Abstract
BackgroundTerpenes are remarkably diverse natural structures, which can be formed via two different pathways leading to two common intermediates. Among those, sesquiterpenes represent a variety of industrially relevant products. One important industrially produced product is β-farnesene as a precursor for a jet fuel additive. So far, microbial terpene production has been mostly limited to known production hosts, which are only able to grow on heterotrophic substrates.ResultsIn this paper, we for the first time describe β-farnesene production by the versatile bacterial host Cupriavidus necator on fructose, which is known to grow hetero- and autotrophically and even in bioelectrochemical systems. We were able to show a growth-dependent production of β-farnesene by expressing the β-farnesene synthase from Artemisia annua in C. necator H16 PHB-4. Additionally, we performed a scale-up in a parallel reactor system with production titers of 26.3 ± 1.3 µM β-farnesene with a fed-batch process.ConclusionsThe β-farnesene production titers reported in this paper are not in the same range as titers published with known heterotrophic producers E. coli or S. cerevisiae. However, this proof-of-principle study with C. necator as production host opens new synthesis routes toward a sustainable economy and leaves room for further optimizations, which have been already performed with the known production strains.
Highlights
Terpenes are remarkably diverse natural structures, which can be formed via two different pathways leading to two common intermediates
The β-farnesene production titers reported in this paper are not in the same range as titers published with known heterotrophic producers E. coli or S. cerevisiae
This proof-of-principle study with C. necator as production host opens new synthesis routes toward a sustainable economy and leaves room for further optimizations, which have been already performed with the known production strains
Summary
Terpenes are remarkably diverse natural structures, which can be formed via two different pathways leading to two common intermediates. The diversity of terpenes is derived from only two different precursor molecules isopentenyl pyrophosphate (IPP) and dimethylallyl pyrophosphate (DMAPP). The natural ability to provide precursors for terpene production [5] as well as its fast growth with high cell densities [6, 7] makes C. necator an attractive host organism to produce different industrially relevant molecules, e.g. terpenes. From FPP, the addition of a terpene synthase results in a onestep enzymatic transformation to the desired natural sesquiterpene. One of those sesquiterpenes is β-farnesene, a terpene converted from FPP by β-farnesene synthase It has been identified as a precursor for the suitable jet fuel substitute farnesane (2,6,10-trimethyl dodecane) and is of special interest [8].
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