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Direct determination of poly(3-hydroxybutyrate) accumulated in bacteria by thermally assisted hydrolysis and methylation-gas chromatography in the presence of organic alkali

Direct determination of poly(3-hydroxybutyrate) accumulated in bacteria by thermally assisted... AbstractThere is considerable interest in the development of simple methods for quantifying production of the biodegradable polyester poly(3-hydroxybutyrate) [P(3HB)] by bacteria. Cells of Cupriavidus necator were grown on agar medium containing different concentrations of glucose (10–25 g/L) as a sole carbon source. Trace amounts (100±5 μg) of dried C. necator cells were directly subjected to thermally assisted hydrolysis and methylation-gas chromatography (THM-GC) in the presence of tetramethylammonium hydroxide (TMAH). On the resulting chromatograms, a series of characteristic peaks, attributed to the THM products from poly(3-hydroxybutylate) accumulated in the bacterial cells, were clearly observed without any interfering component. Based on the peak intensities, the contents of P(3HB) in C. necator were determined precisely and rapidly without using any cumbersome sample pretreatment. Furthermore, the values of the P(3HB) contents coincided overall with those obtained by the conventional method involving solvent extraction followed by gravimetric determination. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Pure and Applied Chemistry de Gruyter

Direct determination of poly(3-hydroxybutyrate) accumulated in bacteria by thermally assisted hydrolysis and methylation-gas chromatography in the presence of organic alkali

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Publisher
de Gruyter
Copyright
©2018 IUPAC & De Gruyter. This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License. For more information, please visit: http://creativecommons.org/licenses/by-nc-nd/4.0/
ISSN
0033-4545
eISSN
1365-3075
DOI
10.1515/pac-2017-0905
Publisher site
See Article on Publisher Site

Abstract

AbstractThere is considerable interest in the development of simple methods for quantifying production of the biodegradable polyester poly(3-hydroxybutyrate) [P(3HB)] by bacteria. Cells of Cupriavidus necator were grown on agar medium containing different concentrations of glucose (10–25 g/L) as a sole carbon source. Trace amounts (100±5 μg) of dried C. necator cells were directly subjected to thermally assisted hydrolysis and methylation-gas chromatography (THM-GC) in the presence of tetramethylammonium hydroxide (TMAH). On the resulting chromatograms, a series of characteristic peaks, attributed to the THM products from poly(3-hydroxybutylate) accumulated in the bacterial cells, were clearly observed without any interfering component. Based on the peak intensities, the contents of P(3HB) in C. necator were determined precisely and rapidly without using any cumbersome sample pretreatment. Furthermore, the values of the P(3HB) contents coincided overall with those obtained by the conventional method involving solvent extraction followed by gravimetric determination.

Journal

Pure and Applied Chemistryde Gruyter

Published: Jun 27, 2018

Keywords: Cupriavidus necator; POLYCHAR-25; poly(3-hydroxybutyrate); tetramethylammonium hydroxide (TMAH); thermally assisted hydrolysis and methylation-gas chromatography (THM-GC)

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