Arabinose is metabolized via a phosphoketolase pathway in Clostridium acetobutylicum ATCC 824
In this report, a novel zymogram assay and coupled phosphoketolase assay were employed to demonstrate that Clostridium acetobutylicum gene CAC1343 encodes a bi-functional xylulose-5-P/fructose-6-P phosphoketolase (XFP). The specific activity of purified recombinant XFP was 6.9 U/mg on xylulose-5-P a...
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| Veröffentlicht in: | Journal of industrial microbiology & biotechnology 2012-12, Vol.39 (12), p.1859-1867 |
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| creator | Servinsky, M. D. Germane, K. L. Liu, S. Kiel, J. T. Clark, A. M. Shankar, J. Sund, C. J. |
| description | In this report, a novel zymogram assay and coupled phosphoketolase assay were employed to demonstrate that
Clostridium acetobutylicum
gene CAC1343 encodes a bi-functional xylulose-5-P/fructose-6-P phosphoketolase (XFP). The specific activity of purified recombinant XFP was 6.9 U/mg on xylulose-5-P and 21 U/mg on fructose-6-P, while the specific activity of XFP in concentrated
C. acetobutylicum
whole-cell extract was 0.094 and 0.52 U/mg, respectively. Analysis of crude cell extracts indicated that XFP activity was present in cells grown on arabinose but not glucose and quantitative PCR was used to show that CAC1343 mRNA expression was induced 185-fold during growth on arabinose when compared to growth on glucose. HPLC analysis of metabolites revealed that during growth on xylose and glucose more butyrate than acetate was formed with final acetate:butyrate ratios of 0.72 and 0.83, respectively. Growth on arabinose caused a metabolic shift to more oxidized products with a final acetate:butyrate ratio of 1.95. The shift towards more oxidized products is consistent with the presence of an XFP, suggesting that arabinose is metabolized via a phosphoketolase pathway while xylose is probably metabolized via the pentose phosphate pathway. |
| doi_str_mv | 10.1007/s10295-012-1186-x |
| format | Article |
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Clostridium acetobutylicum
gene CAC1343 encodes a bi-functional xylulose-5-P/fructose-6-P phosphoketolase (XFP). The specific activity of purified recombinant XFP was 6.9 U/mg on xylulose-5-P and 21 U/mg on fructose-6-P, while the specific activity of XFP in concentrated
C. acetobutylicum
whole-cell extract was 0.094 and 0.52 U/mg, respectively. Analysis of crude cell extracts indicated that XFP activity was present in cells grown on arabinose but not glucose and quantitative PCR was used to show that CAC1343 mRNA expression was induced 185-fold during growth on arabinose when compared to growth on glucose. HPLC analysis of metabolites revealed that during growth on xylose and glucose more butyrate than acetate was formed with final acetate:butyrate ratios of 0.72 and 0.83, respectively. Growth on arabinose caused a metabolic shift to more oxidized products with a final acetate:butyrate ratio of 1.95. The shift towards more oxidized products is consistent with the presence of an XFP, suggesting that arabinose is metabolized via a phosphoketolase pathway while xylose is probably metabolized via the pentose phosphate pathway.</description><identifier>ISSN: 1367-5435</identifier><identifier>EISSN: 1476-5535</identifier><identifier>DOI: 10.1007/s10295-012-1186-x</identifier><identifier>PMID: 22922942</identifier><language>eng</language><publisher>Berlin/Heidelberg: Springer-Verlag</publisher><subject>Aldehyde-Lyases - metabolism ; Analysis ; Arabinose - metabolism ; Bacteria ; Bioassays ; Biochemistry ; Bioinformatics ; Biological and medical sciences ; Biomedical and Life Sciences ; Biotechnology ; Cloning ; Clostridium acetobutylicum ; Clostridium acetobutylicum - enzymology ; Clostridium acetobutylicum - genetics ; Clostridium acetobutylicum - metabolism ; E coli ; Fructose - metabolism ; Fundamental and applied biological sciences. Psychology ; Gene expression ; Gene Expression Regulation, Bacterial - drug effects ; Genes ; Genetic Engineering ; Genetic recombination ; Genetics and Molecular Biology of Industrial Organisms ; Glucose ; Glucose - metabolism ; Inorganic Chemistry ; Life Sciences ; Liquid chromatography ; Metabolism ; Metabolites ; Microbiology ; Pentose Phosphate Pathway ; Physiology ; RNA, Messenger - genetics ; RNA, Messenger - metabolism ; Studies ; Time Factors ; Xylose - metabolism</subject><ispartof>Journal of industrial microbiology & biotechnology, 2012-12, Vol.39 (12), p.1859-1867</ispartof><rights>Springer-Verlag (outside the USA) 2012</rights><rights>2014 INIST-CNRS</rights><rights>Society