Fermenting the future – on the benefits of a bioart collaboration

Abstract In this article we explore the intersection of science and art through a collaboration between us scientists and the bioartists Anna Dimitriu and Alex May, focusing on the interface of yeast biotechnology and art. The collaboration, originally initiated in 2018, resulted in three major artworks: CULTURE, depicting the evolution of yeast and human societies; FERMENTING FUTURES, illustrating a synthetic autotrophic yeast and its link to lactic acid production; and WOOD SPIRIT—AMBER ACID, inspired by the VIVALDI project targeting CO2 reduction to methanol. We emphasize the reciprocal nature of the collaboration, detailing the scientific insights gained and the impact of artistic perspectives on us as researchers. We also highlight the historical connection between art and science, particularly in the Renaissance periods, and underscore the educational value of integrating art into science not only to support public engagement and science dissemination, but also to widen our own perceptions in our research.


Introduction
The connection between science and art is multifaceted and has been a subject of exploration and discussion for centuries.It is c har acterized by shar ed inter ests suc h as observ ation, cr eativity, and the quest for exploring and understanding the unknown.Art and science have often inspired each other, but more than that, the dir ect tr ansfer of scientific knowledge or art practice has enabled br eakthr oughs at certain moments in history, c hanging futur e pr actice.
Peter Weibel, the former director of the Center for Art and Media (ZKM) in Karlsruhe, Germany, claimed that the renaissance periods have created specially vibrant scenes of art-science collaborations (Weibel 2023 ).The exhibition Renaissance 3.0 in the ZKM defines three renaissance periods: the Arab Renaissance, the Italian Renaissance with Florence as its nucleus, and the modern, potentiall y curr entl y ha ppening Renaissance, lar gel y driv en by media art and bioart.Most pr ominentl y, the Flor entine Renaissance has been discussed intensely at the interface of art and science for its incentives to spark collaborations of artists and scientists (Puett and Puett 2016 ), with some stellar figures producing great ac hie v ements in both ar eas, often r eferr ed to as polymaths, skilled individuals educated in a number of different subjects.By refusing barriers and limitations in scientific methods, fields of studies, and the integration of art in kno wledge gain, Alexander v on Humboldt is often regarded as the last Eur opean pol ymath and "Renaissance man" (Gunderman 2014 ).As a bridge to the present, today, pol ymathy is consider ed a pr e v alent tr ait of Nobel laur eates, many of them even describing polymathy as an intentional choice to optimize cr eativ e potential (Root-Bernstein and Root-Bernstein 2023 ).Intriguingl y, using pol ymathy as educational tool by teac hing STEMM (science, technology, engineering, mathematics, and medicine) students arts, crafts and design practices can significantl y impr ov e learning outcomes proving the str ong corr elation between art and science (Root-Bernstein et al. 2019 ).
These records build on analyses of the past and their inter pr etations to w ar ds a future that is emer ging now.Ov er the past fe w years our lab has de v eloped a deep interest in the interface of biosciences and art, often r eferr ed to as bioart.This term was introduced to define the art movement where artists are working with living (micr o)or ganisms or use pr ocesses of life to cr eate their artw ork.In the follo wing w e summarize experiences and learnings from that collaboration, with an outlook to future areas to be explored together.

Our part in bioart
In 2018, we have initiated a collaboration with bioartist Anna Dumitriu and digital artist Alex May, to transform our r esearc h on yeast biotechnology to art, with the initial intention to exhibit the artwork and the process at the International Congress on Yeasts, planned for 2020 in Vienna (Dumitriu et al. 2021 ).Initially, we planned to invite the artists for a residency in our lab to create some yet undefined artworks to show them at the congress location in an area open to the public.The entire endeavor has grown m uc h lar ger since then, including collabor ations with cur ators, gallerists and contemporary art historians, bringing the artwork, the scientific bac kgr ound and our collabor ativ e a ppr oac h to major art galleries in Eur ope.Pr emiering at the Vienna Künstlerhaus, AT, While initiating the collaboration without a distinct intention besides enabling the cr eativ e pr ocess and presenting the outcome, we have realized remarkable feedback on our science and technology, clearl y demonstr ating that this collabor ation is not a one-way from scientific kno who w and stories, technologies and materials to w ar ds the realization of the artwork.
In the following, our collaboration, our perceptions and learnings from it, and feedback to us as scientists will be explained with three examples, by means of three major artworks that emerged from the collaboration.

