{"version":"1.0.0","segments":[{"speaker":null,"startTime":1.916,"endTime":3.558,"body":"Kevin: Welcome to Innovate and React."},{"speaker":null,"startTime":4.096,"endTime":8.056,"body":"You might have heard about bacteriamaking chemicals before like"},{"speaker":null,"startTime":8.318,"endTime":13.957,"body":"insulin, but probably less abouttypical fine chemicals or building"},{"speaker":null,"startTime":13.997,"endTime":15.617,"body":"blocks for the chemical industry."},{"speaker":null,"startTime":16.457,"endTime":22.078,"body":"So this episode will be about howmicroorganisms and biotechnologies can"},{"speaker":null,"startTime":22.397,"endTime":24.897,"body":"turn methanol into value chemicals."},{"speaker":null,"startTime":25.377,"endTime":29.115,"body":"I'm very excited to have Dr.Ronja Rappoldt, the co-founder of"},{"speaker":null,"startTime":29.115,"endTime":30.835,"body":"TENO Bioworks on the show today."},{"speaker":null,"startTime":31.216,"endTime":31.615,"body":"Hi Ronja."},{"speaker":null,"startTime":31.876,"endTime":32.495,"body":"Thanks for joining me."},{"speaker":null,"startTime":33.431,"endTime":34.831,"body":"Ronja Rappold: Hi Kevin,thanks for having me."},{"speaker":null,"startTime":35.531,"endTime":40.731,"body":"Kevin: ja, maybe let's start with whatbrought you to research in biotechnologies"},{"speaker":null,"startTime":41.561,"endTime":44.481,"body":"Ronja Rappold: Yeah, actually,uh, that goes back to high school"},{"speaker":null,"startTime":44.601,"endTime":49.021,"body":"times, like when you are at thisdecision of what you wanna study."},{"speaker":null,"startTime":49.261,"endTime":52.962,"body":"And back then I really lovedall natural sciences, so"},{"speaker":null,"startTime":53.061,"endTime":55.081,"body":"biology, chemistry, and physics."},{"speaker":null,"startTime":55.821,"endTime":58.681,"body":"Um, so I really looked fora university program where I"},{"speaker":null,"startTime":58.722,"endTime":61.142,"body":"could more in all three of them."},{"speaker":null,"startTime":61.941,"endTime":67.302,"body":"Uh, and then I found biotechnology, morespecifically molecular biotechnology."},{"speaker":null,"startTime":67.686,"endTime":68.046,"body":"Kevin: Okay, ja."},{"speaker":null,"startTime":68.386,"endTime":69.526,"body":"For me it was, was the same."},{"speaker":null,"startTime":69.646,"endTime":74.496,"body":"I also liked all the, natural sciencesand then, yeah, I went for chemistry."},{"speaker":null,"startTime":74.798,"endTime":78.025,"body":"But yeah, let's focuson the biotechnologies."},{"speaker":null,"startTime":78.399,"endTime":80.92,"body":"maybe to also introduce the topic a bit."},{"speaker":null,"startTime":81.019,"endTime":85.299,"body":"What is biotechnologies andwhat is it used for usually?"},{"speaker":null,"startTime":85.861,"endTime":90.401,"body":"Ronja Rappold: Yeah, so in generalit is the use of living organisms,"},{"speaker":null,"startTime":90.521,"endTime":94.781,"body":"biological systems or cells tomanufacture products that we need."},{"speaker":null,"startTime":95.642,"endTime":100.481,"body":"Um, so you can recruit microscopicfactories like bacteria, yeast,"},{"speaker":null,"startTime":100.621,"endTime":104.001,"body":"fungi to actually work for us,uh, and produce things that,"},{"speaker":null,"startTime":104.142,"endTime":105.801,"body":"that we want to have produced."},{"speaker":null,"startTime":106.254,"endTime":112.383,"body":"And maybe to give a little bit more of a,broader spectrum so we-- it can be very"},{"speaker":null,"startTime":112.484,"endTime":118.923,"body":"ancient from wild yeast that we use forfermentation for beer and wine since many,"},{"speaker":null,"startTime":118.987,"endTime":120.427,"body":"Kevin: Ja, that everyone knows, ja."},{"speaker":null,"startTime":121.158,"endTime":124.478,"body":"Ronja Rappold: to very modernapproaches, uh, where we use advanced"},{"speaker":null,"startTime":124.557,"endTime":129.557,"body":"genetic engineering tools to reallyfine-tune and edit the genomes"},{"speaker":null,"startTime":129.658,"endTime":133.837,"body":"very precisely to produce, as youmentioned in the introduction, um,"},{"speaker":null,"startTime":133.957,"endTime":135.698,"body":"life-saving drugs such as insulin"},{"speaker":null,"startTime":136.222,"endTime":139.902,"body":"Kevin: Ja, that's, \u00e4h, I found when Iheard about this the first time that,"},{"speaker":null,"startTime":141.103,"endTime":145.423,"body":"well, we need insulin, but it's a verycomplicated molecule, so we cannot easily"},{"speaker":null,"startTime":145.522,"endTime":148.902,"body":"do it, but our body can produce it."},{"speaker":null,"startTime":148.962,"endTime":150.562,"body":"So why not?"},{"speaker":null,"startTime":150.703,"endTime":154.242,"body":"Let's train basically bacteria to do it."},{"speaker":null,"startTime":154.742,"endTime":155.862,"body":"It's very fascinating"},{"speaker":null,"startTime":156.134,"endTime":159.654,"body":"Ronja Rappold: I think that is oneof the superpowers of biotechnology"},{"speaker":null,"startTime":159.815,"endTime":164.708,"body":"because we have, uh, enzymes uh, thathave very specific functions and can do"},{"speaker":null,"startTime":164.783,"endTime":164.903,"body":"Kevin: Mhm."},{"speaker":null,"startTime":165.048,"endTime":168.027,"body":"Ronja Rappold: Specific tasks, andno matter how complex they are."},{"speaker":null,"startTime":168.168,"endTime":172.847,"body":"So producing, very complex moleculesisn't a problem if you find or you can"},{"speaker":null,"startTime":172.886,"endTime":174.966,"body":"design an enzyme to do exactly that."},{"speaker":null,"startTime":175.722,"endTime":177.162,"body":"Kevin: Und they are super efficient."},{"speaker":null,"startTime":177.442,"endTime":178.542,"body":"I guess a molecule,"},{"speaker":null,"startTime":178.698,"endTime":179.078,"body":"Ronja Rappold: They are."},{"speaker":null,"startTime":179.842,"endTime":185.843,"body":"Kevin: molecular, \u00e4h, catalyst researcherI was always a bit jealous of the, \u00e4h,"},{"speaker":null,"startTime":186.082,"endTime":190.645,"body":"yeah, efficiency that you can reach andlow temperatures, with those enzymes."},{"speaker":null,"startTime":190.921,"endTime":192.241,"body":"Like the perfect catalyst."},{"speaker":null,"startTime":192.627,"endTime":193.028,"body":"Ronja Rappold: yeah, it"},{"speaker":null,"startTime":193.107,"endTime":194.727,"body":"is like this low"},{"speaker":null,"startTime":196.919,"endTime":203.974,"body":"great advantages that the biotechnologyhas for production over chemical routes"},{"speaker":null,"startTime":204.065,"endTime":204.215,"body":"Kevin: Mhm."},{"speaker":null,"startTime":204.545,"endTime":210.423,"body":"And, \u00e4hm, have an example maybefor, \u00e4h, yeah, a process that"},{"speaker":null,"startTime":210.482,"endTime":213.383,"body":"is used except for the insulin?"},{"speaker":null,"startTime":214.94,"endTime":217.891,"body":"Ronja Rappold: Yeah, uh, thereis, a few great examples."},{"speaker":null,"startTime":218.132,"endTime":223.404,"body":"Uh, maybe one that is also on industrialscale is what Genomatica did years."},{"speaker":null,"startTime":225.231,"endTime":232.812,"body":"They successfully adapted or engineereda bacterium produce 1,4-butanediol, uh,"},{"speaker":null,"startTime":232.871,"endTime":234.774,"body":"which is used to produce synthetic fiber."},{"speaker":null,"startTime":234.824,"endTime":236.643,"body":"And they do that from sugar."},{"speaker":null,"startTime":237.084,"endTime":241.904,"body":"so they have built a plant in theUS that produces more than 60,000"},{"speaker":null,"startTime":241.964,"endTime":243.886,"body":"tons per biotechnologically  now"},{"speaker":null,"startTime":245.746,"endTime":250.896,"body":"Kevin: So that can turn intothis, \u00e4h, fine chemical."},{"speaker":null,"startTime":251.716,"endTime":255.257,"body":"But, \u00e4h, yeah, when, whentalking about sugar, that's the"},{"speaker":null,"startTime":255.316,"endTime":257.257,"body":"same with all the bioethanol."},{"speaker":null,"startTime":257.896,"endTime":264.136,"body":"It's like you-- a crop that couldtheoretically also be food for then"},{"speaker":null,"startTime":264.177,"endTime":266.296,"body":"the conversion into chemicals or fuels."},{"speaker":null,"startTime":267.677,"endTime":267.916,"body":"So"},{"speaker":null,"startTime":270.176,"endTime":275.436,"body":"be that we are independent of thefossil fuels because we don't need"},{"speaker":null,"startTime":275.496,"endTime":277.956,"body":"the crude oil to make these chemicals."