for Industrial Microbiology and Biotechnology 2012</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c478t-4ee942b3b5935e5cbfee848e212b16c88b6133343b8fc2b3d2580e92f10788893</citedby><cites>FETCH-LOGICAL-c478t-4ee942b3b5935e5cbfee848e212b16c88b6133343b8fc2b3d2580e92f10788893</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s10295-012-1186-x$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s10295-012-1186-x$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,780,784,27915,27916,41479,42548,51310</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=26650634$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/22922942$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Servinsky, M. D.</creatorcontrib><creatorcontrib>Germane, K. L.</creatorcontrib><creatorcontrib>Liu, S.</creatorcontrib><creatorcontrib>Kiel, J. T.</creatorcontrib><creatorcontrib>Clark, A. M.</creatorcontrib><creatorcontrib>Shankar, J.</creatorcontrib><creatorcontrib>Sund, C. J.</creatorcontrib><title>Arabinose is metabolized via a phosphoketolase pathway in Clostridium acetobutylicum ATCC 824</title><title>Journal of industrial microbiology & biotechnology</title><addtitle>J Ind Microbiol Biotechnol</addtitle><addtitle>J Ind Microbiol Biotechnol</addtitle><description>In this report, a novel zymogram assay and coupled phosphoketolase assay were employed to demonstrate that
Clostridium acetobutylicum
gene CAC1343 encodes a bi-functional xylulose-5-P/fructose-6-P phosphoketolase (XFP). The specific activity of purified recombinant XFP was 6.9 U/mg on xylulose-5-P and 21 U/mg on fructose-6-P, while the specific activity of XFP in concentrated
C. acetobutylicum
whole-cell extract was 0.094 and 0.52 U/mg, respectively. Analysis of crude cell extracts indicated that XFP activity was present in cells grown on arabinose but not glucose and quantitative PCR was used to show that CAC1343 mRNA expression was induced 185-fold during growth on arabinose when compared to growth on glucose. HPLC analysis of metabolites revealed that during growth on xylose and glucose more butyrate than acetate was formed with final acetate:butyrate ratios of 0.72 and 0.83, respectively. Growth on arabinose caused a metabolic shift to more oxidized products with a final acetate:butyrate ratio of 1.95. The shift towards more oxidized products is consistent with the presence of an XFP, suggesting that arabinose is metabolized via a phosphoketolase pathway while xylose is probably metabolized via the pentose phosphate pathway.</description><subject>Aldehyde-Lyases - metabolism</subject><subject>Analysis</subject><subject>Arabinose - metabolism</subject><subject>Bacteria</subject><subject>Bioassays</subject><subject>Biochemistry</subject><subject>Bioinformatics</subject><subject>Biological and medical sciences</subject><subject>Biomedical and Life Sciences</subject><subject>Biotechnology</subject><subject>Cloning</subject><subject>Clostridium acetobutylicum</subject><subject>Clostridium acetobutylicum - enzymology</subject><subject>Clostridium acetobutylicum - genetics</subject><subject>Clostridium acetobutylicum - metabolism</subject><subject>E coli</subject><subject>Fructose - metabolism</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene expression</subject><subject>Gene Expression Regulation, Bacterial - drug effects</subject><subject>Genes</subject><subject>Genetic Engineering</subject><subject>Genetic recombination</subject><subject>Genetics and Molecular Biology of Industrial Organisms</subject><subject>Glucose</subject><subject>Glucose - metabolism</subject><subject>Inorganic Chemistry</subject><subject>Life Sciences</subject><subject>Liquid chromatography</subject><subject>Metabolism</subject><subject>Metabolites</subject><subject>Microbiology</subject><subject>Pentose Phosphate Pathway</subject><subject>Physiology</subject><subject>RNA, Messenger - genetics</subject><subject>RNA, Messenger - metabolism</subject><subject>Studies</subject><subject>Time Factors</subject><subject>Xylose - metabolism</subject><issn>1367-5435</issn><issn>1476-5535</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2012</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>8G5</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><sourceid>GUQSH</sourceid><sourceid>M2O</sourceid><recordid>eNp1kE1r3DAQhkVpadJtf0AvQVAKvbiRRpYsHxfTLwj0khyLkLTjRoltbSQ7zebXV2G3TSgUJDSDnpl55yXkLWcfOWPNaeYMWlkxDhXnWlV3z8gxrxtVSSnk8xIL1VSyFvKIvMr5ijEmmwZekiOAtpwajsmPdbIuTDEjDZmOOFsXh3CPG3obLLV0exlzudc4x8EWaGvny192R8NEuyHmOYVNWEZqfQHcMu-G4Eu6Pu86qqF-TV70dsj45vCuyMXnT-fd1-rs-5dv3fqs8nWj56pGLGKccLIVEqV3PaKuNQIHx5XX2ikuhKiF070v3AakZthCz1mjtW7FinzY992meLNgns0YssdhsBPGJRsOZXEFTLKCvvsHvYpLmoo6w7kEBqAbKBTfUz7FnBP2ZpvCaNPOcGYevDd7703x3jx4b-5Kzcmh8-JG3Pyt-GN2Ad4fAJu9HfpkJx_yI6eUZKpsuSKw53L5mn5ieiLxv9N_AzJNmvk</recordid><startdate>20121201</startdate><enddate>20121201</enddate><creator>Servinsky, M. D.