 Thank you very much for giving this opportunity. So I'm just like to add that my background is actually I have a master in food chemistry and engineering and I'm also inside very passionate about the food technology and food production, although I have also chose some path to continue also with the focus with the plant reading so I have PhD in plant reading but I have been working with the plant proteins for most of my life. So here comes, I'm surprised Milenevac we have very similar interests and I think we should collaborate more in the future. Okay, so today my talk is about the hidden structures of the process plant foods and what do I mean with this hidden structures. I think I would like to talk a bit to repeat what Milenevac said and what was said also during two days of the workshop that we should follow actually a researchers industry also as a food consumer as we follow certain food trends. And as you may know, I mean this protein hype protein rich food is actually in focus for several years. People want to eat more healthy foods more to drink more healthy drinks, even as Milenevac showing rapeseed milk or we know very well actually only there's a good example of this representative product that has taken a lot of focus from consumers. As well as we also dream about the food actually will be health. Well, as a medicine for us for for solving some problems, for instance, have certain functionalities, as well as we strive for replace meat and find a good representatives but here I would like also to bring some provocative questions learning from Milenevac talk, because we also talk about the, we need more proteins. And if we compare protein products, how much of protein do we have in meat we have much less protein compared to protein plant protein based foods. And what is the reason what is a why do we need more proteins, we actually from with few proteins we can produce textures and structures that look like meat. So this is actually very simple understanding that we need to learn more about the how we can explore plant proteins and different food products. And as coming from University of Agriculture we also deal with a different. Well, hot topics as a seminal climate is actually can steer very much our food patterns as well as production of food together with sustainability and together with the trend that we also want locally produced and self sustainable products. So, this is a short introduction to what I would like to stress further and yesterday we heard a record from all the pointing out that the whole chain is very important and and and here I would like to bring you for our so called chain. And for our because we want to choose the right crop, we want to choose the right variety even go down to on the variety level, and to do a right processing in order to produce the right food. So then of course, everyone would ask what is right. It depends on the target that depends on the other focus, but in each situation has to be right for specific purpose. So we work with the food protein systems and today and also yesterday we heard a lot that we have differences when we work with the more refined system when there is protein isolate, or if we take the whole flower when we have a many components, including protein starch fibers and so on. To repeat that the genotype, what type of the variety you choose it can play a really different role, what type of outcome or what type of food you can get out. Also, in the way how do you cultivate that crop together. If you aim for specific component of the of the of the plant, the extraction process is really crucial and then we heard a lot, a lot of examples that industrial extraction is really damaging aggregating the protein protein is losing the native properties and you can't can do much with that protein, although native behaviors always more preferred. So how to deal with this challenges in industry how to deal with it today. My tools incorporate processing and if we we try to to play with different additives processing techniques in order to mask or to compensate this structural changes induced by, for instance, different extraction processes. So, the target of our studies have been mainly in focusing on proteins and protein changes, plant protein changes, when we produce different food products, as well as non food. So, we've been working a little bit on how on the structural changes and morphology of starch and fibers. And the interesting part is actually how these components of the plant. They are interacting with each other while they are processed here would like to bring some food examples that we've been working lately and for representative crops that you've probably heard about. And wheat is actually we chose as a model crop we've been working with wheat for very long time we know very well the system the system protein system is very complex is insoluble it aggregates easy and and also cause some processing issues. So, P is a trendy, trendy crop, because it's office nutritional aspects, as well as, as you see the market there's quite many P products that already taking over, although personally I just put another day P pasta and I had actually after cooking it I had to throw away because personally the test taste and smell it just was awful. So I think we have to keep in mind that, well, I think Tommy was also putting in the comments that the taste is really important in this context as well. So, to put up. So here we have also loop in, we have also actually outstanding a crop, but we, well exotic crop with we barely can grow here in Europe, although it comes all the way from South America from Peru, you know, it's a golden crop known by by ancient our ancestors that it contains very attractive nutritional profile and what does it happen with if we process such such cereal into retreat with the high temperatures what does it happen actually with the proteins and other components and loop in. Yes, it's