Cutting-edge Science

R&D in our DNA

Evolva was founded in 2004. Since then it has invested over USD $170 million in R&D. As of year-end 2014, we employed 116 people in R&D, scale-up and production management, and operate some 4,500-square metres of state-of-the-art laboratories. Some of our key technologies include:

Combinatorial genetics

Science_CuttingEdgeScience - Eve in a modern lab explaining the fermentation process

Evolva can create billions of different yeast cells expressing multiple new gene combinations. We have an array of technologies that allow us to rapidly insert and express tens to hundreds of genes in billions of individual yeast cells in a highly combinatorial fashion. This allows us to find those combinations that are necessary to make (biosynthesise) a given ingredient in the most efficient way.

The same approach can create novel pathways that generate diverse small molecules for drug discovery. The genes that we use are either sourced from various species (in compliance with the CBD) or constructed de novo based on online databases or other sequence data.

Screening and analytical technologies

Evolva has an array of advanced screening tools that can select those yeast cells which produce desirable ingredients from a background of a large number of cells. We have both function-led and structure-led screening tools that allow us to rapidly identify which yeasts are making desired ingredients and/or which have acquired desired functions (as a result of making certain ingredients).

Function-led screens are very high throughput and we have used such screens to discover novel molecules with potential utility against cancer and infectious diseases. This approach can also be used to find new functionalities for food ingredients. ­ Structure-led screens use state-of-the art capabilities that combine ultra-high performance liquid chromatography, time-of-flight mass spectroscopy, NMR and informatics. They are primarily used to elaborate production pathways for known ingredients.

Pathway optimisation technologies

Evolva has a number of tools that can improve the efficiency with which yeast produces the desired product, which results in an efficient and responsible use of resources. Once a biosynthetic route has been established, we need to improve it with respect to purity of product, efficiency of feedstock conversion, speed of production and final titre to allow a competitive production of the ingredient.

Evolva has the ability to rapidly assemble multi-step biosynthetic pathways in the yeast genome, allowing us to create the production pathways and optimise gene combinations in a one-step process. We also optimise pathways using a combination of enzyme co-factor balancing, metabolic engineering and pathway flux analysis.

We make our products using yeast (normally bakers yeast), which we then give some extra genes, so that it can do what the original plant or animal does.  Our Stevia product for example is 10 enzymatic steps away from a native yeast metabolite, so we have to put in at least one gene for each of these steps.  These genes are typically ”inspired by” the genes that various plants or animals use for each of the different steps – for example Stevia rebaudiana is by no means the only plant that makes stevia sweeteners and is certainly not the best plant for each step.  We say ”inspired by” rather than ”derived from” since the final task is always to craft a gene that works well in yeast – as just one small example since yeast and plants have different preferences on codon usage the genes should be adapted so the yeast can “read” them well efficiently.

Finally, we have technologies to ensure that a given ingredient will be excreted from the yeast cell to the broth, ensuring a high level of production and ease of product recovery.

Toolbox technologies

Evolva has proprietary technologies that allow us to enhance the properties of ingredients, as well as their economics.  Three particular areas of focus are:

Transporters – By ensuring our ingredients end up in the broth outside the yeast, rather than inside it, we can make more of the ingredient, we can purify it more easily, and we make the regulatory process simpler.   Some ingredients just diffuse out of the yeast cell, but others have to be actively pumped out using transporter proteins.  We have built a significant library of such pumps, so that we can make sure that our yeasts pump out what we want to come out (the final ingredient), and not what we do not want (intermediates that the yeast turns into the final product).

Glycosylation – The process of attaching glucose or other sugars to molecules. Glycosylation allows us to make ingredients (such as stevia and saffron) whose natural properties depend on their glycosylation patterns.  It also allows us to improve the bio-availability of certain ingredients. Finally we can use glycosylation to increase the amount we make of a given ingredient, and to simplify its purification.

Cytochrome P450 enzymes – These enzymes transfer molecular oxygen to -CH, -NH or -SH bonds. Reactions can be very diverse, e.g. hydroxylations, epoxidations, N-, O- and S-dealkylations, deaminations, desulphurations and so on. In particular, we use them to ensure that our ingredient-production pathways are as effective as possible (P450 enzymes are part of most plant metabolite pathways) and to “activate” molecules containing only carbon and hydrogen, in order to diversify basic scaffolds for functionality testing.

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