Discovery of novel geranylgeranyl reductases and characterization of their substrate promiscuity
Abstract Background Geranylgeranyl reductase (GGR) is a flavin-containing redox enzyme that hydrogenates a variety of unactivated polyprenyl substrates, which are further processed mostly for lipid biosynthesis in archaea or chlorophyll biosynthesis in plants. To date, only a few GGR genes have been...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2018-12-01
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| Series: | Biotechnology for Biofuels |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s13068-018-1342-2 |
| _version_ | 1811338020430807040 |
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| author | Corey W. Meadows Florence Mingardon Brett M. Garabedian Edward E. K. Baidoo Veronica T. Benites Andria V. Rodrigues Raya Abourjeily Angelique Chanal Taek Soon Lee |
| author_facet | Corey W. Meadows Florence Mingardon Brett M. Garabedian Edward E. K. Baidoo Veronica T. Benites Andria V. Rodrigues Raya Abourjeily Angelique Chanal Taek Soon Lee |
| author_sort | Corey W. Meadows |
| collection | DOAJ |
| description | Abstract Background Geranylgeranyl reductase (GGR) is a flavin-containing redox enzyme that hydrogenates a variety of unactivated polyprenyl substrates, which are further processed mostly for lipid biosynthesis in archaea or chlorophyll biosynthesis in plants. To date, only a few GGR genes have been confirmed to reduce polyprenyl substrates in vitro or in vivo. Results In this work, we aimed to expand the confirmed GGR activity space by searching for novel genes that function under amenable conditions for microbial mesophilic growth in conventional hosts such as Escherichia coli or Saccharomyces cerevisiae. 31 putative GGRs were selected to test for potential reductase activity in vitro on farnesyl pyrophosphate, geranylgeranyl pyrophosphate, farnesol (FOH), and geranylgeraniol (GGOH). We report the discovery of several novel GGRs exhibiting significant activity toward various polyprenyl substrates under mild conditions (i.e., pH 7.4, T = 37 °C), including the discovery of a novel bacterial GGR isolated from Streptomyces coelicolor. In addition, we uncover new mechanistic insights within several GGR variants, including GGR-mediated phosphatase activity toward polyprenyl pyrophosphates and the first demonstration of completely hydrogenated GGOH and FOH substrates. Conclusion These collective results enhance the potential for metabolic engineers to manufacture a variety of isoprenoid-based biofuels, polymers, and chemical feedstocks in common microbial hosts such as E. coli or S. cerevisiae. |
| first_indexed | 2024-04-13T18:04:40Z |
| format | Article |
| id | doaj.art-8fa81eb620fe4df195291093e38e2f87 |
| institution | Directory Open Access Journal |
| issn | 1754-6834 |
| language | English |
| last_indexed | 2024-04-13T18:04:40Z |
| publishDate | 2018-12-01 |
| publisher | BMC |
| record_format | Article |
| series | Biotechnology for Biofuels |
| spelling | doaj.art-8fa81eb620fe4df195291093e38e2f872022-12-22T02:36:07ZengBMCBiotechnology for Biofuels1754-68342018-12-0111111710.1186/s13068-018-1342-2Discovery of novel geranylgeranyl reductases and characterization of their substrate promiscuityCorey W. Meadows0Florence Mingardon1Brett M. Garabedian2Edward E. K. Baidoo3Veronica T. Benites4Andria V. Rodrigues5Raya Abourjeily6Angelique Chanal7Taek Soon Lee8Joint BioEnergy InstituteTotal Raffinage ChimieJoint BioEnergy InstituteJoint BioEnergy InstituteJoint BioEnergy InstituteJoint BioEnergy InstituteTotal Raffinage ChimieTotal Raffinage ChimieJoint BioEnergy InstituteAbstract Background Geranylgeranyl reductase (GGR) is a flavin-containing redox enzyme that hydrogenates a variety of unactivated polyprenyl substrates, which are further processed mostly for lipid biosynthesis in archaea or chlorophyll biosynthesis in plants. To date, only a few GGR genes have been confirmed to reduce polyprenyl substrates in vitro or in vivo. Results In this work, we aimed to expand the confirmed GGR activity space by searching for novel genes that function under amenable conditions for microbial mesophilic growth in conventional hosts such as Escherichia coli or Saccharomyces cerevisiae. 31 putative GGRs were selected to test for potential reductase activity in vitro on farnesyl pyrophosphate, geranylgeranyl pyrophosphate, farnesol (FOH), and geranylgeraniol (GGOH). We report the discovery of several novel GGRs exhibiting significant activity toward various polyprenyl substrates under mild conditions (i.e., pH 7.4, T = 37 °C), including the discovery of a novel bacterial GGR isolated from Streptomyces coelicolor. In addition, we uncover new mechanistic insights within several GGR variants, including GGR-mediated phosphatase activity toward polyprenyl pyrophosphates and the first demonstration of completely hydrogenated GGOH and FOH substrates. Conclusion These collective results enhance the potential for metabolic engineers to manufacture a variety of isoprenoid-based biofuels, polymers, and chemical feedstocks in common microbial hosts such as E. coli or S. cerevisiae.http://link.springer.com/article/10.1186/s13068-018-1342-2Geranylgeranyl reductaseRedox enzymesIsoprenoidsHydrogenationTerpene biosynthesis |
| spellingShingle | Corey W. Meadows Florence Mingardon Brett M. Garabedian Edward E. K. Baidoo Veronica T. Benites Andria V. Rodrigues Raya Abourjeily Angelique Chanal Taek Soon Lee Discovery of novel geranylgeranyl reductases and characterization of their substrate promiscuity Biotechnology for Biofuels Geranylgeranyl reductase Redox enzymes Isoprenoids Hydrogenation Terpene biosynthesis |
| title | Discovery of novel geranylgeranyl reductases and characterization of their substrate promiscuity |
| title_full | Discovery of novel geranylgeranyl reductases and characterization of their substrate promiscuity |
| title_fullStr | Discovery of novel geranylgeranyl reductases and characterization of their substrate promiscuity |
| title_full_unstemmed | Discovery of novel geranylgeranyl reductases and characterization of their substrate promiscuity |
| title_short | Discovery of novel geranylgeranyl reductases and characterization of their substrate promiscuity |
| title_sort | discovery of novel geranylgeranyl reductases and characterization of their substrate promiscuity |
| topic | Geranylgeranyl reductase Redox enzymes Isoprenoids Hydrogenation Terpene biosynthesis |
| url | http://link.springer.com/article/10.1186/s13068-018-1342-2 |
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