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Application of thin-layer chromatography-flame ionization detection (TLC-FID) to total lipid quantitation in mycolic-acid synthesizing Rhodococcus and Williamsia species

Citation

Nahar, A and Baker, AL and Nichols, DS and Bowman, JP and Britz, ML, Application of thin-layer chromatography-flame ionization detection (TLC-FID) to total lipid quantitation in mycolic-acid synthesizing Rhodococcus and Williamsia species, International Journal of Molecular Sciences, 21, (5) Article 1670. ISSN 1661-6596 (2020) [Refereed Article]


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Copyright 2020 The Authors. Licensed under Creative Commons Attribution 4.0 International (CC BY 4.0) https://creativecommons.org/licenses/by/4.0/

DOI: doi:10.3390/ijms21051670

Abstract

In addition to cell membrane phospholipids, Actinobacteria in the order Corynebacteriales possess a waxy cell envelope containing mycolic acids (MA). In optimized culture condition, some species can also accumulate high concentrations of intracellular triacylglycerols (TAG), which are a potential source of biodiesel. Bacterial lipid classes and composition alter in response to environmental stresses, including nutrient availability, thus understanding carbon flow into different lipid classes is important when optimizing TAG synthesis. Quantitative and qualitative analysis of lipid classes normally requires combinations of different extraction, derivatization, chromatographic and detection methods. In this study, a single-step thin-layer chromatography-flame ionization detection (TLC-FID) technique was applied to quantify lipid classes in six sub-Antarctic Corynebacteriales strains identified as Rhodococcus and Williamsia species. A hexane:diethyl-ether:acetic acid solvent system separated the total cellular lipids extracted from cells lysed by bead beating, which released more bound and unbound MA than sonication. Typical profiles included a major broad non-polar lipid peak, TAG and phospholipids, although trehalose dimycolates, when present, co-eluted with phospholipids. Ultra-performance liquid chromatography-tandem mass-spectrometry and nuclear magnetic resonance spectroscopy detected MA signatures in the non-polar lipid peak and indicated that these lipids were likely bound, at least in part, to sugars from cell wall arabinogalactan. Waxy esters were not detected. The single-solvent TLC-FID procedure provides a useful platform for the quantitation and preliminary screening of cellular lipid classes when testing the impacts of growth conditions on TAG synthesis.

Item Details

Item Type:Refereed Article
Keywords:TLC-FID, Iatroscan rods, NMR, lipid extraction, mycolic acids, Williamsia, Rhodococcus, Antarctic bacteria, lipid analysis, new methods
Research Division:Chemical Sciences
Research Group:Organic chemistry
Research Field:Natural products and bioactive compounds
Objective Division:Expanding Knowledge
Objective Group:Expanding knowledge
Objective Field:Expanding knowledge in the biological sciences
UTAS Author:Nahar, A (Miss Akhikum Nahar)
UTAS Author:Baker, AL (Dr Anthony Baker)
UTAS Author:Nichols, DS (Dr David Nichols)
UTAS Author:Bowman, JP (Associate Professor John Bowman)
UTAS Author:Britz, ML (Professor Margaret Britz)
ID Code:138665
Year Published:2020
Web of Science® Times Cited:2
Deposited By:TIA - Research Institute
Deposited On:2020-04-20
Last Modified:2020-07-24
Downloads:3 View Download Statistics

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