Team:Marburg/Production/Outlook

SynDustry Fuse. Produce. Use.

The Production Line

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Outlook

The Syndustry platform is not only a development in basic research, it has also been developed serving as a production platform making Syndustry attractive to the biotechnological industry. While developing the idea of establishing a production line within the cell via artificial endosymbiosis, we wanted the system to work in a modular fashion as well.

We have chosen to produce terpenoid compounds in our system. Terpenoids give the system flexibility and a variety of different products that can be produced, such as medical compounds, high value biofuels, food compounds and bioplastics [1]. The production of perillyl alcohol serves as an interesting example for the variety of possibilities this system has to offer.

Monoterpenoids based on limonene can be produced in Saccharomyces cerevisiae, which is able to transport these products into the medium via the GcABC-G1 transporter. The production of monoterpenoids based on limonene already gives this system modularity since only a few genes in S. cerevisiae have to be exchanged in order to produce a new bioproduct.

The cytochrome P450 could be exchanged to other production complexes. An example would be the production of carvone, a crucial component of spearmint essential oils, which is highly valuable in food and fragrance industries, also having potential medical applications as well as the ability to function in insect repellents [2][3].

Another possibility might be the introduction of other production steps towards the jet fuel premixture AMJ-700t. This would require strictly regulated expression of the production steps as this fuel precursor consists of 50 % limonene, 40 % farnesene and 10 % cymene [4]. The export of limonene and cymene is achieved by the already implemented GcABC-G1 efflux pump. Farnesene can be exported by the heterologous expression of the MexF efflux pump from Pseudomonas putida [4]. This would grant the ability to export the mixture directly into the media, which makes harvesting and the conversion into the final jet fuel AMJ-700 simpler and more economical.

Two other possible products for this system are the well known taxol and artemisinin emphasizing the tremendous scope of possible terpenoid based compounds that could be produced.

If this system would be used for other more complex compounds a modification would be necessary. Due to the fact that terpenoids tend to be lipophilic it is necessary to increase the organism's tolerance towards this chemical class. This could be achieved by introducing the efflux pumps as well as point mutations that would on the one hand increase the lifespan of the system and on the other hand increasing the bioproduct's yield generated via this system.

Further possibilities could be granted by the production of different chemicals, like fatty acid derived biofuels, however in those cases the production line would require drastic changes in efflux pumps and resistances.

Literature

  1. [1] Jongedijk, Esmer, et al. "Biotechnological production of limonene in microorganisms." Applied microbiology and biotechnology 100.7 (2016): 2927-2938.
  2. [2] de Carvalho, Carla CCR, and M. Manuela R. da Fonseca. "Carvone: Why and how should one bother to produce this terpene." Food Chemistry 95.3 (2006): 413-422.
  3. [3] Lange, Bernd Markus. "Biosynthesis and biotechnology of high-value p-menthane monoterpenes, including menthol, carvone, and limonene."Biotechnology of Isoprenoids. Springer International Publishing, 2015. 319-353.
  4. [4] Brennan, Timothy CR, et al. "Evolutionary engineering improves tolerance for replacement jet fuels in Saccharomyces cerevisiae." Applied and environmental microbiology 81.10 (2015): 3316-3325.