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Report

Teaser, summary, work performed and final results

Periodic Reporting for period 1 - DAFIA (Biomacromolecules from municipal solid bio-waste fractions and fish waste for high added value applications.)

Teaser

Summary

During period 1 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 expected products from both feedstocks, such as gelatine, nucleic acidsâ€¦ 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 gelatine, 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 commercial gelatine (warm and cold). All the process flowsheets have been prepared to fill a data repository. Communication and dissemination are progressing as planned in DAFIA. In the first 18 months of DAFIA, the project start was communicated via partner homepages and newsletters, as well as through articles in magazines and online portals aimed at specific countries and/or sectors.

Work performed

The availability and compatibility of municipal solid wastes (MSW) and marine rest raw materials (MRRM) have been ensured and they have been completely characterized. 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). Poor PDA production was obtained with 100 % hydrolysate medium, so different approaches are being evaluated. The information from the mutational mapping is valuable and can allow further rational engineering to increase lysine and thereby also PDA production levels. For production of adipic acid the strains were tested for xylose utilization in complex media and the possibility to increase xylose utilization was studied. Some of the results described were presented at the Biotechnology for Biofuels and Biochemical congress in USA, Florida in May 2018, where both academia and industries were excited to see the possibility to utilize household waste as a feedstock. The theoretical maximum concentration from unconcentrated waste hydrolysate is expected to be between 4-5 g/L of adipic acid. Future work on adipic acid cell factory includes optimization of adipic acid titers. In the case of cis,cis-muconic acid (CCM) production we are currently investigating different strategies to generate stable CCM production strains. Currently, our best performing stable CCM production strain produces 220mg/L in MSW hydrolysate. We have identified 2 pathways that can realistically be engineered. Initial downstream processing tests (DSP) have been performed on a pure mixture of PDA in water. The pH has an important effect on the efficiency of certain downstream processes.
On the other hand, different components, including gelatine, were used to develop different bio-based flame retardants. The thermal stability of these products was evaluated because the requirements to process polyamides are very exigent and for this reason this aspect is a challenge not easy to solve. Several FRs with different combinations of gelatine were synthesized and thermogravimetric analyses (TGA-FTIR) were done to evaluate their thermal stability profile. Flame retardancy tests were also done. These FRs were compounded with different polyamides in order to find optimal conditions for processing and FR performance. It was seen that it is possible to regulate the amount of gelatine in the FR in order to achieve optimal flame retardancy and compatibility.
Moreover, edible and barrier coatings have been developed from commercial gelatine (warm and cold). This will allow to optimize the process and the best conditions will be the ones applied for the gelatine extracted in the project. Different parameters were taken into account and well as ratio of components. Also, the development of active packaging solutions has just started. Barrier coating based on water-borne fish gelatine has been applied onto polyethylene film substrate by means of gravure roll. Resin coating weight (g/m2) has been determined and permeability tests have been performed.
At this stage, the production of polyamides with commercial monomers has been evaluated through different synthetic routes. The presence of some possible impurities has also been evaluated to study their effect on these approaches and the scaling of polyamide is currently in progress. Compounds of the FR developed with polyamides has been prepared and tested.
The process flowsheets for each process have been prepared to fill a data repository. A platform for the mass and energy balances was set-up and is continuously being updated using the data from the partners and preliminary results were obtained. Concepts for running sensibility analyses on the process models were designed and started being implemented.
Communication and dissemination is progressing as planned in DAFIA. In the

Final results

The main results related with the progress beyond the state of the art are focus on these points:
- Our results 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.
- For production of adipic acid, our results (presented at the Biotechnology for Biofuels and Biochemical congress in USA, Florida in May 2018) have shown an important impact in both academia and industries sector that were excited to see the possibility to utilize household waste as a feedstock.
- We also have tested ability of parent strains to consume adipic acid. Then possible pathways towards adipic acid were determined. The feasible pathway could operate with the maximum theoretical yield of 0.069 g/g.
- In the case of cis,cis-muconic acid (CCM) our best performing stable CCM production strain produces 280mg/L CCM in glucose limited FIT and 220mg/L in MSW hydrolysate.
- Novel bio-based flame retardants including gelatine from MRRM have been developed and patented.