Metabolic engineering of Escherichia coli for production of mixed isoprenoid alcohols and their derivatives
Abstract Background Current petroleum-derived fuels such as gasoline (C5–C12) and diesel (C15–C22) are complex mixtures of hydrocarbons with different chain lengths and chemical structures. Isoprenoids are hydrocarbon-based compounds with different carbon chain lengths and diverse chemical structure...
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BMC
2018-07-01
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| Series: | Biotechnology for Biofuels |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s13068-018-1210-0 |
| _version_ | 1817977095635075072 |
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| author | Bakht Zada Chonglong Wang Ji-Bin Park Seong-Hee Jeong Ju-Eon Park Hawaibam Birla Singh Seon-Won Kim |
| author_facet | Bakht Zada Chonglong Wang Ji-Bin Park Seong-Hee Jeong Ju-Eon Park Hawaibam Birla Singh Seon-Won Kim |
| author_sort | Bakht Zada |
| collection | DOAJ |
| description | Abstract Background Current petroleum-derived fuels such as gasoline (C5–C12) and diesel (C15–C22) are complex mixtures of hydrocarbons with different chain lengths and chemical structures. Isoprenoids are hydrocarbon-based compounds with different carbon chain lengths and diverse chemical structures, similar to petroleum. Thus, isoprenoid alcohols such as isopentenol (C5), geraniol (C10), and farnesol (C15) have been considered to be ideal biofuel candidates. NudB, a native phosphatase of Escherichia coli, is reported to dephosphorylate isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP) into isopentenol. However, no attention has been paid to its promiscuous activity toward longer chain length (C10–C15) prenyl diphosphates. Results In this study, the promiscuous activity of NudB toward geranyl diphosphate (GPP) and farnesyl diphosphate (FPP) was applied for the production of isoprenoid alcohol mixtures, including isopentenol, geraniol, and farnesol, and their derivatives. E. coli was engineered to produce a mixture of C5 and C15 alcohols by overexpressing NudB (dihydroneopterin triphosphate diphosphohydrolase) and IspA (FPP synthase) along with a heterologous MVA pathway, which resulted in a total of up to 1652 mg/L mixture of C5 and C15 alcohols and their derivatives. The production was further increased to 2027 mg/L by overexpression of another endogenous phosphatase, AphA, in addition to NudB. Production of DMAPP- and FPP-derived alcohols and their derivatives was significantly increased with an increase in the gene dosage of idi, encoding IPP isomerase (IDI), indicating a potential modulation of the composition of the alcohols mixture according to the expression level of IDI. When IspA was replaced with its mutant IspA*, generating GPP in the production strain, a total of 1418 mg/L of the isoprenoid mixture was obtained containing C10 alcohols as a main component. Conclusions The promiscuous activity of NudB was newly identified and successfully used for production of isoprenoid-based alcohol mixtures, which are suitable as next-generation biofuels or commodity chemicals. This is the first successful report on high-titer production of an isoprenoid alcohol-based mixture. The engineering approaches can provide a valuable platform for production of other isoprenoid mixtures via a proportional modulation of IPP, DMAPP, GPP, and FPP syntheses. |
| first_indexed | 2024-04-13T22:11:47Z |
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| institution | Directory Open Access Journal |
| issn | 1754-6834 |
| language | English |
| last_indexed | 2024-04-13T22:11:47Z |
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| series | Biotechnology for Biofuels |