attractive you to its ability to replace soil. So research questions actually, well, it's a chemistry what what's playing a chemistry behind the different processes as processes that involve the different additives different processing conditions. And, well, actually what happens on this molecular scale on, I mean the interactions and kind of the structures that are driving the functionality of this focus. This has been quite primarily focused in most of our studies. And to highlight the structures that are on the molecular level we use the synchrotron techniques, primarily small angle and wild bite angle scattering to elucidate differences on on different levels and on these proteins. But how do we start it actually to work with a with a bio based films the protein rich film that is in the cartoon is is made from gluten gluten that has been processed using the chemicals additives, I will come here. We have the graph showing the thick line and the thick spectral line that's showing the gluten together with the with the chemicals that they actually induces intriguing structures induce formation of intriguing structures but we never observed before. So, learning from this system. We actually been actually curious to to understand why these structures are formed in this particular, for instance system, and how these structures are continue forming if we change for instance, different genotype. On your left is a genotype that has more disqualified bonds is a wheat gluten extracted from that type of genotype and on the right side you have the less system having genotype, which is interesting here so you have a different growing inputs to produce these genotypes and we were also interested in how these different growing inputs, beside the temperature there is a nitrogen input, how these growing inputs induce differences on the nano structure. You see a lot of curves and a lot of arrows, I just don't want to go too much into details but the on the left spectra red curve you, you see the black arrows indicating hexagonal structure formation. While the green I was pointing at the lamella structure so in this type of system we observed two different arrangements and in actually gluten films. And that of course is interesting how the structures are correlating with a tensile properties so the one indicated then in the with the bars it shows clearly the very different responding behavior in terms of the elasticity model modulus of these two films. So he has an example showing how genetics can play a role as well as a growing factors. We were one again with a with another known food sample so this is a last known food sample that I wanted to show you we blended in the proteins together with starch we extruded it into composites, and we looked at the how the nano structure of proteins is changing with the composition of the blend. So the bottom curve is showing the nano structure of the gliders and is nice to show here that this hexagonal structure is still remaining through the difference of the blends when the starch is still intact. So what can we learn from that that the proteins in some way acting as a matrix and actually playing a fundamental role and if you introduce touch you still don't break that arrangement in the system. So now we move on to food examples and here I would like to to give the few samples on the porous snacks that we produced for gliding protein slide and as they said we would love to work with gliders because they form this nice structures and he has extreme tomography showing the two different images they are probably difficult to distinguish by I but the difference between them is that one is is actually produced with the no enzyme just a protein another is with the transglutaminase treated protein and this is thanks to collaboration with Steve and we observed that there is in by introducing transglutaminase of course one is expecting that the proteins will get crosslink, but beside that we also observed the porosity differences in porosity and the sizes of the of the bubbles formed so the down curve is showing the higher and bigger sizes of the bubbles. The snacks microstructure we also observed that in this particular samples we also had a different interfacial structures microstructures as you can see pointed out with the yellow arrows no enzyme having the more irregular structure while enzyme treated has kind of sheets holding bubbles together and when we come to the scanning sacks pattern we also compare the effect the enzyme effects as be a six this is specific transglutaminase enzyme is not commercial is inside that we produced in our or in the collaborators lab and we compared how the guidance are impacted by enzyme concentrations the first prime so there is a slight shift in the scattering pattern where they increase in the enzyme concentration if you compare while the glycerol is actually giving no effect the glycerol together with the enzyme is is actually no introducing no structural changes so that the pattern looks rather similar which means that the glycerol in some way is actually contributing to protein aggregation and actually not allowing enzyme to perform its function we want to do another additive which is very interesting linoleic acid effect we wanted to show in the samples but the idea to introduce this additive was to increase the nutritional value of the of the snacks actually the scattering curves showing that no only glide in no additive is actually having the well rather simple simple scattering pattern while there are additive is simply barely differences are between the no enzyme and enzyme having samples again showing that on the if you look at the scattering pattern on the right very clear linoleic acid pattern that is actually no leg acid is not incorporated in the protein network and it's actually sitting as a as a component in the structure loop in frozen cast films we also intrigued houses the structural