Elucidation of Beta-Oxidation Pathways in Ralstonia Eutropha H16 by Examination of Global Gene Expression
Ralstonia eutropha H16 is capable of growth and polyhydroxyalkanoate production on plant oils and fatty acids. However, little is known about the triacylglycerol and fatty acid degradation pathways of this bacterium. We compare whole-cell gene expression levels of R. eutropha H16 during growth and p...
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| Format: | Article |
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American Society for Microbiology
2011
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| Online Access: | http://hdl.handle.net/1721.1/66937 https://orcid.org/0000-0002-1015-1270 https://orcid.org/0000-0002-6671-5987 https://orcid.org/0000-0003-4284-8467 |
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| author | Zeng, Qiandong Holder, Jason W. Mahan, Alison E. Brigham, Christopher J. Budde, Charles F. Rha, Chokyun Sinskey, Anthony J |
| author2 | Massachusetts Institute of Technology. Biomaterials Science and Engineering Laboratory |
| author_facet | Massachusetts Institute of Technology. Biomaterials Science and Engineering Laboratory Zeng, Qiandong Holder, Jason W. Mahan, Alison E. Brigham, Christopher J. Budde, Charles F. Rha, Chokyun Sinskey, Anthony J |
| author_sort | Zeng, Qiandong |
| collection | MIT |
| description | Ralstonia eutropha H16 is capable of growth and polyhydroxyalkanoate production on plant oils and fatty acids. However, little is known about the triacylglycerol and fatty acid degradation pathways of this bacterium. We compare whole-cell gene expression levels of R. eutropha H16 during growth and polyhydroxyalkanoate production on trioleate and fructose. Trioleate is a triacylglycerol that serves as a model for plant oils. Among the genes of note, two potential fatty acid β-oxidation operons and two putative lipase genes were shown to be upregulated in trioleate cultures. The genes of the glyoxylate bypass also exhibit increased expression during growth on trioleate. We observed that single β-oxidation operon deletion mutants of R. eutropha could grow using palm oil or crude palm kernel oil as the sole carbon source, regardless of which operon was present in the genome, but a double mutant was unable to grow under these conditions. A lipase deletion mutant did not exhibit a growth defect in emulsified oil cultures but did exhibit a phenotype in cultures containing nonemulsified oil. Mutants of the glyoxylate shunt gene for isocitrate lyase were able to grow in the presence of oils, while a malate synthase (aceB) deletion mutant grew more slowly than wild type. Gene expression under polyhydroxyalkanoate storage conditions was also examined. Many findings of this analysis confirm results from previous studies by our group and others. This work represents the first examination of global gene expression involving triacylglycerol and fatty acid catabolism genes in R. eutropha. |
| first_indexed | 2024-09-23T13:19:10Z |
| format | Article |
| id | mit-1721.1/66937 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T13:19:10Z |
| publishDate | 2011 |
| publisher | American Society for Microbiology |
| record_format | dspace |
| spelling | mit-1721.1/669372022-09-28T13:21:45Z Elucidation of Beta-Oxidation Pathways in Ralstonia Eutropha H16 by Examination of Global Gene Expression Zeng, Qiandong Holder, Jason W. Mahan, Alison E. Brigham, Christopher J. Budde, Charles F. Rha, Chokyun Sinskey, Anthony J Massachusetts Institute of Technology. Biomaterials Science and Engineering Laboratory Massachusetts Institute of Technology. Department of Biology Massachusetts Institute of Technology. Department of Chemical Engineering Massachusetts Institute of Technology. Engineering Systems Division Sinskey, Anthony J. Sinskey, Anthony J. Holder, Jason W. Mahan, Alison E. Brigham, Christopher J. Budde, Charles F. Rha, ChoKyun Sinskey, Anthony J. Ralstonia eutropha H16 is capable of growth and polyhydroxyalkanoate production on plant oils and fatty acids. However, little is known about the triacylglycerol and fatty acid degradation pathways of this bacterium. We compare whole-cell gene expression levels of R. eutropha H16 during growth and polyhydroxyalkanoate production on trioleate and fructose. Trioleate is a triacylglycerol that serves as a model for plant oils. Among the genes of note, two potential fatty acid β-oxidation operons and two putative lipase genes were shown to be upregulated in trioleate cultures. The genes of the glyoxylate bypass also exhibit increased expression during growth on trioleate. We observed that single β-oxidation operon deletion mutants of R. eutropha could grow using palm oil or crude palm kernel oil as the sole carbon source, regardless of which operon was present in the genome, but a double mutant was unable to grow under these conditions. A lipase deletion mutant did not exhibit a growth defect in emulsified oil cultures but did exhibit a phenotype in cultures containing nonemulsified oil. Mutants of the glyoxylate shunt gene for isocitrate lyase were able to grow in the presence of oils, while a malate synthase (aceB) deletion mutant grew more slowly than wild type. Gene expression under polyhydroxyalkanoate storage conditions was also examined. Many findings of this analysis confirm results from previous studies by our group and others. This work represents the first examination of global gene expression involving triacylglycerol and fatty acid catabolism genes in R. eutropha. Malaysia-MIT Biotechnology Partnership Programme 2011-11-04T16:59:17Z 2011-11-04T16:59:17Z 2010-08 2010-04 Article http://purl.org/eprint/type/JournalArticle 1098-5530 0021-9193 http://hdl.handle.net/1721.1/66937 Brigham, C. J. et al. “Elucidation of -Oxidation Pathways in Ralstonia eutropha H16 by Examination of Global Gene Expression.” Journal of Bacteriology 192 (2010): 5454-5464. Web. 4 Nov. 2011. © 2010 American Society for Microbiology https://orcid.org/0000-0002-1015-1270 https://orcid.org/0000-0002-6671-5987 https://orcid.org/0000-0003-4284-8467 en_US http://dx.doi.org/10.1128/jb.00493-10 Journal of Bacteriology Creative Commons Attribution-Noncommercial-Share Alike 3.0 http://creativecommons.org/licenses/by-nc-sa/3.0/ application/pdf American Society for Microbiology Sinskey |
| spellingShingle | Zeng, Qiandong Holder, Jason W. Mahan, Alison E. Brigham, Christopher J. Budde, Charles F. Rha, Chokyun Sinskey, Anthony J Elucidation of Beta-Oxidation Pathways in Ralstonia Eutropha H16 by Examination of Global Gene Expression |
| title | Elucidation of Beta-Oxidation Pathways in Ralstonia Eutropha H16 by Examination of Global Gene Expression |
| title_full | Elucidation of Beta-Oxidation Pathways in Ralstonia Eutropha H16 by Examination of Global Gene Expression |
| title_fullStr | Elucidation of Beta-Oxidation Pathways in Ralstonia Eutropha H16 by Examination of Global Gene Expression |
| title_full_unstemmed | Elucidation of Beta-Oxidation Pathways in Ralstonia Eutropha H16 by Examination of Global Gene Expression |
| title_short | Elucidation of Beta-Oxidation Pathways in Ralstonia Eutropha H16 by Examination of Global Gene Expression |
| title_sort | elucidation of beta oxidation pathways in ralstonia eutropha h16 by examination of global gene expression |
| url | http://hdl.handle.net/1721.1/66937 https://orcid.org/0000-0002-1015-1270 https://orcid.org/0000-0002-6671-5987 https://orcid.org/0000-0003-4284-8467 |
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