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We already funded many projects that have been highly successful. Please click on the links below to read our case studies.
A major challenge faced by enzyme manufacturers is to produce commercial yields of a wide variety of enzymes in recombinant production strains. Secretion of recombinant enzymes from a production strain is of particular interest for commercial production of enzymes, as it facilitates the downstream processing and product formulation.
It is a challenge to produce a wide variety of different recombinant proteins in any one production strain (e.g. E. coli, B. subtilis, P. pastoris), with a current success rate of these platforms for the production of a commercially viable product of the order of 15-20%.
B. subtilis is an attractive host for recombinant enzyme production since it is a GRAS and QPS host, thereby facilitating regulatory requirements for enzymes for food or pharmaceutical applications. It is also a strong candidate as an organism for protein production since it is known to produce a wide range of recombinant enzymes. It also has the advantage of being able to secrete proteins to the culture medium.
Although there is considerable precedent available in the literature for working with recombinant expression in Bacillus systems, the practical details for performing this work effectively reside typically with academic scientists with the knowledge and skills borne from experience.
The production of the commodity chemical butanol using the acetone-butanol-ethanol (ABE) fermentation of Clostridium species is an important route to producing sustainable lignocellulosic chemicals and biofuels. n-Butanol is a building block chemical in the $115 billion global paints, coatings, adhesives and inks market.
This process, which is the basis for much of the manufacturing by Green Biologics Limited (GBL), would be improved by operating at higher butanol concentrations. Currently, butanol is toxic to the producing bacteria at relatively low concentrations.
For the related fuel ethanol, changes in expression of efflux proteins and alterations in membrane composition have been observed as a result of experimental evolution of strains with increased resistance to this alcohol.
There is a strong commercial interest in biosurfactants – both for sustainability and performance. Mannosylerythritol lipids (MEL) are one example considered to have desirable properties and are of current interest but with commercially limiting yield. They are synthesised from long-chain fatty acids by a number of basidiomycetous yeasts such as Pseudozyma aphidis. An existing process needed improvement, and (as is often the case) it was hypothesised by the industrial partner Croda that much of the flux control lay in transporter reactions involving the uptake of the fatty acid substrates and the efflux of the MEL. Since a genome sequence of P. aphidis (and related species) is available, a bioinformatics approach to seek the transporters based on homology searching and modelling was adopted for this short project.
Solventogenic Clostridia are used by Green Biologics to generate n-butanol from a variety of feed-stocks providing sugars for fermentation. However, n-butanol is expensive to purify from the fermentation broth. The cost of in-situ solvent removal is greatly decreased by fermenting at higher concentrations of n-butanol. The transporter (if any) for export of n-butanol from cells is unknown – it is possible that n-butanol can diffuse across membranes. n-Butanol is also toxic to Clostridia although the mechanism remains largely uncharacterised. Two possibilities are membrane disruption or deleterious effects on membrane proteins. It is possible that by altering the membrane composition or modifying transport activity we will be able to increase the extracellular n-butanol concentration and reduce production costs.