Engineering and manipulation of a mevalonate pathway in Escherichia coli for isoprene production
Isoprene is a useful phytochemical with high commercial values in many industrial applications including synthetic rubber, elastomers, isoprenoid medicines, and fossil fuel. Currently, isoprene is on large scale produced from petrochemical sources. An efficient biological process for isoprene produc...
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| Veröffentlicht in: | Applied microbiology and biotechnology 2019, Vol.103 (1), p.239-250 |
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| creator | Liu, Chun-Li Bi, Hao-Ran Bai, Zhonghu Fan, Li-Hai Tan, Tian-Wei |
| description | Isoprene is a useful phytochemical with high commercial values in many industrial applications including synthetic rubber, elastomers, isoprenoid medicines, and fossil fuel. Currently, isoprene is on large scale produced from petrochemical sources. An efficient biological process for isoprene production utilizing renewable feedstocks would be an important direction of research due to the fossil raw material depletion and air pollution. In this study, we introduced the mevalonate (MVA) pathway genes/acetoacetyl-coenzyme A thiolase (
mvaE
) and MVA synthase (
mvaS
) from
Enterococcus faecalis
(
E. faecalis
); MVA kinase (
mvk
) derived from
Methanosarcina mazei
(
M. mazei
); and phosphomevalonate kinase (
pmk
), diphosphomevalonate decarboxylase (
mvaD
), and isopentenyl diphosphate isomerase (
idi
) from
Streptococcus pneumoniae
(
S. pneumoniae
) to accelerate dimethylallyl diphosphate (DMAPP) accumulation in
Escherichia coli
(
E. coli
). Together with a codon-optimized isoprene synthase (
ispS
) from
Populus alba
(
P. alba
),
E. coli
strain succeeded in formation of isoprene. We then manipulated the heterologous MVA pathway for high-level production of isoprene, by controlling the gene expression levels of the MVA pathway genes. We engineered four
E. coli
strains which showed different gene expression levels and different isoprene productivities, and we also characterized them with quantitative real-time PCR and metabolite analysis. To further improve the isoprene titers and release the toxicity to cells, we developed the extraction fermentation by adding dodecane in cultures. Finally, strain BL2T7P1TrcP harboring balanced gene expression system produced 587 ± 47 mg/L isoprene, with a 5.2-fold titer improvement in comparison with strain BL7CT7P. This work indicated that a balanced metabolic flux played a significant role to improve the isoprene production via MVA pathway. |
| doi_str_mv | 10.1007/s00253-018-9472-9 |
| format | Article |
| fullrecord | <record><control><sourceid>gale_proqu</sourceid><recordid>TN_cdi_proquest_journals_2126732441</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><galeid>A567773970</galeid><sourcerecordid>A567773970</sourcerecordid><originalsourceid>FETCH-LOGICAL-c437t-6bc9ee7d3f3129121b11fb369b4f75c88ce6fd67fd42e0d2e0d8e4fba9f5912a3</originalsourceid><addsrcrecordid>eNp1kEtP7SAYRYm5Ro-PH-DkhuSOq7wKZWjM8ZGYONExUgrnYFqo0Gr899Icr44ckG-y9t5kAXCG0TlGSFxkhEhNK4SbSjJBKrkHVphRUiGO2R-wQljUlahlcwiOcn5BCJOG8wNwSBEVjAu2As_rsPHB2uTDBurQwUEHP869nnwMMDqo4WDfdB-Dniwc9bR91x_QB7jOZltSZus1NLH30MUEfY5jsqGAKXazWTpOwL7TfbanX_cYPF2vH69uq_uHm7ury_vKMCqmirdGWis66igmEhPcYuxaymXLnKhN0xjLXceF6xixqFteY5lrtXR1wTU9Bv92vWX6dbZ5Ui9xTqFMKoIJF5Qwhn-oje6t8sHFKWkz-GzUZc2FEFQKVCi8o0yKOSfr1Jj8oNOHwkgt5tXOvCrm1WJeyZL5-7U_t4PtvhP_VReA7IA8LrJt-vng762fG6yOlQ</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2126732441</pqid></control><display><type>article</type><title>Engineering and manipulation of a mevalonate pathway in Escherichia coli for isoprene production</title><source>Springer Nature - Complete Springer Journals</source><creator>Liu, Chun-Li ; Bi, Hao-Ran ; Bai, Zhonghu ; Fan, Li-Hai ; Tan, Tian-Wei</creator><creatorcontrib>Liu, Chun-Li ; Bi, Hao-Ran ; Bai, Zhonghu ; Fan, Li-Hai ; Tan, Tian-Wei</creatorcontrib><description>Isoprene is a useful phytochemical with high commercial values in many industrial applications including synthetic rubber, elastomers, isoprenoid medicines, and fossil fuel. Currently, isoprene is on large scale produced from petrochemical sources. An efficient biological process for isoprene production utilizing renewable feedstocks would be an important direction of research due to the fossil raw material depletion and air pollution. In this study, we introduced the mevalonate (MVA) pathway genes/acetoacetyl-coenzyme A thiolase (
mvaE
) and MVA synthase (
mvaS
) from
Enterococcus faecalis
(
E. faecalis
); MVA kinase (
mvk
) derived from
Methanosarcina mazei
(
M. mazei
); and phosphomevalonate kinase (
pmk
), diphosphomevalonate decarboxylase (
mvaD
), and isopentenyl diphosphate isomerase (
idi
) from
Streptococcus pneumoniae
(
S. pneumoniae
) to accelerate dimethylallyl diphosphate (DMAPP) accumulation in
Escherichia coli
(
E. coli
). Together with a codon-optimized isoprene synthase (
ispS
) from
Populus alba
(
P. alba
),
E. coli
strain succeeded in formation of isoprene. We then manipulated the heterologous MVA pathway for high-level production of isoprene, by controlling the gene expression levels of the MVA pathway genes. We engineered four
E. coli
strains which showed different gene expression levels and different isoprene productivities, and we also characterized them with quantitative real-time PCR and metabolite analysis. To further improve the isoprene titers and release the toxicity to cells, we developed the extraction fermentation by adding dodecane in cultures. Finally, strain BL2T7P1TrcP harboring balanced gene expression system produced 587 ± 47 mg/L isoprene, with a 5.2-fold titer improvement in comparison with strain BL7CT7P. This work indicated that a balanced metabolic flux played a significant role to improve the isoprene production via MVA pathway.</description><identifier>ISSN: 0175-7598</identifier><identifier>EISSN: 1432-0614</identifier><identifier>DOI: 10.1007/s00253-018-9472-9</identifier><identifier>PMID: 30374674</identifier><language>eng</language><publisher>Berlin/Heidelberg: Springer Berlin Heidelberg</publisher><subject>Air pollution ; Bacteria ; Biological activity ; Biomedical and Life Sciences ; Biosynthesis ; Biotechnological Products and Process Engineering ; Biotechnology ; C5 hydrocarbons ; Coenzyme A ; Diphosphomevalonate decarboxylase ; Dodecane ; E coli ; Elastomers ; Escherichia coli ; Fermentation ; Fossil fuels ; Gene expression ; Genes ; Genetic aspects ; Genetic engineering ; Industrial applications ; Isopentenyl diphosphate ; Isoprene ; Kinases ; Life Sciences ; Metabolic flux ; Metabolites ; Methods ; Mevalonate pathway ; Mevalonic acid ; Microbial genetic engineering ; Microbial Genetics and Genomics ; Microbiology ; Petrochemicals ; Petrochemicals industry ; Phosphomevalonate kinase ; Physiological aspects ; Production processes ; Rubber ; Streptococcus infections ; Streptococcus pneumoniae ; Synthetic rubber ; Thiolase ; Toxicity</subject><ispartof>Applied microbiology and biotechnology, 2019, Vol.103 (1), p.239-250</ispartof><rights>Springer-Verlag GmbH Germany, part of Springer Nature 2018</rights><rights>COPYRIGHT 2019 Springer</rights><rights>Applied Microbiology and Biotechnology is a copyright of Springer, (2018). All Rights Reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c437t-6bc9ee7d3f3129121b11fb369b4f75c88ce6fd67fd42e0d2e0d8e4fba9f5912a3</citedby><cites>FETCH-LOGICAL-c437t-6bc9ee7d3f3129121b11fb369b4f75c88ce6fd67fd42e0d2e0d8e4fba9f5912a3</cites><orcidid>0000-0002-3107-1843</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s00253-018-9472-9$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s00253-018-9472-9$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,776,780,27901,27902,41464,42533,51294</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/30374674$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Liu, Chun-Li</creatorcontrib><creatorcontrib>Bi, Hao-Ran</creatorcontrib><creatorcontrib>Bai, Zhonghu</creatorcontrib><creatorcontrib>Fan, Li-Hai</creatorcontrib><creatorcontrib>Tan, Tian-Wei</creatorcontrib><title>Engineering