},{"speaker":null,"startTime":280.169,"endTime":280.809,"body":"Ronja Rappold: Yeah, definitely."},{"speaker":null,"startTime":280.849,"endTime":285.829,"body":"So I think this is a huge benefit,when we start using fermentation or"},{"speaker":null,"startTime":285.929,"endTime":290.689,"body":"biotechnology in general, that we canbecome independent from, fossil fuels."},{"speaker":null,"startTime":290.856,"endTime":296.876,"body":"The thing is industry chemical produceson a very, very large scale, right?"},{"speaker":null,"startTime":296.936,"endTime":301.486,"body":"We produce more than eight hundred millionmetric tons of base chemicals annually."},{"speaker":null,"startTime":302.216,"endTime":307.216,"body":"if we now think we wanna completelyshift this chemical industry or this,"},{"speaker":null,"startTime":307.315,"endTime":312.034,"body":"this chemical production, to beingmore sustainable, and then we shift"},{"speaker":null,"startTime":312.094,"endTime":315.484,"body":"all of this to sugar, we run intothis problem of this, food versus"},{"speaker":null,"startTime":315.564,"endTime":319.688,"body":"fuel debate, which is one aspect."},{"speaker":null,"startTime":319.762,"endTime":323.902,"body":"the other aspect that I would like tomention is that also when we talk about"},{"speaker":null,"startTime":323.961,"endTime":332.562,"body":"sustainability, the CO2 emission savingswhen we switch to sugar as a, a feedstock,"},{"speaker":null,"startTime":332.641,"endTime":340.267,"body":"as a raw material, are not necessarilyas big as we once believed,  uh, you are"},{"speaker":null,"startTime":340.308,"endTime":342.864,"body":"heavily dependent on industrial farming,"},{"speaker":null,"startTime":343.063,"endTime":343.163,"body":"Kevin: mhm."},{"speaker":null,"startTime":343.84,"endTime":344.3,"body":"Ronja Rappold: this also"},{"speaker":null,"startTime":344.38,"endTime":345.62,"body":"comes with CO2 emissions."},{"speaker":null,"startTime":346.28,"endTime":350.74,"body":"Uh, our own analysis shows thatsugar-based processes save maximum,"},{"speaker":null,"startTime":350.8,"endTime":353.62,"body":"uh, of fifty percent of CO2 emissions,"},{"speaker":null,"startTime":353.721,"endTime":353.8,"body":"uh,"},{"speaker":null,"startTime":353.847,"endTime":354.127,"body":"Kevin: okay"},{"speaker":null,"startTime":354.88,"endTime":358.744,"body":"Ronja Rappold: a fuel-based production ofa chemical with a sugar-based production."},{"speaker":null,"startTime":359.107,"endTime":361.969,"body":"and then you still only accounted for CO2"},{"speaker":null,"startTime":362.42,"endTime":362.62,"body":"Kevin: Mhm."},{"speaker":null,"startTime":362.63,"endTime":362.829,"body":"Ronja Rappold: right?"},{"speaker":null,"startTime":363.53,"endTime":367.85,"body":"there is, there is, much more,to sustainability than just that."},{"speaker":null,"startTime":368.084,"endTime":368.204,"body":"Kevin: Ja."},{"speaker":null,"startTime":368.784,"endTime":372.384,"body":"Land Use and Water Use and all this, ja"},{"speaker":null,"startTime":373.433,"endTime":373.973,"body":"Ronja Rappold: Exactly."},{"speaker":null,"startTime":374.053,"endTime":379.133,"body":"So maybe we can quickly talk aboutthe, the planetary boundary framework"},{"speaker":null,"startTime":379.413,"endTime":383.254,"body":"that developed by the StockholmResilience Centre and the"},{"speaker":null,"startTime":383.313,"endTime":384.834,"body":"team around Johan Rockstr\u00f6m."},{"speaker":null,"startTime":385.693,"endTime":392.501,"body":"Um, so they have generated or come up witha scientific framework to really quantify,"},{"speaker":null,"startTime":392.861,"endTime":397.7,"body":"the safe operating space of the planetEarth, which we need for sustainable"},{"speaker":null,"startTime":397.76,"endTime":400.58,"body":"living, uh, on this planet as humanity."},{"speaker":null,"startTime":400.62,"endTime":406.484,"body":"And you mentioned, land use, freshwateruse are two of these, categories."},{"speaker":null,"startTime":406.865,"endTime":411.359,"body":"There is also nitrogen and phosphorususage, which again, when you,"},{"speaker":null,"startTime":411.907,"endTime":416.648,"body":"look at industrial agriculture,uh, we, we already breached."},{"speaker":null,"startTime":416.869,"endTime":421.442,"body":"and then there is also globalbiodiversity, which we're"},{"speaker":null,"startTime":422.356,"endTime":424.376,"body":"dealing very good in either."},{"speaker":null,"startTime":424.516,"endTime":430.156,"body":"So there we also breach this, uh,category massively actually, um, which"},{"speaker":null,"startTime":430.216,"endTime":435.816,"body":"in the end then circles back to aproblem that we will get in industrial"},{"speaker":null,"startTime":435.896,"endTime":439.856,"body":"agriculture or in agricultural-- in,in the agricultural industry itself"},{"speaker":null,"startTime":440.467,"endTime":444.039,"body":"Kevin: Yeah, and then in the end alsoneed it for food and that, uh, is maybe a"},{"speaker":null,"startTime":444.079,"endTime":446.779,"body":"bit more important than making chemicals."},{"speaker":null,"startTime":447.559,"endTime":452.388,"body":"Although we also need those, but yeah,it's, it's always a difficult, yeah,"},{"speaker":null,"startTime":452.507,"endTime":457.147,"body":"game to see what is more importantwhen we need to find the balance, or a"},{"speaker":null,"startTime":457.267,"endTime":459.247,"body":"different source for the carbon, right?"},{"speaker":null,"startTime":459.943,"endTime":463.163,"body":"Ronja Rappold: Yeah, that's truebecause we, we do not necessarily need"},{"speaker":null,"startTime":463.223,"endTime":469.281,"body":"to use, plant sugars that we couldalso use for feeding, uh, humanity."},{"speaker":null,"startTime":470.181,"endTime":474.839,"body":"there is non-food carbonsources that we could, uh, use."},{"speaker":null,"startTime":474.854,"endTime":480.427,"body":"Glycerol, which is a C3 carbonsource that works super well."},{"speaker":null,"startTime":481.107,"endTime":485.668,"body":"Uh, we can also target agriculturalwaste, forestry waste, and one"},{"speaker":null,"startTime":485.747,"endTime":488.848,"body":"that I'm very excited about, we canalso use captured climate gases."},{"speaker":null,"startTime":490.03,"endTime":493.898,"body":"Kevin: So CO\u2082 that is capturedin a, direct air capture plant,"},{"speaker":null,"startTime":493.978,"endTime":496.238,"body":"for example, or the point source?"},{"speaker":null,"startTime":496.751,"endTime":497.211,"body":"Ronja Rappold: Both work,"},{"speaker":null,"startTime":497.225,"endTime":497.445,"body":"Kevin: Mhm."},{"speaker":null,"startTime":497.571,"endTime":500.531,"body":"Ronja Rappold: So pointsources, days are easier."},{"speaker":null,"startTime":500.632,"endTime":504.152,"body":"They have higher CO2 concentrations,um, than we have in the air,"},{"speaker":null,"startTime":504.938,"endTime":505.119,"body":"Kevin: Ja"},{"speaker":null,"startTime":505.192,"endTime":507.54,"body":"Ronja Rappold: um, hencethey are a bit cheaper."},{"speaker":null,"startTime":507.554,"endTime":509.014,"body":"Kevin: Ja, need less energy, hm"},{"speaker":null,"startTime":509.367,"endTime":509.647,"body":"Ronja Rappold: yeah, there"},{"speaker":null,"startTime":509.687,"endTime":513.868,"body":"is already an, an industrial plantbuilt, so you can, uh, target this and"},{"speaker":null,"startTime":513.907,"endTime":517.588,"body":"maybe couple another industrial plantfor chemical production next to it"},{"speaker":null,"startTime":517.777,"endTime":517.977,"body":"Kevin: Mhm."},{"speaker":null,"startTime":518.637,"endTime":527.337,"body":"So you use the CO\u2082 and how can youincorporate the CO\u2082 \u00e4h, to make chemicals?"},{"speaker":null,"startTime":527.763,"endTime":530.323,"body":"Ronja Rappold: Yeah, so there isvarious different routes to do so."},{"speaker":null,"startTime":531.365,"endTime":538.645,"body":"there is indeed actually organismsthat can metabolize CO2 itself."},{"speaker":null,"startTime":539.431,"endTime":539.711,"body":"Kevin: Ja, okay"},{"speaker":null,"startTime":540.725,"endTime":546.465,"body":"Ronja Rappold: that exists with directCO2 fermentation is that it's a gas"},{"speaker":null,"startTime":546.505,"endTime":549.106,"body":"fermentation approach, which is possible."