Sam Lee, The History, Future and Workings of Synthetic Biology

Flavio00:01

Yes hello and welcome to another exciting episode of bio 2040, where we speak with thought leaders and experienced researchers, entrepreneurs, investors, to uncover the biggest opportunities in the biotech space. And I'm very excited today to have Sam Lee with me, Sam Lee is a very experienced veteran of the chemistry, pharma, health tech, biotech space, and I'm very excited to have you here, Sam. Good day. Good day.

Sam00:31

Pleasure to be here.

Flavio00:33

Awesome. Sam why don't we start off by you giving us a little bit of your background experiences what you've been doing in your career.

Sam00:42

Sure. I'll start from the beginning.

I'm a Canadian. I got my PhD in chemistry, polymer chemistry from the university of Toronto. And my first job was as a research scientist at DuPont. So that was where I started in the division of. Packaging and industrial polymers. So anything that's plastic. And I did that for three years and then I had the pleasure or opportunity to move into the business development side.

And when I did that, I saw much more of the company. Specifically the life science division, which they call the life sciences enterprise that had DuPont pharma, as well as something they called bio based materials. They started to make through synthetic biology, which it wasn't called that at the time fermentation to make a bio based chemical.

That ended up being used to make some fibers. So I found that very interesting. And as a result of that, I quit the company and inside to get into life sciences. The first job I found in that direction was actually in a biopharmaceutical company. It was called NPS pharmaceuticals, which is based in the U S and Toronto.

Which was a company that ended up developing two medicines that are on the market today. Then I went and worked with a cardiologist to start help start his biopharmaceutical company with a medicine he invented for cardiovascular. Did that for a bit. And then I joined a generic pharmaceutical company.

This is the generic pharmaceutical companies are companies that make medicines after they expire, the patents have expired. And this is called Dr. Reddy's Laboratories is one of the largest ones in the world that was based in India, but I spent the next eight years working out of their us office

so it was really quite an interesting range of experiences. And in that process, I just before I ended up back up in Canada, I worked with I became the founding partner with a venture firm called SOSV. That set up an accelerator for life science companies in New York called Indie bio New York.

I spent a little bit of time just before the pandemic started to get that going because it was investing in life science, startups particularly where synthetic biology was a field that they were interested in. So I've had an interesting career as I'm back in Toronto, right now. I'm actually working for BioNTech, which is the company that invented the first COVID m RNA vaccine.

And and that's where I am. But I have the pleasure of meeting you Flavio and continuing this conversation and my interest in synthetic.

Yeah, thanks. Thanks for the introduction. I think at this point, everybody knows, BioNTech which produce these wonderful vaccines that have prevented probably millions of deaths at this point.

Flavio03:31

So that's a, that's an exciting point. We could probably spend an entire episode just talking about that. And then also Indie bio I've been familiar with, it's a very. Sort of incubator. I was more familiar with the one in San Francisco, I think maybe that was the first one. And so you've really had a very interesting and illustrious career across the whole spectrum.

I think that's also why our conversation today is going to be really interesting because you can draw from all these different aspects and experiences. So I think maybe for people to get situated. Would it make sense to a little bit talk about, today we have this field called synthetic biology, but and as you said in your introduction, it wasn't even called like that a while ago.

So does it make sense for us to go and give somewhat of a history? Where did it start and where are we today?

Sam04:20

Sure. So what, where I, my experience, I did not include this when I was giving my introduction is that I did have the opportunity 10 years ago, back in 2010 to 2011 to meet the CEO of a pharmaceutical company. And he contracted me to do some work for him and his family office to look for investments for him. And I was so grateful for that opportunity. He must've seen in me my varied background that I could put a lot of things together, my understanding and experience in life sciences, but also grounded in my original background at DuPont. And so this was to look at things in sustainability and energy. And this was to look for startups that he could invest in or new technologies that him and his family office could invest in. This was just a side job. I had a regular day job. This was something I could do at nights on weekends.

So of course, when you get an opportunity like that you do it. And we started looking at solar energy, for example, at Greentech of various types. And then he came to me and said, why don't you look into synthetic biology? And this was 2011. And I thought, what does that mean? And that's where I started to learn about this space and investigate it.

