Discovery of novel geranylgeranyl reductases and characterization of their substrate promiscuity
| Autor: | Raya Abourjeily, Veronica T. Benites, Edward E. K. Baidoo, Florence Mingardon, Corey W. Meadows, Angélique Chanal, Taek Soon Lee, Andria V. Rodrigues, Brett Garabedian |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Geranylgeranyl pyrophosphate lcsh:Biotechnology 030106 microbiology Saccharomyces cerevisiae Farnesyl pyrophosphate Management Monitoring Policy and Law Applied Microbiology and Biotechnology lcsh:Fuel Industrial Biotechnology 03 medical and health sciences chemistry.chemical_compound lcsh:TP315-360 Geranylgeraniol lcsh:TP248.13-248.65 Lipid biosynthesis biology Geranylgeranyl reductase Renewable Energy Sustainability and the Environment Research Streptomyces coelicolor Farnesol Redox enzymes Chemical Engineering biology.organism_classification Isoprenoids Terpenoid Terpene biosynthesis 030104 developmental biology General Energy chemistry Biochemistry Hydrogenation Biotechnology |
| Zdroj: | Biotechnology for Biofuels Biotechnology for biofuels, vol 11, iss 1 Biotechnology for Biofuels, Vol 11, Iss 1, Pp 1-17 (2018) |
| ISSN: | 1754-6834 |
| Popis: | 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. Electronic supplementary material The online version of this article (10.1186/s13068-018-1342-2) contains supplementary material, which is available to authorized users. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full text from SpringerLink