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

Biocatalysts Ltd是一家位于英国加的夫的中小企业。我们使用微生物发酵技术生产高价值的酶，销售到以下市场应用：(I)用作制药中间体的加工助剂；(Ii)用作食品成分的加工助剂；(Iii)用于生命科学应用中的DNA加工。我们有几个微生物平台，用于生产包含细菌和酵母的重组酶。当我们为新市场开发具有战略意义的酶产品或为客户项目开发新的酶产品时，我们会通过面向制造的设计流程对酶进行勤奋的评估。这是为了最大限度地提高这样的可能性，即如果我们能够在实验室规模制造这种酶，那么我们就能够扩大到大规模的商业可行的发酵和下游过程，以达到成本目标。有些酶比细菌更适合在酵母中表达，我们目前默认的酵母生产平台是In_Pichia Pastoris_。在毕赤酵母中，在学术和商业规模上使用最广泛的表达系统包括使用甲醇作为酶(蛋白质)的高水平表达的诱导剂。在制造背景下大规模使用甲醇充满了经济、健康和安全以及潜在的环境限制。因此，生物催化剂目前仅限于甲醇诱导的毕赤酵母发酵，规模为500升。我们目前正在将我们的制造能力在两年计划内扩大到8,000升。为这样的设施配备能够处理这种发酵规模的甲醇的能力，成本高得令人望而却步。目前我们有3种酶产品在毕加索大规模生产，甲醇诱导生产，因此我们不得不将这项业务出口给欧洲的一家合作伙伴公司。因此，该项目旨在开发一种成本效益高、产量高的无甲醇表达系统，用于在毕赤酵母中进行商业生产规模，以使我们的业务在英国扩大这一领域。“

"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."