| spelling | doaj.art-536edc31ef144476ae0e265a62b225f62022-12-22T02:27:42ZengBMCBiotechnology for Biofuels1754-68342018-07-0111111210.1186/s13068-018-1210-0Metabolic engineering of Escherichia coli for production of mixed isoprenoid alcohols and their derivativesBakht Zada0Chonglong Wang1Ji-Bin Park2Seong-Hee Jeong3Ju-Eon Park4Hawaibam Birla Singh5Seon-Won Kim6Division of Applied Life Science (BK21 Plus Program), PMBBRC, Gyeongsang National UniversitySchool of Biology and Basic Medical Sciences, Soochow UniversityDivision of Applied Life Science (BK21 Plus Program), PMBBRC, Gyeongsang National UniversityDivision of Applied Life Science (BK21 Plus Program), PMBBRC, Gyeongsang National UniversityDivision of Applied Life Science (BK21 Plus Program), PMBBRC, Gyeongsang National UniversityDivision of Applied Life Science (BK21 Plus Program), PMBBRC, Gyeongsang National UniversityDivision of Applied Life Science (BK21 Plus Program), PMBBRC, Gyeongsang National UniversityAbstract Background Current petroleum-derived fuels such as gasoline (C5–C12) and diesel (C15–C22) are complex mixtures of hydrocarbons with different chain lengths and chemical structures. Isoprenoids are hydrocarbon-based compounds with different carbon chain lengths and diverse chemical structures, similar to petroleum. Thus, isoprenoid alcohols such as isopentenol (C5), geraniol (C10), and farnesol (C15) have been considered to be ideal biofuel candidates. NudB, a native phosphatase of Escherichia coli, is reported to dephosphorylate isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP) into isopentenol. However, no attention has been paid to its promiscuous activity toward longer chain length (C10–C15) prenyl diphosphates. Results In this study, the promiscuous activity of NudB toward geranyl diphosphate (GPP) and farnesyl diphosphate (FPP) was applied for the production of isoprenoid alcohol mixtures, including isopentenol, geraniol, and farnesol, and their derivatives. E. coli was engineered to produce a mixture of C5 and C15 alcohols by overexpressing NudB (dihydroneopterin triphosphate diphosphohydrolase) and IspA (FPP synthase) along with a heterologous MVA pathway, which resulted in a total of up to 1652 mg/L mixture of C5 and C15 alcohols and their derivatives. The production was further increased to 2027 mg/L by overexpression of another endogenous phosphatase, AphA, in addition to NudB. Production of DMAPP- and FPP-derived alcohols and their derivatives was significantly increased with an increase in the gene dosage of idi, encoding IPP isomerase (IDI), indicating a potential modulation of the composition of the alcohols mixture according to the expression level of IDI. When IspA was replaced with its mutant IspA*, generating GPP in the production strain, a total of 1418 mg/L of the isoprenoid mixture was obtained containing C10 alcohols as a main component. Conclusions The promiscuous activity of NudB was newly identified and successfully used for production of isoprenoid-based alcohol mixtures, which are suitable as next-generation biofuels or commodity chemicals. This is the first successful report on high-titer production of an isoprenoid alcohol-based mixture. The engineering approaches can provide a valuable platform for production of other isoprenoid mixtures via a proportional modulation of IPP, DMAPP, GPP, and FPP syntheses.http://link.springer.com/article/10.1186/s13068-018-1210-0Isoprenoid mixtureIsopentenolGeraniolFarnesolEscherichia coliNudB |
| spellingShingle | Bakht Zada Chonglong Wang Ji-Bin Park Seong-Hee Jeong Ju-Eon Park Hawaibam Birla Singh Seon-Won Kim Metabolic engineering of Escherichia coli for production of mixed isoprenoid alcohols and their derivatives Biotechnology for Biofuels Isoprenoid mixture Isopentenol Geraniol Farnesol Escherichia coli NudB |
| title | Metabolic engineering of Escherichia coli for production of mixed isoprenoid alcohols and their derivatives |
| title_full | Metabolic engineering of Escherichia coli for production of mixed isoprenoid alcohols and their derivatives |
| title_fullStr | Metabolic engineering of Escherichia coli for production of mixed isoprenoid alcohols and their derivatives |
| title_full_unstemmed | Metabolic engineering of Escherichia coli for production of mixed isoprenoid alcohols and their derivatives |
| title_short | Metabolic engineering of Escherichia coli for production of mixed isoprenoid alcohols and their derivatives |
| title_sort | metabolic engineering of escherichia coli for production of mixed isoprenoid alcohols and their derivatives |
| topic | Isoprenoid mixture Isopentenol Geraniol Farnesol Escherichia coli NudB |
| url | http://link.springer.com/article/10.1186/s13068-018-1210-0 |
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