Metabolic engineering of Clostridium tyrobutyricum for n-butanol production: effects of CoA transferase
The overexpression of CoA transferase (ctfAB), which catalyzes the reaction: acetate/butyrate + acetoacetyl-CoA → acetyl/butyryl-CoA + acetoacetate, was studied for its effects on acid reassimilation and butanol biosynthesis in Clostridium tyrobutyricum (Δack, adhE2). The plasmid pMTL007 was used to...
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| Veröffentlicht in: | Applied microbiology and biotechnology 2015-06, Vol.99 (11), p.4917-4930 |
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| creator | Yu, Le Zhao, Jingbo Xu, Mengmeng Dong, Jie Varghese, Saju Yu, Mingrui Tang, I-Ching Yang, Shang-Tian |
| description | The overexpression of CoA transferase (ctfAB), which catalyzes the reaction: acetate/butyrate + acetoacetyl-CoA → acetyl/butyryl-CoA + acetoacetate, was studied for its effects on acid reassimilation and butanol biosynthesis in Clostridium tyrobutyricum (Δack, adhE2). The plasmid pMTL007 was used to co-express adhE2 and ctfAB from Clostridium acetobutylicum ATCC 824. In addition, the sol operon containing ctfAB, adc (acetoacetate decarboxylase), and ald (aldehyde dehydrogenase) was also cloned from Clostridium beijerinckii NCIMB 8052 and expressed in C. tyrobutyricum (Δack, adhE2). Mutants expressing these genes were evaluated for their ability to produce butanol from glucose in batch fermentations at pH 5.0 and 6.0. Compared to C. tyrobutyricum (Δack, adhE2) without expressing ctfAB, all mutants with ctfAB overexpression produced more butanol, with butanol yield increased to 0.22 − 0.26 g/g (vs. 0.10 − 0.13 g/g) and productivity to 0.35 g/l h (vs. 0.13 g/l h) because of the reduced acetate and butyrate production. The expression of ctfAB also resulted in acetone production from acetoacetate through a non-enzymatic decarboxylation. |
| doi_str_mv | 10.1007/s00253-015-6566-5 |
| format | Article |
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The plasmid pMTL007 was used to co-express adhE2 and ctfAB from Clostridium acetobutylicum ATCC 824. In addition, the sol operon containing ctfAB, adc (acetoacetate decarboxylase), and ald (aldehyde dehydrogenase) was also cloned from Clostridium beijerinckii NCIMB 8052 and expressed in C. tyrobutyricum (Δack, adhE2). Mutants expressing these genes were evaluated for their ability to produce butanol from glucose in batch fermentations at pH 5.0 and 6.0. Compared to C. tyrobutyricum (Δack, adhE2) without expressing ctfAB, all mutants with ctfAB overexpression produced more butanol, with butanol yield increased to 0.22 − 0.26 g/g (vs. 0.10 − 0.13 g/g) and productivity to 0.35 g/l h (vs. 0.13 g/l h) because of the reduced acetate and butyrate production. The expression of ctfAB also resulted in acetone production from acetoacetate through a non-enzymatic decarboxylation.</description><identifier>ISSN: 0175-7598</identifier><identifier>EISSN: 1432-0614</identifier><identifier>DOI: 10.1007/s00253-015-6566-5</identifier><identifier>PMID: 25851718</identifier><language>eng</language><publisher>Berlin/Heidelberg: Springer-Verlag</publisher><subject>1-Butanol - metabolism ; acetates ; acetone ; Acetone - metabolism ; Acids ; Alcohol ; aldehyde dehydrogenase ; Bacteria ; Biodiesel fuels ; Bioenergy and Biofuels ; Bioengineering ; Biofuels ; Biomedical and Life Sciences ; Biosynthesis ; Biotechnology ; Butanol ; butyrates ; Cloning ; Clostridium ; Clostridium acetobutylicum ; Clostridium acetobutylicum - enzymology ; Clostridium acetobutylicum - genetics ; Clostridium beijerinckii ; Clostridium beijerinckii - enzymology ; Clostridium beijerinckii - genetics ; Clostridium tyrobutyricum ; Clostridium tyrobutyricum - enzymology ; Clostridium tyrobutyricum - genetics ; Clostridium tyrobutyricum - metabolism ; Coenzyme A-Transferases - genetics ; Coenzyme A-Transferases - metabolism ; decarboxylation ; Dehydrogenases ; E coli ; Fermentation ; Gene Deletion ; Gene Expression ; gene overexpression ; Genes ; Genetic aspects ; Genetic engineering ; Glucose ; Glucose - metabolism ; Kinases ; Life Sciences ; Metabolic Engineering ; Metabolic Networks and Pathways - genetics ; Metabolism ; Methods ; Microbial Genetics and Genomics ; Microbiology ; mutants ; operon ; Physiological aspects ; Plasmids ; Production processes ; Studies</subject><ispartof>Applied microbiology and biotechnology, 2015-06, Vol.99 (11), p.4917-4930</ispartof><rights>Springer-Verlag Berlin Heidelberg 2015</rights><rights>COPYRIGHT 2015 Springer</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c604t-12489431893556a0cbd5b8de967ff716f95115cd511ca5f8dbd62983c9da8523</citedby><cites>FETCH-LOGICAL-c604t-12489431893556a0cbd5b8de967ff716f95115cd511ca5f8dbd62983c9da8523</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/s00253-015-6566-5$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s00253-015-6566-5$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>315,781,785,27928,27929,41492,42561,51323</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/25851718$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Yu, Le</creatorcontrib><creatorcontrib>Zhao, Jingbo</creatorcontrib><creatorcontrib>Xu, Mengmeng</creatorcontrib><creatorcontrib>Dong, Jie</creatorcontrib><creatorcontrib>Varghese, Saju</creatorcontrib><creatorcontrib>Yu, Mingrui</creatorcontrib><creatorcontrib>Tang, I-Ching</creatorcontrib><creatorcontrib>Yang, Shang-Tian</creatorcontrib><title>Metabolic engineering of Clostridium tyrobutyricum for n-butanol production: effects of CoA transferase</title><title>Applied microbiology and biotechnology</title><addtitle>Appl Microbiol Biotechnol</addtitle><addtitle>Appl Microbiol Biotechnol</addtitle><description>The overexpression of CoA transferase (ctfAB), which catalyzes the reaction: acetate/butyrate + acetoacetyl-CoA → acetyl/butyryl-CoA + acetoacetate, was studied for its effects on acid reassimilation and butanol biosynthesis in Clostridium tyrobutyricum (Δack, adhE2). The plasmid pMTL007 was used to co-express adhE2 and ctfAB from Clostridium acetobutylicum ATCC 824. In addition, the sol operon containing ctfAB, adc (acetoacetate decarboxylase), and ald (aldehyde dehydrogenase) was also cloned from Clostridium beijerinckii NCIMB 8052 and expressed in C. tyrobutyricum (Δack, adhE2). Mutants expressing these genes were evaluated for their ability to produce butanol from glucose in batch fermentations at pH 5.0 and 6.0. Compared to C. tyrobutyricum (Δack, adhE2) without expressing ctfAB, all mutants with ctfAB overexpression produced more butanol, with butanol yield increased to 0.22 − 0.26 g/g (vs. 0.10 − 0.13 g/g) and productivity to 0.35 g/l h (vs. 0.13 g/l h) because of the reduced acetate and butyrate production. The expression of ctfAB also resulted in acetone production from acetoacetate through a non-enzymatic decarboxylation.</description><subject>1-Butanol - metabolism</subject><subject>acetates</subject><subject>acetone</subject><subject>Acetone - metabolism</subject><subject>Acids</subject><subject>Alcohol</subject><subject>aldehyde dehydrogenase</subject><subject>Bacteria</subject><subject>Biodiesel fuels</subject><subject>Bioenergy and Biofuels</subject><subject>Bioengineering</subject><subject>Biofuels</subject><subject>Biomedical and Life Sciences</subject><subject>Biosynthesis</subject><subject>Biotechnology</subject><subject>Butanol</subject><subject>butyrates</subject><subject>Cloning</subject><subject>Clostridium</subject><subject>Clostridium acetobutylicum</subject><subject>Clostridium