Prof. Tobias Erb: Breaking the limits of natural photosynthesis with synthetic biology

Prof. Tobias Erb is synthetic biologist and Director at the Max Planck Institute for terrestrial Microbiology in Marburg, Germany. His team interfaces biology and chemistry and centers on the discovery, function and engineering of CO2-converting enzymes and pathways. Research in Erb’s lab crosses multiple scales: from the molecular mechanisms of carboxylases to their ecological relevance, and from understanding the evolution of natural CO2-fixation to developing new-to-nature solutions, such as synthetic CO2-fixation pathways and artificial chloroplasts.

Frontiers in Science lecture with Prof. Tobias Erb: “Breaking the Limits of Natural Photosynthesis with Synthetic Biology: Designing a New-to-Nature Software for Sustainable Carbon Capture”

A Frontiers in Science Lecture at Dresden University of Technology 👉 https://tu-dresden.de/tu-dresden/profil/exzellenz/veranstaltungen/fris

so welcome everybody thanks for coming um despite of the weather some people are here and we also welcome uh our guests online so um it’s my pleasure to welcome Professor Tobias AB who is here this evening to deliver the fourth lecture of our Frontiers in science series with Frontiers in science we want to make take the current developments within the field of international Cutting Edge research and convey them to a broad multidisciplinary audience within our University public each semester a renowned researcher joins us to discuss their work and its implications for the 21st century challenges in science politics and Society together we explore leading questions theories and methods pushing scientific boundaries no doubt climate change is a persuasive issue that affects every corner of our planet from rising temperatures to extreme weather events the impact of a warming world are undeniable and far-reaching the challenge demands our immediate attention and concerted efforts to mitigate it effects and adapt to its inevitable changes addressing climate change requires Innovative and robust Solutions with technology playing a crucial role among the many technological approaches to mitigating a Chang in climate synthetic biology stands out as particularly promising by engineering biological systems synthetic biology offers novel ways to produce bofs create sustainable materials and even develop new methods for carbon sequestrations and we will hear about this in a minute with tobas AB we are honored to host an acclaimed researcher who is at the Forefront of this exciting field Tobias AB is a director at the max Blan Institute for terrestrial microbiology as well as professor at the Philip University both located in Muk his research group interfaces biology and chemistry and centers on the Discovery function and Engineering of um CO2 converting enzymes and Pathways research in Tobias ABS lab crosses multiple scales from the molecular mechanisms of carbo carox lasis to their ecological relevance and from understanding the evolution of natural CO2 fixation to developing new to Nature Solutions something he’ll talk about in just a moment Tobias AB was awarded a PhD in 2009 at the University of fryberg fryborg and the Ohio State University after a post-doctoral stay at the University of Illinois he headed a junior research group at IAT before relocating to the max plank Institute in maruk where he was promoted director in 2017 Tobias AB has received numerous Awards among them the research Awards of the Swiss and the German Societies of microbi biology and the future Insight award in 2023 he was named one of the 12 upand cominging scientists by the American Chemical Society in 2015 and elected to the European Academy of microbiology in 2019 as and to the European molecular biology organization in 2021 last year he was elected to the National Academy of Sciences leopardina and this year he has made the honor to being awarded the godfried wilham liet prize of the German Research Foundation Dear Professor AB we are delighted to welcome you to T Dron and before we start with your exciting talk I would like to hand over to Professor ton Masha who holds the chair of General microbiology at ton and who will briefly introduce toas ab’s field of research to us and the role it plays at [Applause] theodon yeah so welcome also from my side to beas um when I was asked to give these few words of introduction I was happy and pleased to do so we know each other for some while and so my well my job is to introduce the field that he’s working on and it’s already in the title it says synbio synthetic biology is part of it so that’s the realm we’re about to hear and that is really maybe kind of the latest addition to the many fields of Life Sciences but it started in about 2004 so 20 years of synthetic biology already in and for those that are not familiar I think most people are it’s not a contradiction to say synthetic and biology it really is more in the realm like in chemistry where you have analytical versus synthetic chemistry so it’s applying what you learned in analytical Sciences to design new functionalities and that’s what synthetic biology is to design Implement new functionalities in living systems or to redesign existing um natural system for useful purposes so it’s an engineering discipline it brings together the knowledge from chemistry from biology and the approaches from engineering and it comes in many many different flavors so one of the big toy fields in the beginning was logic gates circuitry people did this in the first decade I would say of this century and this is kind of a tick off so all the logic gates your and gate your or your Norgate can be implemented into cells and bottom up genomics was the next big thing it’s still ongoing and big questions are linked to that what is the minimal functions of Life what are the minimal Gene sets in life that’s what Greg Venta was doing or how to make a better ecoli or how to make a better yeast that’s ongoing projects um or bottom up minimal systems in fact that’s something that is going on pretty strong here ton we had Petra a couple of years back she was doing that she relocated to to Munich many years ago but with I Dora Tang for example or James S there people that do synthetic systems in lipid droplets on lipid membranes to to set up minimal systems that that Sim at life or help to rebuild living systems um but there is also here metabolic engineering and that’s in fact the field that is much closer to what what toas is doing so Tomas Walter for example in bioprocess engineering he’s heading the group of synthetic bio technology and systems biology um and metabolic engineering or the synthetic biology metabolic engineering realm is really trying to bring together the knowledge that we have generated in the last 20 30 years and try to implement it for useful purposes that could be biofuels that could be pharmaceutical compounds arimic AR alic acid is one example from from Jake keasling for example but I think if if you would name two felds in metabolic engineering the sin biow way as the the holy Grails of synthetic biology nowadays it’s either implementing nitrogen fixation into living systems or moving this or to use and apply photosynthesis and carbon fixation and move it into new hose to improve that so these are I think the two big goals that many groups work on and I think when it comes to carbon fixation tobs AB is most probably one of the leading figures worldwide and definitely the leading figure in Germany on that field um try to apply what Nature has evolved try to try to prove what Nature has Evolved first in bacteria later in plants for dealing with big problems like climate change like dealing with carbon dioxide emission and trying to fix that and without further Ado Tobias uh this is what you’re going to talk about you can do it much better than I can so I leave the stage up to you I’m very happy that you’re here and I think we can look forward to a very thrilling talk in the realm of metabolic engineering the sin by way thanks for coming [Applause] and thank you so much for your applause please don’t Applause you don’t know what to expect so be a bit more careful and uh when I was contacted uh to give this lecture I was felt very pleased and also privileged because it’s one of the rare occasions where you can of course talk about science but also develop a little bit more of a vision where science could go in the future future and just as a small spoiler I’m not going to save the world from climate change today probably also not in my lifetime but I could give you Vision where we could move together as a community as scientists maybe as a society and also how the different fields of research can contribute to this big Challenge and what new can actually be created when these different fields meet each other in an inary fashion and so this is why I named my talk breaking the limits of natural photosynthesis with sunion you might realized that I also put a question mark at the end which is different from my original title which was a statement this is a question