},{"speaker":null,"startTime":549.454,"endTime":555.414,"body":"It is just a bit more complicated,uh, on an industrial scale compared to"},{"speaker":null,"startTime":555.555,"endTime":561.135,"body":"liquid fermentation, which is just whatwe usually know and is easier to scale"},{"speaker":null,"startTime":562.472,"endTime":562.512,"body":"Kevin: Ja."},{"speaker":null,"startTime":562.612,"endTime":568.672,"body":"I have come across some methanationprojects where they used CO2 and"},{"speaker":null,"startTime":568.732,"endTime":572.652,"body":"hydrogen and then a bioreactorto make methane out of it."},{"speaker":null,"startTime":573.093,"endTime":577.192,"body":"But that's then more for fueland not really to make chemicals."},{"speaker":null,"startTime":577.992,"endTime":584.894,"body":"And you with your researchthat you also started a company"},{"speaker":null,"startTime":584.974,"endTime":587.458,"body":"with you used methanol, right?"},{"speaker":null,"startTime":587.88,"endTime":591.617,"body":"And why methanol andhow did you get there?"},{"speaker":null,"startTime":592.438,"endTime":596.711,"body":"Ronja Rappold: Yeah, uh, methanol initself can be easily derived from CO2."},{"speaker":null,"startTime":596.878,"endTime":600.796,"body":"so you need renewable energy, and then youcan produce it, completely sustainable."},{"speaker":null,"startTime":602.136,"endTime":605.537,"body":"On the other hand, as I just mentioned,the gas fermentation methanol is a"},{"speaker":null,"startTime":605.597,"endTime":611.558,"body":"liquid, so it can be, stored more easily,it can be transported more easily, and"},{"speaker":null,"startTime":611.678,"endTime":614.878,"body":"also you don't have this explosive risk."},{"speaker":null,"startTime":614.95,"endTime":616.53,"body":"That you have with methane."},{"speaker":null,"startTime":617.591,"endTime":618.61,"body":"Um, yeah."},{"speaker":null,"startTime":618.71,"endTime":625.627,"body":"And then the more indus- industrial,answer to that would be that there is"},{"speaker":null,"startTime":625.728,"endTime":630.408,"body":"already an industry scaling up greenmethanol or renewable methanol, which"},{"speaker":null,"startTime":630.507,"endTime":637.947,"body":"is, um, basically based on the demandof the shipping industry to find new"},{"speaker":null,"startTime":637.967,"endTime":643.482,"body":"fuels that are sustainable So Merck, forexample, is highly focusing on methanol"},{"speaker":null,"startTime":643.702,"endTime":646.242,"body":"already having ships that run on methanol."},{"speaker":null,"startTime":647.003,"endTime":654.205,"body":"Um, there's huge industry push, to reallyupscale the green methanol, supply."},{"speaker":null,"startTime":655.125,"endTime":660.241,"body":"And then maybe as a, as a last one,because I mentioned the natural CO2,"},{"speaker":null,"startTime":660.741,"endTime":666.723,"body":"um, metabolizing bacteria, is alsonatural, methanol eating microbes"},{"speaker":null,"startTime":667.237,"endTime":667.718,"body":"Kevin: Ah, okay."},{"speaker":null,"startTime":668.084,"endTime":670.044,"body":"So they already know how to use it."},{"speaker":null,"startTime":670.794,"endTime":674.721,"body":"Whereas other intermediateproducts might be more challenging."},{"speaker":null,"startTime":674.994,"endTime":675.275,"body":"okay."},{"speaker":null,"startTime":675.633,"endTime":682.053,"body":"And I think another point worth mentioningis that China is also investing a"},{"speaker":null,"startTime":682.133,"endTime":686.733,"body":"lot in the methanol based chemistry."},{"speaker":null,"startTime":687.073,"endTime":692.781,"body":"They mostly doing it with likenormal catalytic processes, to also"},{"speaker":null,"startTime":692.941,"endTime":699.222,"body":"become less dependent on the oil,they built their industry on this."},{"speaker":null,"startTime":699.281,"endTime":704.181,"body":"So yeah, green methanolis scaling definitely."},{"speaker":null,"startTime":704.261,"endTime":710.061,"body":"So it's a very interesting ideato use then the biotechnologies to"},{"speaker":null,"startTime":710.681,"endTime":713.821,"body":"convert that into fine chemicals."},{"speaker":null,"startTime":714.134,"endTime":716.234,"body":"And how did you do this?"},{"speaker":null,"startTime":716.294,"endTime":721.215,"body":"You cannot just use methanol andthen throw it on some bacteria"},{"speaker":null,"startTime":721.274,"endTime":722.674,"body":"and they start eating it, right?"},{"speaker":null,"startTime":723.01,"endTime":729.431,"body":"Ronja Rappold: so the way we approachthis problem is that we wanted to"},{"speaker":null,"startTime":729.57,"endTime":732.17,"body":"combine basically the best of two worlds."},{"speaker":null,"startTime":733.05,"endTime":737.37,"body":"Um, meaning one, that what Imentioned, that there exist naturally"},{"speaker":null,"startTime":737.45,"endTime":739.911,"body":"occurring methanol-eating microbes."},{"speaker":null,"startTime":740.39,"endTime":747.05,"body":"And on the hand, that we alreadyhave bacteria that we use on an"},{"speaker":null,"startTime":747.09,"endTime":750.15,"body":"industrial scale as the one thatI mentioned from Genomatica."},{"speaker":null,"startTime":751.29,"endTime":757.41,"body":"do have an Escherichia coli,um, which is the working horse"},{"speaker":null,"startTime":757.491,"endTime":759.07,"body":"in industrial biotechnology."},{"speaker":null,"startTime":759.75,"endTime":766.985,"body":"And then our approach was to combine thetwo of them, um, and really to rewire,"},{"speaker":null,"startTime":767.372,"endTime":773.791,"body":"the E. Coli to live on methanol, um,which has been compared actually to, to"},{"speaker":null,"startTime":773.851,"endTime":779.914,"body":"a heart trans-transplantation because it,it is super complex, and it requires a"},{"speaker":null,"startTime":780.015,"endTime":788.06,"body":"foreign metabolic system going into a newhost natively doesn't really know what,"},{"speaker":null,"startTime":788.121,"endTime":789.68,"body":"what to do with it and how to use it."},{"speaker":null,"startTime":790.243,"endTime":797.553,"body":"We basically targeted both with the sameapproach, which is engineering the genome"},{"speaker":null,"startTime":797.833,"endTime":804.279,"body":"or the DNA of this bacterium, of this E.coli, so that it basically gets trained"},{"speaker":null,"startTime":804.358,"endTime":811.838,"body":"to first its carbon and its energy fromthe methanol, second, to also completely"},{"speaker":null,"startTime":811.898,"endTime":817.398,"body":"revi-rewire its whole metabolism aroundthis new energy and carbon source"},{"speaker":null,"startTime":818.14,"endTime":820.292,"body":"Kevin: So that, are multiple processes."},{"speaker":null,"startTime":820.372,"endTime":824.972,"body":"It needs to be able to process it, butalso it needs to get the energy somewhere."},{"speaker":null,"startTime":825.112,"endTime":827.412,"body":"Usually that happens with the sugar,"},{"speaker":null,"startTime":827.7,"endTime":827.9,"body":"right?"},{"speaker":null,"startTime":827.99,"endTime":834.79,"body":"And then, when it uses sugar, it getsthe carbon to whatever it produces"},{"speaker":null,"startTime":834.89,"endTime":837.59,"body":"from the sugar and the energy both."},{"speaker":null,"startTime":837.71,"endTime":837.91,"body":"Okay."},{"speaker":null,"startTime":838.285,"endTime":843.17,"body":"Ronja Rappold: Yeah, and so what, whatyou can do is rationally engineer,"},{"speaker":null,"startTime":843.691,"endTime":848.312,"body":"that E. coli can degrade methanol."},{"speaker":null,"startTime":848.951,"endTime":855.771,"body":"For this, what we did is you insertgenes from such a species that is a"},{"speaker":null,"startTime":855.831,"endTime":861.051,"body":"native methylotroph that-- so thatnaturally metabolizes, uh, methanol and"},{"speaker":null,"startTime":861.351,"endTime":863.611,"body":"basically give this toolbox to the E. coli"},{"speaker":null,"startTime":864.338,"endTime":864.518,"body":"Kevin: Okay."},{"speaker":null,"startTime":865.138,"endTime":868.158,"body":"Why can you not use this,\u00e4h, bacterium directly?"},{"speaker":null,"startTime":868.878,"endTime":871.07,"body":"The one that already can use methanol"},{"speaker":null,"startTime":871.612,"endTime":873.492,"body":"Ronja Rappold: Yeah, thisis a super great question."},{"speaker":null,"startTime":873.912,"endTime":877.152,"body":"Uh, you can use it to grow on methanol."