| Autor: |
Orsi E; Bioprocess Engineering, Department of Agrotechnology and Food, Wageningen University and Research, Wageningen, The Netherlands., Folch PL; Bioprocess Engineering, Department of Agrotechnology and Food, Wageningen University and Research, Wageningen, The Netherlands., Monje-López VT; Bioprocess Engineering, Department of Agrotechnology and Food, Wageningen University and Research, Wageningen, The Netherlands., Fernhout BM; Bioprocess Engineering, Department of Agrotechnology and Food, Wageningen University and Research, Wageningen, The Netherlands., Turcato A; Bioprocess Engineering, Department of Agrotechnology and Food, Wageningen University and Research, Wageningen, The Netherlands., Kengen SWM; Laboratory of Microbiology, Department of Agrotechnology and Food, Wageningen University and Research, Wageningen, The Netherlands., Eggink G; Bioprocess Engineering, Department of Agrotechnology and Food, Wageningen University and Research, Wageningen, The Netherlands.; Biobased Products Food and Biobased Research, Wageningen University and Research, Wageningen, The Netherlands., Weusthuis RA; Bioprocess Engineering, Department of Agrotechnology and Food, Wageningen University and Research, Wageningen, The Netherlands. ruud.weusthuis@wur.nl. |
| Abstrakt: |
Rhodobacter sphaeroides is a metabolically versatile bacterium capable of producing terpenes natively. Surprisingly, terpene biosynthesis in this species has always been investigated in complex media, with unknown compounds possibly acting as carbon and nitrogen sources. Here, a defined medium was adapted for R. sphaeroides dark heterotrophic growth, and was used to investigate the conversion of different organic substrates into the reporter terpene amorphadiene. The amorphadiene synthase was cloned in R. sphaeroides, allowing its biosynthesis via the native 2-methyl-D-erythritol-4-phosphate (MEP) pathway and, additionally, via a heterologous mevalonate one. The latter condition increased titers up to eightfold. Consequently, better yields and productivities to previously reported complex media cultivations were achieved. Productivity was further investigated under different cultivation conditions, including nitrogen and oxygen availability. This novel cultivation setup provided useful insight into the understanding of terpene biosynthesis in R. sphaeroides, allowing to better comprehend its dynamics and regulation during chemoheterotrophic cultivation. |