I did that by looking at high-impact journals. And I also did that because of we're looking at startups. I started looking at patents and through that search, that's where I started to see the first iteration or first-generation of companies, which I felt were what one could call synthetic biology companies.

So just to be clear it's a big space. It certainly is now, but even back then, you could define it by, by just about many different criteria. We I broadly categorized it in two broad buckets. One of them would be therapeutics and the other would be non therapeutics. And most of the real-world applications as to where one could go, one could arguably say that there were products in that space in a therapeutic area already.

So I looked at the non therapeutic areas, which is a huge space, but very quickly I narrowed down on a number of companies that were. Being funded at the time. Which we're getting some quite far along with various products and it turned out that these were they would call them building block chemicals or petrochemicals.

And that's when I stumbled upon this connection that these were products that they were developing that were connected to products that otherwise could be made from crude oil. And so that was, it appeared to be the first idea that drove the first generation of companies to, to make products as substitutes or bio based substitutes to petrochemicals.

Flavio07:16

So was that at the time was the idea there to fight climate change was more of a cost driver? We could make a cheaper what were what was the idea and that first sort of wave of synthetic biology in the sustainability area?

Sam07:31

It's a good question. And that's when it at that point in my life and my career, I would have that moment where, when I look back, it all makes sense and it all comes back together because by then I had already left DuPont many years ago.

And at the time I was at DuPont and in business development, there was a big initiative in the DuPont life sciences enterprise. They called it bio-based materials. And for all the scientists who were in the traditional chemistry area we felt, I felt left out because all of the spotlight and career opportunities were going to these biologists and what they were doing was taking fermentation to make a certain compound.

It was called one, 1,3-Propanediol is basically for the organic chemistry. So propane with two alcoholic groups on either end. And that would be the feedstock for polyester. And as I learned the business premise as to why they wanted to do this well, polyester has been around for a long time.

And when in the early days when polyester was first developed there, DuPont had figured out a whole range of different types of polyesters. You could make one of the best polyesters that they could make was actually based on you. This one, three propane dial, the reason they never chose that.

And they ended up using the ethane version of it. The two carbon one is because that was the one that was readily available from crude oil as a feedstock that was much cheaper. And now they found that in order to provide a competitive product let's make 1,3-Propanediol.

We have a way to do that through fermentation and we could make a new polyester fiber that would be much more elastic, much more durable, much stronger and so on. And that was what drove it. So basically the story other than that, you had a performing fabric was that the cost point became accessible because of synthetic biology, which wasn't called synthetic biology at the time, it was just basically a fermentation technology that they through engineering E. Coli was what they started with.

So that was the connection. And then that was back in 1999, 2000 when that product was being developed and as it is on the market in 2011, I began to see that there was a whole bunch of other products made by other companies with that same premise, that there were a lot of products that the, these would be petrochemicals chemicals that are 2, 3, 4 carbons in size that would be produced at petrochemical refineries.

And they would be the feedstock to go into. Other materials downstream, mostly plastic but also a lot of other materials and this first generation of bio based materials that were coming out, these synthetic synthetic biology companies were basically making that as a route alternate as an alternative from petrochemical.

So part of the storyline was to say, this is a sustainable material because they come from. Fermentation, which is basically from sugar rather than petrochemicals.

Flavio10:42

So in this scenario, we were essentially harvesting some kind of sugar. That we have was grown in some field maybe sugar cane or some other source of carbon which then the net CO2 would be zero.

Versus if we, obviously, if we take a petrochemicals, these products are then produced consumed and ultimately oftentimes either end up in waste types or burned, which then adds. CO2 to the atmosphere. Is that the correct way to think about it?

Sam11:12

That's how they expressed it at that they, there was an environmental element to this.

But the interesting thing was here's where the price of oil comes in. And this was, remember, this is thinking that's 10 years, 10 years old. Yeah. So where it is today and the, in the storyline and the business case are different, but it's good to know the past. So this is 2011.

We just had the global financial crisis in 2008. And there was a little, if you just look at the price of oil between through the 1980s, 1990s, it's been relatively flat in us dollars. Let's say about 40, $50. And then it started to go up and 60, 70, 80, 90 over a hundred dollars a barrel.