},{"speaker":null,"startTime":877.352,"endTime":883.132,"body":"The problem comes when you wanna producechemicals from it on an industrial"},{"speaker":null,"startTime":883.232,"endTime":890.874,"body":"scale, because these methylotrophs arenot as well understood as the E. coli"},{"speaker":null,"startTime":890.934,"endTime":895.114,"body":"is, which probably is the understoodorganism that we have on this planet."},{"speaker":null,"startTime":895.934,"endTime":902.075,"body":"we have a lot of genetic engineeringtools to alter the E. coli and to adapt"},{"speaker":null,"startTime":902.114,"endTime":908.914,"body":"it to new scenarios and new industrialsettings, we know that it scales very well"},{"speaker":null,"startTime":909.054,"endTime":915.634,"body":"in, these very, very large bioreactorsthat we need to produce chemicals at the"},{"speaker":null,"startTime":915.694,"endTime":920.234,"body":"level that we actually need, um, to havea, have a shift in the chemical industry."},{"speaker":null,"startTime":920.994,"endTime":924.814,"body":"So in theory, yes, you coulduse a native methylotroph."},{"speaker":null,"startTime":924.907,"endTime":929.608,"body":"in practice, there is many, manychallenges that you or that we"},{"speaker":null,"startTime":929.707,"endTime":934.347,"body":"think with an E. coli we cancircumvent as soon as the E. coli"},{"speaker":null,"startTime":934.407,"endTime":937.427,"body":"has learned to metabolize methanol"},{"speaker":null,"startTime":939.262,"endTime":943.102,"body":"Kevin: And that you did bythe genetic modification?"},{"speaker":null,"startTime":943.567,"endTime":943.907,"body":"Ronja Rappold: Exactly."},{"speaker":null,"startTime":943.987,"endTime":947.968,"body":"So the first step we talked about,uh, which was inserting genes"},{"speaker":null,"startTime":948.207,"endTime":950.347,"body":"that it can metabolize methanol."},{"speaker":null,"startTime":950.948,"endTime":956.308,"body":"The problem is that then basicallythe cell's internal metabolism"},{"speaker":null,"startTime":957.007,"endTime":958.727,"body":"is not fully adapted to that."},{"speaker":null,"startTime":959.568,"endTime":966.907,"body":"Um, so-- And this process is much, muchmore complicated than inserting three or"},{"speaker":null,"startTime":966.968,"endTime":974.607,"body":"four genes, uh, which is also the reasonwhy no one has done that before, um, and"},{"speaker":null,"startTime":974.728,"endTime":980.775,"body":"why this is such a long-lasting goal inacademia to actually manage to do that."},{"speaker":null,"startTime":981.595,"endTime":987.554,"body":"Um, and the approach here at ETH Zurichwith the, with the team in the lab,"},{"speaker":null,"startTime":988.053,"endTime":991.132,"body":"was using basically natural selection."},{"speaker":null,"startTime":991.192,"endTime":995.833,"body":"So basically usingevolution as  little helper"},{"speaker":null,"startTime":996.139,"endTime":996.439,"body":"Kevin: Okay"},{"speaker":null,"startTime":997.079,"endTime":998.078,"body":"Ronja Rappold: um, to, to make this work"},{"speaker":null,"startTime":999.221,"endTime":1003.881,"body":"Kevin: And you can use that becausebacteria, they multiply so fast that you"},{"speaker":null,"startTime":1003.96,"endTime":1011.391,"body":"have multiple generations and then, well,there are mutations, Or how does it work?"},{"speaker":null,"startTime":1011.451,"endTime":1011.772,"body":"And then"},{"speaker":null,"startTime":1012.308,"endTime":1012.508,"body":"Ronja Rappold: Yeah."},{"speaker":null,"startTime":1012.629,"endTime":1020.708,"body":"So basically in, with every generation youaccumulate new mutation, uh, new mutations"},{"speaker":null,"startTime":1020.808,"endTime":1022.188,"body":"just randomly because they happen."},{"speaker":null,"startTime":1022.928,"endTime":1030.292,"body":"Uh, little errors, uh, in copying the DNA,DNA, which can become Very, adventagous"},{"speaker":null,"startTime":1030.54,"endTime":1037.36,"body":"and then you just need to build basicallya laboratory setting around them that"},{"speaker":null,"startTime":1037.46,"endTime":1040.94,"body":"you select for the right mutation"},{"speaker":null,"startTime":1041.278,"endTime":1041.518,"body":"Kevin: Mhm."},{"speaker":null,"startTime":1042.699,"endTime":1044.238,"body":"Okay, how, how can you do that?"},{"speaker":null,"startTime":1045.138,"endTime":1045.278,"body":"that?"},{"speaker":null,"startTime":1045.928,"endTime":1046.408,"body":"Ronja Rappold: so the whole"},{"speaker":null,"startTime":1046.468,"endTime":1048.248,"body":"process, uh, has also a term."},{"speaker":null,"startTime":1048.329,"endTime":1050.408,"body":"It's called adaptive laboratory evolution."},{"speaker":null,"startTime":1051.089,"endTime":1057.146,"body":"And the way, you do that is that youhave these cells that remember how"},{"speaker":null,"startTime":1057.146,"endTime":1060.216,"body":"they can metabolize methanol bit,"},{"speaker":null,"startTime":1060.307,"endTime":1060.527,"body":"Kevin: Mhm."},{"speaker":null,"startTime":1061.006,"endTime":1066.136,"body":"Ronja Rappold: you keep these cells inreally a, um, an excess of methanol."},{"speaker":null,"startTime":1066.296,"endTime":1072.48,"body":"So they always have methanol, liveoff, but you only give them a tiny"},{"speaker":null,"startTime":1072.56,"endTime":1074.84,"body":"bit of an additional feedstock."},{"speaker":null,"startTime":1075.562,"endTime":1076.142,"body":"Kevin: Mhm, mhm."},{"speaker":null,"startTime":1076.66,"endTime":1080.217,"body":"Ronja Rappold: that, uh, waspyruvat-pyruvate, a central metabolite"},{"speaker":null,"startTime":1080.257,"endTime":1082.397,"body":"that comes out of the glycolysis."},{"speaker":null,"startTime":1082.757,"endTime":1086.677,"body":"So basically, when you start with sugar,this is what, what you end up with."},{"speaker":null,"startTime":1087.498,"endTime":1091.977,"body":"Um, and then in this scenario,most cells just die because"},{"speaker":null,"startTime":1092.678,"endTime":1094.498,"body":"they, uh, they don't manage."},{"speaker":null,"startTime":1095.217,"endTime":1101.372,"body":"Um, but those that manage to replicate,as you mentioned, They multiply very fast."},{"speaker":null,"startTime":1101.679,"endTime":1106.719,"body":"Um, and then these random DNA mutationsthat occur at some point randomly"},{"speaker":null,"startTime":1106.779,"endTime":1114.079,"body":"will lead to a beneficial, mutation,and eventually basically a cell will"},{"speaker":null,"startTime":1114.3,"endTime":1118.919,"body":"grow slightly faster than the othersand then will outcompete, uh, the And"},{"speaker":null,"startTime":1118.959,"endTime":1125.879,"body":"then what we did is repeating this ina highly controlled bioreactor over"},{"speaker":null,"startTime":1125.919,"endTime":1130.72,"body":"hundreds of generations, while alwayskeeping this selection pressure on."},{"speaker":null,"startTime":1130.76,"endTime":1134.719,"body":"So always having this hugeamount of methanol and very"},{"speaker":null,"startTime":1134.779,"endTime":1136.48,"body":"limited central metabolite"},{"speaker":null,"startTime":1137.222,"endTime":1137.443,"body":"Kevin: Mhm."},{"speaker":null,"startTime":1137.962,"endTime":1143.623,"body":"So you basically make an environmentwhere only the cells survive that"},{"speaker":null,"startTime":1143.682,"endTime":1148.882,"body":"can live off of methanol because theyhave so little of their normal food"},{"speaker":null,"startTime":1149.115,"endTime":1149.456,"body":"Ronja Rappold: Exactly."},{"speaker":null,"startTime":1150.003,"endTime":1150.902,"body":"Kevin: Okay, interesting."},{"speaker":null,"startTime":1151.462,"endTime":1156.042,"body":"And then you increase the pressureby also giving less and less"},{"speaker":null,"startTime":1156.122,"endTime":1158.825,"body":"of the other, \u00e4hm, material?"},{"speaker":null,"startTime":1159.571,"endTime":1160.212,"body":"Ronja Rappold: Yeah, exactly."},{"speaker":null,"startTime":1160.291,"endTime":1166.131,"body":"So as soon as you see a population arisingthat can deal with the current scenario,"},{"speaker":null,"startTime":1166.291,"endTime":1171.011,"body":"you increase, \u00e4hm, the selection pressureand you remove more or you, yeah, you"},{"speaker":null,"startTime":1171.071,"endTime":1176.992,"body":"rem-remove the, in our case, pyrovat,\u00e4hm, to, to increase the pressure again"},{"speaker":null,"startTime":1177.357,"endTime":1178.097,"body":"Kevin: Hm, okay."