Lipids upgrading and final product characterization

BioSFerA is approaching its last year of project and, with it, the downstream phase that will finally lead to the production of jet and bunker fuels resulting from our innovative biofuels production concept.

The TAG lipids, directly coming out from the fermentation process in the BBEPP’s laboratory, will be sent to the research team of CERTH based in Thessaloniki for the hydrotreatment upgrading. On the first phase, a dedicated TRL 3 hydrotreatment plant of CERTH, will be utilized to optimize the technology. On the second phase, the optimized technology will be scaled up in a TRL 5 hydrotreatment plant of CERTH for the production procedure. Finally, on the third phase, the products from TRL 5 HDT plant, will be fractionated under vacuum in a batch large fractionation unit of CERTH in order to separate the Jet and Bunker fuel fractions.

The downstream step’s trials are going to start within these months to last until the end of the project. CERTH will take care to show its process with video and step-by-step updates. The following pictures give a preview of the CERTH’s laboratory in Thessaloniki and the equipment used in BioSFerA:

TRL3 hydrotreatment plant in CERTH’s premises, Thessaloniki   

TRL5 hydrotreatment plant in CERTH’s premises, Thessaloniki

Large fractionation unit for the separation of the jet and bunker fractions from the total liquid product (Hydrotreatment unit at TRL5, CERTH’s premises in Thessaloniki)


March 2023: the BioSFerA pilot plant integration is starting!

Last updates from the BioSFerA’s pilot plant: after 2400 km over land and sea, the fermentation unit is now arrived in Finland.

The previous months steel mill manufacturer Arcelor Mittal Ghent was host to the Bio Base Mobile Pilot Plant (BBMPP), the fermentation mobile plant designed and constructed by Bio Base Europe Pilot. Now, in the framework of BioSFerA, the objective is to integrate the BBMPP with VTT’s gasification unit. BBEPP and VTT will be responsible of integrating the BBMPP with VTT’s facilities, followed by optimization of the operational parameters of the pilot process. Main objective is the scaling-up of the BioSFerA concept and the production of acetate via the gasification process and the following double-stage fermentation step.

For the rest of the year, the pilot scale process will be operated in collaboration between VTT’s and BBEPP’s engineers, until the finalization of the trials end of 2023.

Curious about the shipping process of the pilot plant? After the uncoupling of the unit from the steel mills off-gases supply and decommissioning of the other utilities, the BBMPP was shipped to Finland for a 2 weeks journey over roads and sea. The unit has just arrived at the Bioruukki site to start the first integration steps.

Some pictures of the BBMPP journey to the VTT’s premises:

BBMPP at Arcelor Mittal’s site during decomissioning.

Bio Base Mobile Pilot Plant arrival at the Bioruukki VTT’s site of installation.


More pictures and videos on the BBMPP’s journey to Finland here.

Alternative Aviation Fuels/Pathways: A Critical Review from CERTH

Recent Advances on Alternative Aviation Fuels/Pathways: A Critical Review” is the new scientific paper published by our coordinator CERTH on the Energies journal (MDPI).

The paper is a critical summary of the current alternative aviation fuels/pathways and a comparative analysis of the dominant technologies considering techno-economic assessment, environmental evaluation, and future projections. Final considerations are also performed on the policy framework aiming to promote and boost all the SAFs in industry. Reading recommended!



Innovation Days 2023 in Greece

A new opportunity for BioSFerA to be introduced to the Greek stakeholders’ community: CERTH participated with its own stand in the Innovation Days 2023 exhibition held in the region of Western Macedonia Greece under the auspices of the Prefecture of Western Macedonia.

From 7th until 9th of February, the exhibition gathered the entire research community of the region as well as top companies of the Greek energy sector. The CERTH team presented the project and its outcomes focusing on future replication in the region. Some pictures of the event:

Presenting BioSFerA at the Biomass Day 2023

Today, the Biomass Day 2023 event is taking place in Athens. The Greek conference is dedicated to the bioenergy role in the energy crisis and gathers public authorities and main scientific community representatives to deal with this topic.