We had the global financial crisis. There was a little blip, went down a bit, but it went back up. So it was at an all time high. And so oil as a feed stock into petrochemical refiners is very high. Oil itself was the burning of it is contributing to carbon dioxide. And so both the price as well as the environmental premise was something that.

And the first-generation in thinking about this problem. Those were undesirable elements and what's going on then is okay. You've got a petrochemical refinery that's being the source of these building block chemicals that you need to. What if we actually use the bio-based source?

So instead of using crude oil, let's just say it turns out to be corn which provides the glucose to do fermentation to make a bunch of materials. It could be Propanediol. It could be succinic acid. It could be isobutanol, butanediol. So these were the types of materials that those first generation of companies made.

They also the thought of making a biofuel. Because that's pretty there's a broad range of different oils you could make from that. So that's what they were doing. And that was possible also because the price of oil was high. So you had this tailwind in the business case, tailwinds are forces, which helped drive your company or in business model.

But it's not a tailwind that's sustainable. If you take that away. A big part of your business case falls apart. But at the time that tailwind was the high price of oil and then there was subsequently in the U S at least there was a lot of government incentives provided for green chemistries that at that time was the Obama administration that provided this.

And those were funds that would go into helping. The green chemicals industry as one way to call it evolve and grow. So that was what constituted, the first-generation the companies they received a lot of investment and at the same time, they also ended up getting products that were being developed and hoping to commercialize.

Flavio13:59

So what happened with these companies? Both the biofuels. I know if I remember correctly, there was a big hype back then, but now it doesn't seem to have gotten as much traction as it could have. I think in Brazil, it's used for example, as a gasoline replacement, but other than that, it hasn't made these broad impact that maybe some people were wishing for.

So what did what happened with biofuels and. What happened with these other uses that you are touching on in and where are we today? Is this now the price of oil has gone back up. Changing the game again, or so maybe if you can touch on the unit economics a bit more in detail, that

could also be interesting.

Sam14:39

Sure. Okay. So biofuels is one segment. These building block chemicals is another segment. If we just look at biofuels as a segment itself, it just basically comes down to price. We've got a lot in terms of the actual value chain or supply chain of how these things are made. You do need to grow the crops that provide the glucose whether that's corn or sugar beet or sugar cane.

You, you have that part, you have to harvest that you have to bring it into a location to concentrate it. That has to go into a fermentor. And so when you compare that with the existing infrared trucks, infrastructure that exists for. With with fuel itself it just is not sustainable unless the price of fuel was extremely high and you got government subsidies once the government subsidies go away which basically happened as the After effects of the global financial crisis were away.

That was one leg that got pulled away. The other was that we never expected that shale oil and the whole fracking technology came into play and it became a major source of cheap oil as well. The price of oil by around 2015, 2016 had dropped. Considerably because of this big supplier of cheap oil that was enabled by another interesting technology, fracking technology that brought it down.

So that, that whole aspect of biofuels was, had basically had its legs kicked out of out of it. And it became unsustainable. It's there, there is a bit still in existence and I wouldn't call it biofuels. It could be ethanol which is added. But that's the remnant of what that part of the industry is.

The other part it's these bio-based chemicals and it's suffered the same effects because your feedstock is very high priced oil at the time, which no longer is the. The other is the, so that's the The input side of it the what your costs are the output is, or is what your market is.

So if it's if it's going into plastic let's say it's polyester, or there has to be a market that's large enough to be able to make your unit economics work. As I indicated, Propanediol was one of the first ones and it's still a product that, that is still made and it's integrated in into the product supply chain.

So that's still sustainable. It's not an industry in itself. It's a product that's made in-house by the company that used to be called DuPont as a product. Succinic acid was one of the other ones that was a product that was made by synthetic biology. It, they first started by fermentation of, e-coli and then also by ferment, they switched to a yeast based fermentation for higher yields.

But still the price could not go down enough. And at most you needed about 40,000 metric tons to even start to get there in unit economics, but you, and you would need to have much higher levels. So the problem there is that the market demand the market size is not high enough. So, and that was the case of a lot of these other products like Butanediol.

So that's when we, when your input and output side are, the economics don't work out, basically those companies don't are not really that viable.

Flavio18:20

I think at this point it might be interesting also for people to realize a little bit more like where these petrochemicals are used.