because it’s still open to me if sinbo can deliver at that scale but I guess we should at least try it and there’s also reasons why we should try it all right so all at the end is about a future I would like to take you with me but before we go into the future I’ll take a step back and look a bit back in the past of this planet or on the planet its current stage and this is just a reminder that this planet is a Le ecosystem and there’s many different species billions of different life forms that inhabit this planet right but it’s also important to note that one species is really actively shaping this planet and that’s very important and this is clear if you for instance look and you know what kind of species I’m referring to it’s human beings it’s us as as humans on this planet there’s just one figure that shows you how much we actually influence this planet this shows actually the accumulated anthropogenic Mass so the mass that was created by mankind over the last years and we have hit round about two or three years ago the point where we have created more mass as Humans Beings than there’s biomass on this planet and this is very interesting to me just as keep in mind as for the reminder for the rest of the talk that’s just one figure but there’s much more and it actually shows quite dramatically how let’s say since the 1950s we have started to really shape this planet in very different different ways so what I show you here is for instance how the population is taking off over the last decades right and this of course comes for instance with an increased water use this comes of course with increased Transportation so all of this is let’s say social trends societal trends at the same time there’s also consequences right and you can see these consequences so what you could see here is for instance how much loss of biodiversity is because we need more space as human beings right you see how much o are cified due to climate change you see how the surface temperature increases and there is probably 20 30 different factors that tell you how human activities influence this ecosystem dramatically and this is called the Great acceleration the great acceleration is that we actually really start to really influence and dramatically change Earth at unprecedented pace and probably the most important most relevant molecule that we are interested in that changes the planet dramatically is this small molecule here called carbon dioxide CO2 and I think we all agree that this is a very potent greenhouse gas and I think we also all know that over the last years decades actually centuries the amount of C2 has also been increasing steadily and this is known from the famous Keeling curve that’s basically recorded yearly how much CO2 is in the atmosphere and just to let you know this is not much it’s actually 400 420 parts per million this is round about 0.04% so in this room here it’s probably couple of bathtubs of CO2 that are finally dispersed that are hanging around in the atmosphere and we all know that these concentrations have increased and we all know that this increase of concentration of C2 have direct impact on climate change because they lead to this greenhouse gas effect which is amplifying and it’s typically the way we look at CO2 being a greenhouse gas being a threat to society to the planet in general but today you would like to turn your view around then it would tell you CO2 can also be a chance why but in principle it’s a simple carbon Source it’s a carbon source that sits there in the atmosphere and in principle we could and should be able to create all our materials that we need all our food all our you can see a lot of chairs made of wood in principle we should be able to capture the CO2 from the atmosphere in sustainable fashion and convert it into useful products and this would of course close the glob and carbon cycle this would immediately lead to a balanced interaction of us human beings with the planet but this sounds like a great Planet I think we Society scientists should be able to do that but the point is we are really bad at it at least in physical chemical terms so there is no single physical chemical process no single Catalyst would allow us to capture CO2 at a global scale sustainable fashion and produce products of our Everyday Use and in fact all our chemical technical efforts are still largely outcompeted by biology and this is sustainable CO2 phys we see here that’s the Green Jungle this is basically light driven carbon capture in a sustainable way so I’m telling you is that plants are very efficient CO2 filters and we actually use this technology already for centuries even for for more than thousands of years and this is just a picture back from maruk where I’m currently located at this is the Amun Borger beckon where you see that we actually can use plants to fix carbon dioxide agriculture right so we can actually make use of this fantastic blueprint which operates at the global scale so it’s fantastic so there a solution to fix CO2 with light converting into biomass we can use this technology and we typically think of plants as being really important figures on this planet that’s true but what also true is that there is also microorganisms and uh I’m happy that tus is sitting here because he loves microorganisms so do I but sometime gets forgotten is as round about 30 maybe even 40% of the carbon dioxide that’s fixed on this planet is fixed by these small microorganisms okay you just don’t see it because it happens very often in the soil and it happens even more often in the oceans and you need a microscope to see CO2 fixation by the way microbes invented CO2 fixation the chloroplast is nothing else than a formo bacterium so if you want to really talk to The Experts those are the experts and I show you one expert here it’s called proclus or CCO cus so those guys fix around about 50 gatons carbon dioxide per year and they produce of course a lot of oxygen and there’s calculations out there that every let’s say fifth or sixth molecule of oxygen you breathe in this room here is produced by these microorganisms so this tells you about the relevance of these small players in the large Global carbon cycle well there’s apparently solution it’s called photosynthesis and so we could say the problem is fixed well it’s not that easy because photosynthesis is sustainable yes it can do CO2 fixation and it actually does it at about 400 gatons of CO2 per year but the problem is that natural photosynthesis is not perfect and not sufficient for us human beings this becomes clear you look for instance at the way how we can capture CO2 through Agriculture and Forestry so there’s round about 10 billion tons of carbon which is round about 40 billion tons of CO2 that we can capture and convert annually through agriculture in a sustainable fashion that’s round about number we can do as human beings with this Old School Technology agriculture Forestry well at the same time we need more energy and we actually create this through releasing additional amounts of CO2 and we see it on the top of it we emit round about 10 10 10 billion CO2 per year from fossil fuels so the global carbon cycle is imbalanced and you can do the math if you want to really now balance This Global carbon cycle you have a growing world population you have more energy demands if you just would use natural photosynthesis to try to bring the back into shape you would need probably two to three Earth until 2050 all right so natural photosynthesis is there but it is not running at the pace that we would need and you could ask what limits actually natural photosynthesis so what is the limiting factor in CO2 fixation through plants or photosynthetic microorganisms and it’s a long-standing question and it boils all down at the end of the day to the actual enzyme the bioch Catalyst the engine that drives carbon dioxide fixation in the plant this is called rios5 bisphosphate oxygenase caroy or short rubisco and that is the Catalyst that grabs CO2 from the atmosphere and puts it into the metabolism of the plant I actually brought it here with me so for each person this room there’s 5 kilogram of Risco so this is probably only 800 grams but it tells how important and how relevant this enzyme is take look at it it’s everything you ate today was fixed to this enzyme and it’s fantastic Catalyst because it can grab atmospheric CO2 but it’s actually also lousy because it’s very slow so an average Risco takes probably 5 to 10 CO2 molecules per second which is surprisingly slow compared to other enzymes we know so it’s slow and there’s another problem the enzyme is also sloppy it can actually only fix CO2 it can also take oxygen from the atmosphere and that is really a downer right because if this happens there’s a process called photorespiration which takes part and the plant actually starts to release CO2 that has fixed before and this error is substantial so robiso has an a rate of round about 20% Which means every fifth time instead of fixing a CO2 robiso picks up the oxygen molecule and this is dramatic as I just told you so you