Abstract
BackgroundConjugated linoleic acid (CLA) has many well-documented beneficial physiological effects. Due to the insufficient natural supply of CLA and low specificity of chemically produced CLA, an effective and isomer-specific production process is required for medicinal and nutritional purposes.ResultsThe linoleic acid isomerase gene from Propionibacterium acnes was expressed in Yarrowia lipolytica Polh. Codon usage optimization of the PAI and multi-copy integration significantly improved the expression level of PAI in Y. lipolytica. The percentage of trans-10, cis-12 CLA was six times higher in yeast carrying the codon-optimized gene than in yeast carrying the native gene. In combination with multi-copy integration, the production yield was raised to approximately 30-fold. The amount of trans-10, cis-12 CLA reached 5.9% of total fatty acid yield in transformed Y. lipolytica.ConclusionsThis is the first report of production of trans-10, cis-12 CLA by the oleaginous yeast Y. lipolytica, using glucose as the sole carbon source through expression of linoleic acid isomerase from Propionibacterium acnes.
Highlights
Conjugated linoleic acid (CLA) has many well-documented beneficial physiological effects
PAI gene codon optimization As the codon usage of the native PAI sequence differs from that preferred by Y. lipolytica, a de novo codonoptimized version of the gene was designed and synthesized
PAI gene was optimized, and as expected, codonoptimization fundamentally enhanced PAI expression (Figure 3) as the translation efficiency was promoted. These results demonstrated that the optimization of Conclusions In this work, we have successfully constructed a de novo CLA biosynthesis system by transforming the oleaginous yeast strain, Y. lipolytica, with the recombinant linoleate isomerase gene from P. acnes
Summary
Conjugated linoleic acid (CLA) has many well-documented beneficial physiological effects. Due to the insufficient natural supply of CLA and low specificity of chemically produced CLA, an effective and isomer-specific production process is required for medicinal and nutritional purposes. In the past three decades, CLA has attracted much attention because of its biologically beneficial functions. These include anti-carcinogenic, anti-atherogenic, anti-diabetic, anti-inflammatory and anti-obesity properties in animal models and humans. Three CLA isomers have been demonstrated to possess beneficial effects individually: cis-9, trans-11 CLA, trans-10, cis-12 CLA and trans-9, trans-11 CLA. CLA isomers occur in meat and dairy products derived from ruminants as a minor component of the lipid isomerase [8,9]. The LA isomerase derived from P. acnes (PAI) is the only LA isomerase with its crystal structure being characterized [10], and had been expressed in Escherichia. coli [11], Saccharomyces. cerevisiae, tobacco seed [12], rice [13] and Lactococcus lactis successively [14]
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