We talked about biofuels but in plastics, but there are other areas as well. Could you give us a quick overview of what are the main other from all the transport and heating, which is, people probably most associated with oil and gas also, where else are petrochemicals used?

Sam18:48

Well, actually predominantly it is plastics. And this is where that was the first generation I would call of synthetic biology companies is that they are making the building blocks for plastics, mostly polyesters. And polyesters are biodegradable. So if it actually breaks down you will have a some sort of closely, hopefully sustainable supply system and.

Degradation of those plastics. The problem is that you're treating these first-generation of companies we're treating their products as commodities. And if one looks at it now eh, today, 10 years, Price of oil is high again, but the the industry and the world has changed and we are no longer, I feel working in a globalized world where products can be shipped anywhere around the world.

We are going to be in markets that are more closed and therefore the production that you make of any material. Plastics for example, and you'd want these plastics to be recyclable or degradable, they would serve a local market. And so the companies that I see evolving today, nevermind synthetic biology for a moment.

These companies make materials that are degradable for disposable wrappers, disposable cutlery, and so on as a very simple example they serve very local markets. And if that's the case, if we go into bio-based materials what one would make are a whole new supply? What I would think is as an idea, I'm not saying that I have the right answer here where I think things are going is that you would have to create a whole new supply chain based on a whole new set of materials.

So right now, what are the kinds of plastics that exist? There's polyethylene. Polyester which is there's many types of polyester, but polyethylene terephthalate is or PET is the most common type of polyester. You would look at different types of polyester. And you would continue to do that.

And basically you're developing a whole new set of materials to replace the plastics that are in existence today. That's one way to do it another or second-generation is that what are the other materials that you could make?

Another set would be. Surfactants

so these are more complex compounds. They are originally come from they don't actually come from some of them. They come from petrochemicals. Most of them come from other sources and including agriculture. The agricultural supply chain this is becoming of interest and they're used in soaps and detergents which is a niche in itself

Another is to make things I one area I like and have been watching for a long time as Palm oil, because Palm oil is used in not just in a whole lot of foods, but in a whole lot of other consumer products, cosmetics.

And they all come from Palm trees, which really come from mostly Indonesia. And that comes at an incredible environmental costs. So where we are with generation to generation three, Is to make a lot more complex systems rather than just building block are petrochemical like compounds.

Flavio22:04

Yeah. So there's a whole new business area really propping up. There's companies like like Gingko Bioworks and many others Amarys that are working on creating. Sometimes they're making less toxic or they're using less toxic waste to make certain industrial compounds that are needed other times they're making you compounds right.

So one example is the flavors and fragrances industry, for example, vanillin, and I think is a classic example. That's the sort of the molecule that gives vanilla it's vanilla taste and it's expensive to grow. It only grows in certain regions of the world, for example, French Polynesia and then it needs to be sourced and treated and all that.

And at the end that the amount of work that goes into making. The vanillin needed is quite great compared to us. If you can have a fermentation based system that makes the exact compound, you need some people also call that precision fermentation then potentially you might have a much more sustainable and also more predictable supply chain.

Maybe I dunno if you have thoughts on that, where, the plants and crops can be subject to. Both to pests climate disaster. And now as we're learning also geopolitics starts playing a role. So instead of having to be dependent on. Molecule being able to be sourced only from a certain region of the world.

We may want to have or countries around the world may want to have a, as you were alluding to earlier their own sort of bio manufacturing hubs, or they can make the desired compounds, where they needed and also be less dependent on both climate, pests and geopolitical perturbations.

Is that something you also see that way or.

Sam23:49

That's right. So Amyris, is, is a very good example of that. That was one of the very first,

Synthetic biology companies out in California, back in the early two thousands, mid two thousands that got founded. And it's been an interesting company to observe because they've been trying to find the right business model for them to stick.

And a very good example of that. That was one of the very first the very first iteration of the company was to create biofuels. They had a way to, to, there was to make a different fatty acids, which are would therefore be oil. That turned out not to be sustainable for reasons which I mentioned it's too high costs compared to what you could actually purchase as regular Fuel from petrochemical sources.