could say the engine of photosynthesis is very slow and it also stutters now you could ask hey we have sbio we have prot in engineering can you make the enzy faster fter and in fact you can make the enzyme faster but then you automatically make the enzyme verse in terms of distinguishing oxygen from CO2 you can try to do the opposite you can try to make the en more specific but then automatically catalysis slows down what I’m telling you is that Risco is trapped in a so-called scientifically speaking perer Optimum where you cannot touch one parameter without negatively affecting the other one and you should think about catalysis really as these two things like speed and specificity and a good example might actually be you typing into a mobile phone you either can type very quickly but in automatic you do a lot of mistakes and the message will be really bad to read or you can try to type very accurately but then you are also very slow and this is exactly what limits catalysis at least in the case of Risco it hits a so-call parito Optimum and this has been really a starting point for our research when you when you really figured it’s really hard to improve this Catalyst why don’t we take a different step why don’t we just let rubisco be rubisco photosynthesis be photosynthesis and think about other ways new ways different ways complete Risco independent ways to fix carbon dioxide a process we we would call synthetic CO2 fixation and build those Pathways from scratch and hopefully hit a new Optimum now you could ask is is it too bold it’s billions of years of photosynthetic Evolution can you actually reinvent and find new ways to fix carbon dioxide and you might want to be humble and actually take a less from nature what I’m showing you here is the space or the biochemical solution space that nature has conquered already during the billions of years Evolution and what what might what you might not know is that besides photosynthesis or photosynthetic CO2 fixation Nature has actually already found other ways of CO2 fixation surprisingly or not surprisingly to me a lot in different microorganisms what you see here is that nature actually has already found six different solutions next to rubisco and that’s the solution that nature has conquered and as I said some of them we already use so I just showed you photosynthesis on the lights on forestry Agriculture and some of the microbial paths we actually also can use already at large scale for instance here shown these aceton methanogens which we can use for instance to convert Co carbon monoxide hydrogen and CO2 high concentrations of steel mills so nature offers different solutions that’s fantastic so me as a biologist I’m looking at at at this landscape and I think it’s really cool how creative Nature has been but me as a synthetic biologist I look at the very same landscape and I think H there’s still a lot of space there’s a little of void and a lot of things that nature has apparently not discovered or touched or maybe just not figured out and this is what we want to do we actually want to go into this empty space and want to build new Solutions new islands in this vast ocean of of possibilities and create those new solutions from scratch so it’s not metabolic Engineering in a sense they try to imp prove something existing it’s really trying to build something fundamentally new to give another example another picture think about how we humans Learn to Fly we actually studied Birds the biological example of flight we extracted the fundamental principles and then we build a plane but the plane we build does not flap it Wings neither does it have or nor does it have feathers but it flies and so that’s exactly what we’re trying to do we try to extract the fun Al principles of how nature captures CO2 converts it into biomass and how it’s powered by light and then we want to build a human-made solution that is as efficient does exactly the same thing but it might look very very differently so how are we going to do this in our lab we have basically laid away a workflow towards this synthetic CO2 fixation it starts all off with just design thinking about how an optimal C2 fixation pathway an optimal photosynthesis an optimal operation system of photosynthesis would look like we evaluate the different designs we then go ahead once we have a good design we try to find the parts and parts are biologically speaking enzymes proteins biocatalysts we put the system together try if it runs try to optimize it in in in different rounds and once you have really robustly operating system we would like to move it inside of a cell and this could be a natural cell or an artificial cell and I will talk about about both of it later in my talk and of course at some point we actually might want to go away from single cells microbes or photosynthetic microorganisms to higher organisms and the Big Goal would of course be also to touch the chloroplast in the plant at the end of the day all right so how do you design these new Cycles the first step how do you think about photosynthesis really radically new what you do is a principle that I have learned as a chemist I’m chemist and a biologist it’s called metabolic radiosynthesis so sit down you start to think and draw potential transformations of metabolites and these are a couple of Cycles we’ve drawn a couple of Cycles we already built in the lab and uh all of these Cycles feature very different reactions important to note is just Theory could just draw it here on this Blackboard that’s actually what we do we sometimes draw these cycles and then we think about those Cycles which of them is a good design or not don’t memorize the different chemicals in there because all of these Cycles are very different in terms of topology and chemistry but they also have a couple of common principles and one principle is that all of these Cycles are as I said Cycles so these pathways are cyclic which means they can go in rounds and rounds and rounds and it can continuously fix CO2 that’s one principle so a cycle to continuous to fix CO2 and the second principle is that all of the cycle so called split reactions reaction where can take off some of the fixed carbon and convert it into a building block building block of life or building block of chemistry now there’s different solutions I showed you catch cycle hopc cycle Theta cycle probably 10 or 20 more Cycles we’ve drawn the question is which of these Cycles is actually a good solution so how do you evaluate your designs and this comes back to chemistry so what we do is we basically have different simple parameters where we look at what would be helpful for a good design and one of course important parameter is kinetics in other words speed so the question how fast can you turn such a cycle and I give you one example for the catch cycle this catch cycle is basically not based on Risco as I said before because it’s a really slow Catalyst this catch cycle is based on a new principle of CO2 fixation is C discovered around about 15 years ago in microorganism it’s called anoa reductases ecrs and those are highly efficient C2 fixing enzymes and you can actually see them that’s a simulation we’ve done with Partners in Chile how good they are in capturing the CO2 from the atmosphere and making a cc Bond directly so that’s the active side of this Catalyst and they are actually 10 to 20 times faster than rubisco which means at the same time you can fix 10 to 20 times more CO2 which is increased space time yield so kinetics matter speed matters but what also matters is thermodynamics or in other words energetic efficiency you want to drive the Porsche fast but you also don’t want to use a lot of gasoline right so exactly that’s what we’re also looking at so how much ATP how much NP do you need to actually operate these Cycles the catch cycle is a good design because the C2 fixation should be 10 to 20 times faster and it only needs half of the ATP that you would need as a naal photosynthesis so this looks like a really great design now comes the more important part and the more exciting and the more challenging part which is you need to go back to the lab and you want to build this system so the evaluation design takes you probably one to two weeks the actual challenge is to find the individual Parts the enzymes the proteins to build the catch cycle where do you find these parts there’s not a shop where I can simply drill it or it can simply make it and I think we are far away from designing enzymes from scratch although we started to do that as a as a as a as a as a scientific Community but what we typically turn to our attention is microorganisms because they are just so diverse in terms of biochemistry and they are so different that you would probably find for each enzyme that you intend to build and to use in your cycle an equivalent in nature just to give you one reference if you have one gram of soil and on this on on this spoon it’s probably seven to eight grams of soil You’ have one billion of these microbes that’s a fantastic diversity look at this spoon this is basically a whole