Development of flow cytometry technique for detection of thinning of peptidoglycan layer as a result of solvent production by <Emphasis Type="Italic">Clostridium pasteurianum</Emphasis> |
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Authors: | M Linhová P Patáková J Lipovský P Fribert L Paulová M Rychtera K Melzoch |
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Institution: | (1) Department of Chemical and Process Engineering, Faculty of Engineering, Universiti Kebangsaan Malaysia, 43600 UKM, Bangi, Selangor, Malaysia;(2) School of Bioscience and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 UKM, Bangi, Selangor, Malaysia |
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Abstract: | Clostridium pasteurianum forms acetic and butyric acids in an initial growth phase, which is a typical feature of clostridial acetone-butanol fermentation
where an initial accumulation of acids is followed by production of solvents 1-butanol, acetone and ethanol. The initiation
of the solvent production coupled with endospore formation leads to decrease of cell-wall thickness; thinner cell wall is
more resistant against solvents and dyes. These changes can be observed by the method based on adaptation of Gram staining.
The cell wall of G+ bacteria allows the entry of hexidium iodide and rhodamine 123, whereas the outer membrane of G− bacteria does not allow the uptake and therefore G+ bacteria are stained with higher fluorescence intensity than G− bacteria. The ratio of fluorescence intensity (FI) to forward scatter (FSC) was determined to correspond to G+ bacteria when clostridia were producing less solvents. The significant drop of the ratio FI to FSC to the level corresponding
to G− bacteria is detected after initiation of solvent production. |
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