Yeast cell surface display of linoleic acid isomerase from Propionibacterium acnes and its application for the production of trans-10, cis-12 conjugated linoleic acid.

Abstract

Conjugated linoleic acid (CLA), a family of geometric and positional isomers of linoleic acid, has many health-promoting properties. Different isomers of CLA may have very different physiological effects. In the current work, we explore the possibility to produce single isomer of CLA by using biocatalysis based on displayed biocatalysts on the yeast cell surfaces. A reporter system used to assess gene expression and protein distribution was established by combining the egfp gene to the N-terminus of Propionibacterium acnes pai gene encoding the linoleic isomerase onto vector pYD1. After induction of the yeast strains containing pYD1::egfp::pai with galactose, strong green fluorescence was observed on the surface of cells, demonstrating that the fusion protein was successfully displayed. Using the engineered strains as whole-cell biocatalyst, trans-10, cis-12 CLA was detected in the reaction mixture. To improve the biocatalytic potential of this system, the first 20 amino codons of pai were modified, and the catalytic reaction conditions were optimized. Optimization of the codon usage resulted in 35% increase of CLA production, and the maximum yield of CLA was observed within 20H in the optimal conditions: pH 7.0, 4mg/mL linoleic acid, 37°C. The system established in the present work can guide the development of biocatalytic strategies to produce trans-10, cis-12 CLA single isomer.