world population on one spoon and there is different species in there and this is a picture that shows you how diverse these microorganisms are they come in very different flavors and every microb has around about in average probably 4,000 th genes we just know half of the of them of their functions and each microb also has unique genes so this biodiversity is just mindblowing to me and this is what we’re trying to do we just screen for the individual parts that we need to build the system through these microbes or we talk to colleagues like for instance torson Masha or for friends to to M Roa who have all these things explored in their favorite micro orms and we sometimes take the parts from them to make a long story short so we had this challenge of finding enzymes to build the first version of The Catch cycle and we actually found those in very different organisms for instance in Racor it’s a classical photosynthetic microorganism that is in the lab we just sneaked and and stole a couple of genes or or proteins from eoli probably also well known as a gut bacterium we took one gene from we actually use genes from methylobacterium it’s an organism we find on the plant surface each plant has methylobacterium on its top on its leaves n pulus which Aon different domain of life we also took some genes we actually had a gene from the human liver it’s not the microorganism I know and we also have even genes from clri which is very close to the basil that torson is working with and we even engineered and took one one enzyme from arabidopsis talana a small plant that we re-engineered to do the reaction we wanted to do so it’s a patchwork pathway but we put all these enzymes together in one endorf cup we actually added ATP NPH the energy chemical energy we could show that the cycle really starts off turning and it starts to convert carbon di oxide into malate malic acid which is a fantastic building block for sales but also for chemical industry so what I was telling you is that we really can think of a new operating system a complete new pathway system for CO2 fixation we can go in the lab we can build the pathway and the pathway operates so this is synthetic biology and we were really super happy to have built this first version until we looked at the kinetics at the speed and we actually realized this cycle is awfully bad and it turns and this is something we faced now several times by building different solutions we very often build something that is somehow functioning somehow doing its job but it’s far away from the optimum it’s very slow it falls apart it’s grappy it’s almost like you start to climb a mountain you’re at the base camp but you still have way to go to hit the top and this exactly what we have to do typically in the lab when we do s bio we have to optimize to improve to build a second a third a fourth a fifth version but why have you to do that so why is the system not better well the point is that all of these enzymes are put together from very different biological backgrounds they don’t have a common history and very often things go wrong and the cycle stops because one enzyme steals a substrate of the other enzyme one enzyme is not specific and branches of a metabol should actually turn in the cycle or just have site and waste products that fall off the cycle so what I’m telling you is even though you think about all the individual enzymes as individual players you try to put a really nice team together the team is not not as good as you think and just because we have European championships I brought a good example so you know it’s not just about thinking each individual player as getting the best person in the team you really need to think about the team being the best team of players what I’m telling you is it’s not an optimization function on the individual Parts but of the whole network of the whole system it’s a different challenge so what can we do how can we improve the system but in principle you can train some enzymes to become better you can train some enzymes to do the right reaction and you ignore the other reaction to hit the gold better that’s one OPP possibility and it’s actually what we did we engineered and redesigned certain enzymes to become better in doing one of the other reaction you can substitute enzymes right you can just swap in or swap out enzymes so that you get a better playe team together and in fact we actually have detoured one reaction you see it here because the original enzyme wasn’t good we substituted it right so that’s also possible and then a team is more than its players you also can actually put in more enzymes supporting stuff that keep primary metabolism up and running and I think that’s a principle that’s very different to a chemical physical audience we build a perfect digital part biy is messy it will always make mistakes biological systems are robust because of additional reactions ongoing in your cells you have primary metabolism but you also have of course a lot of cleanup enzymes that remove toxic metabolites this detoxify things and if you want to build a robust system we actually make it robust by adding such additional support supporting helper enzymes and I think that’s a fundamental difference in building synthetic systems compared to chemical and physical systems okay if you implement all of these things you change Al the turf a little bit you actually see that can improve the catch cycle in its operation by almost 20 fold without changing its topology or its basic chemistry so you go through all these versions version 5.4 it’s already very good it’s much better than the original version it’s at the stage where you are if you take a plant leaf you break the plant leaf and you try to measure CO2 fixation so so this is probably really where we are this slide I’m showing you here took us probably two to three years so that’s just summarizing the work you have to put in to optimize such a system and I’m really glad that Thomas schwander the first author took so much energy to try to optimize and I guess you can also appreciate it’s so complex a system sometimes you don’t understand what’s ongoing and it’s too complicated to oversee all the interactions and this is what has inspired us the last two or three years to think about can we not go a different way in systematically explore the optimum use other ways and uh there it comes into play that it’s important to also have Technologies at hand and one thing we’ve started to do is now pipeting and putting different versions of the cycle together where systematically vary the content the concentration of the individual enzymes and we actually started to go away from the manual labor in the work in in the lab so no cell-based manual work anymore we actually start to use Automation and miniaturization so for instance we have this pipeting robot here which uses ultrasonic sound to move liquids just across no Piper tips no plastic waste fto Pico leaders that are just mooved from A to B with ultrasound contact free super quick and probably more error free than a PhD candidate in our lab I’m sorry to say so the point is this opens up of course paralyzation and allows you to do a lot of different variants of the cycle so you can work in 10 microliters now 384 wells in parallel maybe 1500 Wells and you can do a lot of more essays you can analyze those essays and you can feed it back into a learning algorithm that tries to predict again which of these uh parameters you have changed are important and suggests a new round of experiments if you apply this design buil test learn cycle it’s a typical thing you do in sbio you actually see that within eight rounds of optimization you shoot up by a factor of 60 compared to the first version you have built and this took us only half a year so this gives you rough an idea how fast you can actually speed up if you can generate larger datas and if you actually have the help for instance through machine learning algorithms now where are we now well where stand we for instance compared to chemistry so if you to think about chemistry as general as a field you typically start off of crude oil you start off of uh from from fossil fuels as a resource to do chemistries to do Transformations and chemist you can do one or two Transformations parallel then you have to support to extract your product move into the reaction vessel do the next reaction in a stepwise process you can build up your molecule now the catch cycle is actually different in a sense that it’s 10 to 10 times better in orbe 20 times better because you can already operate 10 to 20 reactions in parallel and it can start off CO2 to make a small organic acid in this case called glyoxalate you can also make malate we can adopt a little bit what the outcome is but overall you basically have now an A system that can do many more reactions in parall factor of 10 more than chemist would be able to do and it can start off CO2 now can we convert this clate in more interesting compounds can you for instance make antibiotic precursors amino acids or Food flavors from it in fact you can do that