Modified recombinant yeast manufacturing process for efficient commercial large-scale production of enzymes.

"Biocatalysts Ltd are an SME located in Cardiff UK. Using microbial fermentation technology, we manufacture high-value enzymes that are sold into the following market applications: (i) used as processing aids in the manufacture of pharmaceutical intermediates (ii) used as processing aids in the manufacture of food ingredients (iii) used for DNA processing in Life Science applications.

We have several microbial platforms that we use to manufacture recombinant enzymes incorporating both bacteria and yeasts. When we develop a strategic enzyme product for a new market or a novel enzyme product for a customer project we perform a diligent assessment of the enzyme with our Design for Manufacture process. This is aimed at maximising the probability that if we can make the enzyme at laboratory scale then we will be able to scale-up to large scale commercially viable fermentations and downstream processes to meet cost targets. Some enzymes are more suitable for expression in yeast than bacteria and our current default yeast manufacturing platform is in _Pichia pastoris_. The most widely used expression system in _Pichia_ at both the academic and commercial scale involves the use of Methanol as an inducer for the high-level expression of the enzyme (protein). Large-scale use of methanol in a manufacturing context is fraught with economic, health and safety and potential environmental constraints. Consequently Biocatalysts is currently limited to methanol-induced Pichia fermentations to a scale of 500 L. We are currently expanding our manufacturing capacity over a 2 year plan to 8,000 L. It is prohibitively expensive to equip such a facility with the capacity to work with methanol at this fermentation scale. Currently we have 3 enzyme products produced at large-scale in _Pichia_ with methanol induction, so consequently we are obliged to export this business to a partner company in Europe. Therefore, this project is aimed at developing a cost effective, high yielding methanol-free expression system for commercial manufacturing scale in the yeast _Pichia pastoris_ to allow our business to expand in this area in the UK."