Glycerol Utilization as a Sole Carbon Source Disrupts the Membrane Architecture and Solventogenesis in <i>Clostridium beijerinckii</i> NCIMB 8052
Efficient bioconversion of abundant waste glycerol to value-added chemicals calls for a wider range of fermentative workhorses that can catabolize glycerol. In this study, we used quantitative gene expression and solvent profiling, qualitative metabolite analysis, and enzyme activity assays to inves...
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MDPI AG
2022-07-01
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| Series: | Fermentation |
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| Online Access: | https://www.mdpi.com/2311-5637/8/7/339 |
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| author | Eric Agyeman-Duah Santosh Kumar Bhavana Gangwar Victor C. Ujor |
| author_facet | Eric Agyeman-Duah Santosh Kumar Bhavana Gangwar Victor C. Ujor |
| author_sort | Eric Agyeman-Duah |
| collection | DOAJ |
| description | Efficient bioconversion of abundant waste glycerol to value-added chemicals calls for a wider range of fermentative workhorses that can catabolize glycerol. In this study, we used quantitative gene expression and solvent profiling, qualitative metabolite analysis, and enzyme activity assays to investigate the factors that limit glycerol utilization as a sole carbon source by <i>Clostridium beijerinckii</i> NCIMB 8052. <i>C. beijerinckii</i> NCIMB 8052 did not produce acetate, acetone and butanol on glycerol. Congruently, the genes encoding the coenzyme A transferase subunits (<i>ctfAB</i>) and bifunctional acetaldehyde-CoA/alcohol dehydrogenase (<i>adhE</i>) were down-regulated up to 135- and 21-fold, respectively, at 12 h in glycerol-grown cells compared to glucose-grown cells. Conversely, NADH-dependent butanol dehydrogenase A (<i>bdhA</i>) was upregulated 2-fold. Glycerol dehydrogenase (<i>gldA</i>) and dihydroxyacetone kinase (subunit <i>dhaK</i>) were upregulated up to 5- and 881-fold, respectively. Glyceraldehyde-3-phosphate dehydrogenase (<i>gapdh</i>) showed mostly similar expression profiles at 12 h on glucose and glycerol. At 24 h, <i>gapdh</i> was downregulated 1.5-fold, while NADP<sup>+</sup>-dependent <i>gapdh</i> was upregulated up to 1.9-fold. Glycerol-grown cells showed higher or similar activity profiles for all solventogenic enzymes studied, compared to glucose-grown cells. Butyraldehyde (3 g/L) supplementation led to the production of ~0.1 g/L butanol, whilst butyrate (3.5 g/L) supplementation produced 0.7 and 0.5 g/L acetone and butanol, respectively, with glycerol. Further, the long chain saturated fatty acids cyclopentaneundecanoic acid, methyl ester and hexadecanoic acid, butyl ester were detected in glucose- but not in glycerol-grown cells. Collectively, growth on glycerol appears to disrupt synthesis of saturated long chain fatty acids, as well as solventogenesis in <i>C. beijerinckii</i> NCIMB 8052. |
| first_indexed | 2024-03-09T10:19:07Z |
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| institution | Directory Open Access Journal |
| issn | 2311-5637 |
| language | English |
| last_indexed | 2024-03-09T10:19:07Z |
| publishDate | 2022-07-01 |
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| spelling | doaj.art-3e901ff1680c49088e69bc46a7bb7fc02023-12-01T22:08:24ZengMDPI AGFermentation2311-56372022-07-018733910.3390/fermentation8070339Glycerol Utilization as a Sole Carbon Source Disrupts the Membrane Architecture and Solventogenesis in <i>Clostridium beijerinckii</i> NCIMB 8052Eric Agyeman-Duah0Santosh Kumar1Bhavana Gangwar2Victor C. Ujor3Fermentation Science and Metabolic Engineering Group, Department of Food Science, Babcock Hall, University of Wisconsin-Madison, 1605 Linden Drive, Madison, WI 53706, USAFermentation Science and Metabolic Engineering Group, Department of Food Science, Babcock Hall, University of Wisconsin-Madison, 1605 Linden Drive, Madison, WI 53706, USAFermentation Science and Metabolic Engineering