Enzymes in Distilling | novonesis

Enzymes in Distilling | novonesis

Dermott Dowling

Here we share an excellent keynote presentation by Dr Jan from novonesis from the 20th Anniversary Australian Distillers Conference 28-30 April 2024.  Dr Jan shares insights into how enzymes are naturally occurring proteins found in nature and how they are widely used in Distilling around the world.  Topics covered in this keynote presentation and technical QandA include:

 Audio File from Presentation

Transcript to accompany Presentation

I may have complained that, there weren't that many technical topics and here I am. So, but that's good though. I know. I appreciate the the organisers actually the feedback. My name is Jan Kristensen. I've worked with enzymes for about 20 years. So it's a topic I'm very passionate about and it's great to to see so many people interested in hearing about it.

I work for a company called Novenesis. Up until recently, we were known as Novozymes but we have recently gone through a large merger with a company called Chr Hansen, which is why we now have a new company, which we may not have come across before. We are in the world of the area of Bio Innovation and biosolutions.

And although we do many different things. Enzymes continue to be the bulk of what we do, we are the world's largest producer of enzymes, enzymes that are used in more than 30 different Industries. If you're stealing being one of them, we Supply enzymes to the majority of Distillers worldwide, larger distilleries, and an increasing number of craft distilleries as well including here in here in Australia.

I was born in Denmark, I was exported to Australia about six years ago, you'll have to excuse the accent, and I properly became an Australian citizen not too long ago. Um, so yeah. Um, today it's going to be a mini crash course in enzymes. For some of you, it may be more of a refresher.

But basically I'm going to talk about what enzymes are how we produce them, what they use for? What are some of the benefits? Um, I'm going to talk a bit about starch because what it's all about is converting that starch into fermentable sugars that we can use for Spirits for production.

Let's talk about the the different types of science for different types of enzymes that we're using distilling, how they're used. Um, and I hope to maybe also dispel a couple of myths. Sometimes I hear people who tell me, oh, we use Malt now. Still, they have enzymes in them.

We don't need to add anything else or even some people say, oh, and those enzymes. You produce, they're artificial. We're making a natural process. We don't, we don't want those. Um, so let's see. Um, a little bit about what enzymes are they are many, many different things. They are Nature's tools and they are essentially what make living sales work.

So every single living cell, in the world inside you, in leaves, and everything are full of thousands events on there. What make the sales work? They are also. They speed up biological processes, they're basically responsible for them. They're also catalysts and I'll talk a bit more about that. So the basically they speed up chemical reactions, very specific chemical reactions.

So enzymes are highly specific. It's not like you have a Wonder enzyme that can do everything, they're very specific in terms of reaction, and, and substrate. Enzymes are pure proteins. That means that they're fully biodegradable. There's nothing synthetic about it. Um, so the breakdown in amino acids. Just log the enzymes that are already found in the field.

Um, lastly in industrial enzymes are made from microorganisms which also a little bit about So just to recap, what a catalyst is, a catalyst is something that speaks up a reaction without being used of itself. Um, in the case of starch, if we were to convert starch into glucose without any enzymes, you would have to impart a lot of heat, maybe some chemicals onto the system that reaction is not.

Well, it is going to occur by itself but it will take years to break down the starch without anything. So by having an enzyme in the system, what you're doing is that you're basically lowering what we call the activation energy. So now, Reaction is going to run a lot smoother, a lot faster, without the enzyme being used of itself.

One enzyme can catalyse tens of thousands of reactions per second. They're amazing little things.

Informativity. Um, so again each enzyme has its own specific reaction profile so it's not like it's not like a chemical. Uh, so when we use enzymes there's a number of things that we want to take into account. They typically work at a certain pH range, certain temperature range. We want to consider the enzyme dosage and the contact time which are closely related, sometimes you may want to use high enzyme doses and if you have a shock retention time, if you have a longer retention time, you can combine using the lower inside usage.