we’ve done this already and just one slide I don’t want to dive into the details just want to give you one example how we were actually able to produce the antibiotic precursor aryin for more concrete six step which is the precursor without the sugar Moy attached by coupling more than 50 reactions to each other and it’s not only coupling the reactions again you have to optimize you have to finely tune the enzymes and you have to design new Pathways to make all this happening but in principle it’s possible so I told you already how good we are and of course we are proud of it right and we also are really proud that the system has started to work after 10 years of uh of let’s say laboratory Evolution human design but of course we are still far away from what cells can deliver if you think about cells like TOS is working with right living cells they can operate a thousand maybe 2,000 different reactions in parallel that is just amazing to me how you can control in time and space all this different reactivities and so this inspired us to think even Beyond this 50 reaction think about more in the context of a living cell that’s also something you doing in Dron for instance the physics of life you start to extract those principles how life operates and it’s more than just chemistry think about the chloroplast or photosynthetic M organism it is light powered so it can convert energy that comes from the outside it can actually do its metabolism to sustain and to grow and to divide and it can self-replicate it can self optimize so those are all the function functionalities that such a system offers and it’s far away from the small enzymes that we have in our in in our endorf cup and so the idea would be can we actually now also power the system instead of providing ATP and dph can we use light or other sources of energy can we structure it in three dimensions can you eventually also put information in in there can we actually get a self- sustaining system and these are all the things we are currently exploring in our lab so maybe the first question can we simply power the system with light so can we take the synthetic catch cycle we’ve designed in the lab and can we just couple it to the native photosynthetic Machinery so can we for instance take spinach that we buy in Market in marook on the market can we grind up the spinach extract the photosynthetic membranes and shine light on photosynthetic membranes and use the ATP and NPH to actually power this synthetic catch cycle and interesting this is not possible right that’s very interesting to us so what we figured is that the catch cycle synthetic design and the Natural Energy Machinery of the photosynthetic cell cannot talk to each other what you have to do is you have to go back and find the bottlenecks the road blocks you have to swap out one or two enzymes and only then those two systems operate with each other what I’m telling you what I’m telling you is that even in cells individual modules energy regeneration metabolism do not evolve independent of each other they are finely tuned and synchronized with each other stochiometry is play a role or chemical chemistries toxicity all right okay so this is just about powering such system but you can even think bigger and broader think about building eventually also a an envelope an artificial cell around this metabolic core this metabolic heart this metabolic operating system so how do you do that well if you think about cells at the end of the day it’s nothing else than water droplets surrounded by an oil or membrane and again hanging around in water it’s nothing else than a vinegret if you want to say so right so it’s nothing else than small droplets water in oil in water droplets or water in oil Min droplets that you can actually create if you for instance mix vinegar together with water so of course we don’t do that to make these small droplets what we use is a technology called microfic and this allows you to actually produce thousands of small droplets water and oil droplets per second and you can use these micropets as empty cells if you want to say so cell envelopes and you can start to load those envelopes with different Technologies and techniques and you can for instance put in photos cenic membranes can for instance also put in uh put in the enzymes of the catch cycle and then you end up at something that we would call an artificial chloroplast A system that contains this uh these uh these photosinc membranes shown in green it is like cell size bit bigger than a yeast and if you shine light on this system it actually starts to power up and it starts to generate energy and what you see in blue is ndph if you turn off the light the ndph can be used to fix carbon dioxide so you can really charge decharge charge decharge charge decharge the system round about for two hours and it can fix CO2 and you control this activity in time and space just like a real chloroplast all right so I told you already a little bit how we now move away from this natural system towards a blended system where you have some natural Parts some synthetic parts and we have this artificial chloroplast it’s not very stable but it actually operates and you can control it it’s like a small cell cell like organel but of course you can think bigger especially a place like here where you have Also let’s say hardwired physics or Material Sciences you could think of moving up this ladder towards a bit more hardwired World think about where we can get energy from we can for instance use electricity and would it be actually possible to interface biochemistry with electricity would it be possible to power CO2 fixation directly with electricity it is something we’ve also started to think about recently so what you need to run the cycle is on the oneand reduxx reactions you can directly use electrons but you also need ATP and ATP is the one compound in your body in every living system that provides the cell with energy so if you move around you need a lot of ATP every person in room probably has 70 kilograms of ATP synthesized per day just by walking around and being being happily happy human being so the question is how can you generate ATP can you generate ATP uh in in in in a different way than nature does it because nature does it in a very complicated fashion it actually uses charge separation at membranes and many many different complexes so nature creates a chemo electrical gradient across the membrane and this gradient is harvested by ATP synthase a rotating protein complex that then produces the ATP I don’t dare to build such a system this is way too complicated for me but I can think a synthetic biologist again I can think think is there other ways could we directly interface for instance proteins with electricity or with the technical electrical world can we use a minimal set of components maybe no membranes at all and can we actually use this as a core an energy module to power chemical reactions and what we found is actually a way how to do that and this is shown here it’s a c protein called a it’s a if you want to say so an adapter between the biological and the chemical or technical electrical world is a protein which can accept electrons low potential electrons and the protein has iron sulfur clusters in inside and those are Su are like wires Nano wires that can accept electrons and the electrons can move from the outside all the way to the reactive center of the protein and this is where chemistry happens and you can for instance reduce an acid to an alahh that’s is pretty hard typically for an enzyme without activation but if you have low potential electrons this enzyme is exactly in very efficient add it and what you can do now is you can basically put the enzy on an electrode you can feed it with electricity it would reduce acids to alahh and then you can close a cycle by just oxidizing again alahh to acids and this process you can release ATP and you can use this ATP for instance to make starch out of glucose or starch precur out of glucose you can even power transcription translation systems in vro with this system here what I’m telling you is this is probably one of the most exciting discoveries and and designs for me that you really can use electricity to power and produce the the molecule of life from uh the the power molecule of the cell directed from electricity all right I talked a lot about artificial cells because that’s also a strong part here in in d uh this kind of bottom up construction of minimal systems to some extent but what about natural cells so can we for instance take the whole operating system instead of building an complicated cell around this metabolic operating system can we for instance simply take this operating system and bring it back in the context of a natural cell so can we take the catch the Theta the HC cycle for that matter and can we boot this software inside of this billion years old hardware and you might appreciate that this is a big challenge because cells have billions of your Evolution and the question is are this able to learn new things it’s like as human beings are