Isomerase were approached in 2020 by a biotech company looking to set up a biocatalytic cascade process, which relied on three separate enzyme activities and removal of one background competing activity present in standard Escherichia coli strains.
• Solution: Bioengineering, enzyme expression in E.coli with parallel process development
We inactivated the gene in E.coli expression strains coding for the competing activity, then expressed genes coding for the three enzyme activities both in independent strains and together in one strain (independent or simultaneous expression). A scalable high-density fermentation with optimised downstream processing (DSP), including Tangential Flow Filtration (TFF) was developed and used to generate initial batches of 100s grams of target enzymes to enable initial development of the biocatalytic process.
Further work was then carried out to improve the stability of the enzymes on storage, with a range of stabilising agents trialled and one taken forward. A Technical Transfer (tech transfer) document was generated and used to support transfer of the strains and process to a scaled enzyme manufacturer on behalf of the client.
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