BioSFerA H2020, project’s update and meeting in Madrid

For the first time, after two years of project, the BioSFerA Consortium meets in person in Madrid. The first physical meeting took place last 3rd – 4th May at the Center for Biological Research Margarita Salas (CIB) of our partner CSIC, the Spanish National Research Council. The two-days meeting was also the opportunity to visit the CIB-CSIC’s laboratories, where a large number of researchers conduct their researchs on biotechnology, included the experiments for BioSFerA.

 

Concerning the CSIC’s activities, the research centre is continuing its trials currently dedicated to the genetic modification of selected acetogenic bacterial and oleaginous yeast strains: based on genetic engineering strategies, CSIC has created 8 recombinant Moorella strains with different acetate production rates. Results are still not fully satisfying, so that CSIC will move forward to better understand the recombinant bacterial behaviour, considering extreme fermentation conditions and syngas contaminants. On the other hand, genetic modifications on several genes involved in lipid metabolism are also in progress for Y. lipolytica, the oleaginous yeast used in the triacylglycerides (TAGs) production phase from acetate.

 

Meanwhile, the investigation of the gas & liquid fermentation processes for the parameters’ optimization are still ongoing. The activities were conducted on pressurized bioreactors of 1 L and 10 L dimensions for CARTIF and BBEPP respectively and using realistic data on the VTT’s syngas mix. They operated in different conditions to explore the factors influencing the acetate production, such as the use of different media or the application of a continuous fermentation mode with a membrane for cell recycling. A general finding from these tests is that increased biomass concentration can lead to higher volumetric acid acetic production. In addition, increased pressure was applied to overcome gas transfer limitations. Further investigations on the oleaginous yeasts’ fermentation step are also proceeding: BBEPP presented the results so far of acetate fermentation trials in the 7 L fed-batch bioreactor applying a pH-static feeding strategy, whereby different trials were performed using the both wild-type and genetically modified Yarrowia strains. CARTIF achieved tests using a fed-batch fermenter and two different continuous feeding strategies (high constant and gradually increased flow rate), but both the feeding and the cell recycling strategies need to be optimised to further increase TAGs production. First experiments were also performed by ENVIPARK with its steam explosion pilot plant as first step in the downstream process for the TAGs recovery. These first trials were conducted using a broth sample provided by BBEPP and literature parameters for an initial setting.

 

Even the preparations for the pilot process activities are advancing: VTT updates the Consortium with the successful accomplishment of the Factory Acceptance Test and the last steps to conclude the gasification unit, while BBEPP informed about the finalisation of the Bio Base Mobile Pilot Plant (BBMPP) construction. A draft timeline of the work has been presented, defining the transportation of the BBEPP’s mobile unit to the VTT site at the end of the year 2022 and the launch of the pilot process integration within 2023.

 

Within the progresses of the project’s activities, it is time for the scaling-up of the BioSFerA’s concept, starting with the DFB (Dual Fluidized Bed) gasification system as first block to help its functionality and transferability at industrial level. The techno-economic, environmental and social analysis has started as well. RINA, and the other partners involved in the activity, are preparing the basis for the future assessments related to the LCA/LCC analysis (CERTH), and the social-LCA too (NTUA). One reason of discussion was the current energy crisis and the impact of the COVID-19 outbreak on the expected techno-economic performance indicators, which will need to take into account these aspects.

 

Finally, the dissemination activities continue to follow the project’s progresses and advance with new activities, such as the release of the official BioSFerA’s newsletter at the end of May and the update of the website with the news related to the pilot process. On the 4th May afternoon, after the meeting, a workshop on “Advanced Technologies for Green Molecules Production” was also organised in hybrid format in synergy with other two projects COS2MOS and Life Biomass C+, giving evidence of the first project’s results and its contribution to the biotechnology research development. After this fruitful meeting, the Consortium expects to meet again in October, this time in Belgium at the BBEPP’s headquarter.

The BioSFerA Consortium at CSIC premises during the 4th GA meeting

VTT EXPLAINS: THE SYNGAS PROCESS AND ITS INTEGRATION WITH THE FERMENTATION UNIT

Interview with Ville Nikkanen

The syngas production from different biomass feedstocks is the first process step characterising the concept proposed by BioSFerA. Thanks to the exploration of these biofeedstocks from all over Europe, the improvement of the gasification process allows to increase the value of the BioSFerA’s technology according to sustainability and circularity principles. As initial stage, it is also crucial to have a good operative and performance control for the following gas fermentation steps and downstream process to achieve the final biofuels as expected.