And then they started to pivot in a number of different ways. One was in pharma ingredients. Another was like you said in, in fragrances these would be fine chemicals. Each one of those, it could be an industry is an industry in itself. And so this is where a one way I thought about it was let's just look, let's just look at the evolution of the chemical industry which began long before the industrial revolution you had companies that started making fine chemicals which went into pharmacy pharmaceutical ingredients you had from that company.

These were all based in Germany and Switzerland companies that went into specialized in flavors and fragrances. You had companies that ended up going into dyes and pigments and so all of those industries are still I would have thought of viable as approaches to the first generation and second generation of synthetic biology companies instead of using traditional chemical routes, most of which comes through petrochemicals go through synthetic biology route and we ended up we've been seeing that.

But now we're starting to see more advanced versions of that. So Amyris, I think never ended up catching on because they never caught the right business model. So the way I would position it is that generation one are these building block industrial chemicals that, that these companies attempted to.

And their failure is that those are your, those are commodities and you've positioned them competing against petrochemicals. Version 1.1 I would say is to start to produce these specialty chemicals and build a whole new supply chain. And that's a possibility if there's some entrepreneur out there that wants to have this vision of doing it, I think that's a very interesting premise.

Version two is instead of using petrochemicals as a, as an input, why don't we consider some other carbon source, such as carbon dioxide or methane, and we're starting to see companies like that where your feedstock is not a sugar, which is extremely expensive. You've got to go through the agricultural supply chain is secure that, but basically hooked your chemical your company up to an industry that's supplying either carbon dioxide or methane and or natural gas, some form of natural gas, and we're starting to see that. And that's driven by okay. If you have that input, what can the organisms that you've engineered. Make that's high value. So that's another generation.

There are other companies that can make dyes most of these dyes, again, come from fine chemicals through a petrochemical routes. Many of these dyes are not biodegradable, they end up polluting riverways and your sewer systems with these intensely colored, sometimes carcinogenic dyes.

And so there are companies that are thinking of, can we look at dyes that are sourced from organisms, which are not just therefore sustainable, but also not so toxic. And that's another set of companies. We have companies that are using synthetic biology to, to make compounds that really are valuable.

So one of the ones I found that was very interesting was ways to make human milk oligosaccharides. These are sugars found in human breast milk and sugars in breast milk are unique to. the the mammal. So the, one you find it in humans is different from what you find in Whales and what you find in marsupials and so on.

And there were many companies that had a technology to engineer E. Coli to make these there's about. Over 200 of these human milk oligosaccharides. And why is the baby eating these things? And nature and evolution must have designed it in a certain way.

And there have been studies to show if you're actually having breast milk with these human milk oligosaccharides there, you do have lower incidents of infection. You do have a better growth improved brain development marginally. And it would be best for infant formulas too, to actually have these ingredients in them as well.

But you can't make these they're very complicated by chemical routes. You'd have to use a biological route. And there was a company in Germany that, that figured it out. Two very brilliant brothers, Jennewein Biotechnologie, what it's called and that's an example I love as a citing as an example of a synthetic biology company that made a very useful, very high value product.

And just a year ago they were acquired by a major specialty chemical company for I think, 400 million Euro. So hats off to them. That's a win that's a very good example of a, of an application that, that actually is very useful.

Flavio29:27

Yeah. So I love what you were describing, how, like the version 1.0, these sort of building block chemicals really turns out that you too closely competing with petrochemicals.

Oil, actually, is an incredibly cheap material in compared to many other for the amount of versatility did you can do and the amount of energy that's stored inside, right? So it's a sort of very hard to compete with that. And then now we're learning that, as I guess our tool becomes better, our tools become better.

Be in the synthetic biology field, the sequencing costs have come down. We have better assays. We have high throughput assays to test different to really do cell engineering, cell lines and so on. And so we can start engineering more complex pathways. I think that's what you're talking about now to make more complex molecules, oligosaccharides or proteins that are larger proteins that also maybe have a post-translational modifications and so on.

So it's really a whole new field that's emerging now as both the wet lab and dry lab come better. There's a whole new field of things that they're opening up. And I certainly love the example you just mentioned with the human breast milk. I think that would be a boon. If these infant formula all had these sort of healthier compounds.

Yeah I think that's a very interesting way to think about everything. Maybe for people we can, get slightly technical here in just explain a little bit more the synthetic biology we've talked about, how does it actually work? What do people how do you go from an E. Coli you find in nature to actually, making the compound.