and manipulation of a mevalonate pathway in Escherichia coli for isoprene production</title><title>Applied microbiology and biotechnology</title><addtitle>Appl Microbiol Biotechnol</addtitle><addtitle>Appl Microbiol Biotechnol</addtitle><description>Isoprene is a useful phytochemical with high commercial values in many industrial applications including synthetic rubber, elastomers, isoprenoid medicines, and fossil fuel. Currently, isoprene is on large scale produced from petrochemical sources. An efficient biological process for isoprene production utilizing renewable feedstocks would be an important direction of research due to the fossil raw material depletion and air pollution. In this study, we introduced the mevalonate (MVA) pathway genes/acetoacetyl-coenzyme A thiolase (
mvaE
) and MVA synthase (
mvaS
) from
Enterococcus faecalis
(
E. faecalis
); MVA kinase (
mvk
) derived from
Methanosarcina mazei
(
M. mazei
); and phosphomevalonate kinase (
pmk
), diphosphomevalonate decarboxylase (
mvaD
), and isopentenyl diphosphate isomerase (
idi
) from
Streptococcus pneumoniae
(
S. pneumoniae
) to accelerate dimethylallyl diphosphate (DMAPP) accumulation in
Escherichia coli
(
E. coli
). Together with a codon-optimized isoprene synthase (
ispS
) from
Populus alba
(
P. alba
),
E. coli
strain succeeded in formation of isoprene. We then manipulated the heterologous MVA pathway for high-level production of isoprene, by controlling the gene expression levels of the MVA pathway genes. We engineered four
E. coli
strains which showed different gene expression levels and different isoprene productivities, and we also characterized them with quantitative real-time PCR and metabolite analysis. To further improve the isoprene titers and release the toxicity to cells, we developed the extraction fermentation by adding dodecane in cultures. Finally, strain BL2T7P1TrcP harboring balanced gene expression system produced 587 ± 47 mg/L isoprene, with a 5.2-fold titer improvement in comparison with strain BL7CT7P. This work indicated that a balanced metabolic flux played a significant role to improve the isoprene production via MVA pathway.</description><subject>Air pollution</subject><subject>Bacteria</subject><subject>Biological activity</subject><subject>Biomedical and Life Sciences</subject><subject>Biosynthesis</subject><subject>Biotechnological Products and Process Engineering</subject><subject>Biotechnology</subject><subject>C5 hydrocarbons</subject><subject>Coenzyme A</subject><subject>Diphosphomevalonate decarboxylase</subject><subject>Dodecane</subject><subject>E coli</subject><subject>Elastomers</subject><subject>Escherichia coli</subject><subject>Fermentation</subject><subject>Fossil fuels</subject><subject>Gene expression</subject><subject>Genes</subject><subject>Genetic aspects</subject><subject>Genetic engineering</subject><subject>Industrial applications</subject><subject>Isopentenyl diphosphate</subject><subject>Isoprene</subject><subject>Kinases</subject><subject>Life Sciences</subject><subject>Metabolic flux</subject><subject>Metabolites</subject><subject>Methods</subject><subject>Mevalonate pathway</subject><subject>Mevalonic acid</subject><subject>Microbial genetic engineering</subject><subject>Microbial Genetics and Genomics</subject><subject>Microbiology</subject><subject>Petrochemicals</subject><subject>Petrochemicals industry</subject><subject>Phosphomevalonate kinase</subject><subject>Physiological aspects</subject><subject>Production processes</subject><subject>Rubber</subject><subject>Streptococcus