Considering the Russia-Ukraine war and global geopolitical conflicts, it is always increasingly difficult to operate bearing in mind the decarbonization and climate neutrality European goals, as set by the Green New Deal and the Fitfor55 plan. The conference guides the discussion on the active role of the bioenergy in our economy to answer some questions: how can the contribution of bioenergy in the expansion and diversification of EU energy sources be easily and consistently ensured through the “RePowerEU” project? Is there a possibility in Greece and the EU of a more substantial energy use of biomass to alleviate the unsustainable energy costs of industries, commercial companies /services and households?

BioSFerA partecipates at the debate having our coordinator Konstantinos Atsonios (CERTH) as representative and handling a presentation on “Gasification technologies for the production of liquid and gaseous biofuels and their contribution to sustainable transport”.

For the full presentation (in greek) CLICK HERE.


Biomass Conversion and Biorefinery Journal: a new paper on BioSFerA

The CERTH team published a new scientific paper resulting from the development of the BioSFerA’s concept. The publication, entitled “Design considerations of an integrated thermochemical/biochemical route for aviation and maritime biofuel production“, describes the conceptual design of the novel integrated thermochemical-biochemical BtL process performed in our project.

The process design has been analysed considering three different operational scenarios examined and assessed mainly through overall performance indicators: carbon utilization (CU), energetic fuel efficiency (EFE), liquid fuel mass yield, and overall energetic/exergetic efficiency. As outlined in the study, design considerations and their impact on process efficiency were performed within the assumed scenarios, including parameters such as internal/external hydrogen securement via pressure swing adsorption (PSA)/water electrolysis and oxy-/air-acetate fermentation as well as autothermal/allothermal operation of the catalytic reformer. Aim of the study is also the definition of the key process specifications and the evaluation of the proposed concept compared to other competitive technologies.

To have a full overview of the BioSFerA process design, explore the entire article, free available in the Biomass Conversion and Biorefinery Journal (Springer) and in our Publications section.


Updates from BioSFerA H2020: biological process status and forward project’s steps

Another important General Assembly meeting was held last October within the BioSFerA Consortium.  On this occasion, the partners met in Ghent (Belgium), at the Bio Base Europe Pilot Plant’s headquarter: last project updates and an interesting tour at the biobased process plant made well successful the meeting at BBEPP.

At this stage of the project, the biological process plays a significant role for all the following steps. Several partners contribute to the lab-scale optimisation to fully comply the further piloting activity and the overall BioSFerA concept achievement. Starting from the microorganisms’ investigation, CSIC updates the Consortium with its work to improve both acetogenic bacteria and oleaginous yeast strains on their production rates and contaminants resistance through a metabolic engineering strategy. On one hand, the Spanish research centre is working to optimize the overproduction of enzymes in the acetogenic bacteria Moorella, by increasing the copy number of the genes introduced in the strains using plasmids. Furthermore, they have genetically modified the oleaginous yeast to create an obese strain overexpressing synthetic DGA genes (diacylglyceride acyltransferase) and deleting a gene involved in the fatty acids degradation metabolism. This Y. lipolytica yeast strain has been tested in different media with single or combined substrates (glucose-acetate-glycerol). The results showed better performances in all the cases for the GMO yeast strain than the wild type. Next months, CSIC plans to test new obese strains and perform other experiments to increase the production of TAGs with higher C14 content.

First lab-scale results begin to be achieved in the gas&liquid fermentation processes, where BBEPP and CARTIF are mainly involved. They are currently testing the Moorella strains considering different gas composition and other process parameters to understand the biomass behaviour and its productivity. From the BBEPP side, the syngas fermentation trials are currently under operation using 1 L batch fermentation and 10 L continuous bioreactors aiming to increase the process performance; meanwhile, CARTIF is conducting the gas fermentation tests on 1 L pressurized bioreactor, discovering the glucose as possible co-substrate useful to increase biomass and acetate production.

For what concern the liquid fermentation, BBEPP successfully cultivated Y. lipolytica on the gas fermentation effluent in shake flasks, thus confirming the BioSFerA concept. Furthermore, a series of bioreactor trials were performed to assess the TAG accumulation potential of the Y. lipolytica strain modified by CSIC on acetic acid (AA) and glucose. In parallel, CARTIF also completed fed-batch and continuous optimization trials testing different substrates, such as AA substrate, AA and glycerol or only glucose. The parameters still need to be optimised to reach the final target set at 60 % TAGs accumulation and 50-100 g/L TAGs. CARTIF will continue the trials focusing on the AA/glycerol substrate and improving concentrations and other process parameters.