Uh, also substrate Inhibitors of things like that. You also want to want to take into consideration. So how do we produce enzymes? So we have a team of very lucky scientists that get to travel around the world looking for microorganisms. They collect fungi and bacteria. Bring them home in our labs and catalogue them.

We have a library with hundreds of thousands of microorganisms and then they look at what enzymes can these microorganisms produce. Sometimes we come up with something that could be relevant for distilling and brewing. And so we take it into our production, although sometimes we may have to take that enzyme and put it into a production organism in order for us to actually produce enough of it.

So, what we do is we do a very, very tightly controlled fermentation, not unlike how you would ferment a mash, but we're very tightly controlled in terms of In terms of pH and temperature and things like that and during that fermentation the microorganism is going to hopefully produce a lot of the enzyme that we're interested in.

So if we separate the enzyme from the production, strain we formulated and So eventually we have a product either liquid product or granulated product. It's important to note that when we separate it, we separate it completely from the production strand. So an enzyme is a protein, it has no DNA in the end product and that also means that an enzyme cannot be a GMO, it cannot be a genetically modified organism because it's a protein, there's no DNA in it.

That's one misconception that I sometimes come across. Or so if we sort of start backwards, if we want to make ethanol, obviously we need some sugar source. Ferments use less we start out with tuba as a sugar cane, we use something that contains starch. So from an enzymatic, viewpoint all the different substrates sort of falling into the same class.

They are all starch bases. So when I speak today it it basically it's not just whisky it, not just gin or vodka. It's about converting the starch into Spirits. A bit of a refresher on starch. Starch is the Energy storage mechanism. It's basically a launchbox of energy that the plant gives the seed to that the seed can grow and become a new plant.

Um, it's a partially crystalline granule and I'll get a little more into why that is important. It's water insoluble at ambient temperature. However, if we increase the temperature in the starches water, it's going to swell and your lab noise upon heating, We'll talk more about that. And again, obviously, the single building block of starch is the proteins that we want to get to so that we increment it Now, the gel organisation is when the starch starts taking up order and as well.

You can see up here, we have a microscope image of starch at 40 degrees. You can see the starch granules as we increase the temperature. They take up water and they become less crystalline, and they swell in size. But that happens, it also means that enzymes will have accessibility accessibility into it.

Without the gelatinization. Enzymes cannot break down the starch. At least not directly learning setups. So, the gel optimisation of our substrate is really, really important. And it's also important to remember that this geology optimisation happens across a number of temperatures, like, your range of temperature depending on what starches that we're using.

What, what grain Um, what also happens is we get an increase to like solubilization and we also see the viscosity increase in our mesh And so in order to start breaking it down, we basically need two different enzymes. The first one is an alpha amylase. It works at high temperature and it cleaves the gelatinized starch into what we call dextrins.

So dextrins are shorter shang chains of glucose. That's the the first step. Now, these dextrins unfortunately are not fermentable except for the db2s and the maltose and that's what the next time. So, I'm coming to the picture and that's the glucoamylase. So the glucoamylase will cut these dextrins into maltose and glucose.

So, converting the dextrins into fermentables of course. If we sort of look at a stepwise, Um, at least from from my enzyme perspective, it's about gelatinous and starch. Then when dramatically hydrolyzed societextrins with lipidextrins into preventable sugars, you ferment the sugars into ethanol and you're just Should be relatively simple.

Um, A few words about malt. So, malt has been used. Probably, for thousands of years, for spirit production, for beer production. And if you ever came up with the idea of of using malt, instead of grain was probably a coincidence. But the reason that it works so much better is that once the the grain starch, contains the enzymes, it needs to break down that lunchbox of energy that it has with it.

And in order to do that, it starts expressing enzymes itself. So, the big difference between the grain and the malt is that the malt contains enzymes and that's why we've been using Malt for so long. So the question is whcih enzymes is malt containing inside? Why would we even add extra enzymes on?

What am I doing here? The beta amylase that is found in the malt does come with some limitations. It is not as thermostatable as the products that we produce. Uh, and also there's only a certain amount of it. And obviously, if we want to use a grain instead of a malt for gin or vodka, even for grain whisky.

