interview
The story of eXoZymes: Interview with Tyler Korman and Paul Opgenorth
1,212 views
Interviews with eXoZymes co-founders, Tyler Korman, PhD - and - Paul Opgenorth, PhD, sharing how the basic idea - liberating the enzymes from the confines of working in cells - was hatched in Jim Bowie's lab at UCLA, before being spun out as Invizyne Technologies in 2019 (rebranded to eXoZymes in 2025), including some of the projects and challenges along the way.
View transcript
There was an aha moment where Jim came out of his office one day and said, let's just get rid of the cells because this all doesn't work all that great. And, you know, we were probably just naive enough and talented enough to say, okay, well, let's do it. And the rest is history. And then it was up to me and Tyler to actually design these systems, how all of this concert of enzymes and co-factors producing and consuming and consuming these co-factors would actually work cell-free. The things that I was doing on the enzymology side and the enzyme engineering, Paul could then take and put them together and make it work. Or figure out why it was working because that happened as well. I had worked engineering cells for years before I went cell-free. And I was aware of the teams that it would take to get proof of concepts. And just moving cell-free, we were able to rapidly design and prototype these systems in one-tenth the time and one-tenth the manpower that it took to get a similar result in cells. So it was actually taking it from a concept of, hey, what if we got rid of the complexities of life to actually thinking about it in terms of a chemical reaction, a chain of chemical reactions that are working continuously in the same pot. And how do you actually make that into a self-sustaining chain reaction? We stumbled upon a system that worked very, very well. To the point that it worked so well, we thought it had failed initially. Because usually when you're working with enzymes, when things precipitate and crash out, that's usually a bad sign. The first time we set the system up, we actually thought that our enzymes were precipitating, but it actually turned out it was the bioplastic. So it was kind of a nice surprise when we ran the GC and it turned out we were making bioplastic. Isobutanol has a similar story. We made so much of that that it also crashed out enzymes and it smelled of solvent. So isobutanol is an interesting compound because it's audacious. Biofuels in and of themselves, just the scale and the magnitude, and that's something that we understand. But it also demonstrates the power of cell-free and these other technologies because if you can make a ton of it, because you have to make a ton of it, then it shows that cell-free can be a viable platform for almost anything. The production of isobutanol is actually really complicated in cells because it itself is toxic. So at a certain level of production, you're actually going to kill your organisms. What we've done is we've gotten rid of the organisms. So all we have to do is worry about, can we produce isobutanol with our enzymes? Enzymes, while they are very complex, they're actually known entities and you can engineer them pretty efficiently for stability. And I think marching up this landscape towards enzyme stability is going to make our enzymatic pathways work that much better. Also, integrating the DSP in the whole enzymatic process is that next step that we are taking to commercialize isobutanol. The work started under a Department of Energy grant on the cell-free front to convert sugars into something useful. Isobutanol was a good energy molecule. And the first system we demonstrated worked well, worked the same as what could be done in cells, but it also suggested that there was significant headroom beyond what you could do in cells. And so we spent a couple years, Paul and I, developing and improving the system to a point where we are now orders of magnitude better than what you could get in cells. And we continue to push the limits here at Invisign even further, both on, not just on the system and enzyme engineering, but in the reaction engineering, isolation, all the aspects of what you would need for something to be now commercial. So the cannabinoid project is interesting because that actually spun out from a project based on terpene production. Um, we had a pathway that was like 36 enzymes that converted glucose into terpenes. Terpenes are actually very interesting because they're very toxic to cells, which is how we got into it. But we thought about how can we actually make a more value added product. And that turned out that the penultimate step in the production of terpenes is the production of this very high value compound GPP that we, we decided that if we could append that to different small molecules, we would have a higher value compound. So one of those compounds that we settled on was, um, CBG. From start to finish, I think the design to testing to demonstrating it worked took less than three months. Um, and so once we showed we could make cannabinoids, then we, expanded that system to, not just one cannabinoid, but multiple. and then since here at Invizyne, we've expanded that even further to, to really completely, expand the scope of things that you can make. Some of the challenges with, with Invisign, as you can maybe imagine, taking all these enzymes outside of the cell, you still have to produce them, right? So getting industrial level production of enzymes and cofactors is something that you just don't have to think about when you're in cells. Um, so that's another level of logistics and complication. But once you solve that, it's so much easier to actually do the production without the complications of life, right? And that is part of this engineering solution that we've kind of made here in order to show that a quick proof of concept, but then also taking that proof of concept, to industrial levels of production. A lot of credit needs to be given to Jim for having this idea of let's go cell free. And once he had that idea, we were off to the races where we trying to figure out what systems to put together cell free and how we would do it. And, you know, over the course of 12 to 18 months, Paul and I tested a lot of different systems, to really focus on how you make cell free function. really well, not just one or two steps, but multiple steps and how you do that as fast as possible. We raised some money spun out 2019. It was me and Paul and eventually John in a, you know, small little space and we've grown to what it is today. So yeah, it's been great.