Genome-wide screening for improved fatty alcohol production in yeast using a biosensor

Wu, Yuying; Si, Tong

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https://hdl.handle.net/2142/90230 Copy

Description

Title

Genome-wide screening for improved fatty alcohol production in yeast using a biosensor

Author(s)

• Wu, Yuying
• Si, Tong

Contributor(s)

Huimin, Zhao

Issue Date

2016

Keyword(s)

• fatty alcohol
• biosensor
• Saccharomyces Cerevisiae
• directed evolution
• fluorescence-activated cell sorting

Abstract

Fatty alcohols have wide applications in producing surfactants, lubricants, cosmetics and pharmaceuticals. Compared with traditional petrochemical approaches, synthesis of fatty alcohols using microbial fermentation from renewable feedstocks is considered more environmentally friendly. Here, we performed a high-throughput screening for genetic mutations to enhance 1-hexadecanol production in the bakers yeast. To facilitate this screening, we constructed a genetic sensor, which could convert the intracellular concentrations of lipids (key precursors of 1-hexadecanol biosynthesis) into fluorescence signals of the green fluorescence protein (GFP). By combining this sensor with a genome-wide reduction-of-function library, three mutant strains were isolated with enhanced GFP fluorescence. As expected, these strains exhibited increased lipid contents and 1-hexadecanol production than the wild-type strain, which was confirmed by quantifying fermentation products using gas chromatography mass spectrometry. In addition, the underlying mutations were identified using DNA sequencing, and their impact on target gene expression was revealed using quantitative polymerase chain reaction (qPCR). Together, we demonstrated the genetic sensor as a powerful tool in high-throughput screening, and the isolated mutant strains could be potentially employed for synthesizing other classes of lipid-derived chemicals.

Type of Resource

text

Language

en

Permalink

http://hdl.handle.net/2142/90230

Copyright and License Information

Copyright 2016, Wu and Si

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