Isomerase were approached in 2018 by a biotech company looking to set up a biocatalytic process, based on literature evidence of a potential enzyme activity in a number of filamentous fungi.


• Solution: bioinformatics-led heterologous expression in Pichia pastoris


We sourced the strains cited in the literature from both the Isomerase and public strain collections and set up analytical, fermentation and biotransformation protocols. This enabled us to confirm the presence of relevant biocatalytic activities in each of the strains. The fungal strains produced a complex mixture of products due to multiple related enzyme activities. Therefore, we genome sequenced a selection of strains and analysed the assembled genomes to identify the genes coding for the enzymes carrying out the specific biotransformation the client required.


A shortlist of candidate fungal genes were identified and each of these was generated by DNA synthesis and expressed in Pichia pastoris. Each of the Pichia strains was assessed for the required enzyme activity and one was identified which carried out the correct biotransformation. This lead candidate was taken forward for development and scale-up.

 

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fungal enzyme id and characterisation

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