Process of producing monoterpenes

Abstract

The present invention relates to a process of producing a monoterpene and / or derivatives thereof. The process comprises the steps of: a) providing a host microorganism genetically engineered to express a bacterial monoterpene synthase (mTS); and b) contacting geranyl pyrophosphate (GPP) with said bacterial mTS to produce said monoterpene and / or derivatives thereof. The present invention also relates to a microorganism for use in producing a monoterpene and / or derivatives thereof and a recombinant microorganism adapted to conduct the step of converting geranyl pyrophosphate (GPP) into a monoterpene and / or derivatives thereof by expression of a bacterial mTS. It was shown to produce 1,8 cineole using 1,8 cineole synthase and to produce linalool using linalool synthase, both from Streptomyces clavuligerus.

Description

FIELD OF THE INVENTION[0001]The present invention relates to a process of producing monoterpenes. In particular, the present invention relates to a process of producing monoterpenes, for example in a host microorganism, by the action of a bacterial monoterpene synthase.BACKGROUND TO THE INVENTION[0002]Terpenoids (also called isoprenoids) are the most abundant and largest class (>75000) of natural products. Most commonly found in plants, their biological roles are multitude ranging from species to species communication to intracellular signalling and defence against predatory species. They have a wide range of applications and are used in pharmaceuticals, herbicides, flavourings, fragrances and biofuels. Due to the broad commercial interest for terpenoids, efforts to synthesize them by synthetic biology routes have gathered pace in recent years.[0003]Terpenoid substrates are synthesized from the C5 isoprene building blocks, namely dimethylallyl pyrophosphate (DMAPP) and isopenteny...

AI Technical Summary

Benefits of technology

The invention is based on studies that show that using certain bacterial enzymes to produce monoterpenes in Escherichia coli results in high yields and purity of the monoterpenes. This is surprising because previous studies have shown that plant-derived enzymes were not as efficient in producing monoterpenes. The inventors have also discovered that bacterial enzymes are better at converting a specific molecule, called GPP, to monoterpenes. This results in higher yields and purity of monoterpenes compared to using plant enzymes. Therefore, the invention provides a way to improve the yield and purity of monoterpenes in microorganisms.

Problems solved by technology

However, the resulting monoterpene yields are relatively low.

However, the use of such plant mTS has associated disadvantages.

This limited solubility has proved to be a bottleneck in the production of monoterpenes as the majority of the GPP molecules in the host cell are not utilised for the synthesis of monoterpenes, resulting in low product yields / titres.

Lack of robustness also makes plant mTS less attractive targets for protein engineering.

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