Journal Article

Metabolic engineering for ricinoleic acid production in the oleaginous yeast Yarrowia lipolytica

Beopoulos, A, Bordes, F, Marty, A, Nicaud, Jean-Marc, Verbeke, J, Guicherd, M, Bressy, M

2014

Overview

Although there are numerous oleochemical applications for ricinoleic acid (RA) and its derivatives, their production is limited and subject to various safety legislations. In an effort to produce RA from alternative sources, we constructed a genetically modified strain of the oleaginous yeast Yarrowia lipolytica. This strain is unable to perform β-oxidation and is invalidated for the native triacylglycerol (TAG) acyltransferases (Dga1p, Dga2p, and Lro1p) and the ∆12 desaturase (Fad2p). We also expressed the Ricinus communis ∆12 hydroxylase (RcFAH12) under the control of the TEF constitutive promoter in this strain. However, RA constituted only 7 % of the total lipids produced by this modified strain. By contrast, expression of the Claviceps purpurea hydroxylase CpFAH12 in this background resulted in a strain able to accumulate RA to 29 % of total lipids, and expression of an additional copy of CpFAH12 drove RA accumulation up to 35 % of total lipids. The co-expression of the C. purpurea or R. communis type II diacylglycerol acyltransferase (RcDGAT2 or CpDGAT2) had negative effects on RA accumulation in this yeast, with RA levels dropping to below 14 % of total lipids. Overexpression of the native Y. lipolytica PDAT acyltransferase (Lro1p) restored both TAG accumulation and RA levels. Thus, we describe the consequences of rerouting lipid metabolism in this yeast so as to develop a cell factory for RA production. The engineered strain is capable of accumulating RA to 43 % of its total lipids and over 60 mg/g of cell dry weight; this is the most efficient production of RA described to date.

• Publisher: Springer Verlag,HAL CCSD,Springer-Verlag,Springer Berlin Heidelberg,Springer,Springer Nature B.V
• Subject: Accumulation / Acid production / Applied Genetics and Molecular Biotechnology / beta oxidation / Biomedical and Life Sciences / Biosynthesis / Biotechnology / Claviceps / Claviceps - enzymology / Claviceps - genetics / Claviceps purpurea / Derivatives / diacylglycerol acyltransferase / Engineering / Enzymes / enzymology / Fatty acids / Gene Deletion / Gene Expression / Genes / Genetic aspects / Genetic engineering / genetics / Life Sciences / Lipids / Manufacturing engineering / Metabolic Engineering / Metabolic Networks and Pathways / Metabolic Networks and Pathways - genetics / Metabolism / Metabolites / Methods / Microbial Genetics and Genomics / Microbiology / Microorganisms / Molecular Sequence Data / Oilseeds / Production processes / Recombinant Proteins / Recombinant Proteins - genetics / Recombinant Proteins - metabolism / Reproduction / Ricinoleic acid / Ricinoleic Acids / Ricinoleic Acids - metabolism / Ricinus / Ricinus - enzymology / Ricinus - genetics / Ricinus communis / Seeds / Sequence Analysis, DNA / Strain / Studies / triacylglycerols / Yarrowia / Yarrowia - genetics / Yarrowia - metabolism / Yarrowia lipolytica / Yeast / Yeast fungi / yeasts
• ISBN: 0003290936000