infections</subject><subject>Streptococcus pneumoniae</subject><subject>Synthetic rubber</subject><subject>Thiolase</subject><subject>Toxicity</subject><issn>0175-7598</issn><issn>1432-0614</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2019</creationdate><recordtype>article</recordtype><sourceid>BENPR</sourceid><recordid>eNp1kEtP7SAYRYm5Ro-PH-DkhuSOq7wKZWjM8ZGYONExUgrnYFqo0Gr899Icr44ckG-y9t5kAXCG0TlGSFxkhEhNK4SbSjJBKrkHVphRUiGO2R-wQljUlahlcwiOcn5BCJOG8wNwSBEVjAu2As_rsPHB2uTDBurQwUEHP869nnwMMDqo4WDfdB-Dniwc9bR91x_QB7jOZltSZus1NLH30MUEfY5jsqGAKXazWTpOwL7TfbanX_cYPF2vH69uq_uHm7ury_vKMCqmirdGWis66igmEhPcYuxaymXLnKhN0xjLXceF6xixqFteY5lrtXR1wTU9Bv92vWX6dbZ5Ui9xTqFMKoIJF5Qwhn-oje6t8sHFKWkz-GzUZc2FEFQKVCi8o0yKOSfr1Jj8oNOHwkgt5tXOvCrm1WJeyZL5-7U_t4PtvhP_VReA7IA8LrJt-vng762fG6yOlQ</recordid><startdate>2019</startdate><enddate>2019</enddate><creator>Liu, Chun-Li</creator><creator>Bi, Hao-Ran</creator><creator>Bai, Zhonghu</creator><creator>Fan, Li-Hai</creator><creator>Tan, Tian-Wei</creator><general>Springer Berlin 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and manipulation of a mevalonate pathway in Escherichia coli for isoprene production</title><author>Liu, Chun-Li ; Bi, Hao-Ran ; Bai, Zhonghu ; Fan, Li-Hai ; Tan, Tian-Wei</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c437t-6bc9ee7d3f3129121b11fb369b4f75c88ce6fd67fd42e0d2e0d8e4fba9f5912a3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2019</creationdate><topic>Air pollution</topic><topic>Bacteria</topic><topic>Biological activity</topic><topic>Biomedical and Life Sciences</topic><topic>Biosynthesis</topic><topic>Biotechnological Products and Process Engineering</topic><topic>Biotechnology</topic><topic>C5 hydrocarbons</topic><topic>Coenzyme A</topic><topic>Diphosphomevalonate decarboxylase</topic><topic>Dodecane</topic><topic>E coli</topic><topic>Elastomers</topic><topic>Escherichia coli</topic><topic>Fermentation</topic><topic>Fossil fuels</topic><topic>Gene expression</topic><topic>Genes</topic><topic>Genetic aspects</topic><topic>Genetic engineering</topic><topic>Industrial applications</topic><topic>Isopentenyl diphosphate</topic><topic>Isoprene</topic><topic>Kinases</topic><topic>Life Sciences</topic><topic>Metabolic flux</topic><topic>Metabolites</topic><topic>Methods</topic><topic>Mevalonate pathway</topic><topic>Mevalonic acid</topic><topic>Microbial genetic engineering</topic><topic>Microbial Genetics and Genomics</topic><topic>Microbiology</topic><topic>Petrochemicals</topic><topic>Petrochemicals industry</topic><topic>Phosphomevalonate kinase</topic><topic>Physiological aspects</topic><topic>Production processes</topic><topic>Rubber</topic><topic>Streptococcus infections</topic><topic>Streptococcus pneumoniae</topic><topic>Synthetic rubber</topic><topic>Thiolase</topic><topic>Toxicity</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Liu, Chun-Li</creatorcontrib><creatorcontrib>Bi, Hao-Ran</creatorcontrib><creatorcontrib>Bai, Zhonghu</creatorcontrib><creatorcontrib>Fan, Li-Hai</creatorcontrib><creatorcontrib>Tan, Tian-Wei</creatorcontrib><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>ABI/INFORM Collection</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 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C)</collection><collection>Biological Science Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>ProQuest Central (New)</collection><collection>ProQuest One Academic (New)</collection><collection>ProQuest Health & Medical Research Collection</collection><collection>ProQuest One Academic Middle East (New)</collection><collection>ProQuest One Health & Nursing</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 Applied & Life Sciences</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central Basic</collection><jtitle>Applied microbiology and biotechnology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Liu, Chun-Li</au><au>Bi, Hao-Ran</au><au>Bai, Zhonghu</au><au>Fan, Li-Hai</au><au>Tan, Tian-Wei</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Engineering