are you able to learn new language are those cells able to learn a new operating system system and how you going to do this so what we do is we basically create incentives for the cell we go into the cell metabolism we look what the cell needs and we actually create dependencies let’s take a simple cell like eoli that needs for instance a compound shown in yellow which is called saqu that’s a chemical a chemical compound a metabolite that Eola needs to make a cell wall or Lysine for instance now just block every pathway that synthesizes suxin COA just stop Metabolism from producing this compound the cell is dead it cannot grow anymore that’s a strong or it’s a killer basically it’s if you knock out all these genes so the cell is dead until you provide the cell with a new way to actually make this compound and this is what we’ve been doing we actually brought in parts of the catch cycle not the full catch cycle but part of the reaction sequence and it can actually produce now it sucks Nua through CO2 fixation starting from crotonate and the cell can start to grow again and that’s one of the first examples how you can actually put part of the catch cycle inside of a living cell and it actually fixes C2 and provides the cell with some biomass now that’s a starting point to do real Evolution it’s also a starting point to not only make lysine maybe or the cell wall from CO2 but eventually make the whole biomass from CO2 and that’s the way we actually want to use now in the future living cells to optimize our systems and to get them onboarded into into into the back ground of natural cells okay so what I’ve been showing you doesn’t save the world from from climate change it’s too small of droplets or too fragile or far away from being already fixing so much CO2 that you can actually make a difference to this planet but we actually have started to think how to build such system and we’ve actually shown you can build new solutions to the to the space and that’s the mod of the max Blanc society which says inside M preced application so aspiration is not necessary to have it in two or three years we think just broader and more Visionary hopefully and this also keeps us doing these things in the lab if I have another five minutes I would just try to also tell you these instan we’ve gained can tell us also about real world solutions that we can create now and for that I want to come back to the initial slide where I talked about how bad photosynthesis is and about the process of photorespiration that is actually limiting rubisco and I told you about this Vision to really reinvent photosynthesis or C2 fixation build a complete new to Nature solution it’s almost like you would have a diesel car a diesel engine and then you would build an electrical Car electrical engine so a completely different design but think about what you can also do in synthetic bi you can for instance help to boost the diesel engine you can make a turbo diesel you can basically try to boost photosynthesis not replacing it but using the same principles and just try to improve photosynthesis and improve photorespiration and this is something we can do already now so what does it look like that’s again a bit more technical I’m sorry for that this is how nature fixes carbon dioxide naturally photos photosynthesis it’s a Calvin cycle Risco is involved and Risco fires and Risco misfires and it makes this compound called two phosphoglycolate which is a photorespiration starting point this is toxic to the cell needs to be detoxified which requires a lot of energy which takes many many different steps which releases ammonia and more importantly as I said before which releases CO2 at Large scale and this is limiting photosynthesis to some extent now take again our thinking of a synthetic biologist think about this awfully complicated pathway that even loses CO2 on the way of mitigating photo respiration how would I design that as a as a human being or as a synthetic biologist I think we should not spend much much energy to regenerate this molecule to phosphoglycolate I think we should have a short pathway sequence and I think we should also not release CO2 and this is what has inspired us to think about a new pathway in photo respiration that starts off converting two phosphoglycolate into glycolate activate this compound through cooo sorry for the chemistry then you actually fix CO2 you don’t lose CO2 you fix CO2 to make a compound that’s called tcoa and that’s how we named the pathway taco t pathway you need two more steps and you’re back in the Calvin cycle it’s simply four steps much shorter it’s probably the shortest possible sequence to regenerate two phosphoglycolate and it needs just a minimum amount of energy this is shown here on this slide how many steps how many rred how many ATP you need it’s on the left lower side just compared natural photosynthesis should be a super short and super energy efficient pathway very cool the problem is each of these individual reactions does not exist in nature and the most challenging one is actually this C 2 fixing step it’s an enzyme that we didn’t know before all right so how do we go about that what we do typically is we screen we screen enzymes that can fix CO2 and we try to find an enzyme that shows a little bit of activity and this is shown here so we want a carox glycol COA and we found a propy COA carox that could accept glycol we solved the structure and we look at the Active side and that’s a starting point for us if you have the active side you can shape the active side so that your real substrate fits much better in so really take you take it by the heart in synthetic B by reineer on the active side of the enzyme and just to give an impression I think we had something about 0.1 turnovers per second okay really really almost nothing to measure in the we had to measure overnight to see activity of the enzyme but once you have the structure once you start to redesign the enzyme you can actually start to improve it was around about probably five six seven eight mutations we had to introduce so this is an old slide we have probably tested around about 20,000 variants we have a mixture of engineering by looking rational design and also just High throughput screens of random M mutants and what you see here is we actually improved the enzyme B about factor of 500 and we increase also its energetic efficiency and reduce it by about about 30 fold so what I’m telling you is you can actually create a new to Nature enzyme that has parameters that are comparable to existing carox lases we did it also for the other enzymes this was the biggest challenge and we could put this pathway together and we actually managed to get this path already into plants this was with the help of Andreas vber dorf and what we see right now is that these plants seem to grow better especially under ambient C2 fixation where the photorespiration is a challenge and they seeming to grow a little bit better still on the high CO2 concentrations so we have an interesting phenotype we still have to confirm if this is AC or if this is other stuffff that’s happened when you introduce it into plants but at least that’s one of the first examples where we have artificial pathway complete new to Nature design and you can even go inside of a more complex organism I think that’s the future for us at least in the lab all right so that’s the almost the end of my presentation I hope I could show you how we started off just by thinking about photorespiration photosynthesis and then we actually designed new Solutions we moved from the lab individual enzymes to path that we can operate in vitro optimize with machine learning automation then go into cellular environments natural artificial cells and in some cases even into chloroplast what we still miss are of course principles how to organize in three in in in three dimensions in time and space principles that for instance you also investigate in D like liquid face separation or scaffolding we also miss a lot of tools to turn on this pathes regulatory elements that’s something also torson is doing but we probably also need in the future we want to build complex designs complex regulations Cascades circuits so all of this still we need to do and we need to of course also optimize our ways how to bring stuff into the chloroplast but we at least have ideas that there are solutions out there it’s worthwhile to develop these Technologies what I also told you is today that the climate crisis is a global Challenge and there will be not one single Silver Bullet you need to think in different aspects of your life Transportation energy carbon capture and for each of these individual parts will be IND idual different solutions It Won’t Be One Carpet it will be many different solutions but I believe that SBO also really has the potential to offer new Solutions on the biology side because we also need biology to address climate change and we also need probably synthetic biology because natural photosynthesis is not sufficient I also told you that we actually use genetically modified organisms