28th October 2015Back
The customer used a wild type bacterial dehydrogenase in one of their biosensors for a medical application. The challenge was to produce a recombinant version of their existing enzyme with exactly the same activity specification. The recombinant enzyme was to have a higher specific activity and the production method scalable to accommodate increased order sizes.
Application: The customer used a wild type bacterial dehydrogenase in one of their biosensors for a medical application.
Customer Challenge: “To produce a recombinant version of the existing enzyme with exactly the same activity specification. The recombinant enzyme should have a higher specific activity and the production method must be scalable to accommodate increased order sizes.”
Approach: A codon-optimised version of the gene was synthesised and cloned into a E. coli expression vector (IP free). After transforming into a suitable E. coli host, positive clones were identified and expression of soluble, active enzyme was confirmed. The key parameters for optimal expression of soluble, active enzyme were determined at 1 mL scale using 48-well parallel microfermentations.
The resulting fermentation yielded 20-fold more active, soluble enzyme than the non-induced control.
Standard enzyme characterisation test showed that the recombinant enzyme was indistinguishable from the wild type enzyme. Expression levels were confirmed at 3 L in one of Biocatalysts’ high yielding, scalable E. coli fermentations and acceptability of the recombinant enzyme was confirmed by the customer after application tests.
- Cloning and optimisation of enzyme expression levels – 5 weeks
- Enzyme characterisation – 1 week
- Production of samples – 2 weeks
- Customer application tests – 6 weeks
Benefits to customer
- Cost effective production methods
- Additional profit
- No batch-to-batch variation