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Single cell assessment of yeast metabolic engineering for enhanced lipid production using Raman and AFM-IR imaging

Overview of attention for article published in Biotechnology for Biofuels, April 2018
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Title
Single cell assessment of yeast metabolic engineering for enhanced lipid production using Raman and AFM-IR imaging
Published in
Biotechnology for Biofuels, April 2018
DOI 10.1186/s13068-018-1108-x
Pubmed ID
Authors

Kamila Kochan, Huadong Peng, Bayden R. Wood, Victoria S. Haritos

Abstract

Biodiesel is a valuable renewable fuel made from derivatized fatty acids produced in plants, animals, and oleaginous microbes. Of the latter, yeasts are of special interest due to their wide use in biotechnology, ability to synthesize fatty acids and store large amounts of triacylglycerols while utilizing non-food carbon sources. While yeast efficiently produce lipids, genetic modification and indeed, lipid pathway metabolic engineering, is usually required for cost-effective production. Traditionally, gas chromatography (GC) is used to measure fatty acid production and to track the success of a metabolic engineering strategy in a microbial culture; here we have employed vibrational spectroscopy approaches at population and single cell level of engineered yeast while simultaneously investigating metabolite levels in subcellular structures. Firstly, a strong correlation (r2 > 0.99) was established between Fourier transform infrared (FTIR) lipid in intact cells and GC analysis of fatty acid methyl esters in the differently engineered strains. Confocal Raman spectroscopy of individual cells carrying genetic modifications to enhance fatty acid synthesis and lipid accumulation revealed changes to the lipid body (LB), the storage organelle for lipids in yeast, with their number increasing markedly (up to tenfold higher); LB size was almost double in the strain that also expressed a LB stabilizing gene but considerable variation was also noted between cells. Raman spectroscopy revealed a clear trend toward reduced unsaturated fatty acid content in lipids of cells carrying more complex metabolic engineering. Atomic force microscopy-infrared spectroscopy (AFM-IR) analysis of individual cells indicated large differences in subcellular constituents between strains: cells of the most highly engineered strain had elevated lipid and much reduced carbohydrate in their cytoplasm compared with unmodified cells. Vibrational spectroscopy analysis allowed the simultaneous measurement of strain variability in metabolite production and impact on cellular structures as a result of different gene introductions or knockouts, within a lipid metabolic engineering strategy and these inform the next steps in comprehensive lipid engineering. Additionally, single cell spectroscopic analysis measures heterogeneity in metabolite production across microbial cultures under genetic modification, an emerging issue for efficient biotechnological production.

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The data shown below were collected from the profile of 1 tweeter who shared this research output. Click here to find out more about how the information was compiled.

Mendeley readers

The data shown below were compiled from readership statistics for 46 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 46 100%

Demographic breakdown

Readers by professional status Count As %
Student > Ph. D. Student 14 30%
Researcher 8 17%
Student > Master 5 11%
Other 3 7%
Student > Postgraduate 2 4%
Other 6 13%
Unknown 8 17%
Readers by discipline Count As %
Agricultural and Biological Sciences 10 22%
Biochemistry, Genetics and Molecular Biology 8 17%
Chemistry 6 13%
Engineering 3 7%
Physics and Astronomy 2 4%
Other 5 11%
Unknown 12 26%

Attention Score in Context

This research output has an Altmetric Attention Score of 1. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 13 April 2018.
All research outputs
#10,225,835
of 12,802,184 outputs
Outputs from Biotechnology for Biofuels
#725
of 977 outputs
Outputs of similar age
#207,006
of 274,108 outputs
Outputs of similar age from Biotechnology for Biofuels
#10
of 10 outputs
Altmetric has tracked 12,802,184 research outputs across all sources so far. This one is in the 11th percentile – i.e., 11% of other outputs scored the same or lower than it.
So far Altmetric has tracked 977 research outputs from this source. They receive a mean Attention Score of 4.4. This one is in the 15th percentile – i.e., 15% of its peers scored the same or lower than it.
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