acetobutylicum - enzymology</subject><subject>Clostridium acetobutylicum - genetics</subject><subject>Clostridium beijerinckii</subject><subject>Clostridium beijerinckii - enzymology</subject><subject>Clostridium beijerinckii - genetics</subject><subject>Clostridium tyrobutyricum</subject><subject>Clostridium tyrobutyricum - enzymology</subject><subject>Clostridium tyrobutyricum - genetics</subject><subject>Clostridium tyrobutyricum - metabolism</subject><subject>Coenzyme A-Transferases - genetics</subject><subject>Coenzyme A-Transferases - metabolism</subject><subject>decarboxylation</subject><subject>Dehydrogenases</subject><subject>E coli</subject><subject>Fermentation</subject><subject>Gene Deletion</subject><subject>Gene Expression</subject><subject>gene overexpression</subject><subject>Genes</subject><subject>Genetic aspects</subject><subject>Genetic engineering</subject><subject>Glucose</subject><subject>Glucose - metabolism</subject><subject>Kinases</subject><subject>Life Sciences</subject><subject>Metabolic Engineering</subject><subject>Metabolic Networks and Pathways - genetics</subject><subject>Metabolism</subject><subject>Methods</subject><subject>Microbial Genetics and Genomics</subject><subject>Microbiology</subject><subject>mutants</subject><subject>operon</subject><subject>Physiological aspects</subject><subject>Plasmids</subject><subject>Production processes</subject><subject>Studies</subject><issn>0175-7598</issn><issn>1432-0614</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNp9kltrFDEUx4Modq1-AF90wBd9mJrLJJP0bVm8FCqCrc8hk8uQMpusSQbstzfTqZcVkcAJJ_z-J-ckfwCeI3iGIOzfZggxJS1EtGWUsZY-ABvUEdxChrqHYANRT9ueCn4CnuR8AyHCnLHH4ARTTlGP-AaMn2xRQ5y8bmwYfbA2-TA20TW7KeaSvPHzvim3KQ5zjV7XzMXUhLbmKsSpOaRoZl18DOeNdc7qku_kcduUpEJ2Nqlsn4JHTk3ZPrvfT8H1-3fXu4_t5ecPF7vtZasZ7EqLcMdFRxAXhFKmoB4MHbixgvXO9Yg5QRGi2tSoFXXcDIZhwYkWRnGKySl4vZatXX2bbS5y77O206SCjXOWiHEkoMCwq-irv9CbOKdQm7uj6sMyTH5To5qs9MHFOpReisptRyojGF-uPfsHVZexe69jsM7X8yPBmyNBZYr9XkY15ywvrr4cs2hldYo5J-vkIfm9SrcSQbn4QK4-kNUHcvGBpFXz4n64edhb80vx8-MrgFcgH5YPt-mP6f9T9eUqcipKNSaf5dcrXIHqLM4pgeQHAN_ERQ</recordid><startdate>20150601</startdate><enddate>20150601</enddate><creator>Yu, Le</creator><creator>Zhao, Jingbo</creator><creator>Xu, Mengmeng</creator><creator>Dong, Jie</creator><creator>Varghese, Saju</creator><creator>Yu, Mingrui</creator><creator>Tang, I-Ching</creator><creator>Yang, Shang-Tian</creator><general>Springer-Verlag</general><general>Springer Berlin Heidelberg</general><general>Springer</general><general>Springer Nature B.V</general><scope>FBQ</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>ISR</scope><scope>3V.</scope><scope>7QL</scope><scope>7T7</scope><scope>7WY</scope><scope>7WZ</scope><scope>7X7</scope><scope>7XB</scope><scope>87Z</scope><scope>88A</scope><scope>88E</scope><scope>88I</scope><scope>8AO</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>8FL</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BEZIV</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FRNLG</scope><scope>FYUFA</scope><scope>F~G</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K60</scope><scope>K6~</scope><scope>K9.</scope><scope>L.-</scope><scope>LK8</scope><scope>M0C</scope><scope>M0S</scope><scope>M1P</scope><scope>M2P</scope><scope>M7N</scope><scope>M7P</scope><scope>P64</scope><scope>PQBIZ</scope><scope>PQBZA</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>Q9U</scope><scope>7X8</scope></search><sort><creationdate>20150601</creationdate><title>Metabolic engineering of Clostridium tyrobutyricum for n-butanol production: effects of CoA transferase</title><author>Yu, Le ; Zhao, Jingbo ; Xu, Mengmeng ; Dong, Jie ; Varghese, Saju ; Yu, Mingrui ; Tang, I-Ching ; Yang, Shang-Tian</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c604t-12489431893556a0cbd5b8de967ff716f95115cd511ca5f8dbd62983c9da8523</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>1-Butanol - metabolism</topic><topic>acetates</topic><topic>acetone</topic><topic>Acetone - metabolism</topic><topic>Acids</topic><topic>Alcohol</topic><topic>aldehyde dehydrogenase</topic><topic>Bacteria</topic><topic>Biodiesel fuels</topic><topic>Bioenergy and Biofuels</topic><topic>Bioengineering</topic><topic>Biofuels</topic><topic>Biomedical and Life Sciences</topic><topic>Biosynthesis</topic><topic>Biotechnology</topic><topic>Butanol</topic><topic>butyrates</topic><topic>Cloning</topic><topic>Clostridium</topic><topic>Clostridium acetobutylicum</topic><topic>Clostridium acetobutylicum - enzymology</topic><topic>Clostridium acetobutylicum - genetics</topic><topic>Clostridium beijerinckii</topic><topic>Clostridium beijerinckii - enzymology</topic><topic>Clostridium beijerinckii - genetics</topic><topic>Clostridium tyrobutyricum</topic><topic>Clostridium tyrobutyricum - enzymology</topic><topic>Clostridium tyrobutyricum - genetics</topic><topic>Clostridium tyrobutyricum - metabolism</topic><topic>Coenzyme A-Transferases - genetics</topic><topic>Coenzyme A-Transferases - metabolism</topic><topic>decarboxylation</topic><topic>Dehydrogenases</topic><topic>E coli</topic><topic>Fermentation</topic><topic>Gene Deletion</topic><topic>Gene Expression</topic><topic>gene overexpression</topic><topic>Genes</topic><topic>Genetic aspects</topic><topic>Genetic engineering</topic><topic>Glucose</topic><topic>Glucose - metabolism</topic><topic>Kinases</topic><topic>Life Sciences</topic><topic>Metabolic Engineering</topic><topic>Metabolic Networks and Pathways - genetics</topic><topic>Metabolism</topic><topic>Methods</topic><topic>Microbial Genetics and Genomics</topic><topic>Microbiology</topic><topic>mutants</topic><topic>operon</topic><topic>Physiological aspects</topic><topic>Plasmids</topic><topic>Production processes</topic><topic>Studies</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Yu, Le</creatorcontrib><creatorcontrib>Zhao, Jingbo</creatorcontrib><creatorcontrib>Xu, Mengmeng</creatorcontrib><creatorcontrib>Dong, Jie</creatorcontrib><creatorcontrib>Varghese, Saju</creatorcontrib><creatorcontrib>Yu, Mingrui</creatorcontrib><creatorcontrib>Tang, I-Ching</creatorcontrib><creatorcontrib>Yang, Shang-Tian</creatorcontrib><collection>AGRIS</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Gale In Context: Science</collection><collection>ProQuest Central (Corporate)</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Access via ABI/INFORM (ProQuest)</collection><collection>ABI/INFORM Global (PDF only)</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>ABI/INFORM Global (Alumni Edition)</collection><collection>Biology Database (Alumni Edition)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Science Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ABI/INFORM Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Business Premium Collection</collection><collection>Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Business Premium Collection (Alumni)</collection><collection>Health Research Premium Collection</collection><collection>ABI/INFORM Global (Corporate)</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Business Collection (Alumni Edition)</collection><collection>ProQuest Business Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>ABI/INFORM Professional Advanced</collection><collection>ProQuest Biological Science Collection</collection><collection>ABI/INFORM Global</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Science Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biological Science Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>ProQuest One