title: Genetic engineering of Yarrowia lipolytica for enhanced production of trans-10, cis-12 conjugated linoleic acid.
abstract: BACKGROUND
  Conjugated linoleic acid (CLA) has been extensively studied for decades because of its health benefits including cancer prevention, anti-atherogenic and anti-obesity effects, and modulation of the immune system. We previously described the production of trans-10, cis-12 CLA in Yarrowia lipolytica by expressing the gene coding for linoleic acid isomerase from Propionibacterium acnes (pai). However the stable strain produced CLA at about 0.08% of dry cell weight (DCW), a level of production which was not high enough for practical applications. The goal of the present study was to enhance production of CLA by genetic engineering of Y. lipolytica strains.
  
  
  RESULTS
  We have now co-expressed the delta 12-desaturase gene (FADS12, d12) from Mortierella alpina together with the codon-optimized linoleic acid isomerase (opai) gene in Y. lipolytica, expressed under the control of promoter hp16d modified by fusing 12 copies of UAS1B to the original promoter hp4d. A multi-copy integration plasmid was used to further enhance the expression of both genes. Using glucose as the sole carbon source, the genetically-modified Y. lipolytica produced trans-10, cis-12-CLA at a level of up to 10% of total fatty acids and 0.4% of DCW. Furthermore, when the recombinant yeast was grown with soybean oil, trans-10, cis-12-CLA now accumulated at a level of up to 44% of total fatty acids, which represented 30% of DCW after 38.5 h of cultivation. In addition, trans-10, cis-12-CLA was also detected in the growth medium up to 0.9 g/l.
  
  
  CONCLUSIONS
  We have successfully produced trans-10, cis-12-CLA with a titre of 4 g/l of culture (3.1 g/l in cells and 0.9 g/l in culture medium). Our results demonstrate the potential use of Y. lipolytica as a promising microbial cell factory for trans-10, cis-12-CLA production.
authors.forename: Baixi
authors.initials: B
authors.lastname: Zhang
authors.forename: Haiqin
authors.initials: H
authors.lastname: Chen
authors.forename: Min
authors.initials: M
authors.lastname: Li
authors.forename: Zhennan
authors.initials: Z
authors.lastname: Gu
authors.forename: Yuanda
authors.initials: Y
authors.lastname: Song
authors.forename: Colin
authors.initials: C
authors.lastname: Ratledge
authors.forename: Yong Q
authors.initials: YQ
authors.lastname: Chen
authors.forename: Hao
authors.initials: H
authors.lastname: Zhang
authors.forename: Wei
authors.initials: W
authors.lastname: Chen
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D044943:Fatty Acid Desaturases / Q000235:genetics / Q000378:metabolism; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D005818:Genetic Engineering; D007535:Isomerases / Q000235:genetics / Q000378:metabolism; D007536:Isomerism; D044243:Linoleic Acids, Conjugated / Q000096:biosynthesis*; D020094:Mortierella / Q000201:enzymology / Q000235:genetics; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011425:Propionibacterium acnes / Q000201:enzymology / Q000235:genetics; D025062:Yarrowia / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D005656:Fungal Proteins; D044243:Linoleic Acids, Conjugated; D044943:Fatty Acid Desaturases; D007535:Isomerases