We either have to rely on using the limited beta amylase and whatever model we included in the process, or we have to rely 100% on the enzymes for running a grain-based process. So even if you have a process where malt is part of the equation using external enzymes will have or can have a number of benefits.

Uh, you may see a more complete mashing and a better viscosity break. Uh, as much as we can, partially fully replace them all with grain so there's a potential cost serving in here. Um, If you lower the viscosity, you're going to have a more efficient solid separation in filtrations, and your processing is going to be easier.

Again if you have a more complete mashing you don't leave any starch or any dextrins behind so you're going to see high conversion and tall yields. Not always desirable but in some processes it may you may have faster limitations if you're interested in that. And we're going to talk a bit about energy reduction, or energy, and water consumption reduction, as well.

So if we dive in a little more and look at the front end of our process, now I'm not going to talk about a specific process, but we use our Alpha amylase in in the beginning. Um, If we have a process where we also want to use malt then having a two-step process where we start at a lower temperature and increase it.

So basically both the mashing and liquid fashion can be an advantage. The alpha amalyase families, that we Supply, for example, Liquoflow Go 2X, Has an ideal temperature of about 85-87 degrees for it to work, as fast as possible. And as efficiently as possible, So there are many ways of building new process depending on what you're doing.

And and and that's part of what we do. I love going getting out and working with our customers in optimising the process, and then, basically making things run as efficiently and investmentally as possible. There is another class of enzymes that you can also obey and also need to add up front and there is the viscosity we're using it.

So I'll get back to this in just a minute. If we look at the uh sort of a little later in the process, we talked about the glucamylase, it could be a product like Saczyme Go 2X which is the primary product that we Supply here in Australia. Um there's a couple of different ways of using that again depending on the process.

If you have a Distillery, which I would refer to as a Brewhouse Distillery basically, you have solids separation lautering in the mashing before you start your fermentation, then you want to add the amylase. Once you have cooled off your Mash, but before you boil it, Whereas, if you have a process where you ferment on the grain on the malt, then typically we add the Glucoamylase at the beginning of fermentation.

And basically, let it work throughout the fermentation. That has actually an added benefit because it means that your sugar concentration is never going to be really really high. And that's going to reduce any de-stress That you have in there. So again, different ways of using games forms and we like to be involved with helping you across with the students.

This this up here is basically a sort of a diagram of a plant cell and we have our starch granules in the middle here and they are surrounded by a cell wall. Depending on the grain that you work with these cell walls will contain different polymers. And these polymers will include things like beta-glucans and cylin and cellulose rapidins sometimes they even structure of proteins.

Depending on what grain it is, these polymers can create a lot of viscosity. Like, if you're doing a ride motion, I'm sure you've noticed how viscous it can become especially if you're working in household slowly. Um, These polymers. We sometimes we refer to them as being non-slash polymers. And although they are similar to dextrins, they are not broken down by amylase.

So we need, basically, we need to be to vulcanized we need silentized cellulose, and so on to, to break this down, Um, they say the proof is in the pudding. So I brought a photo of two ride mashes with the same water to express ratio and I'm pretty sure you can see, which one of them has had enzyme added.

It's a very significant effect. You can have on the viscosity. Why, why is that viscosity so important? Well, the less water you add to your system, but that's what you need to eat. But also the less water, you need to get out of your, this system in your distillation, so that can save you a lot of energy.

It saves you, a lot of costs. It may even impact the capacity of your Distillery. The figure here shows you that if you're able to lower your solid content of your match by seven percent, You may save as much as 25% of your energy input. The other thing is the ability to run a smooth and stable process to lower the viscosity the easier, it is to pump, the less fouling, you're going to see the more efficient solar separation, the more efficient infiltration.

So using a product called Viscoferm can really, really help your processing before.

Getting to the summary. Enzymes, we talked about their proteins that act as catalysts, they're very specific. So we need at very specific enzyme for each step in our process. We use the enzymes to efficiently converge start into the fermentable sugars, and we can also use them to lower that mesh viscosity that we just discussed.