},{"speaker":null,"startTime":1178.717,"endTime":1183.617,"body":"So you definitely need this selectionpressure for something to happen."},{"speaker":null,"startTime":1184.077,"endTime":1188.818,"body":"Because I also, when I read up aboutthis a bit, \u00e4h, I was asking myself:"},{"speaker":null,"startTime":1188.897,"endTime":1193.237,"body":"When they mutate so much, how canyou keep them stable in the end?"},{"speaker":null,"startTime":1193.837,"endTime":1198.077,"body":"But then you basically don'thave this, \u00e4h, pressure anymore."},{"speaker":null,"startTime":1198.157,"endTime":1202.538,"body":"You just have your constantmethanol environment without"},{"speaker":null,"startTime":1202.577,"endTime":1204.117,"body":"the pressure to change anymore."},{"speaker":null,"startTime":1204.217,"endTime":1204.417,"body":"Okay."},{"speaker":null,"startTime":1204.784,"endTime":1205.444,"body":"Yeah, interesting."},{"speaker":null,"startTime":1206.078,"endTime":1206.579,"body":"Ronja Rappold: Exactly."},{"speaker":null,"startTime":1206.981,"endTime":1212.501,"body":"Kevin: So now you have trained thebacteria to live off of methanol and"},{"speaker":null,"startTime":1212.941,"endTime":1216.541,"body":"how do you select what they produce?"},{"speaker":null,"startTime":1216.802,"endTime":1220.421,"body":"Because in the end you want tohave some target chemicals and"},{"speaker":null,"startTime":1220.461,"endTime":1222.882,"body":"not just random byproducts."},{"speaker":null,"startTime":1223.758,"endTime":1224.438,"body":"Ronja Rappold: Yeah, exactly."},{"speaker":null,"startTime":1224.498,"endTime":1230.538,"body":"So after we managed to have themethylotrophic growth, uh, we"},{"speaker":null,"startTime":1230.579,"endTime":1236.499,"body":"basically could approach it as everyother production problem, which means"},{"speaker":null,"startTime":1236.958,"endTime":1241.798,"body":"you continue to engineer the cell,but in the downstream metabolism."},{"speaker":null,"startTime":1242.025,"endTime":1242.206,"body":"Kevin: Mhm"},{"speaker":null,"startTime":1243.098,"endTime":1243.199,"body":"Ronja Rappold: what"},{"speaker":null,"startTime":1243.259,"endTime":1245.558,"body":"we did was producing lactic acid."},{"speaker":null,"startTime":1246.123,"endTime":1251.323,"body":"and for that, you basically need oneadditional enzyme for the cell to"},{"speaker":null,"startTime":1251.363,"endTime":1254.774,"body":"produce it, which you can easily insert."},{"speaker":null,"startTime":1256.304,"endTime":1260.185,"body":"Kevin: So that is not done bythe laboratory evolution, that"},{"speaker":null,"startTime":1260.225,"endTime":1263.324,"body":"is done by, \u00e4h, the old school"},{"speaker":null,"startTime":1263.761,"endTime":1265.301,"body":"biotechnology, ja"},{"speaker":null,"startTime":1265.833,"endTime":1266.034,"body":"Ronja Rappold: yes,"},{"speaker":null,"startTime":1268.122,"endTime":1272.372,"body":"Yeah, so basically what you do, you,you insert, instruction for how to"},{"speaker":null,"startTime":1272.622,"endTime":1275.526,"body":"produce lactic acid into the E. coli."},{"speaker":null,"startTime":1276.386,"endTime":1279.335,"body":"you give it a trigger, andthen you, you can produce it,"},{"speaker":null,"startTime":1279.593,"endTime":1285.431,"body":"Kevin: Okay, and then when you have theready bacterium that can live off of"},{"speaker":null,"startTime":1285.511,"endTime":1290.855,"body":"methanol and produce, in this case, lacticacid, to make a process out of it, you"},{"speaker":null,"startTime":1290.935,"endTime":1295.97,"body":"have to increase the amount it producesas much as possible from the fuel."},{"speaker":null,"startTime":1296.243,"endTime":1297.623,"body":"How is that possible?"},{"speaker":null,"startTime":1298.752,"endTime":1299.512,"body":"Ronja Rappold: Yeah, exactly."},{"speaker":null,"startTime":1299.772,"endTime":1304.419,"body":"Um, so what you can do is youcan streamline the metabolism."},{"speaker":null,"startTime":1304.619,"endTime":1307.419,"body":"Also, again, you tweak the DNA"},{"speaker":null,"startTime":1307.479,"endTime":1307.539,"body":"a"},{"speaker":null,"startTime":1307.841,"endTime":1308.021,"body":"Kevin: Mhm."},{"speaker":null,"startTime":1308.779,"endTime":1308.859,"body":"Ronja Rappold: you"},{"speaker":null,"startTime":1308.919,"endTime":1310.739,"body":"make some enzymes more efficient."},{"speaker":null,"startTime":1310.996,"endTime":1314.647,"body":"and then when you're in a bioreactor,what you can do is You can make the"},{"speaker":null,"startTime":1314.688,"endTime":1322.027,"body":"surrounding setting as as possible,so that it always has all nutrients,"},{"speaker":null,"startTime":1322.127,"endTime":1328.088,"body":"\u00e4hm, at the right time, \u00e4hm, so that itthen can produce as efficient as, as it"},{"speaker":null,"startTime":1328.127,"endTime":1328.307,"body":"gets"},{"speaker":null,"startTime":1328.63,"endTime":1333.769,"body":"Kevin: What are there, \u00e4hm, metricsthat you can change to make,"},{"speaker":null,"startTime":1333.909,"endTime":1335.87,"body":"\u00e4h, the process produce more?"},{"speaker":null,"startTime":1336.274,"endTime":1342.314,"body":"Like when you have a given bioreactorand you put your bacterium in there, does"},{"speaker":null,"startTime":1342.354,"endTime":1349.115,"body":"it already have enough capacity directlyfrom the point when you started, okay,"},{"speaker":null,"startTime":1349.174,"endTime":1350.954,"body":"it can produce the lactic acid now?"},{"speaker":null,"startTime":1351.094,"endTime":1356.375,"body":"Or did you have to do other changesto have it in a process basically?"},{"speaker":null,"startTime":1359.084,"endTime":1361.684,"body":"Ronja Rappold: You still needto adapt it further, um, to"},{"speaker":null,"startTime":1361.745,"endTime":1363.984,"body":"produce or to have a higher titer"},{"speaker":null,"startTime":1364.07,"endTime":1364.251,"body":"Kevin: Mhm."},{"speaker":null,"startTime":1365.51,"endTime":1366.9,"body":"Wa-was ist der titer?"},{"speaker":null,"startTime":1367.229,"endTime":1370.569,"body":"Ronja Rappold: It is, uh, basicallythe, amount you produce per"},{"speaker":null,"startTime":1371.003,"endTime":1371.443,"body":"Kevin: Ah, okay."},{"speaker":null,"startTime":1372.229,"endTime":1373.409,"body":"Ronja Rappold: of volumein your bioreactor"},{"speaker":null,"startTime":1374.404,"endTime":1377.503,"body":"Kevin: And, \u00e4hm, whatdoes have to change there?"},{"speaker":null,"startTime":1378.312,"endTime":1382.352,"body":"Ronja Rappold: One of the challengesto then increase, the production of,"},{"speaker":null,"startTime":1382.532,"endTime":1387.805,"body":"for example, lactic acid or any other,chemical is that in the bioreactor"},{"speaker":null,"startTime":1387.925,"endTime":1391.465,"body":"you need to have, um, basicallythe perfect environment for the"},{"speaker":null,"startTime":1391.505,"endTime":1393.965,"body":"bacteria to be able to produce it."},{"speaker":null,"startTime":1394.445,"endTime":1399.265,"body":"Meaning, um, the oxygen transfer rate mustbe adapted, the nutrients must be adapted."},{"speaker":null,"startTime":1399.685,"endTime":1404.744,"body":"So all the things that this bacterianeed to thrive and, um, to have enough,"},{"speaker":null,"startTime":1405.015,"endTime":1411.454,"body":"basically molecular Lego buildingblocks to then produce, um, specific"},{"speaker":null,"startTime":1411.515,"endTime":1414.454,"body":"chemical that you instructed it to"},{"speaker":null,"startTime":1414.515,"endTime":1414.895,"body":"produce"},{"speaker":null,"startTime":1414.965,"endTime":1415.124,"body":"Kevin: Mhm."},{"speaker":null,"startTime":1416.044,"endTime":1416.884,"body":"Na, interesting."},{"speaker":null,"startTime":1417.544,"endTime":1424.465,"body":"And, \u00e4hm, to make this bacterium inthat way was basically, \u00e4h, the basis."},{"speaker":null,"startTime":1424.765,"endTime":1429.544,"body":"And then you started withyour colleagues TENO Bioworks."},{"speaker":null,"startTime":1430.105,"endTime":1434.262,"body":"what brought you there that you said:now we can actually start a company"},{"speaker":null,"startTime":1434.383,"endTime":1436.439,"body":"and it's worth to found a startup?"},{"speaker":null,"startTime":1436.665,"endTime":1437.285,"body":"Quick break."