VTT, as an expert in the gasification process, has carried out the bench-scale test at TRL4 with 5 different feedstocks, necessary to proceed with the second step of fermentation. These preliminary analyses are also important for the scale-up activity consisting into the integration of the BBEPP’s mobile gas fermentation unit with the syngas process and having as main results the construction and run of a pilot process plant at TRL5.

Mr. Ville Nikkanen, which has coordinated the VTT activity on the syngas production, explains more in detail the path taken and the future development concerning the syngas plant.

 

What tests have you conducted so far and what results have they produced?

Preliminary tests at bench-scale have been conducted to provide data describing the syngas composition produced by the feedstock used for the biological research and the following double-stage fermentation. The bench-scale tests were conducted in a bubbling fluidized bed gasifier (BFB), showed in figure 1, with continuous flows of feedstock, bed material (sand and dolomite) and gasification agents (H2O and O2). A hot gas filter and reformer units were used for eliminating particles and tars from the produced syngas. For removing trace impurities and for improving the syngas quality, a water scrubber and adsorber beds after the reformer unit were tested. The gas quality were analysed by using different online and offline gas analysis methods. During the experiments, the main parameters influencing the final syngas composition are the feedstocks and the feed gasses (O2, CO2, H2O, N2) and the used processing units. The achieved results show the reforming unit is required in the gas fermentation process in order to remove the harmful components (e.g., naphthalene and benzene). Moreover, gasses that are fed into the gasifier and reformer (O2, N2, H2O, CO2) and their volume flow influence the syngas composition more than the selected feedstock. Finally, water samples collected after the reformer unit seem to be suitable for gas fermentation. The long-term accumulation of impurities and the effect of this will be studied later in the project in piloting runs expected to be carried out in 2023.

 

How does the different feedstock affect the functioning of the system?

5 different feedstocks in the form of pellets have been tested: forest residue, bark, straw, sunflower husk and olive pruning. Higher concentrations of H2S, COS, HCN, and NH3 are measured with feedstock that have high sulfur (S) and nitrogen (N) content, such as straw, sunflower husk, and olive prunings. This might lead to larger requirements for the gas cleaning in the downstream, if these gaseous components inhibit the growth of microbes. Also, ash sintering can cause more operational problems in the gasifier if straw, sunflower husk, or olive prunings are used as a feedstock in the pilot process. Based on those bench-scale gasification tests all of the feedstocks could be potentially selected for the piloting runs, specifically because the reformer helps to average the syngas composition. Then the main criteria for the feedstock selection are their availability, cost, and sustainability.

 

What is the added value in integrating the syngas unit with the fermentation unit? What kind of technical limits must be overcome?

The integration between the syngas and fermentation units allows to join the best characteristics of each technology in the right combination in a single process. The thermochemical and biochemical approaches offset each other in some steps, reducing costs and enhancing the efficiency of the overall process. For example, the costs of sulfur (H2S) and nitrogen (NH3) removal could be potentially minimized as the microbes seem to be very tolerant toward these components. However, some specific components such as hydrogen cyanide (HCN, COS) can be potentially very inhibiting and toxic even at very low concentrations leading to a need to remove these components selectively. Sure, it is also a challenge to find the right syngas composition to feed the microorganisms, the conditions to exploit the technology according to the available feedstock, and from a bench to a pilot-scale until TRL5. One of the main technical parameters is the monitoring and minimisation of the inhibiting components in short and long-term operation activity. An ultra-cleaned syngas unit helps to control these data and a continuous syngas flow is required to minimise the production of unwanted substances from not standard operation.

 

What are the next steps and expected results to reach?

Last improvements to optimise the syngas production are focused on the minimisation of the operational risks in the piloting phase. Therefore, the main activity is the identification of the most critical impurities and, specifically, the impurity tolerance to better control the risks and select the only the necessary process units, reducing costs and enhancing the overall efficiency. In parallel, an action plan has been settled with BBEPP, in charge of the mobile gas fermentation unit. The main next steps and the shipping of the components at Bioruukki in Finland have been defined, planning to start with the integration operations in March 2023 to have the process completed by the end of March 2024. The pilot-scale gasifier can be fed with maximum 100 kg of biomass per hour. The aim of these piloting tests is to produce a realistic gas mixture for the gas fermentation and run the process continuously for hundreds of hours to verify the performance in a realistic environment and to scale up the gas fermentation step in an integrated process. Until now, we have worked together with BBEPP to identify some starting parameters to join the two units, such as the minimum gas pressure at the compressor inlet and also safety and automation requirements.

