Periodic Reporting for period 3 - DAFIA (Biomacromolecules from municipal solid bio-waste fractions and fish waste for high added value applications.) Reporting period: 2020-01-01 to 2020-12-31 Summary of the context and overall objectives of the project A huge advance in the project progress has been achieved focus on the exploitation of MSW and MRRM as feedstocks for high value products. In the first stages the availability and compatibility of selected raw materials and their fractions have been ensured and they have been completely characterized. Different technologies have been applied to isolate the gelatine and nucleic acid extraction from fish-processing residues. Several strains of bacteria have been applied to produce the desired bio-monomers (1,5-pentanediamine, muconic acid and adipic acid) from MSW hydrolysates.On the other hand, different components, including fish bones, were used to develop different bio-based flame retardants and they have been compounded in commercial polyamides including bio-polyamides. Moreover, edible and barrier coatings have been developed from high purity extracted gelatine. All the process flowsheets have been prepared to fill a data repository for environmental and economic analysis. Communication and dissemination activities through articles in magazines, articles and online portals aimed at specific countries and/or sectors has been very frecuent. Technology Transfer related to products and chemical building blocks developed has been carried out between the consortium. The exploitable results in DAFIA can be divided: coatings and barriers from fish residues, flame retardants from fish residues, monomers produced by fermentation and bio based flame retardant polyamides. Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far Different technologies have been applied to isolate the desired components from both feedstocks and preservation conditions for MSW have been evaluated too. Several strains and bacteria have been used to produce the desired bio-monomers (1,5-pentanediamine, muconic acid and adipic acid). Different approaches to improve the productivity of PDA were the hydrolisate is the source of carbon are being evaluated, and a better performance is observed. The production of AA strains was tested for xylose utilization in complex media. The theoretical maximum concentration from unconcentrated waste hydrolysate is expected to be between 4-5 g/L of adipic acid, but even after cell factory the titers achieved were very low, so this line has been rejected. In the case of CCM production we are currently investigating different strategies to generate stable CCM production strains. Final CCM production strain produces 20.8g/L in MSW hydrolysate at 50L scale. Downstream processing tests (DSP) have been performed on fermentation broth and purified by three different methods. So far the best result allowed to achieve a total yield of 66.4% with a purity of 95.5 of de diacid. Different components, including gelatine and cod bones, were used to develop different FR. The thermal stability of these products was evaluated because the requirements to process polyamides are very exigent. The formulation of the FR has been optimised and their thermal stability profile studied. Flame retardancy tests were also done and compounded with different PA (PA66 & PA11) in order to find optimal conditions. It was seen that it is possible to have good retardancy properties when 25% FR is added to PA11.Moreover, edible and barrier coatings have been developed from extracted gelatine as alternative to EVOH. Different parameters were taken into account and well as ratio of components. This allowed to optimize the process and the best conditions in order to obtain OTR values comparable to the commercial barrier packaging. Also, the development of active packaging solutions has been performed. Barrier coating based on water-borne fish gelatine has been applied onto polyethylene/polypropylene and polyethylenterephtale film substrate by means of gravure roll in a semi-industrial lamination equipment. Resin coating weight (g/m2) has been determined and permeability tests have been performed. The scale-up process has been optimised in order to ensure good oxygen permeability. The production of PA56 with commercial AA and synthetised PDA from the project has been evaluated through different synthetic routes, achieving similar results to the commercial counterparts. Compounds of the FR developed with PA were made and tested in commercial PA66 and PA11. In relation to the modification of muconic acid, high conv.(98%) and select.(86%) achieved with amine based catalysts when converting dimethyl maleate. Working with dimethyl muconate is more challenging as lower activity is shown and higher yields of by products are obtained. Economic analysis and LCA have point out the challenges to overcome to increase the TRL of DAFIA developments.Conferences: 20Oral Presentation: 9Role-up, poster leaflet: 17Workshop: 3Presentation: 17Newsletter: 5Book chapter: 6S.Journal: 7Magazine: 11PublicAtions: 7Patents: 2 Progress beyond the state of the art and expected potential impact (including the socio-economic impact and the wider societal implications of the project so far) The main results related with the progress beyond the state of the art are focus on these points:- For lysine and PDA enzymatic production can supply valuable information to be added to the novel information from the mutational mapping and can allow further rational engineering to increase lysine and thereby also PDA production levels. Utilisation of hydrolysate from MSW was not been done before. DAFIA has demonstrated that the production of high purity >99% PDA is possible by waste valorisation and strain engineering. - For production of adipic acid has been discontinued due to the better performance of the fermentation of the PDA and CCM.- In the case of CCM, stable CCM production strain produces 20.8/L CCM using MSW hydrolysate as a carbon source was validated at 50L scale. The literature showed much lower titers(9g/L) than the ones found at DAFIA. - Novel bio-based flame retardants including cod bones & gelatine from MRRM have been developed and patented. Their application in PA66 & PA11 led to a good flammability properties. This technology has not been applied before to such a high temperature processing polymers.- Bioactive coatings containing gelatine and nisin has shown good performance, and the production at semi-industrial level of films with good barrier properties, comparable to the commercial OTR values for EVOH, has been obtained. Dip coatings showed also good antioxidant properties.- The synthesis of PA56 in solventless conditions shows a promissing alternative route to the current commercial products.Exploitation workshops about the KERs were held between NNFCC and individual partners, in the form of 1-to-1 phone calls. Based on the outcome of these calls, the Exploitation Plan working document was updated by NNFCC. DAFIA has many potential KERs, the results are divided into 3 groups: fish residue valorization, microbial transformation of waste, and polymer production. Business plans are being written for the most promising KERs. Based on exploitation workshops with the partners, 4 KERs were selected for 4 business plans. All IP ownership is clear in the project and no IP issues are foreseen. FTO searches have been carried out.Economic analysis showed that taste-neutral gelatine has a potential market. PDA economics showed that more investigation is needed in DSP.LCA analysis demonstrated that energy consumption is one of the challenges to overcome in terms of carbon footprint in MSW hydrolysate, FR synthesis and bio-based coatings. In the case of gelatine extraction the critical point is the use of solvents and for PDA and PA synthesis the use of commercial raw materials schematic-fractionation-of-salmon-skins-extraction-of-gelatine.jpg
