Using eXoZymes' tech transfer package, Cayman Chemical advanced the protocol from a one-liter setup to a 100-liter pilot run, operating the reaction, downstream processing, and analysis independently. During the run, the team encountered real-world conditions - including pH variation and precipitation - which often cause enzyme-based processes to fail. Despite this, the reaction continued to perform reliably and delivered strong results, ultimately producing more than 500 grams of pharma-grade N-trans-caffeoyltyramine (NCT) at 99.6% purity, a result that is difficult to obtain in biomanufacturing without extensive purification steps. Additionally, the 99% conversion of feedstock to end product highlights a level of efficiency that is challenging to achieve and maintain in traditional biomanufacturing.

For Cayman Chemical, the outcome stood out based on prior experience as highlighted by Patrick Westcott, Director of Catalog Chemistry Production at Cayman Chemical: “I’ve performed a lot of cell-based enzymatic biocatalysis over the years; it is a critical component of what we do at Cayman. One common aspect of this work is a drop in efficacy during scale-up. Anytime you move up in scale orders of magnitude, a process changes enough that it is considered a different process altogether.”

Westcott, continues, “During the experiment, from the first sample that we took the analysis showed over a 99% conversion rate. There was some concern because I thought that that maybe was a little bit too high. So, we decided to take a second sample to see if it maintained a similar conversion rate. And we were a little surprised to find out that we actually did achieve this 99% conversion in our first 100-fold scale up of their process. That was very much an aha moment for us!”