</creator><creator>Germane, K. L.</creator><creator>Liu, S.</creator><creator>Kiel, J. T.</creator><creator>Clark, A. M.</creator><creator>Shankar, J.</creator><creator>Sund, C. 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J.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c478t-4ee942b3b5935e5cbfee848e212b16c88b6133343b8fc2b3d2580e92f10788893</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2012</creationdate><topic>Aldehyde-Lyases - metabolism</topic><topic>Analysis</topic><topic>Arabinose - metabolism</topic><topic>Bacteria</topic><topic>Bioassays</topic><topic>Biochemistry</topic><topic>Bioinformatics</topic><topic>Biological and medical sciences</topic><topic>Biomedical and Life Sciences</topic><topic>Biotechnology</topic><topic>Cloning</topic><topic>Clostridium acetobutylicum</topic><topic>Clostridium acetobutylicum - enzymology</topic><topic>Clostridium acetobutylicum - genetics</topic><topic>Clostridium acetobutylicum - metabolism</topic><topic>E coli</topic><topic>Fructose - metabolism</topic><topic>Fundamental and applied biological sciences. 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D.</au><au>Germane, K. L.</au><au>Liu, S.</au><au>Kiel, J. T.</au><au>Clark, A. M.</au><au>Shankar, J.</au><au>Sund, C. J.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Arabinose is metabolized via a phosphoketolase pathway in Clostridium acetobutylicum ATCC 824</atitle><jtitle>Journal of industrial microbiology & biotechnology</jtitle><stitle>J Ind Microbiol Biotechnol</stitle><addtitle>J Ind Microbiol Biotechnol</addtitle><date>2012-12-01</date><risdate>2012</risdate><volume>39</volume><issue>12</issue><spage>1859</spage><epage>1867</epage><pages>1859-1867</pages><issn>1367-5435</issn><eissn>1476-5535</eissn><abstract>In this report, a novel zymogram assay and coupled phosphoketolase assay were employed to demonstrate that
Clostridium acetobutylicum
gene CAC1343 encodes a bi-functional xylulose-5-P/fructose-6-P phosphoketolase (XFP). The specific activity of purified recombinant XFP was 6.9 U/mg on xylulose-5-P and 21 U/mg on fructose-6-P, while the specific activity of XFP in concentrated
C. acetobutylicum
whole-cell extract was 0.094 and 0.52 U/mg, respectively. Analysis of crude cell extracts indicated that XFP activity was present in cells grown on arabinose but not glucose and quantitative PCR was used to show that CAC1343 mRNA expression was induced 185-fold during growth on arabinose when compared to growth on glucose. HPLC analysis of metabolites revealed that during growth on xylose and glucose more butyrate than acetate was formed with final acetate:butyrate ratios of 0.72 and 0.83, respectively. Growth on arabinose caused a metabolic shift to more oxidized products with a final acetate:butyrate ratio of 1.95. The shift towards more oxidized products is consistent with the presence of an XFP, suggesting that arabinose is metabolized via a phosphoketolase pathway while xylose is probably metabolized via the pentose phosphate pathway.</abstract><cop>Berlin/Heidelberg</cop><pub>Springer-Verlag</pub><pmid>22922942</pmid><doi>10.1007/s10295-012-1186-x</doi><tpages>9</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Journal of industrial microbiology & biotechnology, 2012-12, Vol.39 (12), p.1859-1867 |
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| subjects | Aldehyde-Lyases - metabolism Analysis Arabinose - metabolism Bacteria Bioassays Biochemistry Bioinformatics Biological and medical sciences Biomedical and Life Sciences Biotechnology Cloning Clostridium acetobutylicum Clostridium acetobutylicum - enzymology Clostridium acetobutylicum - genetics Clostridium acetobutylicum - metabolism E coli Fructose - metabolism Fundamental and applied biological sciences. Psychology Gene expression Gene Expression Regulation, Bacterial - drug effects Genes Genetic Engineering Genetic recombination Genetics and Molecular Biology of Industrial Organisms Glucose Glucose - metabolism Inorganic Chemistry Life Sciences Liquid chromatography Metabolism Metabolites Microbiology Pentose Phosphate Pathway Physiology RNA, Messenger - genetics RNA, Messenger - metabolism Studies Time Factors Xylose - metabolism |
| title | Arabinose is metabolized via a phosphoketolase pathway in Clostridium acetobutylicum ATCC 824 |
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