GMO yeast instead of hops. The BohemiaBio team is changing the world of brewing and sustainability.

How and when was the BohemiaBio team founded?

I learned about the iGEM Meetup competition from Klára Hlouchová from Charles University’s Faculty of Science in November 2024. The idea intrigued me, so during that December, I found a few similarly enthusiastic people and in January 2025, Klára and I started developing the project with our small group. At that time, we called ourselves “iGEM Prague”. Later, we found out that the team name could not contain the word “iGEM”, so we came up with a new name: BohemiaBio. Since then, our team has grown and we recruited our newest member about two weeks ago. It is a continuous process; today, the team has 15 students plus several instructors (doctoral students) and three principal investigators (PIs, who are the leaders of the research groups, guarantors, and mentors responsible for results and research).

Where did you get the inspiration for the project?

We talked to members of the Brno iGEM team who came to give us a lecture in Prague: Miroslav Rasputinský and Matúš Grieš. They showed us their project and that’s when the first ideas started to emerge. We also drew inspiration from Tomáš Pluskal’s group, which deals with the biochemistry of plant metabolites. We wanted to start with something traditional related to the Czech Republic, and beer was the ideal choice because it connects yeast and hops, the two “research areas” with which we are all familiar.

How does the Beer without Hops project differ from other biotechnologies used in brewing beer?

Most of the biotechnologies I know focus on yeast breeding: crossing and selecting mutants. There is a start-up in the US that modifies yeast but not to replace hops. They do this to improve aroma or taste while still using hops for bitterness. We decided to start from scratch and found that 50-80% of hops are added precisely for bitterness, which is the most difficult substances to extract, because the molecules are non-polar and poorly soluble in water. Furthermore, with global warming, the properties of hops are changing, which causes inconsistent taste. Our goal is to create yeast that would produce bitterness on its own so that it is consistent. Then hops could be used only for taste and aroma, which would significantly reduce the quantities of hops used in beer production.

Can the taste of hops be fully replaced?

With current technology, probably not, but it depends on the time horizon. By the end of October, when we have to finish the project, we will be happy if we create yeast that produces at least some of the substances responsible for bitterness. If we worked on it for 2-3 years, we could probably replace almost everything. And maybe in 10 years, hops could be replaced completely.

Is this the same yeast that produces ethanol from sugar?

We are using genetically modified model yeasts that are not commonly used in brewing but are used in research. These yeasts produce terpenes and their precursors, which are the basis for the substances responsible for bitterness. If we can do it in a model, we plan to transfer the pathway to regular brewing yeasts.

What specific compounds do your yeast produce?

We are currently producing florisovalerophenone, a precursor to α- and β-acids (molecules that cause bitterness). The goal is to produce humulone and lupulone and their isomers. We also want to produce xanthohumol and desmethylxanthohumol, which provide antimicrobial activity and foam stability.

How do you measure bitterness?

Because it is a GMO, we cannot taste or smell anything. We measure using mass spectrometry on an Orbitrap in Tomáš Pluskal’s group, where our colleague Josh Smith helps us with the experiments.

Have you ever tried making beer in its final form?

We have our first test batch in a control medium; it's still far from real beer. We’ve detected the production of precursors, but we’re missing the last two enzymatic steps to form humulone and lupulone.

How do you measure taste and aroma when you can’t taste or smell? Do you have any sensors?

We don’t have sensors; we are mostly biochemists and analysts with no experience with measurement technology. Food and beer quality experts recommended NMR and mass spectrometry for quantification and comparison with commercial beers. Complex bioassays are not realistic for us given the time pressure we have to complete the project.

How specifically does genetic engineering work in yeast and how does it differ from modification in plants in agriculture?

Electroporation is a modern method that uses a short, strong electrical impulse to introduce DNA into cells, even those with a solid cell wall, such as yeast. In practice, this involves preparing electrocompetent cells, mixing them with DNA, applying an electrical impulse, regenerating the cells, and selecting the transformed ones.

Plant transformation is done differently.  For example, using the gene gun method, chemically inert gold or tungsten particles carry DNA and are shot into plant cells, where they penetrate the cell wall and allow the DNA to enter. The transformed cells are then selected and the entire plant is regenerated. The mechanism is therefore different, although the goal is the same: to introduce a foreign gene into the organism.

