Solving The Enzyme Immobilization Problem

A Deep Dive Into EnginZyme

Hey There 👋

Welcome back to the newsletter’s monthly feature. I hope the weekend was enjoyable. I spent it enjoying the spring weather and writing this article. Before I spoke to Karim, the founder of EnginZyme, I was skeptical of what they were doing, but after our conversation I was really excited. If only I was an accredited investor. Maybe someday.

This issue of The Polymerist is sponsored by:

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Why You Should Care About Immobilized Enzymes

If we want to solve issues around global climate change and secure the future of the planet it will involve a massive undertaking in changing how we produce the materials and services that power modern human life. The heavy industries that produce our steel, chemicals, and machinery all utilize an enormous amount of energy and raw materials.

These industries are starting to do their part in reducing their carbon footprint. The New York Times had a great story on the topic as did the Wall Street Journal and Eastman is betting over a billion dollars on it. If you want to go in and really try to change an industry you might as well join it, become the CEO of a company, and execute on your vision. Ilham Kadri is a good example if I was going to pick a role model.

Or you could be in graduate school, come up with a great idea in a lab, raise money and start a company.

When I wrote The Chemical Company Of The Future and used Solugen as an example the founders have a clear vision on how they will use enzymes and plant based feedstocks to make carbon negative chemicals. Even just writing that sentence feels audacious, “carbon negative chemicals” is the type of bold thinking we need. Even if Solugen cannot get all of their chemicals carbon negative, maybe they can be carbon neutral or at a minimum have a fraction of the carbon footprint of the incumbents. 

For those of you who didn’t read my article on Solugen I recommend that you do it, but later, after you’ve finished reading this one. The summary of it is that Solugen is making enzymes cheaper than they have been before by metabolic engineering microbes via CRISPR and using those enzymes to make traditional chemicals. Enzymes are nature’s catalyst and we can think of a catalyst as the philosopher’s stone, the alchemists’ holy grail, which can turn lead into gold. If you replace lead with biomass or carbon dioxide and gold with a target molecule of interest you have the chemical industry of the future. Solugen is trying to bring the future to us today and the big chunk of money they raised in 2021 will help them with that vision.

Enzymes typically work at room or biological temperature. The enzyme lactase for instance breaks down lactose into glucose and galactose and most humans have this enzyme. Those who do not we consider to be lactose intolerant. Enzymes also help break down polyesters and Corbion for instance is betting big on their enzymes degrading polylactide or PLA (the polymer of lactic acid) and they also happen to be a large global producer of PLA too. Enzymes can also degrade polyethylene terephthalate (PET), bisphenol A (BPA), and they can also be used to synthesize polyesters. Enzymes are also used in the conversion of carbon dioxide to starch, we could literally feed people with waste carbon dioxide. Enzymes are the most promising and exciting catalysts that could be studied right now. 

One of the problems that the Solugen founders and I discussed was one of enzyme immobilization. Traditionally, enzyme production has been expensive and one way to drive down costs is to immobilize or trap those enzymes in a porous support where they can be reused over and over again. The catalytic converter in your car is similar where the platinum and palladium catalysts are fixed and turn carbon monoxide into carbon dioxide and we can think of a typical catalytic converter as a fluidized fixed bed continuous reactor (as long as the engine is running).

Here is a summary of the problem I wrote in late 2021 about enzyme immobilization from my previous article: 

Immobilization is also not that easy and can incur significant costs. The other issue with heterogeneous catalysis can also be one of mass transport where the catalysts need to come into contact with all of the reactants and can lead to longer reaction times.

In our conversation we also touched on enzyme immobilization can lead to lower enzyme reaction efficiency (enzyme activity) due to aggregation on the substrate (e.g. multiple layers) or the active site of the enzyme being unavailable due to how the enzyme immobilized on the support. If I think back to graduate school the two main pathways to enzyme immobilization are to physically immobilize such as just trying to stick them on a porous substrate or chemically bonding them to a substrate utilizing a highly reactive group such as an isocyanate (think brown gorilla glue which is pMDI) or epoxide rings (think unreacted epoxy resin). 