For meshing, the loop of fraction we're using the alpha amylase such as liquid fluids converts, the gelatinized starch into maltodextrins. Next up is as we saw, viscosity reduction. As we just saw, it can be really important, especially if you work with Right. And also, if you are used for things, like umalted rye, for sacrification.

We use glucoamylase to convert the dextrins into fermentable sugars. And as we saw, Addition of enzymes can be used for grain, both unprocessed and malt, and growing past. One example I've run in and a couple of times is say, you are a distiller somewhere in Australia. You want to use a Heritage Barley that has been grown locally.

But they only produce a small volume. It may be hard to find them. Also is willing to turn that into a malt. Using enzymes, could allow you to use Heritage grain like that in your, in your process even though you normally work with with them all, So I think that's a good example of where sort of enzymes, I'd like to refer to them as being a powerful toolbox for the stores.

So if you are to just take one thing home from my little talk today, is that enzymes? Essentially provide a toolbox that can help the starch conversion processes. Um so and and we spoke about some of these potential benefits easier processing higher yields. Energy savings and and saving money. 

Happy to talk and take some questions. So, if you produce risk and you're using, yeah. Obviously, the Scots approved on that. You can't do it. Yeah. Because Very good in this year. Is just intuitive and you're trying to follow the things in that and you make a mistake and then you end up with two sugars in your fermentation.

Can you just add? Take your glucoamylase and just Chuck the fermentation at that stage? Yes, absolutely. Can you tuck it in in your second slide, should it be about 75 degrees Celsius? You could? But the, the longer retention time, the better obviously, because it's going to take the enzymes some time and especially with a relatively low temperature fermentation, That affects the kinetics of the enzyme.

So the sooner the earlier during fermentation where you can add the enzyme the better Uh, if you added leg in the Run you would need a high enzyme to to get the same effect basically. What is the is there any temperature in what's going to analyse with DNA? Um, the optimum temperature is actually as high as 60 to 65 degrees.

So if you have a process while you separate the solids, that is the temperature. I would run it for a couple of hours. Alternatively, if you add it at the beginning of fermentation, then you have instead of then, maybe you keep it in there for five days. So, sort of Optimum temperature, 66.5 degrees you can get by using it for a few hours or you can actually use lower temperature, your lower dosage.

But for much longer time, Uh, please use it for a few hours. Do you mean Chuck it in and that's it for two hours or do you mean if it's a longer frame and you want to have more? You just keep adding more out of the second base or there's one person being home.

So, if you add it and what I refer to as a sacrification, so then then I think there are some distillers that will boil their water and that's going to inactivate it. If you don't boil it, the inside is going to continue working through our fermentation. So you're sort of you're better off doing that, but then, of course, that comes with some other risks.

Um, there is also for large distilleries, they have a very specific addition profile of their enzymes. They will typically even ferment on, on the grain. But instead of just pouring in all the glucoamylase at the beginning of fermentation, they may sort of dribble it in over the first 24 hours again to make sure that your glucose Peak doesn't get too high because as soon as that gets over five percent, your yeast is actually going to be stressed a little bit and the more stretching useless and more girls are all you're going to see and the slower, your kinetics are going to be So again that's something we can help with but yes your Google analyse and dosage profile can actually have have an effect.

Absolutely. So, if we use Saczyme Go 2X a single mesh. Um, It's a glucose in fermentation. Um, but if you use them just straight on barley, it's a Maltose fermentation. So is there a slight of difference? Do you think between a glucose and the fermentation and a multicellular penetration?

I would say it's not a glucose only. It it produces both glucose and maltose. So you're going to have a mix but there's definitely going to be less maltose in there compared to. If it's malt only Um I have never heard of anyone saying there should be a, I don't think that yes, whether it immense glucose and maltose.