Okay. So there's many different ways to do it, but I'll just cite the and that's why the field is so broad. And I'll just cite the industrial biochemicals that bio-based chemicals that we started this conversation with where originally the upstream the feedstock is sugar glucose.

And there's a number of pathways and these are ones that our fundamental to two organisms as to how to they metabolize in this case, sugar for energy. And as I, what I noticed is that when I looked at the patents, the evolution or the timeline of what, when these companies and when these things came out closely matched the, how far you are going down each of these pathways.

Sam31:50

So the very first one is the glycolysis pathway. Which takes sugar and breaks it or builds that up into Phosphoenolpyruvate and Pyruvate which, which are energy sources that go on into other pathways within the organism. And when you look at that Phosphoenolpyruvate is one that can be engineered further. By that, the pathways as to where it goes to make succinic acid. And so that was one of the first approaches and early companies using technology based on engineering, the organism to take sugar. And as it goes through the the glycolysis pathway gets to phosphoenolpyruvate ends up making succinic acid.

Now if one takes the glycolysis pathway further, it goes into a number of other pathways which are possible. And this concept of metabolic engineering is about how do you engineer the organism to steer the direction of the pathway to maximize the yield while keeping the organism highly productive.

So another direction it could go is this aromatic amino acid pathway. So after you go through the glycolysis pathway, if you steer it towards the aromatic amino acid pathway, And, aromatic is basically means benzene type compounds and where you go then may allow you to make benzene-like compounds and that's useful to make terephthalic acid, which is a building block, for example for polyester.

And so there was a one or two companies that actually started doing that because they managed to the professors that did that work managed to find an optimal metabolic engineered system to do that. Another direction you could go is after the glycolysis pathway, you could go towards the mevalonate pathway and that's gets interesting because you it goes into a lot of other different products.

You could make fatty acids from. You can make 1,3 Propanediol, which is the example I personally saw in DuPont. Fatty acids, those were in fact that's where Amyris, their, their expertise was the mevalonate pathway. And I think the professors originally came from UC Berkeley and developed the expertise in that to create a system and a set of products that, that developed that could be used to develop these fatty acids. And then furthermore, after that, as you go further downstream, you could end up engineering a lot of other pathways towards the they're called phosphates drain your phosphate, farnesyl phosphate, pyrophosphate.

And it's so on. And that leads to a whole range of more complex compounds. And when you get that far down, you're starting to make compounds that could go into flavor, instruct precursors flavors and so on. So that's the first generation of these industrials chemicals. And they're all driven by how you optimize that.

And they're all done in e-coli. Eventually they will be switched to yeast because you get a more productive system. And that's one way.

Flavio34:54

Okay. Yeah. So it seems like just to summarize a bit more from a layman's perspective, you essentially have sugar coming from essentially, either corn or sugar beet or some other type of plant.

And in this sugar gets turned into various compounds via pathways, which are really just enzymes modifying these compounds from one step to the next. And you have these very complex branches of pathways things can flow into. And then the idea is here, if I understood this correctly.

So finding out that the product you desire to make finding out chemically, where are you in? Close in the natural wild type sort of pathway and then engineering adding enzymes or modifying enzymes and so on to steer the flux, the metabolic flux into the direction that you desire and it, and ideally have at the end of the organism that, has a hijacked pathway now making the product that you want, and then you, there's a whole bunch of downstream processing that needs to happen after that. But is that's the high-level goal here and then synthetic biologists companies like Amirys and Gingko and others have specialized. In doing these metabolic engineering tasks there's a whole bunch of high throughput screening going on sequencing understanding the natural pathways and in testing various biological components swapping out promoters and enzymes and so on to optimize the yield and the.

titer your that your host can make, whether it's E. Coli, yeast or other, even more complex hosts like filamentous fungi and so on are starting to be used.

Sam36:40

Yeah, exactly. You define that quite well. That's, what's going on. And it's there's a lot of research ultimately to create an organism, which you could use to drive this. Now one thing, I think that's important is that many technologies cannot produce a product on their own. You need a lot of parallel technologies. And I think one of the big problems is that this is taking place in an organism and you got to extract the product out of that. On the engineering side, I think a cell-free system is something that will be needed in the future to support a more efficient production.