},{"speaker":null,"startTime":1437.548,"endTime":1441.898,"body":"If you enjoy this chemistry content,please consider following Innovate"},{"speaker":null,"startTime":1441.898,"endTime":1445.488,"body":"and React on whatever platform you'relistening to keep the show on your feed."},{"speaker":null,"startTime":1445.86,"endTime":1451.168,"body":"If you want to do even more, youcan tip a one-time amount or become"},{"speaker":null,"startTime":1451.168,"endTime":1455.118,"body":"a member of the show with the BuyMe A Coffee link in the show notes."},{"speaker":null,"startTime":1455.542,"endTime":1457.502,"body":"Thank you for your timeand back to the episode"},{"speaker":null,"startTime":1458.205,"endTime":1462.865,"body":"Ronja Rappold: Yeah, so I think thevery first moment was when actually"},{"speaker":null,"startTime":1462.925,"endTime":1467.645,"body":"this evolution experiment generatedthis methylotrophic E. coli."},{"speaker":null,"startTime":1468.546,"endTime":1471.024,"body":"Um, that was a huge, eureka moment."},{"speaker":null,"startTime":1471.085,"endTime":1476.005,"body":"And then the second one, more goinginto the industrial application, where"},{"speaker":null,"startTime":1476.064,"endTime":1481.564,"body":"then also this commercialization aspectcame into play, was when we transferred"},{"speaker":null,"startTime":1482.604,"endTime":1488.324,"body":"the whole concept from shake flasksfrom the lab into a small bioreactor"},{"speaker":null,"startTime":1488.404,"endTime":1491.404,"body":"and seeing that not only the E."},{"speaker":null,"startTime":1491.464,"endTime":1495.724,"body":"coli is thriving, but alsothe production actually scaled"},{"speaker":null,"startTime":1495.784,"endTime":1502.704,"body":"proportionally with the scale of thereactor, so the volume it can live in."},{"speaker":null,"startTime":1503.384,"endTime":1507.824,"body":"Um, so basically that the biologynot collapse, although it was in a"},{"speaker":null,"startTime":1507.884,"endTime":1512.612,"body":"different environment,  confirmed forus that actually using this industrial"},{"speaker":null,"startTime":1512.652,"endTime":1517.152,"body":"workhorse from which we came from,this whole idea of, like, really going"},{"speaker":null,"startTime":1517.193,"endTime":1519.752,"body":"this tedious long path of teaching E."},{"speaker":null,"startTime":1519.792,"endTime":1525.407,"body":"coli to utilize methanol will workout because In the first, scale"},{"speaker":null,"startTime":1525.448,"endTime":1530.948,"body":"up step, managed to see this, thisincrease in production already"},{"speaker":null,"startTime":1532.269,"endTime":1535.349,"body":"Kevin: Ja, that's, \u00e4h,basically the, a good basis."},{"speaker":null,"startTime":1536.2,"endTime":1537.9,"body":"now what, what are your next steps?"},{"speaker":null,"startTime":1538.04,"endTime":1542.28,"body":"You scale this, \u00e4h, processor how do you go further?"},{"speaker":null,"startTime":1543.075,"endTime":1546.775,"body":"Ronja Rappold: So one of this, uh,ongoing steps is, for one of the"},{"speaker":null,"startTime":1547.234,"endTime":1552.174,"body":"products that we showed in our proofof concept, itaconic acid, really"},{"speaker":null,"startTime":1552.315,"endTime":1555.194,"body":"increase the titer and the productivity."},{"speaker":null,"startTime":1555.374,"endTime":1560.155,"body":"So how much of this chemical we c-we can produce in a certain time."},{"speaker":null,"startTime":1561.781,"endTime":1568.12,"body":"Kevin: And then show to other customersthat, \u00e4h, they can have basically the"},{"speaker":null,"startTime":1568.16,"endTime":1574.71,"body":"same, quality and the same product theyare used to just from biological pathways."},{"speaker":null,"startTime":1575.915,"endTime":1577.515,"body":"Ronja Rappold: Yeah, justin a very sustainable,"},{"speaker":null,"startTime":1577.565,"endTime":1577.785,"body":"Kevin: Mhm."},{"speaker":null,"startTime":1579.255,"endTime":1579.695,"body":"Ronja Rappold: method."},{"speaker":null,"startTime":1580.395,"endTime":1586.122,"body":"then one thing that, we also highlightand that we will test more in the"},{"speaker":null,"startTime":1586.442,"endTime":1593.324,"body":"future is that so far, the lab workbasically, uh, is performed with usual"},{"speaker":null,"startTime":1593.525,"endTime":1599.484,"body":"gray methanol, so the one that you canjust buy from a chemical supplier, labs."},{"speaker":null,"startTime":1600.265,"endTime":1605.274,"body":"Um, we had first experiments with greenmethanol, which looked very promising."},{"speaker":null,"startTime":1605.733,"endTime":1611.8,"body":"And then one of the other things that,we were basically claiming to do is"},{"speaker":null,"startTime":1611.86,"endTime":1617.001,"body":"that we can also use raw methanol sothat we don't need this last step,"},{"speaker":null,"startTime":1617.161,"endTime":1619.2,"body":"um, of purification in the synthesis."},{"speaker":null,"startTime":1619.326,"endTime":1619.506,"body":"Kevin: Mhm."},{"speaker":null,"startTime":1620.24,"endTime":1623.681,"body":"Ronja Rappold: also their firstexperiments indicate that this"},{"speaker":null,"startTime":1623.76,"endTime":1628.54,"body":"actually, uh, works very well andthat would further down the line,"},{"speaker":null,"startTime":1629.42,"endTime":1631.4,"body":"help us massively to reduce costs."},{"speaker":null,"startTime":1631.856,"endTime":1635.237,"body":"Kevin: also if someone can producegreen methanol and they don't have"},{"speaker":null,"startTime":1635.317,"endTime":1640.336,"body":"to purify it as much to sell it,then this will also be better for"},{"speaker":null,"startTime":1640.376,"endTime":1642.376,"body":"them to sell a raw product basically."},{"speaker":null,"startTime":1642.749,"endTime":1642.789,"body":"Ronja Rappold: Ja,"},{"speaker":null,"startTime":1643.568,"endTime":1648.284,"body":"Kevin: and then, \u00e4hm, yeah, if thebacteria can, use raw methanol,"},{"speaker":null,"startTime":1648.325,"endTime":1654.615,"body":"is this, \u00e4h, in comparison to acatalytic process, \u00e4h, an advantage"},{"speaker":null,"startTime":1654.716,"endTime":1658.68,"body":"or do you think, \u00e4h, the bacteria candeal better with these impurities?"},{"speaker":null,"startTime":1659.546,"endTime":1661.006,"body":"Ronja Rappold: Yeah, I thinkthey can definitely deal"},{"speaker":null,"startTime":1661.046,"endTime":1662.947,"body":"better with the impurities."},{"speaker":null,"startTime":1663.187,"endTime":1668.086,"body":"Uh, we have, have had the list ofimpurities that they encountered in the"},{"speaker":null,"startTime":1668.472,"endTime":1668.612,"body":"Kevin: Mhm."},{"speaker":null,"startTime":1668.707,"endTime":1668.887,"body":"Ronja Rappold: we've"},{"speaker":null,"startTime":1669.816,"endTime":1675.897,"body":"and we know that there is contaminantsthat for certain catalysts, uh,"},{"speaker":null,"startTime":1676.237,"endTime":1677.636,"body":"would have been a huge problem"},{"speaker":null,"startTime":1678.302,"endTime":1678.802,"body":"Kevin: Ja, okay."},{"speaker":null,"startTime":1678.862,"endTime":1686.082,"body":"So it's not only the aspect that youcan use green methanol to produce some"},{"speaker":null,"startTime":1686.163,"endTime":1690.943,"body":"chemicals, but you can also make theprocess a bit linear because you don't"},{"speaker":null,"startTime":1691.042,"endTime":1696.063,"body":"need as many purification steps in themiddle and thus save costs probably."},{"speaker":null,"startTime":1696.659,"endTime":1697.259,"body":"Ronja Rappold: So far it looks"},{"speaker":null,"startTime":1697.279,"endTime":1697.539,"body":"like it,"},{"speaker":null,"startTime":1697.669,"endTime":1698.569,"body":"Kevin: ja, sounds great."},{"speaker":null,"startTime":1699.043,"endTime":1702.323,"body":"Where do you see biotechnology evolving?"},{"speaker":null,"startTime":1703.539,"endTime":1709.099,"body":"Ronja Rappold: So I think, uh, that the21st century actually will be defined by"},{"speaker":null,"startTime":1709.139,"endTime":1711.709,"body":"biotechnology and by synthetic biology."},{"speaker":null,"startTime":1712.049,"endTime":1717.779,"body":"We had amazing advan-advancements,uh, coming in the recent years."},{"speaker":null,"startTime":1717.849,"endTime":1723.729,"body":"Um, I think CRISPR means somethingfor everyone now, uh, which already"},{"speaker":null,"startTime":1723.729,"endTime":1730.008,"body":"in itself is that a biotech, or atechnology that we use in the lab is so"},{"speaker":null,"startTime":1730.008,"endTime":1734.918,"body":"well-known, um, is very, very excitingfor, for a biotechnologist like me."