Abstract
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.
Highlights
The purple non-sulfur bacterium Rhodobacter sphaeroides is a metabolically versatile microorganism capable of growing, e.g., aerobically and anaerobically, photoautotrophically and heterotrophically [28, 43]
All the data focused on production of coenzyme Q10 (CoQ10, C 59H90O4)
Because of its versatile metabolism, the natural terpenoid producer R. sphaeroides offers an interesting platform for studying chemoheterotrophic production of these compounds and their regulation
Summary
The purple non-sulfur bacterium Rhodobacter sphaeroides is a metabolically versatile microorganism capable of growing, e.g., aerobically and anaerobically, photoautotrophically and heterotrophically [28, 43]. Bioprocess Engineering, Department of Agrotechnology and Food, Wageningen University and Research, Wageningen, The Netherlands. Laboratory of Microbiology, 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 sphaeroides has been explored with respect to photoheterotrophic hydrogen biosynthesis [21] and chemoheterotrophic terpene production [5, 27]. Terpenes form the largest class of natural products with regard to their structural diversity and are synthesized by all organisms [11]. Interest for their application in the bio-economy is increasing. They are used in the pharma and nutraceutical sectors [1, 20, 35, 39, 48], and they show potential for application in the polymer industry [2] and as advanced biofuel [9, 53]
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