I can't imagine that would affect the flavour of protein. I've never heard of a dead list. Okay, my guess. Is that the pathway inside the ears? Probably the same. Um, I'm not a yeast expert. I imagine that the yeast might actually reduce the motors into cookbooks Before it starts converting.

So you've got a stuck ferment and go through the process and work out what's going on. Yeah, with enzymes assist, if you're still at, you know, multiplicity. Um, it depends. The stock fermentations tend to be more related to the yeast and Bud. Obviously, the new needs to be something for the yeast to grow.

So, if you're going to be re-pitching, you want a certain Level of glucose and maltose in there. And that's where the gliogramolites could help. Uh but obviously without a happy ease, the reap which in the yeast, the glucose is not going to ferment by itself. That? So in addition to some other questions but if you've got um, add the enzyme Can presence to that.

I mean, influence and happiness apart from the other fact that you're going to get

Not just the fact that anyone's presence is that Dalton County East Works. And so forth. But is that thing there will produce out of opinions that doesn't its mobile. Yeah, no. So the the enzyme doesn't doesn't enter the yeast cell and and so essentially the yeast doesn't know that there is an enzyme there.

It just it takes up the timals and the glucose. So, yeah, so it's not like it's gonna feel that I'm going to be stressed because there's tons of in some circling around in the system. It doesn't it doesn't get into the cell much, much too big. I mean, there are a lot of A lot of polysaccharides contributing to the viscosity.

Yes. And you have data as to how that treatment enzyme earlier, Reacts with the different polysaccharides to reduce reduce viscosity. In other words, could you possibly select? And the reduction of a particular polysaccharide based on maybe temperature or pH or something like that. Um, I'm not worth the product that we're using.

You would basically need a model component product to do that. So what you're saying is the viscous firm is a, is a mixture. Yes of Viscoferm mixture. Yeah. Exactly. Cylinase or cellular and cellulose. Yeah. Yeah. So it comes out of certain ratio that we have found Uh, it was actually developed specifically for Rye, but we found that.

It's very good all-around for any small brand. The coronary to that thing is that are you in a position to alter your mixture of enzymes to selectively. Um, yes. And we do have other products. So many of our products are spin-offs from the biofuel industry, because for us, that's a huge industry.

Um, and so we have products that are optimised for wheat, for example, That's how that's what I was getting at this. Say perhaps you have a particular product that is optimised for say. Yeah, and that would be the risker firm actually. And we also have a week because week there's a big thing in Europe but we don't have for barley and triticale and so on because developing a new product is something that takes years for us.

And so there's a huge investment in that and and as much as I had to admit, it distilling is a relatively small industry compared to our biofuel industry. And again, that's why we sort of adopt our products from, but yeah. Theoretically, you could do that and we do have a number of products that we could, uh, that we could play around.

Yen you mentioned that, you know, new plants, generate your enzymes and so you said they're not genetically modified. What happens. If the plant that you actually neutralising to break the enzyme is genetically modified So, Yeah, that that's um it's kind of a long explanation. There are essentially two different classes of enzymes.

There are those that we call Classic enzymes, but we've gone out. We found a microorganism. We use it as So we've fermented and separated and that's what we use to primarily do. What often happens is that when we find a new enzyme, it may be a really, really good enzyme, but that microorganism may not produce nitrofit.

That. Turn into being a really really expensive product to produce. If we make a huge fermentation and it produces just a couple kilogrammes of enzymes. That'll be really expensive. So what we typically do with those is we Or well, you have some very clever people in our lapse, they will find the gene that is responsible for that enzyme, they cut out that Gene and they put it into one of our well-known production organisms.

So now it reduces the exactly the same protein, exactly the exact same enzyme but it just produces more effort. It's more efficient way, more efficient. That means that we can produce a product at a reasonable price that we can actually do so. Those products are not GMO but they are GM derived.

Um, and There are places in the world. Where there are companies that are not keen on using this. I think in Brewing, for example, if you saw 15, 20 years ago, vast majority of our products would Classic Products. But in order to keep pushing the envelopes to come up with better products, we gradually moved to the GM derived.