},{"speaker":null,"startTime":1735.598,"endTime":1736.358,"body":"Kevin: Ja, definitely"},{"speaker":null,"startTime":1736.378,"endTime":1739.028,"body":"Ronja Rappold: it willimplicate healthcare, the food"},{"speaker":null,"startTime":1739.028,"endTime":1740.538,"body":"industry, the chemical industry."},{"speaker":null,"startTime":1740.538,"endTime":1746.028,"body":"So really think that we will makemassive progress with, with biotech."},{"speaker":null,"startTime":1746.518,"endTime":1750.628,"body":"Um, and I think you can also seeit reflected in policy papers,"},{"speaker":null,"startTime":1750.928,"endTime":1752.408,"body":"um, published around the globe."},{"speaker":null,"startTime":1752.408,"endTime":1758.428,"body":"So, um, Europe last year publishedtheir Biotech Act I, which was mostly"},{"speaker":null,"startTime":1758.428,"endTime":1763.178,"body":"focused on healthcare, but the BiotechAct II is coming up, where they"},{"speaker":null,"startTime":1763.178,"endTime":1766.628,"body":"really wanna talk about industrialbiotechnology and biomanufacturing."},{"speaker":null,"startTime":1767.518,"endTime":1774.018,"body":"we saw it in China's, uh, five-yearplan that was published in spring."},{"speaker":null,"startTime":1774.631,"endTime":1778.358,"body":"They also put biomanufacturingas a strategic national priority."},{"speaker":null,"startTime":1778.741,"endTime":1781.717,"body":"we see it in Japan, we see itin countries in the Middle East."},{"speaker":null,"startTime":1781.987,"endTime":1788.497,"body":"Also, Germany has it on their high techagenda as of the six key technologies"},{"speaker":null,"startTime":1788.507,"endTime":1791.397,"body":"next to quantum computing and fusion."},{"speaker":null,"startTime":1792.027,"endTime":1792.387,"body":"Uh,"},{"speaker":null,"startTime":1792.603,"endTime":1792.784,"body":"Kevin: ja"},{"speaker":null,"startTime":1793.187,"endTime":1796.577,"body":"Ronja Rappold: I think this isall really, really exciting and"},{"speaker":null,"startTime":1796.647,"endTime":1798.237,"body":"I'm super happy to be part of this"},{"speaker":null,"startTime":1799.819,"endTime":1802.399,"body":"Kevin: Ja, definitely alot of potential there."},{"speaker":null,"startTime":1802.899,"endTime":1806.159,"body":"And, \u00e4h, where do yousee Tenos role in this?"},{"speaker":null,"startTime":1807.637,"endTime":1813.377,"body":"Become a big chemical producer orhow is, how do you see TNO evolving?"},{"speaker":null,"startTime":1814.055,"endTime":1817.984,"body":"Ronja Rappold: Yeah, so the,the goal we have is, I think,"},{"speaker":null,"startTime":1818.105,"endTime":1819.23,"body":"as you said, it's quite big."},{"speaker":null,"startTime":1819.503,"endTime":1826.563,"body":"it's being the technology provider forchemical production for chemicals that"},{"speaker":null,"startTime":1826.643,"endTime":1829.563,"body":"can be produced using m- biotechnology."},{"speaker":null,"startTime":1830.363,"endTime":1836.264,"body":"So having this platform technology nowwhere we have this, this bacterial strain"},{"speaker":null,"startTime":1836.344,"endTime":1842.883,"body":"that really can transform basically CO2via the step of methanol into a variety"},{"speaker":null,"startTime":1842.943,"endTime":1847.724,"body":"of products, which we have shown already,with then in the future years of more"},{"speaker":null,"startTime":1847.763,"endTime":1854.843,"body":"development of this strain and adaptationto more, chemical products, we really"},{"speaker":null,"startTime":1854.963,"endTime":1861.445,"body":"wanna become Yeah, this technologyprovider, um, for also large and small"},{"speaker":null,"startTime":1861.465,"endTime":1862.985,"body":"chemical companies around the world"},{"speaker":null,"startTime":1863.479,"endTime":1868.606,"body":"Kevin: So provide the technology forother, companies to then produce,"},{"speaker":null,"startTime":1868.807,"endTime":1871.047,"body":"so not become the producing entity."},{"speaker":null,"startTime":1872.107,"endTime":1872.307,"body":"Okay,"},{"speaker":null,"startTime":1872.404,"endTime":1872.484,"body":"Ronja Rappold: Yeah,"},{"speaker":null,"startTime":1872.567,"endTime":1872.966,"body":"Kevin: interesting"},{"speaker":null,"startTime":1873.724,"endTime":1873.805,"body":"Ronja Rappold: it,"},{"speaker":null,"startTime":1873.844,"endTime":1877.805,"body":"always is the problem withlarge scale production."},{"speaker":null,"startTime":1877.924,"endTime":1878.884,"body":"You need a lot of steel in"},{"speaker":null,"startTime":1878.944,"endTime":1879.004,"body":"the"},{"speaker":null,"startTime":1879.311,"endTime":1879.43,"body":"Kevin: Ja."},{"speaker":null,"startTime":1879.684,"endTime":1879.785,"body":"Ronja Rappold: And"},{"speaker":null,"startTime":1881.344,"endTime":1886.884,"body":"so many companies that have theseindustrial plants that have the"},{"speaker":null,"startTime":1886.924,"endTime":1888.485,"body":"capacity to do fermentation."},{"speaker":null,"startTime":1889.184,"endTime":1895.09,"body":"And we basically now only need totrain our strain to use the specific"},{"speaker":null,"startTime":1895.15,"endTime":1901.15,"body":"product this company wants, and thensecond, um, change production in this"},{"speaker":null,"startTime":1901.21,"endTime":1907.291,"body":"plant, and then you make best useout of what is existing already and"},{"speaker":null,"startTime":1907.37,"endTime":1909.99,"body":"what we can bring in as, as novel"},{"speaker":null,"startTime":1911.052,"endTime":1915.513,"body":"Kevin: So it could be a drop in solutionwhere you use the already existing"},{"speaker":null,"startTime":1915.573,"endTime":1921.593,"body":"reactors and they can just use yourbacteria to produce whatever they want"},{"speaker":null,"startTime":1923.082,"endTime":1923.522,"body":"Ronja Rappold: Exactly."},{"speaker":null,"startTime":1923.563,"endTime":1927.782,"body":"With a small caveat or with a smallchallenge that they need to be"},{"speaker":null,"startTime":1927.842,"endTime":1932.223,"body":"adapted a little bit for methanolfermentation, because most of them"},{"speaker":null,"startTime":1932.282,"endTime":1938.722,"body":"will work with sugar or glycerol orother feedstocks, with few adaptations,"},{"speaker":null,"startTime":1938.902,"endTime":1940.423,"body":"methanol fermentation is possible."},{"speaker":null,"startTime":1941.503,"endTime":1944.523,"body":"Um, but it's a complete differentpicture than building a  whole new"},{"speaker":null,"startTime":1944.622,"endTime":1944.903,"body":"plant"},{"speaker":null,"startTime":1944.968,"endTime":1946.148,"body":"Kevin: yeah, yeah, definitely."},{"speaker":null,"startTime":1947.248,"endTime":1953.572,"body":"And, um, another topic that, uh, wouldbe interesting for me is you as, uh, one"},{"speaker":null,"startTime":1953.612,"endTime":1959.793,"body":"of the co-founders of this biotechnologycompany that will produce chemicals or"},{"speaker":null,"startTime":1959.973,"endTime":1962.032,"body":"deals with company that produce chemicals."},{"speaker":null,"startTime":1962.092,"endTime":1968.293,"body":"You, um, really have to be at theintersection of so many different"},{"speaker":null,"startTime":1968.372,"endTime":1973.032,"body":"disciplines, biology, chemistry,but also business and finances."},{"speaker":null,"startTime":1973.732,"endTime":1974.752,"body":"How do you manage that?"},{"speaker":null,"startTime":1975.23,"endTime":1978.13,"body":"Ronja Rappold: Yeah, I think firstthis is one of the things that"},{"speaker":null,"startTime":1978.23,"endTime":1979.89,"body":"excites me most about this whole"},{"speaker":null,"startTime":1979.97,"endTime":1980.491,"body":"adventure."},{"speaker":null,"startTime":1980.52,"endTime":1980.7,"body":"Kevin: Mhm."},{"speaker":null,"startTime":1981.43,"endTime":1981.79,"body":"Ronja Rappold: Um,"},{"speaker":null,"startTime":1982.15,"endTime":1987.131,"body":"coming out of the science, I thinkfor a scientist, I already had"},{"speaker":null,"startTime":1987.15,"endTime":1990.67,"body":"to deal with a lot of differentstakeholders during my time as a phD."},{"speaker":null,"startTime":1990.696,"endTime":1990.836,"body":"Kevin: Mhm."},{"speaker":null,"startTime":1991.551,"endTime":1991.81,"body":"Ronja Rappold: Um,"},{"speaker":null,"startTime":1992.17,"endTime":1996.27,"body":"back then it was scientistsout of very different fields,"},{"speaker":null,"startTime":1996.57,"endTime":1999.13,"body":"plus clinicians plus patients."