dynamics is changing by introducing different additive so if you look at the a picture showing no additive just looping protein isolate beside to mention that this looping protein isolate has been produced by dry fractionation although with idea to to make the highest possible looping isolate later to be used in different foods while in B image you see the introduction of the transglutaminase in serious introduction of transglutaminase and lecithin and indeed you see a linoleic acid and actually enzyme so there is quite a dynamic change in morphology of these films as well as in the microstructure and with the scanning electron microscopy we were able to to see that the structure works was extremely fragile as you could see is a bit messy image on a in a case but you could able we were able to distinguish the pores of the lamellas as well as the actually specifically formed bridges between lamellas in the lecithin case so it's again and what actually we can learn from the microstructure we can learn the the certain level of the how ingredients together with the with the proteins interact but on the nano level which is shown in this graph we compared the impact of the nanostructure of these different additives and well in the glycerol case we we see that there is a well commercial in the first black curve indicating the commercial enzyme a slight impact while in the in the following curves we see no really not not big influence of the glycerol and enzymes used by the lecithin case we with the enzyme we see the the violet top curve we see some structures some nano structures formed due to enzyme and protein while in the case of the transglutaminase and linoleic acid we see very clear pattern of linoleic acid profound well actually formed and actually buried in the structure so this gives us more understanding how we can play with different systems and in this case in the case that I showed it was with the lupin but as well as I think playing with it more refined system is giving more clarity while here we move on to more complex sample as it is we have the quinoa on the first image you have quinoa seed following image we have flour and that flour was extruded into the so called Peruvian actually national food is actually pellets or flakes and these flakes were dried in the high temperatures so we're curious to see what's happening with the proteins and the nutritional components during processing and the proteins the top graph is showing the from HPLC the green graph is showing the protein pattern as it is extracted from the flour and it shows a different type of proteins while the proteins very clearly aggregate and there in the curve below that after processing so the extrusion although was not very high temperatures but still impact severely the protein this actually could be we compared the microstructure of the proteins another protein sorry the extrudates and the one on the on your left is actually extrude as it comes the one on your right is extrude that it has a post processed with the temperature which means that he has been drying at the high temperature for very short time from outside you see the color differences from inside or actually just color differences but they look very similar from inside as you could see from this microstructure they look entirely different the one on the left is very compact on the one on the right is very porous from nutritional profiles one of the questions is what what is happening actually if we use the high extrusion temperatures how the this super serial nutritional profile is affected and by our big surprise that only methionine and and cysteine have been reduced while in a while other amino acids they still kept quite quite good numbers after after extrusion so this is rather well I think it's important information to learn that you still can can do a play with the processing and rather high temperatures but you still don't affect in in severely the nutritional composition again another well similar information has been obtained with of these samples we looked all the with the extreme tomography and I think here is only new information is from the profile picture that that you can see the structure and distribution through longitude direction the the sample on your right so what is happening on the nano structure level of these process serials and here actually is is pretty complex pretty complex spectra to to interpret for the protein part because we can't tell much about proteins but at least we can tell about the starch so the the top down to curves representing the grain and flower while the other top three curves representing processed materials so so the extruded extruded dried and extruded grinded material and there you have a different examples of different starch crystallinity and we look closer what happens with this starch crystallinity dynamics we can actually understand from the study that crystallinity degree of crystallinity is changing so here we have a crystallinity which is around not around very specific very exactly 29% so if we apply extrusion we have a decrease in crystallinity as it's obvious decrease in the peaks and we have a decrease up to 8% but if we drive this material if we drive this food we have a even further decrease and further shifts in crystallinity of starch and we moved to to other very brief example of the P protein and an idea with this was to also biofractionation method to produce protein rich fraction fiber rich fraction blend them together with a process using the temperature here we have the initial I think it has been met has not been mentioned during the during the this workshop that the importance of the particle size of the importance of the beside the purity of the fractions importance of the particle size is playing very fundamental role how these ingredients are blended further in the food and we kind of got the fraction from our collaborators from University of