And these days I would say about 95% of what we sell in brief. For example, a GM derived product. Where things got a little more tricky is that. We may also sometimes take that Gene and modify the loop. But if we found a really great enzyme but we wanted to work at a lower pH or the higher temperature, we may tweak it a little bit before we put it into our production opinion.

Then that you're not just talking about a gm engineered product, but also a protein engineered product. And that's where it becomes a little more grey area in food because in Australia, FSANZ requires that if you use something that's protein engineered, if the enzyme is in the end product, you may be required to label it, which no one is interested in doing.

But again, the dosages of the enzyme are so small and they are all removed through during filtration. So you don't actually need to do the labelling. Uh, but it does sometimes provide that we sit down with our customers and talk through about how it works.

So I'm curious you're in Scotland and you've got an audience, a lot of Single Malt Whisky producers. And you're talking about enzymes. A lightened presentation, just talking to the group what what sort of feedback do they would like to do it. They see it was beneficial but I I haven't had many of those discussions with with Scottish people about it but I know that there are restrictions on.

I don't believe you can use enzymes at all when it comes to Scottish Whisky. Yeah. And of course, being an SM guy. I think that's a restricting restriction and curiously we kind of see. So Scotland now I think is the only country where you can't use an enzyme, a single model, German is the only country in the world where you can't use enzymes in brewing.

In beer production, France is the only country in the world where you can't use enzymes in wine production. And I would love for Australia to be sort of pushing the envelope when it comes to New World whisky. And and again see the enzymes as a toolbox. Okay. Yeah. That that makes sense.

Make sure that your fermentation is super dry. There's no nothing left behind. Yeah, I'm finishing gravity's on the ride Ashes. We've sort of not seen it in Vlog at about 1008 for the finishing gravity. Um, just a couple of way that we've done. Um, It is very white with the 20 made before about dosing the glutenlays.

Across the first few hours implementation. It's an interesting one. I'm just not sure how we can operationalise that one, but I'll talk about it. But where should we see our finishing gravities? I'm 100 by our matches. So my background is in my field, so when you're talking gravity, I'm used to total solace in percent.

So Yeah, that's a good question. And and it may even depend on what kind of Rye you're using. Um, I think it looks like you guys are already doing a really good job with the Viscoferm. Um, it's about optimising, the temperature and of course the longer the retention time.

You have the better. It's going to be Um, so yeah, I'm sorry. I probably don't have a concrete figure for you. Yeah. So from a process, you can speak. So you've seen videos, it goes from being, you know, forage to, you know, just because of what those monumental polysaccharides that the biscoffers breaking down is just getting that sense of like what that represents in terms of you know anything over one zero zero zero as a.

Yeah. Um, through fermentation and we're sort of yeah it was trying to keep experimenting how important or because one is somewhere where we can get some further information, right? Yeah. I mean it's something that we can look into and maybe look at some of our other products. Um you could even start looking at something like a protease uh, The project is the only one though, where if you overdose it you may start.

There is a risk that you might actually affect the flavour profile. So that's why you need to be a little cautious. And one of the reasons I didn't include it today, Uh, but there are many other enzymes than the ones we talked about today. So some something we can look at

That work with.

Not a whole lot, unfortunately. We Supply alpha amylase to the cane processing industry. To basically break down a little bit of starch. That is found in in the cane sugar because that improves their processing. But apart from that we don't unfortunately we used to have a product which was a mixture of various enzyme activities.

But it was pruned due to lack of sales, unfortunately. So yeah, not a whole lot. What happens if you overdose? I'm just going to cost you too much money. Hear that one. Yeah. But but apart from that no. And and that's also when I talk to someone who said a gamma process and they say, how much enzyme should I put in there.

I recommend actually starting out by overdosing to find out what, what is the maximum effect you can get and then you slowly dial it back to you, find sort of that sweet spot where you have close to the maximum performance. But, at the lowest upset, But they shouldn't be, uh, except with the exclusion after protease, you should not see any negative effects.

All right. Thank you very much.