},{"speaker":null,"startTime":2000.17,"endTime":2003.866,"body":"so there is already a very differentapproach to the same problem."},{"speaker":null,"startTime":2005.107,"endTime":2009.634,"body":"And now I think the challenge isthat first we need to translate"},{"speaker":null,"startTime":2009.674,"endTime":2014.834,"body":"this quite complex biology thespecific languages of the people."},{"speaker":null,"startTime":2015.915,"endTime":2022.754,"body":"so probably the salesperson ofcompany doesn't really, uh, wanna know"},{"speaker":null,"startTime":2022.875,"endTime":2025.225,"body":"about Adaptive laboratory evolution."},{"speaker":null,"startTime":2026.344,"endTime":2028.265,"body":"wanna know, is it a drop-in chemical?"},{"speaker":null,"startTime":2029.084,"endTime":2029.404,"body":"buy it?"},{"speaker":null,"startTime":2029.484,"endTime":2030.204,"body":"What is the price?"},{"speaker":null,"startTime":2031.064,"endTime":2037.595,"body":"And this is, uh, what over the last monthwas a wonderful learning experience that"},{"speaker":null,"startTime":2037.735,"endTime":2044.404,"body":"people are open, that they are interested,but they wanna also know the parameters"},{"speaker":null,"startTime":2044.443,"endTime":2048.644,"body":"that are important for them, and thisis our task to pr- to provide those"},{"speaker":null,"startTime":2049.334,"endTime":2054.653,"body":"Kevin: Yeah, it's basically a translatorbetween different words and I think this"},{"speaker":null,"startTime":2054.713,"endTime":2058.774,"body":"is really interesting also to interactwith so many different disciplines."},{"speaker":null,"startTime":2059.213,"endTime":2063.19,"body":"Because back in the days where likeeveryone was in their own world, like"},{"speaker":null,"startTime":2063.25,"endTime":2068.11,"body":"the chemists just had their chemistryproblems and the biology people their"},{"speaker":null,"startTime":2068.171,"endTime":2076.47,"body":"biology and yeah, that's really in therecent time that we have to collaborate"},{"speaker":null,"startTime":2076.55,"endTime":2080.35,"body":"with as many different people as possible."},{"speaker":null,"startTime":2080.39,"endTime":2085.628,"body":"And then this really brings outnew, ideas, new technologies, and"},{"speaker":null,"startTime":2085.668,"endTime":2088.608,"body":"this can then also impact the future"},{"speaker":null,"startTime":2089.095,"endTime":2095.295,"body":"Then, yeah, we can come to an end of thisdiscussion with the closing questions."},{"speaker":null,"startTime":2095.415,"endTime":2098.155,"body":"What is your favoritescientific discovery?"},{"speaker":null,"startTime":2098.275,"endTime":2102.016,"body":"Usually I ask about the chemicaldiscovery, but since you're"},{"speaker":null,"startTime":2102.596,"endTime":2106.935,"body":"not a chemist, uh, I go for thebroader scientific discovery."},{"speaker":null,"startTime":2108.233,"endTime":2109.553,"body":"Ronja Rappold: Thankyou for adapting then."},{"speaker":null,"startTime":2111.474,"endTime":2116.393,"body":"So, uh, the question, uh,because I love to talk about GFP."},{"speaker":null,"startTime":2116.874,"endTime":2120.473,"body":"Uh, it's, it's the abbreviationstands for green fluorescent protein"},{"speaker":null,"startTime":2120.667,"endTime":2120.827,"body":"Kevin: Mhm."},{"speaker":null,"startTime":2121.433,"endTime":2125.773,"body":"Ronja Rappold: I think since my firstsemester in bachelor's, I just love that."},{"speaker":null,"startTime":2126.514,"endTime":2132.75,"body":"Um, because as biologists, we do useit, basically to light up the inside of"},{"speaker":null,"startTime":2132.869,"endTime":2133.249,"body":"cells."},{"speaker":null,"startTime":2133.263,"endTime":2133.503,"body":"Kevin: Mhm."},{"speaker":null,"startTime":2134.569,"endTime":2138.41,"body":"Ronja Rappold: what you can do is youexcite it with a specific wavelength,"},{"speaker":null,"startTime":2138.729,"endTime":2142.109,"body":"and then under a fluorescentmicroscope, you can basically"},{"speaker":null,"startTime":2142.209,"endTime":2144.81,"body":"see really inside a living cell."},{"speaker":null,"startTime":2145.25,"endTime":2150.47,"body":"So you can fuse it with a protein ofinterest, and then you basically can"},{"speaker":null,"startTime":2150.55,"endTime":2154.314,"body":"live track the, the molecular adventure"},{"speaker":null,"startTime":2154.364,"endTime":2155.024,"body":"Kevin: interesting."},{"speaker":null,"startTime":2155.085,"endTime":2155.185,"body":"Ja"},{"speaker":null,"startTime":2156.03,"endTime":2156.851,"body":"Ronja Rappold: inside the cell."},{"speaker":null,"startTime":2158.651,"endTime":2164.27,"body":"um, since my bachelor thesis, I havebeen fascinated, uh, by that, and will"},{"speaker":null,"startTime":2164.29,"endTime":2167.63,"body":"never forget how it was the first timethat I saw this actually happening."},{"speaker":null,"startTime":2168.612,"endTime":2172.152,"body":"Kevin: Yeah, that sounds reallycool to really see with your"},{"speaker":null,"startTime":2172.212,"endTime":2173.612,"body":"own eyes what's happening."},{"speaker":null,"startTime":2174.232,"endTime":2175.452,"body":"Yeah, incredible."},{"speaker":null,"startTime":2175.992,"endTime":2181.272,"body":"I mean, everyone probably knows fromschool these, \u00e4h, when you have a plant"},{"speaker":null,"startTime":2181.372,"endTime":2186.313,"body":"cell under a microscope and you seethe different organelles, but there"},{"speaker":null,"startTime":2186.393,"endTime":2190.912,"body":"you can follow your changes, I guess."},{"speaker":null,"startTime":2191.535,"endTime":2192.935,"body":"Or, or what do you use the"},{"speaker":null,"startTime":2193.355,"endTime":2193.835,"body":"protein for?"},{"speaker":null,"startTime":2194.582,"endTime":2199.154,"body":"Ronja Rappold: you can basicallyinvestigate very, very specific molecular"},{"speaker":null,"startTime":2199.214,"endTime":2204.575,"body":"events inside this living cell, and Ithink this is just super fascinating"},{"speaker":null,"startTime":2204.883,"endTime":2205.023,"body":"Kevin: Ja."},{"speaker":null,"startTime":2205.683,"endTime":2206.893,"body":"And it glows."},{"speaker":null,"startTime":2207.72,"endTime":2211.13,"body":"Ronja Rappold: An it glows..That is, that is the cool part."},{"speaker":null,"startTime":2211.27,"endTime":2215.41,"body":"And you can win, \u00e4h, imagecompetitions with wonderful microscopy."},{"speaker":null,"startTime":2215.507,"endTime":2216.087,"body":"Kevin: Oh cool"},{"speaker":null,"startTime":2216.83,"endTime":2220.17,"body":"Ronja Rappold: images with that whichthen, sometimes can finance your,"},{"speaker":null,"startTime":2220.429,"endTime":2221.789,"body":"your next scientific conference."},{"speaker":null,"startTime":2223.107,"endTime":2224.126,"body":"Kevin: That sounds interesting."},{"speaker":null,"startTime":2224.426,"endTime":2230.546,"body":"Yeah, \u00e4hm, maybe we can link, \u00e4h, oneof these pictures in the, in the show"},{"speaker":null,"startTime":2230.646,"endTime":2233.827,"body":"notes to make that also visible to you."},{"speaker":null,"startTime":2234.786,"endTime":2239.066,"body":"And yeah, I will also link inthe show notes the publications"},{"speaker":null,"startTime":2239.826,"endTime":2242.786,"body":"from Ronja and her coworkers."},{"speaker":null,"startTime":2242.866,"endTime":2246.566,"body":"And then, \u00e4h, yeah, thank youfor joining me and telling me all"},{"speaker":null,"startTime":2246.626,"endTime":2248.246,"body":"about this biotechnology stuff"},{"speaker":null,"startTime":2249.613,"endTime":2250.654,"body":"Ronja Rappold: Thankyou for having me, Kevin"},{"speaker":null,"startTime":2251.039,"endTime":2252.669,"body":"Kevin: Also, thank youvery much for listening."},{"speaker":null,"startTime":2253.179,"endTime":2256.539,"body":"Check out the show notes forfluorescence microscope images"},{"speaker":null,"startTime":2257.079,"endTime":2259.089,"body":"and the links to the publications."},{"speaker":null,"startTime":2259.398,"endTime":2263.328,"body":"if you would like to talk about yourresearch in the podcast, contact me via"},{"speaker":null,"startTime":2263.328,"endTime":2268.984,"body":"email to contact@innovateandreact.comor connect with me on LinkedIn."},{"speaker":null,"startTime":2269.297,"endTime":2271.907,"body":"Until next time, keepinnovating and stay curious."}]}