Figure 1 – Syngas process scheme: dual fluidized bed gasifier,
hot-gas filter and catalytic reformer unit.

 

Report Deliverable 3.1 – Bench-scale gasification tests at TRL4

Scientific Paper Activated Carbons for Syngas Desulfurization: Evaluating Approaches for Enhancing Low-Temperature H2S Oxidation Rate

The importance of the microorganisms’ study in the BioSFerA concept

Interview with José Luis García

Spanish Nation Research Council (CSIC), as one of the largest European public research institution with high expertise in the biotechnology field, gives its contribution to BioSFerA on biological processes at laboratory scale thorough its institute, the Center for Biological Research Margarita Salas (CIB) (Madrid). Its main activity is the research of the best microorganisms, playing a key role in the double stage fermentation characterising the BioSFerA process.

Exploring the micro level and the microorganisms’ interaction with the surrounding environment is crucial when moving processes from the laboratory to pilot scale level. Few questions to go deepen the CSIC activity and the biological research in laboratory with José Luis García, Research Professor at CIB (CSIC) and leader of the Environmental Biotechnology group:

 

As project partner, how CSIC contribute to the project and what is the innovative aspect led by your research activity?

Our main contribution to BioSFerA is to develop a new collection of system metabolic engineering tools that can allow to modify the acetogenic bacteria and the oleaginous yeasts in order to obtain robust bacteria and yeast that could render higher production yields of acetate and lipids than the natural wild type strains. Using these tools and considering the lessons learned during the implementation of the processes at industrial scale it will be possible to propose other modifications to improve even further the production yields.

 

Your work is focused on bacteria and their interaction with organic matter and the surrounding environment. What are the main limitations hindering bacterial activities to consider for their performance, specifically in the BioSFerA processes?

It’s important to consider the final objective of the process, which in BioSFerA is to transform the syngas obtained from biomass gasification into acetate that will be further used to produce lipids. Analysing our model, we can split it into two main steps: in the first one, we use acetogenic bacteria to transform syngas into acetate, whereas in the second step we use oleaginous yeasts to transform acetate into lipids. To develop an industrial efficient process the first problem that we have to handle is how to increase the bacterial production of acetate from syngas. This can be achieved by combining the production of enough microbial biomass from syngas and an efficient transformation of syngas into acetate. In addition, we have to take care of possible syngas contaminants that are toxic for the bacterial acetogenic strains and that can affect bacterial growth or viability. Hence, we have performed a screening of different acetogenic bacteria that are able to grow at high cell densities, that produce high amounts of acetate, and that are resistant or tolerant to some contaminant/impurities present in syngas.

Example of Y. lipolytica strains growing
on YPD (rich liquid medium) agar plates.

 

Considering the acetate production, how the syngas composition from the different biomass can affect the fermentation process?

As mentioned above, in BiosFerA we are traying to transform the residual biomass into syngas: a mixture of carbon dioxide, carbon monoxide and hydrogen, that is obtained by a process named gasification. The syngas, as result of a gasification process, depends on the origin and properties of the biomass which influence the presence of the contaminants and the syngas composition. Usually, the biomass gasification not only produces syngas, but also other compounds like hydrogen sulphide, hydrogen cyanide, benzene, ammonia, methane, etc. This means the production of acetate will be affected by the proportion of the main gases (CO2, CO, H2), that varies depending of the biomass and the gasification conditions and that are responsible of the bacterial growth and the acetate yields, and also by the presence of toxic unwanted contaminants. Therefore, it is important to find the most appropriated syngas composition as well as to eliminate as much as possible the unwanted contaminants to a level that can not affect bacterial growth or acetate production. 

 

What kind of approach have you followed in these research activities and how is the work coordinated with the other partners? What are the advantages of having such a broad, interdisciplinary team as BioSFerA?

