Improved production of fatty acids by Saccharomyces cerevisiae through screening a cDNA library from the oleaginous yeast Yarrowia lipolytica
Artikel i vetenskaplig tidskrift, 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

Författare

Shuobo Shi

Chalmers, Biologi och bioteknik, Systembiologi

H. C. Ji

Chalmers, Biologi och bioteknik

Verena Siewers

Chalmers, Biologi och bioteknik, Systembiologi

Jens B Nielsen

Chalmers, Biologi och bioteknik, Systembiologi

FEMS Yeast Research

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

Vol. 16

Ämneskategorier

Biologiska vetenskaper

Styrkeområden

Energi

Livsvetenskaper och teknik

DOI

10.1093/femsyr/fov108