and manipulation of a mevalonate pathway in Escherichia coli for isoprene production</atitle><jtitle>Applied microbiology and biotechnology</jtitle><stitle>Appl Microbiol Biotechnol</stitle><addtitle>Appl Microbiol Biotechnol</addtitle><date>2019</date><risdate>2019</risdate><volume>103</volume><issue>1</issue><spage>239</spage><epage>250</epage><pages>239-250</pages><issn>0175-7598</issn><eissn>1432-0614</eissn><abstract>Isoprene is a useful phytochemical with high commercial values in many industrial applications including synthetic rubber, elastomers, isoprenoid medicines, and fossil fuel. Currently, isoprene is on large scale produced from petrochemical sources. An efficient biological process for isoprene production utilizing renewable feedstocks would be an important direction of research due to the fossil raw material depletion and air pollution. In this study, we introduced the mevalonate (MVA) pathway genes/acetoacetyl-coenzyme A thiolase (
mvaE
) and MVA synthase (
mvaS
) from
Enterococcus faecalis
(
E. faecalis
); MVA kinase (
mvk
) derived from
Methanosarcina mazei
(
M. mazei
); and phosphomevalonate kinase (
pmk
), diphosphomevalonate decarboxylase (
mvaD
), and isopentenyl diphosphate isomerase (
idi
) from
Streptococcus pneumoniae
(
S. pneumoniae
) to accelerate dimethylallyl diphosphate (DMAPP) accumulation in
Escherichia coli
(
E. coli
). Together with a codon-optimized isoprene synthase (
ispS
) from
Populus alba
(
P. alba
),
E. coli
strain succeeded in formation of isoprene. We then manipulated the heterologous MVA pathway for high-level production of isoprene, by controlling the gene expression levels of the MVA pathway genes. We engineered four
E. coli
strains which showed different gene expression levels and different isoprene productivities, and we also characterized them with quantitative real-time PCR and metabolite analysis. To further improve the isoprene titers and release the toxicity to cells, we developed the extraction fermentation by adding dodecane in cultures. Finally, strain BL2T7P1TrcP harboring balanced gene expression system produced 587 ± 47 mg/L isoprene, with a 5.2-fold titer improvement in comparison with strain BL7CT7P. This work indicated that a balanced metabolic flux played a significant role to improve the isoprene production via MVA pathway.</abstract><cop>Berlin/Heidelberg</cop><pub>Springer Berlin Heidelberg</pub><pmid>30374674</pmid><doi>10.1007/s00253-018-9472-9</doi><tpages>12</tpages><orcidid>https://orcid.org/0000-0002-3107-1843</orcidid></addata></record> |
| fulltext | fulltext |
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| ispartof | Applied microbiology and biotechnology, 2019, Vol.103 (1), p.239-250 |
| issn | 0175-7598 1432-0614 |
| language | eng |
| recordid | cdi_proquest_journals_2126732441 |
| source | Springer Nature - Complete Springer Journals |
| subjects | Air pollution Bacteria Biological activity Biomedical and Life Sciences Biosynthesis Biotechnological Products and Process Engineering Biotechnology C5 hydrocarbons Coenzyme A Diphosphomevalonate decarboxylase Dodecane E coli Elastomers Escherichia coli Fermentation Fossil fuels Gene expression Genes Genetic aspects Genetic engineering Industrial applications Isopentenyl diphosphate Isoprene Kinases Life Sciences Metabolic flux Metabolites Methods Mevalonate pathway Mevalonic acid Microbial genetic engineering Microbial Genetics and Genomics Microbiology Petrochemicals Petrochemicals industry Phosphomevalonate kinase Physiological aspects Production processes Rubber Streptococcus infections Streptococcus pneumoniae Synthetic rubber Thiolase Toxicity |
| title | Engineering and manipulation of a mevalonate pathway in Escherichia coli for isoprene production |
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