20.500.12556/RUP-23554 Spectroscopic insights into nutrient-controlled biofilm formation in Aureobasidium pullulans as a basis for engineered living materials Fungal biofilms represent complex extracellular systems with potential application in engineered living materials (ELMs), yet their chemical composition and structural variability under defined environmental conditions remain poorly understood. Biofilms of Aureobasidium pullulans strains grown on different solid media were investigated using a multimodal spectroscopic approach, including FT-IR, FT-NIR, and Raman spectroscopy. Nutrient-rich media (PDA, SDA, YNB) produced biofilms with largely conserved chemical profiles across strains, whereas nutrient- poor SNA medium induced pronounced changes in polysaccharide, protein, and lipid components, as well as in matrix organization and hydration. Principal component analysis and multiblock data fusion revealed that growth medium is the primary driver of spectral variation, with strain identity exerting a secondary influence. Vibrational bands corresponding to hydroxyl (−OH) and aliphatic (−CH) groups were identified as key contributors to differentiation, reflecting changes in the composition of Extracellular Polymeric Substances (EPS). Raman spectroscopy provided complementary molecular information, such as the presence of characteristic protein and polysaccharide vibrations, which helped confirm the biochemical composition of the biofilms and supported the observations from IR and NIR analyses. Results show that the control of nutrient composition and growth conditions enables precise tuning of biofilm properties, including hydration, matrix density, and functional polysaccharide content. Biofilm composition is primarily influenced by nutrient composition rather than by the strain’s morphological features. This is important because it indicates that research on living coating formulation can focus on supplying nutrients that promote EPS production and biofilm development, rather than on selecting strains with particular morphologies. These findings provide a foundation for understanding the environmental control of A. pullulans biofilm formation and are relevant to the future development of engineered living coatings, where environmental conditions can be leveraged to influence biofilm composition, structure, and its functional properties Glivne biofilme predstavljajo kompleksni zunajcelični sistemi z velikim potencialom za uporabo v inženirsko zasnovanih živih materialih (ELM), vendar sta njihova kemijska sestava in strukturna variabilnost v opredeljenih okoljskih pogojih še vedno slabo raziskani. Biofilme sevov glive Aureobasidium pullulans, gojenih na različnih trdnih gojiščih, smo proučevali z multimodalnim spektroskopskim pristopom, ki je vključeval spektroskopijo FT-IR, FT-NIR in Ramanovo spektroskopijo. Na s hranili bogatih gojiščih (PDA, SDA, YNB) so biofilmi različnih sevov izkazovali pretežno ohranjene kemijske profile, medtem ko je gojenje na s hranili revnem gojišču SNA povzročilo izrazite spremembe v polisaharidnih, beljakovinskih in lipidnih komponentah ter v organizaciji in hidraciji matriksa. Analiza glavnih komponent in večblokovno zlivanje podatkov sta pokazala, da je gojišče glavni dejavnik spektralne variabilnosti, medtem ko ima identiteta seva sekundarni vpliv. Vibracijski pasovi, povezani s hidroksilnimi (−OH) in alifatskimi (−CH) skupinami, so bili opredeljeni kot ključni dejavniki razlikovanja in odražajo spremembe v sestavi zunajceličnih polimernih snovi (EPS). Ramanova spektroskopija je zagotovila dopolnilne molekularne informacije, kot je prisotnost značilnih vibracij beljakovin in polisaharidov, kar je pripomoglo k potrditvi biokemijske sestave biofilmov ter podprlo ugotovitve analiz IR in NIR. Rezultati kažejo, da nadzor sestave hranil in pogojev gojenja omogoča natančno uravnavanje lastnosti biofilmov, vključno s hidracijo, gostoto matriksa in vsebnostjo funkcionalnih polisaharidov. Na sestavo biofilma vpliva predvsem sestava hranil in ne morfološke značilnosti seva. To je pomembno, saj kaže, da se lahko raziskave formulacij živih premazov osredotočijo na zagotavljanje hranil, ki spodbujajo nastajanje EPS in razvoj biofilma, namesto na izbiro sevov s specifičnimi morfološkimi značilnostmi. Te ugotovitve predstavljajo osnovo za razumevanje okoljskega uravnavanja tvorbe biofilmov vrste A. pullulans in so pomembne za prihodnji razvoj inženirsko zasnovanih živih premazov, pri katerih je mogoče z okoljskimi pogoji vplivati na sestavo, strukturo in funkcionalne lastnosti biofilma. spectroscopy fungi engineered living materials biofilm Raman spectroscopy FTIR FT-NIR spektroskopija glive inženirski živi materiali biofilm Raman spektroskopija FTIR FT-NIR true false true Angleški jezik Slovenski jezik Članek v reviji 2026-08-31 15:56:27 2026-08-31 15:56:27 2026-09-01 03:07:32 0000-00-00 00:00:00 2026 0 0 str. 1-14 iss. Vol. 2026 0000-00-00 PostprintKoncna NiDoloceno apc 0000-00-00 0000-00-00 0000-00-00 62 1520-5827 10.1021/acs.langmuir.6c01932 289513475 RAZ_Cernosa_Anja_2026.pdf RAZ_Cernosa_Anja_2026.pdf 1 E9BED65EFA893C66628C7586719BFB76 e8bdae924c4981000f252d7699d2e11e98553083f5297506b9b865d234e7def6 b027e0c4-a543-11f1-9e8d-005056ac49c0 https://repozitorij.upr.si/Dokument.php?lang=slv&id=34968 https://pubs.acs.org/langd5/article/doi/10.1021/acs.langmuir.6c01932/5383264/Spectroscopic-Insights-into-Nutrient-Controlled 1 828d17cd-a543-11f1-9e8d-005056ac49c0 https://repozitorij.upr.si/Dokument.php?lang=slv&id=34967 Inštitut Andrej Marušič 0 0 0