Last stage of the biological process is the recovery and purification of the TAGs, which is currently under investigation by BBEPP. During the meeting, a sample of purified microbial oil produced by Y. lipolytica was showed to the Consortium to better understand each step development. A sample of the obtained oil will be also sent to KPRT for further analysis, while a broth sample will be available for ENVIPARK to perform the steam explosion and advances with the downstream process step.


Progress from the piloting: last September BBEPP and VTT met to define next steps and integration process between the two gasification&fermentation units. A HAZOP study was conducted, risks were listed, together with mitigation measures, as well as the following BBMPP’s shipping and the preliminary operational conditions. BBEPP is also taking care of the fermentation scaling-up activities, running trials on 150 L fermenter with Y. lipolytica and glucose as substrate to set up the process parameters. The gas&fermentation piloting phase will start in April 2023 and will be finished latest by October 2023.


Continue the BioSFerA model concept development: the EMMS model for the DFB (Dual Fluidized Bed) and the full chain model at commercial scale are performed by CERTH with SFW and partners collaboration. The techno-economic analysis will consider the entire value-chain accounting for capital and operational costs and biofuel minimum selling price. A sensibility analysis will be performed from the base-case scenario in order to identify hotspots for cost optimization. At the end, at least 3 scenarios in different EU countries will be analysed to verify the versatility of the model at territorial level. The analysis will be completed with the LCC and LCA results.

A first market assessment was also present by GF, showing the competitor market analysis of different biofuel processes. Likewise, the evolution of pricing over the years, as well as forecasts for future pricing, were discussed for conventional and alternative technologies. According to GF some serious impacts could be originated by the Ukrainian war on biofuel production, due to European imbalance of energy supply and offer. From a legal point of view, KPRT rather reports possible further revision of a large number of European policies and directives associated with the Fit-for-55 package.


The Consortium is also very active from the dissemination side and the partners participated to several events during last months. Worth mentioning, in November ENVIPARK took part at the international fair on circular economy ECOMONDO (Italy) with an institutional stand and its projects, included BioSFerA. The project was even presented during a workshop organised by GLAMOUR, another European project with an active collaboration.

After more than two project’s years, BioSFerA has reached some relevant results and the project is well advancing. Some important challenges still need to be faced, but the activities respect the project schedule and the Consortium is hopeful to easily accomplish them, thanks to the high-level and cooperative partnership. Further updates will be released after the meeting planned next May 2023 at VTT’s premises in Finland.

The BioSFerA Consortium at BBEPP premises during the 5th GA meeting

The biological pathway for the fuels of the future

Interview with Jose M Sanz Martin, CARTIF

A central step to fully comply with the successful operation of the BioSFerA model is to ensure the highest yield of the fermentation steps by optimizing the different parameters that influence the biomass growth and its productivity.

In the project, CARTIF is one of the partners with an important role in the biological process for the lipids production at lab-scale. Thanks to its experience as Applied Research Centre Foundation in terms of R&D and technology transfer activities, CARTIF is leading the biological step referred to the project’s WP3 activity. Here, few questions to Jose M Sanz Martin, Researcher at CARTIF in the Agrifood and Sustainable Processes Division, to better understand the BioSFerA biological process development at lab-scale and the double-stages fermentation process that leads to the acetate before and after to the TAGs lipids production:

What are the last updates on the optimisation of the acetate fermentation process parameters for C14 and C16-18 TAGs production?

We are currently working on finding the best bioreactor strategy to favour lipid accumulation and for this we are testing nitrogen-limited conditions, continuous feeding of acetate as a main carbon source and recycling of cells using hollow fibre membranes. We are also using an engineered Yarrowia lipolytica strain developed by CSIC, which has improved triacylglycerides (TAGs) productivity. The results are promising and all this leads us to increase lipid production with respect to the wild-type strain.