In a very early steps, we have collected all the information available in the literature dealing with the metabolism, molecular biology and processes of the organisms of interest, this is acetogenic bacteria and oleaginous yeasts. Then, we have selected the best strains and performed a screening of their properties using different culture conditions in order to test which are the best acetate producers from syngas and lipid producer from acetate. After selecting the best performing microorganisms, i.e. the acetogenic bacteria Moorella thermoacetica and the oleaginous yeast Yarrowia lipolytica, we have used systems metabolic engineering approaches to identify the target genes that have to be deleted or overexpressed, to channelling the metabolism to the production of the metabolite of interest. After modifying the strains, they have been initially tested at CIB-CSIC at lab scale using shake flask cultures and then transferred to CARTIF and BBEPP for fermentations in bioreactors at high volume levels. At this point, the interdisciplinary nature of the different groups involved in BioSFerA project allows us to find the best operational conditions to produce the compounds of interest at pilot and industrial scales.

 

Acetate metabolism in acetogenic bacteria.
Target genes are indicated in colours.

Lipid (TAG) metabolism in oleaginous yeasts.
The main target genes are indicated in red.

 

So, first you analyse the organism’s metabolism to identify the best strains and then you guide them in the metabolic process of the desired substances, through genetic modifications. What is the value added to use GMO microorganisms?

GMO microorganisms are designed for different purposes, but in principle they are all aimed at increasing the performance of processes. The yield of the process can be increased by redirecting the carbon flux to the production of the desired product and at the same time by avoiding the formation of unwanted by-products. Additionally, as it is the case of lipids, we can also modify the microorganism eliminating the degradative pathways of lipids and fatty acids, facilitating that the decided final product is accumulated inside the cell, without the possibility of its further degradation. The GMOs constructed in BiosFerA project are oriented to increase the production of acetate from syngas in the case of acetogenic bacteria, and to increase the production of lipids from acetate in the case of oleaginous yeast. If required, the strains can be also modified to acquire other properties, for instance the resistance or tolerance to specific toxic contaminants.

Electroporator used to create OGMs

 

Detailed information on the biological research at laboratory scale carried out by the BioSFerA’s partners are available in the Deliverable 3.2Deliverable 3.3.

Further explore through the opinion-Editorial Letter by CSIC – Integrating greenhouse gas capture and C1 biotechnology: a key challenge for circular economy.

BioSFerA at the BIOCON-CO2 Final Symposium

On 14th – 15th of June, BioSFerA will be hosted at the final event of its sister project BIOCON-CO2, a H2020 project which worked on the development and validation of a versatile platform capable of using biological processes to transform raw CO2 waste from the iron, steel, cement and electric power industries into value-added chemicals and plastics.

A two-days event as opportunity to present their results and go deepen the topics of carbon capture and carbon and biological process valorisationa as well. Together with other European funded projects and professionals, BioSFerA will present its progresses and research on its innovative double-stage fermentation technology to a wide audience gathering figures from the industry, science and policy fields.

To know more about the single sessions, discover here the draft agenda.

REGISTRATION ARE OPEN (hybrid event)

Talking about BioSFerA in the magazine Nuova Energia

A new article for our project: BioSFerA is mentioned in the Italian specialized magazine Nuova Energia dedicated to the innovation and last results on the energy field.

The contribution written by the partner ENVIPARK and entitled “Scarti vegetali e rifiuti per decarbonizzare navi e aerei” gives a short presentation of the project and how it can positively impact on the society and contribute to the energy transition and decarbonisation of the maritime and aviation transports.

 

 

 

 

 

 

 

 

 

 

 

READ THE ENTIRE ARTICLE

 

BIOSFERA CONSORTIUM MEETS IN MADRID

Last 3rd – 4th of May 2022, the BioSFerA Consortium met for the first time at the CIB-CSIC headquarter in Madrid.

After two years of online work, the 11 partners joined in a physical General Assembly. The face-to-face meeting has paid off, increasing the collaboration and ideas among the partners.

Productive discussions came out within the Consortium showing the interconnection among the research activities and how open debate and opinions sharing can improve the project’s final results.

In the afternoon of the 4th of May, BioSFerA also led a workshop dedicated to the “Advanced technologies for green molecules production”. The event was organised in hybrid format with the collaboration of other two projects working on bioprocess research, CO2SMOS and LIFE BIOMASS C+. Interesting speech came also by Margarita De Gregorio, Director of the Spanish industrial platform BIOPLAT and representative of the Spanish and European bioeconomy strategy, brining important insights on bioproducts and their role on the climate crisis action.

