Abstract
BackgroundAn advantageous but challenging approach to overcome the limited supply of petroleum and relieve the greenhouse effect is to produce bulk chemicals from renewable materials. Fatty alcohols, with a billion-dollar global market, are important raw chemicals for detergents, emulsifiers, lubricants, and cosmetics production. Microbial production of fatty alcohols has been successfully achieved in several industrial microorganisms. However, most of the achievements were using glucose, an edible sugar, as the carbon source. To produce fatty alcohols in a renewable manner, non-edible sugars such as xylose will be a more appropriate feedstock.ResultsIn this study, we aim to engineer a Saccharomyces cerevisiae strain that can efficiently convert xylose to fatty alcohols. To this end, we first introduced the fungal xylose utilization pathway consisting of xylose reductase (XR), xylitol dehydrogenase (XDH), and xylulose kinase (XKS) into a fatty alcohol-producing S. cerevisiae strain (XF3) that was developed in our previous studies to achieve 1-hexadecanol production from xylose at 0.4 g/L. We next applied promoter engineering on the xylose utilization pathway to optimize the expression levels of XR, XDH, and XKS, and increased the 1-hexadecanol titer by 171 %. To further improve the xylose-based fatty alcohol production, two optimized S. cerevisiae strains from promoter engineering were evolved with the xylose as the sole carbon source. We found that the cell growth rate was improved at the expense of decreased fatty alcohol production, which indicated 1-hexadecanol was mainly produced as a non-growth associated product. Finally, through fed-batch fermentation, we successfully achieved 1-hexadecanol production at over 1.2 g/L using xylose as the sole carbon source, which represents the highest titer of xylose-based 1-hexadecanol reported in microbes to date.ConclusionsA fatty alcohol-producing S. cerevisiae strain was engineered in this study to produce 1-hexadecanol from xylose. Although the xylose pathway we developed in this study could be further improved, this proof-of-concept study, for the first time to our best knowledge, demonstrated that the xylose-based fatty alcohol could be produced in S. cerevisiae with potential applications in developing consolidated bioprocessing for producing other fatty acid-derived chemicals.Electronic supplementary materialThe online version of this article (doi:10.1186/s12934-016-0423-9) contains supplementary material, which is available to authorized users.
Highlights
An advantageous but challenging approach to overcome the limited supply of petroleum and relieve the greenhouse effect is to produce bulk chemicals from renewable materials
The xylose utilization pathway was selected from our previous study [29], which included a xylose reductase (XR) from Candida shehatae, a xylitol dehydrogenase (XDH) from Candida tropicalis and a xylulose kinase (XKS) from Pichia pastoris
We found that the growth rates of most of the promoter-engineered strains were reduced to some extent, and the 1-hexadecanol production for most of the recombinant strains was not significantly improved
Summary
An advantageous but challenging approach to overcome the limited supply of petroleum and relieve the greenhouse effect is to produce bulk chemicals from renewable materials. With the development of metabolic engineering and synthetic biology, microbial production of fatty alcohols from renewable feedstock has been achieved successfully in both Escherichia coli [4, 9] and. There has been an increasing interest on developing yeast, such as S. cerevisiae, as a cell factory for fatty acidderived biofuel production. Through over-expression of acetyl-CoA carboxylase (ACC) and fatty-acyl-CoA synthases (FAS), the engineered S. cerevisiae strain produced 98.0 mg/L total fatty alcohol from 20 g/L glucose in batch culture in minimal medium [3]. By manipulating the structural genes in yeast lipid metabolism, tuning the regulation of phospholipid synthesis, and increasing the supply of key precursors, 1-hexadecanol was produced at 1.1 g/L using glucose as the carbon source in a fed-batch fermentation [10]
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