Copenhagen we found it well we looked at the microscope and we're very surprised they call this fiber fraction fiber rich fraction although it was filled with a lot of starch after fractionation process and this type is was very well acting as a filler in the matrix in in the process process food and of course it was contributing to different functional properties to this stretchness as you could see the protein isolate sample was was much more elongated compared to the fractions where the proteins and fibers were blended in while hardness was no influence at all so this information can help us understand what is happening on the on the micro structure on the secondary structure using FTI are method and we show that if you have the only proteins we have a domination of the intermolecular better sheet structure formation versus alpha helix and this goes on for all the say this is a protein and this is in the blends and then we have actually only fiber where is no such secondary structures are presented so it's another additional information to understand the system and it's rather helpful although for for sex part I still put the question mark is one of the could be possibilities to understand what happens with the protein fiber interaction but we should not forget we have starch there we have a shift in distances for the protein isolate or protein isolate we have a lower distances while we have a blend we have a high distances how much we can learn from protein fiber interaction interpretation is still rather rather complex so we need to learn a lot how to interpret this data again to to break down it's like like Lego right I mean you you you build some you have something built and then you have to to put it down to the pieces and then try to to work with the piece and add piece by piece and try to build the understanding a structure so to conclude to conclude for instance Kinoa is extrusion well has been extrusion processing using the trend temperatures between 69 and 95 so surprisingly fairly good temperatures to to to play around and still not damage for instance nutritional aspects and still get the the product well processed although that we can't avoid the the the using temperatures and can't avoid protein cross linking and aggregation so we have to live with it and and and that's what we we have to keep in mind as well as the porosity you can play around with the proteins and different additives you can play around also with the porosity and different type of morphology we've been observed by we observed that with a scanning electron microscopy and ray tomography as well as decrease if we want to to play with the starch crystallinity it decrease it this is one of the examples and I didn't mention that but the starch types has been changed during processing so they the type a turn into type V starches and that was observed using the wax technique and extrusion is still good processing versus roasting and still nutritional attractive so future prospects I think is good to to to highlight that is controlling the structure of plant components is is important is could be used as a tuning a functional element in producing food as well as a deep understanding a plant components using the techniques synchrotron techniques together in combination with the other methods I think could be a very good approach to understand and more explore protein protein protein starch and protein fiber interactions as well as produce new yeah new ways of trying to to tune food for specific properties so I would like to thank collaborators colleagues funders max for laboratory for providing being time and industry partners as well as you for your attention so thank you very much thank you Ramona we have time for a very quick question if anyone has before break we are two minutes over any thought I'll just ask you a quick question Ramona you started the presentation it also goes a little bit to what Milena was saying that we I understood that you said that native behavior of proteins are the preferred behavior but you do see that you get the difference after extrusion but the nutritional was not so bad and I think also Milena said that sometimes we have to destroy so to say the protein because we have to have a heat systems could you elaborate a little bit more on what you mean with the problem problems of native or non native proteins. Yeah I mean that was also talk to think during the well in Milena's talk and I think in several other talks that for instance spray drying is very hash processing methods and you never get the protein in native food so you simply introduce changes and that's what we explored it in the lab you know to work with the glutinous is rather easy because you can easy wash it by hand it flew from flower so you just use simply make it make a matrix, make a dough and then you wash under the water and then you we analyze that the structure and we compared with the industrially produced gluten and when we saw much more native properties of gluten the one that was produced in the lab, meaning that proteins are more prone to interact and form the the polymers the polymers are important for functionality while and it's the same with the potato protein is the same with other proteins so but they understand as well as the industry can't throw away machines they have to use the machines that they use but probably they have to play with a bit with tuning and trying I mean I cannot give the answer that well we have to use it to throw away and use the native way of making proteins but I think it's good to keep in mind what kind of functions you would have with a protein if you would not introduce this well modifications during extraction and processing so let's say trade off I mean trade off depending on the on the yeah on the focus as well so yeah you have to find some somewhere compromise and I think that's what is industry struggling and doing and yeah thank you thank you