4th GA BioSFerA Meeting

Advanced technologies for green molecules production workshop’s panelists

BioSFerA
Newsletter

BioSFerA is ready to annouce its project newsletter.

WHY A NEWSLETTER?

Starting our third year of research work, we have decided to promote the project with a dedicated newsletter available every 4 months where go deepen on BioSFerA progresses and results, share news and opportunities related to biofuels in maritime and aviation industry.

Wheter you are a researcher, sectorial company or policy makers you might be interested to find some specific articles where better understand the BioSFerA technology potentialities from its development until its final application.

To discover more SUBSCRIBE HERE

 

Valuable mid-term results for BioSFerA H2020

At nearly half of its way, the BioSFerA project is aligned with the planned activities and some first interesting results highlight the technical value of the BioSFerA concept. Achieved the potential assessment of its replicability across Europe at commercial scale, as well as the identification of the overall value chain, progresses follow from the biological treatment tests of the syngas performed at lab scale.

More in detail, Moorella and Clostridium strains have been already identified as the wild type promising species, showing high resistance to the syngas contaminants for both wild and genetically modified type. Currently CSIC is working on the recombination strains phase, planning to perform tests in real conditions and apply modifications in the yeast to produce C14-chain length fatty acids.

Encouraging results for scale-up activities are also obtained in the gas fermentation tests, where BBEPP performed experiments in bioreactor using glucose as a carbon source, as well as the oxygen tolerance tests in serum bottles. On its side, CARTIF confirms the Moorella Thermoacetica wild-type strain 2955 as the best anaerobic bacterial strain for gas fermentation trials in bioreactors and identify the so called Fröstl medium, characterized by the absence of yeast extract, as the medium with best performance. Next steps expect to work on the effects of the contaminant’s accumulation in a 1.5-litre pressurized bioreactor with a gas recirculation system (CARTIF) and on a continuous bioreactor of 10 litre capacity where a cell recycle system is installed (BBEPP).

For the optimization of TAGs production from acetate fermentation, processes in bioreactor with the select wild type strains W29 or YB-392 of oleaginous yeast Yarrowia Lipolytica have been explored. The experiments completed by CARTIF in batch mode show a high total lipid content and titer achievement, to be improved in the next steps focusing on the cell recycling and joint feeding strategy of N and C nutrients in the substrates, with the double ability to increase cell growth, prevent lipid inhibition and citrate formation. The better performance observed with the increase in the C/N ration of the feed have been confirmed by BBEPP tests, which is now moving further working on the development of TAG identification, quantification method together with the installation of a 7-litre continuous bioreactor with a cell recycle system. Preliminary tests on lipid purification analysis also involve ENVIPARK with a first approach in laboratory on a Y. Lipolytica bacteria culture sample provided by BBEPP.

Besides the lab-scale tests, VTT is working on the strategy for syngas cleaning modification in the pilot campaign and GoodFuels presented a preliminary analysis on crucial parameter for BioSFerA fuels market penetration to meet specific standards and products specifications.

On the communication side, the BioSFerA dissemination results is proceeding with the creation of a ResearchGate project profile, giving visibility through papers and activities shared with the scientific community. The official BioSFerA newsletter is also under development to disseminate the innovative BioSFerA concept to a wide public. Although the consortium has never met during this first year and a half, the team is focused on the project’s goals and showed productive. Good feedback also come from the European Officer after the acceptance of the 1st Periodic Report and its positive evaluation on the promising results and works lead until now by the BioSFerA project.

BioSFerA meets refining industry leaders at the Downstream 4.0 Summit

Mr. Maarten Van Haute from Q8 Research partecipates at the Downstream 4.0 Summit, the international summit gathering more than 20 globlal industry leaders to accelerate refining business conversion towards 2050 goals, by adopting and deploying new energy transition trends with technology and production developments. A good opportunity for discussion and presentation of the research development of BioSFerA project.

The summit was also the occasion for the publication of a scientific article for the specialised magazine Biofuel International, September/October issue: “Highlighting biofuels production from syngas fermentation – Setting the course for sustainable aviation and marine fuels”.

BioSFerA project presented at ECOS 2021 Conference

CERTH team has successfully participated to the ECOS 2021 Conference presenting the paper ‘Aviation & maritime biofuels production via a combined thermochemical/biochemical pathway: A conceptual design and process simulation study’.  The presented paper contained a first orientation of the suggested concept from start-to-end, as developed within the first year of the project.

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