How is your cooperation with colleagues from Charles University’s Faculty of Science? What is the structure of the team?

The team is composed of about half UCT Prague students and half from Charles. One colleague and I also work at IOCB. Our cooperation is excellent; we have facilities at both universities, a dedicated space, and team members complement each other well. At the beginning, we worked in Professor Demnerová’s lab under the leadership of Dr. Kamila Zdeňková. Thanks to her, we already have our first results. We learned a lot from her and were able to apply it to our work. I thank her very much on behalf of the entire team. Now we work in the practical training laboratories at the Faculty of Science, Charles University, which our principal supervisor, Klára Hlouchová, arranged for us.

What did the Best Presentation award and other awards at the Frankfurt Meetup mean to you?

It was a great encouragement and success. Our presentation, in the form of a dialogue, was very well received. We established new contacts and opened up opportunities for future cooperation. This motivates us to continue and prepare the best possible result for the Grand Jamboree in Paris.

How would you describe the atmosphere and experience of the iGEM competition?

iGEM ​​is not just about competition; it’s mainly a friendly environment full of interactions and discussions. In Frankfurt, many judges and participants came to give us positive feedback, asking additional questions that inspired us to expand the project. We all consulted and supported each other. iGEM is about mutual help and inspiration.

What is your strategy for presenting at the Grand Jamboree 2025 in Paris?

We plan to use the dialogue format again, perhaps expanded to include more people, to make the story more vivid and engaging for the audience. We have had good experience with this concept both in Frankfurt and in various promo videos.

What results do you have from the laboratory so far?

We have reached about half of the metabolic pathway for the production of hop α-acids. The biggest achievement is the confirmation that the intermediate is released from the yeast into the medium, which is key for the final production in beer. The transformation and measurement methods work. Now we are fine-tuning the details.

How do you address issues of biosafety, genetic stability, and potential release into the environment?

We use a plasmid system with selection pressure. Yeast needs a plasmid to produce uracil; without it, it will die. We plan to integrate the genes into the genome, which will increase stability. We are consulting with experts on biosafety and we do not plan to release them into the environment. The resultant beer will be filtered so that GMOs do not get into it.

What impact do you expect on the Czech brewing industry and on sustainable food production?

The impact in the EU will be limited due to regulations, but the project has great potential in the US, Brazil, and Asia, since they are all more open than the EU to GMOs. Our technology should reduce hops consumption and energy requirements and increase production efficiency.

Are you planning to implement your ideas in an industrial context or a start-up?

It depends on our results. If we are successful, we are considering a start-up that would produce modified yeast for international markets, since this would be more complicated in the EU.

How do you finance your project?

The main funding sources are contributions from IOCB and the Charles University Faculty of Science’s Endowment Fund, which jointly finance travel and competition fees for students. The IOCB Tech Endowment Fund contributed significantly to our registration fees, without which we would not have been able to participate in the competition. We also received support from a law firm, Craft Legal, for which I would like to extend thanks. We have received tremendous help from our leaders: Klára Hlouchová, Tomáš Pluskal, and Kamila Zdeňková, who provide us with logistics.

What surprised you during your participation in iGEM?

People say that iGEM is demanding, and it really is; I underestimated it a bit. The first surprise was how much hard work the competition actually requires. The second, more pleasant surprise was when I found out how many people were willing to help, whether at UCT Prague or at other institutions. I would like to thank Ms. Kovaříčková from UCT Prague’s Technology Transfer Department. She also helped us finance the trip to Frankfurt and the Grand Jamboree. Without her willingness to assist, we would not have achieved such great results. I also want to give a shout out to Michal Dvořák, who helped us a lot in finding the right people at the beginning of the process. The biggest surprise for me, however, is how quickly our team is learning and growing. My colleagues started without much experience in molecular biology, and after a few months, they are already designing and conducting their own experiments. I am grateful to have such an amazing team.

What will happen to the project after it’s finished?

I would like to continue developing the project idea. We plan to participate in the next iGEM and we are looking for new ideas and new people. If readers are interested in the iGEM concept, I would be happy if they contact me.

What are your personal ambitions for the future?

Finish my Bachelor degree, continue with a Master degree, then a doctorate, or gain experience in the biotechnology industry. I am considering starting a start-up, either with this project or with another one.