Metabolic engineering of the moss Physcomitrella patens to produce the sesquiterpenoids patchoulol and α/β-santalene

Abstract

The moss Physcomitrella patens, has been genetically engineered to produce patchoulol and β-santalene, two valuable sesquiterpenoid ingredients in the fragrance industry. The highest yield of patchoulol achieved was 1.34 mg/g dry weight. This was achieved by non-targeted transformation of the patchoulol synthase and either a yeast or P. patens HMGR gene under the control of a 35S promoter. Santalene synthase targeted to the plastids yielded 0.039 mg/g dry weight of α/β santalene; cytosolic santalene synthase and 35S controlled HMGR afforded 0.022 mg/g dry weight. It has been observed that the final yield of the fragrance molecules is dependent on the expression of the synthase. This is the first report of heterologous production of sesquiterpenes in moss and it opens up a promising source for light-driven production of valuable fragrance ingredients.

Efficient phototrophic production of a high-value sesquiterpenoid from the eukaryotic microalga Chlamydomonas reinhardtii

“…However, a similar study by the same research group recorded biomass of ~1 g CDW L -1 in 14 d for the moss cultivated under similar conditions (Pan et al, 2015 (Zhan et al, 2014). However, our analyses were conducted in 400 mL scale-up compared to shake flask cultivations of the moss.…”

“…However, heterologous terpenoid production from photosynthetic cyanobacteria have rarely been reported over a milligram per litre (Lai and Lan, 2015), with the sole exception of β-phellandrene production in Synechocystis described above in Section 3.4 (Formighieri and Melis, 2015). Three previous studies have used photosynthetic microbial systems specifically for sesquiterpenoid production: the moss Physcomitrella patens (Zhan et al, 2014), the cyanobacteria Synechococcus PCC 7002 (Davies et al, 2014), as well as Anabaena sp. PCC 7120 (Halfmann et al, 2014).…”

“…Some photosynthetic hosts such as cyanobacteria and eukaryotic moss tissue culture have already been demonstrated to be amenable to heterologous terpenoid expression (Anterola et al, 2009;Davies et al, 2014;Halfmann et al, 2014;Zhan et al, 2014). Eukaryotic microalgae as photosynthetic production platforms, however, are notoriously difficult to genetically engineer and there are limited reports on synthetic metabolic modifications of these organisms (Cordero et al, 2011;Kaye et al, 2015).…”

Optimization of Biochemical Screening Methods for Volatile and Unstable Sesquiterpenoids Using HS-SPME-GC-MS

“…Head Space-Solid Phase Micro Extraction (HS-SPME) fibers are used in many areas of volatility studies and are an obvious choice for fast screening of volatile sesquiterpenoids on GC-MS [5,[14][15][16][17]. Continued studies have led to a variety of different fiber coating materials, and combinations thereof, which are optimal for different kinds of volatiles [18,19].…”

Cytochromes P450 for Terpene Functionalisation and Metabolic Engineering

“…The moss Physcomitrella patens represents a plant system uniquely suited to highvalue terpenoid production due to a natural tolerance to accumulation of terpenoids, a simple endogenous terpenoid profile, and established genetic tools for genome editing by homologous recombination [6,133]. In contrast to higher plant species, P. patens lacks gibberellin diterpene phytohormones but accumulates ent-kaurene intermediates of the gibberellin pathway.…”