Culture
CULTURE tells a story of evolution of yeast and human societies.We humans see ourselves as a species that is able to change natur e ar ound us by impacting the envir onment of other life forms for our purpose.At the same time we tend to neglect the ability of other species to do so as w ell, ho w e v er yeast also might have played a similar role during the neolithic era.Its ability to ferment carbohydrates to ethanol and carbon dioxide has inspired humans to br e w beer and bake leav ened br ead, inducing a demand for cereal crops which, over time, motivated them to settle and cultiv ate cr ops (Dietric h et al. 2019 ).The earliest indications of very first human settlements, connected with brewing and baking, are found in Göbeklitepe in Turk e y (Dietrich et al. 2012 ) and Raqefet Cave in Israel (Liu et al. 2018 ).
Discussions on the historical connections of humans and y easts w ere initiated b y resear ch in our lab on the evolution of fermentation in yeasts.Some years ago we found a genetic master regulator of glycolysis and fermentation in Komagataella phaffii (formerly known as Pichia pastoris ) (Ata et al. 2018 ), and we investigated its potential role in the evolution of fermentative traits in yeasts .T he artists planned to span an arc from the prehistoric connections of yeasts and humans, mirroring the cultural evolution of human settlement with the metabolic evolution of yeast fermentation.These detailed discussions have definitely changed our scientific perspective on our object of study, yeasts.With the first attempts to bake bread with this engineered potential "missing link" of yeast metabolic evolution we realized that it can ferment glucose, but not maltose, the k e y structural subunit of starch, and it could not leav en br ead.In Sacc harom yces cerevisiae maltose is transported into the cell where an intracellular maltase hydr ol yzes maltose into glucose.Her e, we used another a ppr oac h and expressed an extracellular maltase using markerfree CRISPR-Cas9 technology.We could demonstrate the success of strain engineering as we could bake bread with the new K. phaffii str ain, whic h was then used as a material for the cr eation of the sculpture (Fig. 1 ).

Fermenting Futures
Some years ago, we created a synthetic autotrophic yeast by re-engineering K. phaffii to assimilate CO 2 instead of methanol (Gassler et al. 2020 ).After further metabolic engineering this strain can also produce organic acids (as chemical building blocks) by the conversion of CO 2 .A special attention went to lactic acid as the monomer of polylactic acid (Baumschabl et al. 2022 ).For the narr ativ e of the sculpture FERMENTING FUTURES we connected this work with another project where we produce lactic acid from glucose with engineered S. cerevisiae .The sculpture is composed of a glass vessel connected with a bundle of silicon tubes, resembling a bior eactor wher e the yeasts ar e cultiv ated in the labor atory.3D printed models of yeast colonies cover the flask, one of them made of the biodegradable polymer polylactic acid (PLA) that was polymerized in our laboratory containing the yeast-produced lactic acid (Fig. 2 ).The vessel rests on a plinth of chestnut wood-the tr ee species wher e the first isolate of Pichia pastoris (now named K omagataella spp.) w as found.A major intention of the artists w as to produce PLA in the lab from a mix of lactic acid produced from glucose but notably also lactic acid made from CO 2 with the autotr ophic str ain.We had an inter est in this pol ymerization pr ocess but as we are no polymer chemists we had refrained from practically implementing it.In retrospect, we have probably avoided the hassle of engaging in a technology we felt uncomfortable with, being non-experts.As there was a need now but no protocol in place, we consulted our colleagues and assembled se v er al methods (Moon et al. 2000, Auras et al. 2010 ) to adapt them to our needs and technical setup for the successful polymerization of lactic The plinth supporting the sculpture was to be made of horse c hestnut wood-the tr ee species wher e the first sample of K. phaffii was isolated from.The literature describes it as the edible chestnut ( Castanea sativa ), ho w ever a look at the original paper by A. Guilliermond ( 1920 ) a ppear ed a ppr opriate.It r e v ealed a tr anslation error that was transcribed further until recently: Guillermond describes the tree as a marronnier which is tempting to be translated as the edible chestnut (which is, ho w ever, called châtaignier in Fr enc h).Having r e v ealed this little mystery of the origin of the y east w e had to solve the puzzle where to get horse chestnut wood fr om, whic h is not tr aded, differ ent to the edible chestnut.

Wood spirit-Amber acid
The third artwork discussed here was inspired by the European Union Horizon 2020 project VIVALDI (grant number 101000441) that aims to reduce CO 2 of industrial off-gas electrochemically to methanol, and to upcycle methanol to organic acids with the metabolic activity of K. phaffii .Succinic acid is one of the target products of VIVALDI.It is known as a base chemical for organic synthesis that can be converted into a variety of products.Anna Dumitriu was inspired by the traditional names of substrate and product to name the artwork WOOD SPIRIT-AMBER ACID: methanol was first produced by dry distillation of wood, and succinic acid was first pr epar ed by distillation of amber.Wood is also a likely source of methanol for the yeasts living on it.
Besides the definition of sources of the substrate, we had to answer the application of succinic acid in chemical technology.Succinic acid is often described as a base chemical, and applications for pol ymers, r esins and solv ents ar e indicated.The artist, howe v er, asked for defined products that are made of succinic acid, to select material for the artwork from that.This made it evident to us , although ha ving a r ough pictur e of the potential a pplication of our target product, that we have not taken the time to research its actual use in detail.One quick answer is nylon: many types of nylon (or polyamides) are produced, defined by the types of precursors (dicarboxylic acids + diamines, or aminocarboxylic acids), and the chain lengths of these monomers.Succinic acid is a precursor of nylon PA 5.4, where the 4 denotes the chain length of the acid precursor.
Questions continued for other products out of succinic acid, and after some further r esearc h we found two r ather ne w types of synthetic pigments which were developed in the 1970 s and introduced to artists' palettes recently: quinacridone pigments, featuring golden to magenta hues, and the diketop yrrolop yrrole family with a colour range from orange to violet (Christie andAbel 2021 , 2022 ).Succinic acid, as many other potential products of industrial biotechnology, is a business-to-business product not dir ectl y known to end consumers.We have to acknowledge that we as r esearc hers working on the production of the precursors were not familiar with the details of the application, and the r esearc h for the artw ork taught us a lot bey ond our area of expertise.The artwork is a necklace made of nylon that w as dy ed with quinacridone pigments, including some amber beads and pendants containing culture liquids and yeast colonies (Fig. 3 ).A positive side effect of preparing these pendants was learning how to conserv e a gar cultur es for visual inspection by embedding them in e po xy r esin, a tec hnique we ar e using now also for science fairs.