Group, Department of Food Science, Babcock Hall, University of Wisconsin-Madison, 1605 Linden Drive, Madison, WI 53706, USAFermentation Science and Metabolic Engineering Group, Department of Food Science, Babcock Hall, University of Wisconsin-Madison, 1605 Linden Drive, Madison, WI 53706, USAEfficient bioconversion of abundant waste glycerol to value-added chemicals calls for a wider range of fermentative workhorses that can catabolize glycerol. In this study, we used quantitative gene expression and solvent profiling, qualitative metabolite analysis, and enzyme activity assays to investigate the factors that limit glycerol utilization as a sole carbon source by <i>Clostridium beijerinckii</i> NCIMB 8052. <i>C. beijerinckii</i> NCIMB 8052 did not produce acetate, acetone and butanol on glycerol. Congruently, the genes encoding the coenzyme A transferase subunits (<i>ctfAB</i>) and bifunctional acetaldehyde-CoA/alcohol dehydrogenase (<i>adhE</i>) were down-regulated up to 135- and 21-fold, respectively, at 12 h in glycerol-grown cells compared to glucose-grown cells. Conversely, NADH-dependent butanol dehydrogenase A (<i>bdhA</i>) was upregulated 2-fold. Glycerol dehydrogenase (<i>gldA</i>) and dihydroxyacetone kinase (subunit <i>dhaK</i>) were upregulated up to 5- and 881-fold, respectively. Glyceraldehyde-3-phosphate dehydrogenase (<i>gapdh</i>) showed mostly similar expression profiles at 12 h on glucose and glycerol. At 24 h, <i>gapdh</i> was downregulated 1.5-fold, while NADP<sup>+</sup>-dependent <i>gapdh</i> was upregulated up to 1.9-fold. Glycerol-grown cells showed higher or similar activity profiles for all solventogenic enzymes studied, compared to glucose-grown cells. Butyraldehyde (3 g/L) supplementation led to the production of ~0.1 g/L butanol, whilst butyrate (3.5 g/L) supplementation produced 0.7 and 0.5 g/L acetone and butanol, respectively, with glycerol. Further, the long chain saturated fatty acids cyclopentaneundecanoic acid, methyl ester and hexadecanoic acid, butyl ester were detected in glucose- but not in glycerol-grown cells. Collectively, growth on glycerol appears to disrupt synthesis of saturated long chain fatty acids, as well as solventogenesis in <i>C. beijerinckii</i> NCIMB 8052.https://www.mdpi.com/2311-5637/8/7/339<i>Clostridium beijerinckii</i> NCIMB 8052glycerol fermentationsaturated long chain fatty acidssolventogenesisbutanol production |
| spellingShingle | Eric Agyeman-Duah Santosh Kumar Bhavana Gangwar Victor C. Ujor Glycerol Utilization as a Sole Carbon Source Disrupts the Membrane Architecture and Solventogenesis in <i>Clostridium beijerinckii</i> NCIMB 8052 Fermentation <i>Clostridium beijerinckii</i> NCIMB 8052 glycerol fermentation saturated long chain fatty acids solventogenesis butanol production |
| title | Glycerol Utilization as a Sole Carbon Source Disrupts the Membrane Architecture and Solventogenesis in <i>Clostridium beijerinckii</i> NCIMB 8052 |
| title_full | Glycerol Utilization as a Sole Carbon Source Disrupts the Membrane Architecture and Solventogenesis in <i>Clostridium beijerinckii</i> NCIMB 8052 |
| title_fullStr | Glycerol Utilization as a Sole Carbon Source Disrupts the Membrane Architecture and Solventogenesis in <i>Clostridium beijerinckii</i> NCIMB 8052 |
| title_full_unstemmed | Glycerol Utilization as a Sole Carbon Source Disrupts the Membrane Architecture and Solventogenesis in <i>Clostridium beijerinckii</i> NCIMB 8052 |
| title_short | Glycerol Utilization as a Sole Carbon Source Disrupts the Membrane Architecture and Solventogenesis in <i>Clostridium beijerinckii</i> NCIMB 8052 |
| title_sort | glycerol utilization as a sole carbon source disrupts the membrane architecture and solventogenesis in i clostridium beijerinckii i ncimb 8052 |
| topic | <i>Clostridium beijerinckii</i> NCIMB 8052 glycerol fermentation saturated long chain fatty acids solventogenesis butanol production |
| url | https://www.mdpi.com/2311-5637/8/7/339 |
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