Jul 15, 2021
By Pae Wu
OzoneBio: Emissions-free chemicals brought to life by zombie cells

I have this feeling, but limited means to confirm it, that deep in the archives of Russian scientific literature lies a treasure trove of valuable new technologies, just waiting to spring forth and solve big problems.

It turns out that secret treasure trove landed on our doorsteps a stone’s throw away in Nova Scotia, Canada.  Khorcheska Bhatyrova and Anna Khusnutdinova are two biochemistry Ph.D.s who are taking on greenhouse gas emissions and the petrochemical industry with their invention, the zombie cell.

OzoneBio is producing an emissions-free, low-cost adipic acid produced from waste feedstocks to make the world’s only bio-nylon 6, 6 that can compete in the open market. No green premiums needed here. The zombie cell technique invented by co-founders Khorcheska Batyrova and Anna Khusnutdinova eliminates the need for costly metal catalysts. This adipic acid is scalable, cost competitive with petroleum-derived products, and can be made from wood-derived aromatics.

How did you come up with the idea for the zombie cells?

Everyone knows that all living organisms consist of enzymes, biological molecules that perform all kinds of chemical reactions inside living cells.  We thought, what if we can make this enzyme work robustly in our target reactions. We were lucky to have great profs in Russia who passed to us their knowledge and skills. Back in Russia we discussed with our profs the idea of what will happen if we make enzymes so stable and active so they would perform like chemical catalysts in the reactions, that is how we invented zombie cells.

Why is it you’re taking on nylon 6, 6? What’s so special about it?

Nylon 6, 6 is used everywhere from car parts to running shoes, it is used also to make your favorite yoga pants.  

Our first product is adipic acid (one of two key building blocks of Nylon 6, 6).  Currently, adipic acid is made only from oil, and it is one of the most toxic production processes in the world.  It leads to at least 60 million tons per year of greenhouse gases.  Especially nitrous oxide, which should scare us all, a gas that is 300 times more toxic than CO2 and stays in the atmosphere for 115 years destroying our only protection from UV radiation — the ozone layer.  That is why we named our company Ozonebio.

Seems limited to only make nylon 6, 6.

Alone, the fact that we already can make the powder of emission-free adipic acid on the bench is super exciting.  Adding in the fact that at scale we can make it cost competitive with the petroleum-derived adipic acid is revolutionary. 

We have made a complex family of enzymes work in our zombie cells.  This makes us a platform company.  That family of enzymes can be used to produce plastic precursors, fragrances or drugs. 

Why is your process so much less expensive than other bio-processes for adipic acid?

Using our zombie cells there is no need to use expensive manipulations to purify enzymes and there is no enzyme degradation.  No need to express helper proteins to ensure your product comes out the cells, [and] we discover[ed] the way to make cells porous just enough.  In addition, zombie cells together with our cheap helper chemical allow us to replace the expensive protein cofactors and Pt/Pd catalysts with hydrogenation facilities

And we have no need to keep cells happy and in a comfortable environment to make them produce your product. 

Keeping cells happy, this is where the capex and opex in biofermentation drives up cost and energy requirements in traditional biomanufacturing?

Yes, exactly.  Plus, we can use challenging / toxic feed stocks that [would otherwise] simply kill alive cells. And our feedstocks are waste products.

Circular and emissions-free and cost-competitive!


It seems like you want to save the world one yoga pant at a time.  Why the urgency?

For over a decade, industry was looking for the replacement of expensive NAD(P)H cofactors and Pt/Pd catalysts using a cheap, robust approach. We could stabilize enzymes inside the cells without degradation and found a replacement for expensive cofactors, that is when I realized it is the time for the fast actions.  Thankfully, IndieBio is the place where things get done super-fast and efficiently.  That awesome team took our company to the completely different level in just 4.5 month! [Editor’s note: your check’s in the mail.]

This is a very difficult problem that many before you have failed to solve in a way that can compete on cost.  What possessed you to go after such a hard problem?

Throughout our careers in academia we used to work on complex enzymes trying to make them work, either adjusting reaction conditions or engineering enzymes. Same happened in this case, it was one of the complex enzymes we were working on, and in that case an uncommon technique worked to solve the problem of this enzyme stability and function.  That’s how we invented the zombie cells approach.  We made one of the most complicated and sensitive families of enzymes in the world to work without any extra manipulations.

How did you meet?

We met in 2007 in the Institute of Basic Biological Problems, in Russia, where we both got our PhD. There we worked on another group of complex enzymes that are responsible for hydrogen production. 

How did you two decide to start this company?

We started our company because I realized that it is time for us to be independent and start our own journey.  Anna and I had become experienced enough to identify the direction of our research and decide how to proceed.

Nova Scotia isn’t the first place that comes to mind for building a start-up, tell us a little bit about your journey there.

In Canada, the pandemic made many [start-up spaces] disappear.  Nova Scotia responded quickly to our request and they had the facility we needed to succeed through the IndieBio program.  We moved here 2 months ago, and during these two months we were able to make our first batch of the first in the world emissions-free adipic acid.  We only regret we did not move here earlier, right away after we were accepted to IndieBio, I think now we would already have Nylon 6, 6 chips made. But we are working on it now!

How did you feel the first time you produced adipic acid using your process?

We felt immediately that there is so much more work we need to do, and we kept going!

Quick word association exercise, what is your first reaction to the following phrase: You can’t do that.

Anna: Why?

Khorcheska: I prefer to decide it on my own, whether I can or can’t do it 😊