Zymvol’s team designed and implemented a novel enzymatic synthesis featuring an integrated co-factor regeneration system, allowing continuous in situ recycling of the cofactor rather than one-time use.
Pharmaceutical
Breaking IP barriers in API production
The Challenge
Navigating a Crowded Patent Landscape
Freedom-to-operate (FTO) can often be a hurdle in biocatalytic production. An API manufacturer approached Zymvol with a critical bottleneck: they required a novel R-selective transaminase for a key project, but the existing landscape was heavily restricted by over 10 different patents.
Without a novel, non-infringing enzymatic solution, the manufacturer faced the prospect of shutting down the project entirely.
Structural model highlighting mutation positions in the target transaminase.
Image of the engineered transaminase showing the location of introduced mutations (dark blue) in relation to the PLP cofactor (yellow).
The Solution
To overcome these IP barriers in biocatalysis, Zymvol implemented a two-phase computational approach combining Enzyme Discovery and Enzyme Optimization:
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- Enzyme Discovery
We began by computationally screening over 400,000 sequences, from which we managed to narrow down a selection of 16 top performers for laboratory validation. Out of the 16 sent to the lab, 6 enzyme hits were discovered. - Enzyme Optimization
Upon discovering 6 active hits from the initial 16, we set out to bridge the performance gap between the natural hits and the industrial biocatalysis requirements. Our team conducted 3 enzyme engineering rounds, testing only 300 mutants in the lab, a fraction of the effort required by traditional directed evolution methods.
- Enzyme Discovery
Performance summary of top enzyme variants.
Comparison of wild-type and optimized mutants over 3 rounds of engineering. Under high substrate load (300 mM), initial optimization rounds (1–2) successfully pushed conversion from 10% to 34%. Engineering priorities shifted in Round 3 to center on improving stereoselectivity to 99.8% e.e. (R).
Results & Impact
- Outperforming commercial standards
The enzyme engineering campaign yielded a lead candidate that significantly surpassed existing benchmarks. Experimental validation confirmed that Zymvol’s top hit delivered 10x higher activity than the leading patented enzyme.
- Superior enantiomeric purity
Beyond activity, the precision of the engineered enzyme was significantly higher. While the leading patented enzyme achieved only 70% enantiomeric excess (e.e.), Zymvol’s best hit reached 99% e.e., meeting the stringent chiral purity standards required for high-quality API production. - Revitalizing “dead-end” projects
By delivering a high-performing, novel enzyme in just 3 rounds of engineering, Zymvol enabled the client to bypass 10+ IP barriers and recover a project that was previously headed for cancellation.
Performance Highlights10x
Activity
Our engineered hit outperformed the best patented enzyme available.
99%
Enantiomeric excess
Achieved near-perfect stereoselectivity, compared to only 70% from existing patented solutions.
3
Engineering rounds
A highly accelerated timeline, going from 400k sequences to a final optimized lead after testing only 300 mutants.
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