbiology / methods*
  • Industrial Oils* / analysis
  • Industrial Oils* / microbiology
  • Metabolic Engineering / methods
  • Microalgae / enzymology
  • Microalgae / genetics*
  • Microalgae / metabolism
  • Saccharomyces cerevisiae / enzymology
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae / metabolism

Substances

  • Biofuels
  • Fatty Acids
  • Diacylglycerol O-Acyltransferase
  • Diacylglycerol Cholinephosphotransferase