But the steps you've described is exactly what's going on. In the case of human milk oligosaccharides, as I described, that was in fact finding the right enzyme and where the patents are concerned. It's whether you choose that enzyme from one organism and you patent that or an enzyme from another organism and you insert it into your E. Coli or ultimately a yeast or something in other organism that's where you get intellectual property position.

And have the engineered organism produce your human milk oligosaccharide that way. There are many different types of human milk oligosaccharides, some are more complex than others. And of course the more complex ones require more enzymes and you'd have to find the genes for that and figure that out.

And that's still a work in progress. It's that's still very interesting.

Flavio37:59

You mentioned the cell-free systems. Can we dig into a little bit about that? What are the main advantages and then maybe also current challenges with establishing cell-free systems to

make compounds?

Sam38:09

Quite simply it's because when you create the product, it's still in the cell, you've got to actually extract.

There, there are ways to do that. And the simplest is is basically have a transporter that exports it out of the cell. And in that way, that's how you catch, you get a labeling of non-GMO at least for food ingredients, for example. And, but the thing is if it was not in the cell itself then you don't have for example, an E. Coli, there may be the potential pyrogens, these are toxins that could, if you're going to rupture the cell, could present a safety issue for human consumption and so on. Finding a cell-free system is engineering a number of requisite enzymes. And whether they're in immobilized onto beads or done in other ways, there's many ways to do this.

There's many different approaches to it. A lot of people say, oh, there's people already doing it but having a cell-free system to create proteins is different than having a cell-free system to create a specialty carbohydrates or cell-free system to make fatty acids. So there's a lot of opportunities there.

Flavio39:15

So I haven't thought about this very deeply. So just, can you draw the line maybe between like classical more chemical synthesis and then the cell-free system. And then I guess the bio-, cell-based system, where, what are the clearest distinctions and what is need, I guess I'm trying to figure out for the cell-free system.

What is needed to make it work, that's maybe challenging. Why hasn't it

proliferated more?

Sam39:43

If we look at just the chemical side of it and again, this is where my pharma background comes in useful, is that you can only create relatively simple chemical structures through synthesis.

Now those are still fairly expensive and time-consuming to do because they're done in big chemical reactors and they're done in several sequential stages. And where that industry is going nowadays is to go through continuous processing. So that you're not cleaning out vessels, you're not transporting the products of reaction, A plus B to C, and then you have another vessel C plus D plus E and so on to make F you're just going to run this through a number of columns and other systems to ultimately put, have the product made at the end of that.

You can do that for relatively simple compounds. But when you start to make proteins, when you start to make complex carbohydrates when you start to make a specialty fatty acids, I think that's where pure synthetic chemistry is going to be a lot more costly, and that's where you would want a biological system because enzymes can do that much more exquisitely.

They can do that the yield and then the rate of reaction much faster. And that's where you'd want a biological system to do that. Now right now, that takes place in cells because the cell has to produce the enzyme as well as any necessary co-factors to actually do that in a cell-free system, that's the genius or the innovation that's required to liberate all of these processes from a cellular system.

Flavio41:18

So just to understand what the would the enzymes required in the cell-free system? They would've probably still been made biologically a cell at some point, then you would how do I imagine this? It's a big soup of different enzymes and then you add like you're starting products to it, or how do I imagine this working?

Sam41:40

Yeah, correct. So that's, many enzymes are made through, through a cell based system, originally, that's another opportunity in synthetic biology is there a way to make enzymes more effectively, more efficiently, cost-effectively and there are companies that do that as well.

Flavio41:59

Okay. Are there any cell-free systems that are already up and running today in a large scale? Or is it, is this still in the earlier phases?

Sam42:10

It's early. So there's lots of opportunities there. And I'm stunned at how quickly the field is evolving. I went to I, one of the jobs I had earlier was to actually look for companies that had technologies like this, and I would go to certain conferences and see who is actually presenting at the investor pitches to see if it's interesting.

And I would flag these companies. I would go after them as a corporate development kind of approach. And what surprises me is that, about one year after I approached these companies, they're getting major rounds of financing from various investors. Which shows how quickly they're going. Because if you're running a startup company, you know how long it takes.

But these are getting into series A