so we have to talk about that we have to go out and we have to talk to the to the uh to the public and we you discuss risks but also benefits you have to trade you have to trade sometimes if you want to save the climate or if you are afraid of genetic modified organisms and we have a lot of experience we have a lot of experience with GMOs so there might be solution but of course we need to convince politicians and we to convince also society that there is a safe technology maybe or to be more precise that each technology has its certain risks AI also has its risk a car has a risk even if I walk up the stairs that’s risk right you need to be able to balance those risks you need to make fact based decisions and last but not least I think it’s also important to involve younger generation and what I’m really happy about is that some stuff that we have designed in the Labis already in certain textbooks and this provides you with a great opportunity to provide especially young school kids within Vision that science can provide new Solutions as much as we were able to overcome the corona pandemic with I would say synthetic biology some of the nuclear tides were synthetic right so here is also new technology we might actually be able to use and the world is not necessarily Gray I think if we wrap ourselves together we can show again that there is technology can help us to overcome this challenge which is a global challenge all right last but not least I think you’re not politicians I can quickly glance over it but I think there’s also a lot of potential in s bio maybe it would be wise to think also strategically as a new field as torsson just phrased it chemistry analytic synthetic the last century was the synthetic chemistry it has brought a lot of wealth to Germany think about all the big companies BSF buyer right what we have built on synthetic chemistry maybe we can also build things on synthetic biology and the markets are there and they will be there and if it’s only 1% of the predicted Market which is 44 450 billion euros not too bad all right so that’s the end of my talk I would like to thank you again for inviting me I hope I could elucidate a little bit I hope I didn’t uh I didn’t disappoint you that I didn’t bring uh the super machine that saved the plant from global warming but it could give hopefully some inspiration that biology is cool to do and that technical biology is also something very cool thank you so much for your attention of course I have to thank a great team I highlighted only couple of people doing my talk it’s a fantastic team a national team and it’s a lot of different uh different disciplines that come together in our lab I’m of course grateful for the funding by the maxl society but also different funding sources and of course my colleagues have been collaborating with I thank you for your attention and I’m happy to answer questions thank you ah okay yeah thank you so much for this very exciting talk so the future is open um do you have questions yeah first of all thank you for your talk very interesting and also motivating um I have a question about the workflow when you talked about the Retro syn synthesis because I was wondering how did you yeah made up the reactions when you after that you searched for the enzymes so how do you know that these reactions you created on scratch really exist because it I mean it’s more work to do if you yeah you know what I mean I think right so the absolutely correct so I mean we not just hoping in in in in blank space of course we have starting points and I I think the the catch cycle for inst was built around CCR and we know what’s what reactions CCR or ECR does so it takes croa and it makes e so this was a block we knew this reaction existed it was highly efficient and then we build around the cycle the taco pathway is different the tath starts off with a concrete metabolite and you want to go back to something and there you need to design a reaction sequence so that is a different kind of design so we have also ways how to look and design things as I said so either you start from a very good CO2 fixing reaction you build the cycle around it or you start from a from a substrate and want to go to a product and then you start to find the enzymes in one case it’s easier right catch cycle is easier because you have the C2 fixing reaction at hand Taco cycle has a different challenge right but you’re absolutely right that’s the way we look at things so yeah okay yeah that works um you mentioned that enzyme that uh you can interface with um electronic circuits um so what is is this something that only this enzyme can do and if yes why can it do that and of course not I mean a lot of enzymes that do have uh they do have um Iron sulfur cluster you can feed from the outside right you know this better than I do so it’s all about the chain of of iron sulfur clusters inside of the protein right so you can for instance every protein has a fer toxin which is a small protein that can be loaded with iron can accept electrons can do it the challenge for us was here how to create an enzyme that can generate ATP right and so this is a specific enzyme because it can reduce an acid to an alahh and use very low potential electrons and has so much energy and power that you can actually create an an alahh from an acid if you go back you can release one ndh one and one ATP so this is where we go there’s thousands of other enzymes that can simply use for instance electrons just to do simple reduxx reaction but here you need to create such a big electric energy difference that you are sufficient enough to make an ATP which is a lot and the electrons come from okay so you can use yeah yeah fair enough so what we did in the beginning is we simply took for instance uh titanium citate to feed the electrons from from the outside just to test if the cycle is operating and then we adopted and put the put the put the put the uh the protein on the electrode directly and it can actually directly accept it you can actually use the same principle to make and regenerate ndph so you can for instance take uh it’s called fnr ferrodoxin Uh NPH reductase you can also put this enzyme on the electrode it will reduce n and this n can float around in the in the in the environment to to actually be used by enzymes so this kind of electrochemistry is prototyped but I think challenge here was really to make ATP would have another question but no you can you can ask questions that’s fine my train doesn’t leave so okay I quickly keep going um completely different question I mean you alluded to that at the very end that um structural organization is of course key in any system in by in the cell um so how do how much so far you don’t really take care of it it seems like and do you think this is is really something to invest in and especially if you think about nature does that does that many algae use a carbon concentrating mechanism where really funnel a CO2 directly to the to the Risco to avoid this uh competitor oxygen fantastic question so is there something you’re working on in this direction I’m not ignoring it I’m just incapable of doing it so this tells you how good synthetic biology is I mean so the the point is what you see here I just pulled off the slide here look at the glow blast you know have this membrane Stacks you have them Parts where the where starch is stored and you know one trick of cells is to separate chemistry which are not compatible of each other and we have not used this principle necessarily yet we have started to investigate look at face separation for instance to store some stuff inside of of let’s say an own content say to do certain chemistries there we also think about maybe equipping the membranes we have with channels to let only certain things through and really if you think about a cell you said is correct think about a UK carotic cell a bit more complicated right so you do have the nucleus Source information you have of course the mitochondria to make the ATP of the Chlor you have the chloroplast to fix the carbon and it’s also Source sync you know you can push a lot of more through if you create a source sync principle so all of this is not implemented and this I need the help of my colleagues I need the help of cell biologists for instance right to tell me how to structure complexes or create metabolons enzymes that have active sides facing facing each others and part of the of of of of of the of the success or not success of Building Systems is because we still have already too many chemistries interacting with each other and I would love to actually build cages who would love to actually separate certain chemistries from each other but this will take another 10 15 years and maybe this is a direct transition to my question which is of course I mean what you achieved is is is massive but you still you’re not in the cell really right so I mean what what do you what do you see as as the challenges or where do I mean you mentioned two or three already but what are the the challenges you you’re going for in terms of moving these reaction