Did you find any barriers and if yes, how are you trying to dealing with them?

Regarding the gas fermentation, one of the key aspects to improve the process is the choice of a suitable working strain, as well as the establishment of process conditions that allow for higher productivity. In addition, in order for the gases to be used efficiently by the microorganisms, it is very important that they are solubilised properly in the culture medium. To solve these problems, we have selected a strain of the acetogenic bacterium Moorella thermoacetica that is capable of producing a large amount of acetate from syngas, and we have improved the growth conditions by using bioreactors that allow us to control the flow rate of the gas fed as well as to increase the working pressure to favour its solubility in the medium.

As for acetate fermentation with Y. lipolytica, although acetate is a promising cost-effective and suitable carbon source for microbial fermentation, its use as the sole carbon source to enhance lipid synthesis is not very favourable from a metabolic point of view. To overcome this problem, we have confirmed that feeding a co-substrate such as glycerol can help to improve biomass production and drive metabolic flux towards TAGs production.

1.5 L continuous fermentations for TAGs production (installation of cell recycling system and continuous feeding strategy). Detail of the growth of Y. lipolytica from acetate+glycerol.


What are the last promising results?

In relation to acetate fermentation with Y. lipolytica, although we have not yet achieved the expected objectives of lipid production, we observed that the addition of glycerol during the fermentation process together with a nitrogen starvation strategy and limited stress conditions greatly favours the accumulation of TAGs by facilitating their metabolic biosynthesis. From here, we are conducting tests to find the most suitable concentration of glycerol to maximise this lipid accumulation.


For the overall biological production process of lipids from syngas at lab scale, several partners are involved in its optimisation and in different way. As leader of this activity, how this collaboration can impact the research and results?

I believe that the collaboration between all the partners involved in this work package is very beneficial for the progress of the project, as we can share the large amount of work that needs to be done according to the experience and capabilities of each partner. For example, VTT’s expertise in gasification has allowed us to select the best process conditions to achieve a syngas suitable for fermentation with microorganisms.  CSIC has obtained genetically modified strains of Moorella and Yarrowia that will maximise the productivity of the process, which is essential for the next stage of scaling up. In terms of bioprocess optimisation, BBEPP has focused more on optimising the anaerobic fermentation conditions, while CARTIF has focused on aerobic fermentation. Finally, ENVIPARK has contributed its expertise in the further processing of the lipids obtained in the fermentation broth.

In addition, the sharing of all the work done and the close collaboration that we have maintained throughout the project has allowed us to clarify many doubts and resolve the technical difficulties that we have encountered. It is certainly very gratifying to be able to work on this topic with some of the best researchers in Europe.

CSIC poster on the microbial lipids research in BioSFerA

Last 2-4 November, CSIC participated at the 6th Congress of Applied Synthetic Biology in Europe.

An important opportunity to show the last results about BioSFerA through the presentation of a poster entitled “Lipase overexpression in Yarrowia lipolytica for direct biofuel production“. It describes the research work led by our partner CSIC in the framework of the project: final goal is to build a Y. lipolytica obese strain expressing a robust lipase for the in situ production of biodiesel. The engineered strain will be able to produce TAGs from glycerol and to produce a lipase (either intracellularly or surface displayed), which after cell lysis with organic solvent and methanol will perform the direct transesterification of TAGs into FAMEs (biodiesel).

Download the poster here.

BioSFerA at the ECOMONDO Exhibition

From 8th to 11th November, BioSFerA will take part at the ECOMONDO Exhibition, International fair dedicated to the circular economy and the ecological transition with base in Rimini (Italy).

An important annual event for circular economy experts from several sectors to meet each other and receive updates about the last technological innovations. BioSFerA will be there in different ways:

  • virtual stand, from our partner’stand Environment Park, search for BioSFerA in its projects’ list;
  • in-presence with some project’s representatives from Environment Park, scheduling B2B meetings from the virtual stand with each person;
  • project presentation at the INDUSTRIAL&REPLICATION WORKSHOP, 9th November, 2-6 pm, organised by GLAMOUR project. The event aims to give last updates on the clean technologies for second generation biofuels, gathering experts and International research projects. Here the link to register at this hybrid event and the agenda.


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