Conclusions
Crossing these boundaries between art and science and following the creation of an artw ork, w as not only fun and exciting for us as r esearc hers, but encounter ed us also with ne w experimental tec hniques, a ppr oac hes and mindsets .Despite ha ving obvious differ ences, thr ough this collabor ation we experienced the numerous similarities of a ppr oac hing art and science.During the actual r esearc h, we scientists ar e guided by our experimental outcomes, and data must be e v aluated as objectiv el y as possible, while art making and perception can be m uc h mor e emotional, subjectiv e and influenced by opinions.But at the same time, passion and curiosity of exploring the world, creating new knowledge , concepts , and ways of thinking are the foundations and driving forces of both art and science.Re v ealing the scientific truth about yeasts makes us scientists as excited or mesmerized as seeing an art piece .Furthermore , being in volved in a bioart project, where art and science trul y meet, cr eated a deeper understanding of our research and expanded our perspective and knowledge about the y east w orld and especially the connection betw een y easts and humans.
Supporting the artistic process was not the end of our interest but we were keen to see what kind of emotions and perspectives the resulting artworks would spread.As the outcome of our collabor ation gr e w m uc h lar ger than anticipated and r esulted in a br oad wav e of positiv e public r esponse, we came in touch with the art museum scene and the organization of exhibitions, and w e w ere inv olved in science talks, visitor discussions and guiding tours through the art show.Engaging with visitors was more important as we did not aim to advertise our r esearc h or bias the artists with our way of thinking.Just as hoped for, we could observe the impact of the collaboration both on our research, and on the artwork unfold by different modes of exhibition, approaching visitors in a variety of different wa ys .
In addition, this tr ansdisciplinary collabor ation pushed our work in new directions including the search for new materials and techniques, or by just gaining a new perspective on our own r esearc h.Howe v er, it also opened new and important questions concerning the display of bioart and the possible contributions of the biosciences to that: can bioart be living and/or deteriorable, or should it be stable and "dead", to be suited for museums?How can bioart be conserved and regenerated, and is this desired at all?Will there be a de v elopment of a bioart material canon, based on either li ving, inacti v ated, or dead material?These ar e just some topics for new collaborations where life sciences will have a k e y role.

Ac kno wledgements
We thank Anna Dumitriu and Alex May for the inspiring collabor ation, and Sonja Sc hac hinger and Wolfgang Giegler who enabled the primary exhibitions of the artwork.Further thanks go to the curators of the international exhibition houses who sho w ed the F ermenting Futures series , especially to Oana Romocea for premiering Wood Spirit-Amber Acid.

Figure 1 .
Figure 1.(A) CULTURE installation symbolizing the connection of yeast's fermentation evolution and cultural evolution of human throughout the history.© Left Studio, Vienna.(B) A "Pichia " bread fermented by the CRA1 overexpressing strain.(C) Baked bread using whole-grain Einkorn wheat, one of the oldest domesticated wheat species , and lea vened with the fermenting K. phaffii strain.This bread loaf represents bread baked by the first human settlers in the Fertile Crescent in the Middle East, and was crushed to cover the sculpture.

Figure 2 .
Figure 2. (A) FERMENTING FUTURES sculpture resembling a laboratory bioreactor containing lactic acid-producing yeast.(B) Closeup of a PLA-form containing yeast-produced lactic acid.(C) Technical setup including a rotation evaporator for polymerizing lactic acid to PLA.

Figure 3 .
Figure 3. (A) WOOD SPIRIT-AMBER ACID is a sculptural necklace narrrating the story of the sustainable production of succinic acid from methanol using Komagataella phaffii .(B) Pendants containing engineered yeast cells that were embedded in epoxy resin.(C) Glass beads were filled with fermentation supernatants containing yeast cells and succinic acid.