Improved production of fatty acids by Saccharomyces cerevisiae through screening a cDNA library from the oleaginous yeast Yarrowia lipolytica
Journal article, 2016

Biological production of fatty acid (FA)-derived products has gained increasing attention to replace petroleum-based fuels and chemicals. FA biosynthesis is highly regulated, and usually it is challenging to design rational engineering strategies. In addition, the conventional 'one sample at a time' method for lipid determination is time consuming and laborious, and it is difficult to screen large numbers of samples. Here, a method for detecting free FAs in viable cells using Nile red staining was developed for use in large-scale screening. Following optimization of the method, it was used for screening a cDNA library from the oleaginous yeast Yarrowia lipolytica for identification of genes/enzymes that were able to enhance free FA accumulation in Saccharomyces cerevisiae. Several novel enzymes resulting in increasing FA accumulation were discovered. These targets include a GPI anchor protein, malate dehydrogenase, glyceraldehyde 3-phosphate dehydrogenase, FA hydroxylase, farnesyltransferase, anoctamin, dihydrolipoamide dehydrogenase and phosphatidylethanolamine-binding protein. The best enzyme resulted in a 2.5-fold improvement in production of free FAs. Our findings not only provide a novel method for high-throughput evaluation of the content of free FAs, but also give new insight into how enzymes from Y. lipolytica may increase the production of fatty acids in S. cerevisiae.

fatty acids

cDNA library

Yarrowia lipolytica

neutral lipid-content

carboxypeptidase-y

codon usage

escherichia-coli

transgenic arabidopsis

microbial-production

nile red fluorescence

triacylglycerol synthesis

Saccharomyces cerevisiae

sodium-chloride

high-throughput screening

ethyl-ester production

Author

Shuobo Shi

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

H. C. Ji

Chalmers, Biology and Biological Engineering

Verena Siewers

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Jens B Nielsen

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

FEMS Yeast Research

1567-1356 (ISSN) 1567-1364 (eISSN)

Vol. 16 1

Subject Categories

Biological Sciences

Areas of Advance

Energy

Life Science Engineering (2010-2018)

DOI

10.1093/femsyr/fov108

More information

Created

10/8/2017