Why isn’t immobilization that easy and why would we want to immobilize? 

I didn’t really dive into that issue in late 2021, but the perfect immobilization strategy for enzymes would be to have them adhere to the surface of a support medium that is stable when in contact with water, heat, and solvents. The enzyme activity when immobilized would be maintained at 100% and there would only be a mono-layer of enzymes on the support—maximum efficiency.

While heterogeneous catalysis does have some drawbacks such as mass transfer effects it does tend to do well in continuous flow reactors for large scale commodity operations. If we think about Solugen’s free enzyme approach it lends itself well to batch reactors, kind of like making a soup and then just filtering out the enzymes after the soup is finished. A large part of the specialty chemicals industry is built on doing large scale batch reactions. There are significant quality control benefits when it comes to batch reactors and traceability potential which is phenomenal when trying to do root cause failure analysis.

A Message From My Sponsor

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Origin’s first plants are taking shape in Sarnia, Ontario, Canada and Geismar, Louisiana, USA. Once operational they’ll start fulfilling the $5.6 billion worth of signed offtake agreements and capacity reservations from companies like PepsiCo, Ford, Nestle Waters, Danone, Mitsubishi Gas Chemical, and Solvay.  They are looking to hire passionate chemical industry professionals who believe in doing something different. If you’re interested in working for or with Origin, head to their website. The top 3 types of positions right now are:

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Commodity Chemicals

If we think about large scale production of commodities that involve single or two step reactions, heterogeneous catalysis is a star player because reactants can go into one side of a continuous reactor and product comes out the other after the required time, temperature, and contact with a catalyst. Here are some examples where heterogeneous catalysis plays a big role:

1. Synthesis of high density polyethylene or polypropylene

2. Synthesis of ammonia via Haber-Bosch process

3. Production of hydrogen from natural gas via steam reforming

4. Catalytic conversion in the exhaust system of a car (e.g. carbon monoxide to carbon dioxide)

Enzymes can’t really do those things (at least not yet), but if we could get perfect immobilization it would help drive costs down, and lower cost catalysts that work without needing to significant energy are a perfect tool for carbon footprint reduction. With CRISPR and perfect immobilization we could have an enzymatic catalysis revolution on our hands in the next few years. Building a better way to enzyme immobilization is a “if you build it, they will come” moment. Rather than emerging from a cornfield I suspect companies would be trying to break down the door to whoever solved the problem first in an effort to get samples.

It just so happens that I spoke to a company who reports to have solved this problem.

EnginZyme

I got a chance to speak with the EnginZyme CEO Karim Engelmark Cassimjee last month. Karim started EnginZyme from a discovery he had in the lab at Stockholm University. Karim was working on enzyme immobilization on glass with specific surface chemistry and when he measured enzyme activity of his immobilization he got 100%. After re-running the experiments eight times he knew he had something worth patenting and the beginnings of a company. This is the second start-up I’ve interviewed based in Sweden (is there something in the food?)

EnginZyme is creating immobilized enzymes that the chemical and food industry can use to make chemical transformations less wasteful and more energy efficient. Karim views the problems of the chemical industry as one of incumbency. The way we have always done things is often with heavy metal catalysts and high temperatures. If we were to swap our heavy metal catalysts with enzymes those high temperatures wouldn’t be needed and we could run chemical reactions at a fraction of the temperatures and at a fraction of the energy. If we use waste biomass or carbon dioxide we can think of chemicals as being carbon negative.

EnginZyme is focused on creating specific surface chemistries on porous glass supports that can do perfect enzyme immobilization. Karim told me the binding is non-destructive and they can typically achieve 30% mass loadings of enzymes on their surface modified glass supports and still maintain a pure monolayer of enzymes. Karim and his team have yet to encounter an enzyme they cannot immobilize and they employ a range of coatings on their porous glass supports from hydrophobic to hydrophilic. This means an immobilized enzyme from EnginZyme is going to be significantly more active than current incumbent products and thus become even more effective catalysts.