Business</collection><collection>ProQuest One Business (Alumni)</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central Basic</collection><collection>MEDLINE - Academic</collection><jtitle>Applied microbiology and biotechnology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Yu, Le</au><au>Zhao, Jingbo</au><au>Xu, Mengmeng</au><au>Dong, Jie</au><au>Varghese, Saju</au><au>Yu, Mingrui</au><au>Tang, I-Ching</au><au>Yang, Shang-Tian</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Metabolic engineering of Clostridium tyrobutyricum for n-butanol production: effects of CoA transferase</atitle><jtitle>Applied microbiology and biotechnology</jtitle><stitle>Appl Microbiol Biotechnol</stitle><addtitle>Appl Microbiol Biotechnol</addtitle><date>2015-06-01</date><risdate>2015</risdate><volume>99</volume><issue>11</issue><spage>4917</spage><epage>4930</epage><pages>4917-4930</pages><issn>0175-7598</issn><eissn>1432-0614</eissn><abstract>The overexpression of CoA transferase (ctfAB), which catalyzes the reaction: acetate/butyrate + acetoacetyl-CoA → acetyl/butyryl-CoA + acetoacetate, was studied for its effects on acid reassimilation and butanol biosynthesis in Clostridium tyrobutyricum (Δack, adhE2). The plasmid pMTL007 was used to co-express adhE2 and ctfAB from Clostridium acetobutylicum ATCC 824. In addition, the sol operon containing ctfAB, adc (acetoacetate decarboxylase), and ald (aldehyde dehydrogenase) was also cloned from Clostridium beijerinckii NCIMB 8052 and expressed in C. tyrobutyricum (Δack, adhE2). Mutants expressing these genes were evaluated for their ability to produce butanol from glucose in batch fermentations at pH 5.0 and 6.0. Compared to C. tyrobutyricum (Δack, adhE2) without expressing ctfAB, all mutants with ctfAB overexpression produced more butanol, with butanol yield increased to 0.22 − 0.26 g/g (vs. 0.10 − 0.13 g/g) and productivity to 0.35 g/l h (vs. 0.13 g/l h) because of the reduced acetate and butyrate production. The expression of ctfAB also resulted in acetone production from acetoacetate through a non-enzymatic decarboxylation.</abstract><cop>Berlin/Heidelberg</cop><pub>Springer-Verlag</pub><pmid>25851718</pmid><doi>10.1007/s00253-015-6566-5</doi><tpages>14</tpages></addata></record> |
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| ispartof | Applied microbiology and biotechnology, 2015-06, Vol.99 (11), p.4917-4930 |
| issn | 0175-7598 1432-0614 |
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| source | MEDLINE; SpringerNature Journals |
| subjects | 1-Butanol - metabolism acetates acetone Acetone - metabolism Acids Alcohol aldehyde dehydrogenase Bacteria Biodiesel fuels Bioenergy and Biofuels Bioengineering Biofuels Biomedical and Life Sciences Biosynthesis Biotechnology Butanol butyrates Cloning Clostridium Clostridium acetobutylicum Clostridium acetobutylicum - enzymology Clostridium acetobutylicum - genetics Clostridium beijerinckii Clostridium beijerinckii - enzymology Clostridium beijerinckii - genetics Clostridium tyrobutyricum Clostridium tyrobutyricum - enzymology Clostridium tyrobutyricum - genetics Clostridium tyrobutyricum - metabolism Coenzyme A-Transferases - genetics Coenzyme A-Transferases - metabolism decarboxylation Dehydrogenases E coli Fermentation Gene Deletion Gene Expression gene overexpression Genes Genetic aspects Genetic engineering Glucose Glucose - metabolism Kinases Life Sciences Metabolic Engineering Metabolic Networks and Pathways - genetics Metabolism Methods Microbial Genetics and Genomics Microbiology mutants operon Physiological aspects Plasmids Production processes Studies |
| title | Metabolic engineering of Clostridium tyrobutyricum for n-butanol production: effects of CoA transferase |
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