moving these Cycles really in whatever system that’s a good question so if I think about solution I think in waves the first wave is just Bo photosynthesis I think that’s doable and I think we have good chance the bigger challenge is to get these more complex systems up and running um in Eola certain modules work but ecola is not a chloroplast and to jump from from Eola into a a plant is also just crazy so what we’ve been really working on is using unic algae and to build up a clor blast toolbox and uh I really think this is something that that will be helpful because you have a small mic organism division Times short generation times can multiply paralyze and uh then of course we need genetic regulation right to address and to express things in the chloroplast so we have a basic toolbox now that we can use clonas but it would be nice to have orthogonal parts for instance right all of this regulation and so it’s again a mixture of building up Technologies multiply paralyze do all these transformations in in in in the unicellular algae maintaining this algae but at the same time also having the genetic tools right so you know this is again like why should it care about circuits if you don’t have something to to you’re aiming for to develop these complicated circuits and why should I build metabolisms if I don’t have any genetic regulation at hand and I think this is where we meet again each other I think this is the next step tools combined with Pathways and the more complicated it gets the more complicated will be and I’m not sure for instance if orthogonality is important or not um we’ve seen seen this in cells orthogonality can be can be a liability or it can be an asset you never know what kind of effect the one thing has or doesn’t have right so all of these things are super interesting for me but also so hard to address and this is also why we actually sign so many cycles because some of them will just not work in eoli or will just not work in the chloroplast because we have overseen that one pabol will not be functional right so this is why we also think about this more parallelization of path is yeah so mic micro algae is currently your preferred Direction not like cyano bacteria to go to the bacterial synthetic organism yeah currently it’s it’s it’s it’s clamy for us and we have a little bit of cacas in the lab and but the point also is that we have of course a great environment Felix won is now here in maror who has a lot of expertise so it also depends on your environment what kind of organism you’re working with I you know better than I do right yeah yeah thank you also uh from my side question from a former biologist I might say I used to be one not a microbiologist so when you’re talking about like boosting uh photosynthesis how about adopting or adapting photosynthesis because there was a recent paper in nature I’m not sure if you’re aware that in the canopy of the Amazon uh it’s becoming so hot that photosynthesis stops working so bold question can you do can you engineer something that photosynthesis can also work in like hotter conditions well micro orgasms solved the challenge already I think because you have thermophilic algae for instance or micro algae cob bacteria um you know it’s it’s it’s it’s it’s a multifaceted thing so uh of course it will happen automatically but you know the speed how this happens is probably decisive Factor right and you always trade something um so so my answer would be for the wild I mean I don’t dream of releasing organisms to the environment is probably not what’s going to happen and but it’s interesting for me to dive back into into this challenge of what does it need to adopt right and studying it and you can look at it and trying to to see how over 10 20 years things will evolve or you can try to do it in the lab so as a matter of fact we also touch again a little bit robiso from different reasons we should go back in time because Risco is a very interesting historical angle which is the enzyme started off by the way I need to get it back later on it started off a billion years of evolution without any oxygen and only then oxygen came so now if you replay if you rewake and re resurrect billion year old riscos which we’ have done now in the lab together with evolutionary biologist and directly confront the enzyme with oxygen maybe you can redirect The evolutionary evolutionary path of in way of it and so these are things we are currently doing and I think that’s also again like it’s a mixture of sbio but also looking back into what hobi has solved and which path it is taken it’s more like very often you think about B being creative finding new Solutions but the older thing gets the more complex it is and the hard it gets to go back in time it’s like bureaucracy at universities so they are just too complex to you know and this tend to grow and to become even more complex because every person has a certain certain function in there and I think that’s what you see if you see a protein which is two billion years old it’s hard to go back without losing function there certain interconnections which are hard to to do and so you learn something if you break it down into easier parts and you evolve it again I think that would be one solution to but again it doesn’t help right now but it gives an idea of what the landscape is and what path up blocked or open thank you another question there have you only focused on uh creating artificial pathway cycles and if you do so what is again the advantage of only focusing on cycles and not yeah on a linear pathway creating an artificial linear pathway yeah that’s a very good question so the the the point is that you need harder chemistries to build carbon molecules if if you have a cyc as as follows I will actually I can use a Blackboard it’s cool want to do it here chalk fantastic always wanted to do that you know if you get carbon in for that matter need carbon is always red for me you throw it onto a compound to make a cc c bond okay and then you kind of reduce it along and then you can take the fixed carb out so that’s a chemistry where you make a cc Bond first and then it’s easier to handle this molecule if you think about linear Pathways you need to reduce a CO2 molecule which is much harder and very often requires anerobic conditions not good here in this atmosphere okay so this is why we actually think about Cycles about these carbon cycl because it is much more compatible with uh with uh with with transformation of CO2 easy way otherwise you can of course reduce right then you get it up here but that is problematic under C conditions and you still need to make a c on performation do them later on but in principle it’s about this anerobic and the investment of the energy you have to do does it make sense so maybe I asked the last questions um sure how about uh computational modeling would this help speed up um your experiments uh just to predict or build like a digital twin of um a chlorop plas that is also very good question so I want answer in two or three different layers um even pathway design now can be automated so there is tools that just browse through all the enzymes listed 40,000 different enzymes and it’s actually automated automated pathway pathway generation software this one lacks still the enzymes that think of could Bridge certain certain metabolites okay wow party starts second point is digital twins perfect but what we for instance lack still is a great understanding of concentrations you know compartmentalization just as we talked about if you break a cell you get let’s say .5 Millar COA all across but within the chloroplast it’s probably you know so we actually lack ways to really get the information we need as Biers and that’s again and maybe the last the last important thing we think also about enzymes doing Transformations a Tob but of course enzymes are also taught and trained to not do other reactions and typically enzymologists tend to measure the one reaction they want to measure but all the others things not and this also makes it hard to predict how system will react I think this is our that we have to face a digital twin would be perfect the cool thing in B is you can Model A lot if you take the right parameters but if it’s then predictive very often not thank you so much last question do condensates be play a role here that um compartmentalizes so that that is that is something we are keen on doing we actually also played around a little bit with doing that and I think that we be part of the solution would be also to have very interesting com condensates as as as a way to structure certain parts of metabolism or to concentrate certain metabolites uh we have already tapped a bit into rubisco and con condensates right and we actually see that they speed up for instance C2 fixation so there is cool things that we would like to explore in the future maybe with th I’m not sure yeah okay there are obviously no further questions thank you very much again for this exciting talk and inspiring talk and uh we hope um we will see you again here thank you thank [Applause] you e