In comparison Karim told me that the current incumbent leader in the space, Novozymes, doesn't have a wide range of immobilized enzyme catalysts available. Further, a great enzyme immobilization right now in the industry is a 10% loading with significantly reduced activity whereas EnginZyme sees 10% loading as quite low and they typically acheive enzyme activity anywhere from 50-100%.

Karim told me that EnginZyme is seeking to generate revenue by selling their catalysts, licensing their technology and manufacturing process, and building their own plant so that they can be vertically integrated. Right now, EnginZyme is tolling their enzyme fermentation, but they are developing their own strains that produce their enzymes. I think at least for right now this is a very efficient use of the capital that they do have and as they start to generate more revenue they can use their profits to build out their business. 

Two Use Cases

Karim and his team are already working with Tetra Pak to improve food & beverage production, such as turning food waste into products. Their first project is to explore the potential of making something more useful out of acid whey. If you’ve ever made yogurt you know that you need a lot of milk to make a cup of yogurt and the byproduct there is acid whey. I’m personally a big fan of grass fed Greek yogurt, but my yogurt habit produces a lot of acid whey (not your whey from GNC). Acid whey is a thin, watery substance that can severely damage ecosystems if it makes it to groundwater so you can’t just pour it down the drain. EnginZyme aims to use their immobilized enzyme technology to create something useful out of acid whey like a fiber (Bioextrax is doing something similar with Keratin).

Another example that Karim gave me was in production of a prebiotic disaccharide named Kojibiose, which has a sweet taste, but has significantly less calories and can even produce a delayed glucose response. Karim spoke about immobilizing enzymes that were recently discovered that could produce Kojibiose from readily available starting materials such as sucrose or lactose and yielding 300 grams per liter of the once difficult to produce sugar. Stevia, aspartame, and agave, move over, there is a new sweetener coming soon.

Cost To Produce and Total Addressable Market

When I asked about costs to produce these immobilized enzymes Karim told me it’s about $0.10-1.00 per kilogram. To give you an idea of what large scale industrial polymers cost, prices right now are at a minimum a dollar per pound. The fact that a high value reusable enzyme catalyst can be produced at such low prices is astounding. I don’t have pricing from Novozymes, but a good way to get an estimate of how low cost the EnginZyme product is go to the Millapore Sigma website, type in Novozymes, and then divide all of those prices by about 10 to get an idea of what Millapore Sigma might be paying for them. Granted, not all enzymes are equivalent in terms of costs to produce, but even if EnginZyme can get costs down by 20-50% I suspect we might have a biocatalytic revolution on our hands in the next five years.

Forbes ascribes that there is a $4 trillion dollar gold rush going on right now in synthetic biology, but I believe the real high value is going to be captured by companies that can be paid to take waste and produce high value products such as LanzaTech or companies that can produce significant deflationary value when compared to the current chemical industry. I’m all for fermentation, but if you can show me another option that has high throughput yields at a fraction of a cost the choice is easy.

Karim believes that technology should ultimately be deflationary for making products by a reduction in cost, energy, and waste. Making the stuff we use and consume everyday cheaper, less energy intensive, and with less waste is going to be better for everyone and the planet. You can do well by doing good.

Karim and his team are aiming to raise additional funds sometime this year and in the future and believes that going public would be the best option for EnginZyme. There are so many applications for immobilized enzymes and going public to me sounds like the best option. Karim views his company as a chemical engineering services firm and not a synthetic biology company, which I thought super interesting and reminds me a bit of the model that Johnson Mathey or Axens have been employing. Just because you can make chemicals doesn’t mean that you might be best suited to make chemicals. 

Sometimes providing the tools so that others can realize their own companies is more valuable.

EnginZyme currently employs about 55 people and Karim hopes to add about 30 more people this year. If you are an investor looking to get on the cap table or a customer who wants to trial EnginZyme’s technology you can reach out to Karim at karim@enginzyme.com.

Tony