Cocultivation of fungi and algae can result in a mutualistic or antagonistic interaction depending on the species involved and the cultivation conditions. In this study, we investigated the growth behavior and enzymatic activity of two filamentous white-rot fungi (Trametes versicolor and Trametes pubescens) and two freshwater algae (Chlorella vulgaris and Scenedesmus vacuolatus) cocultured in the presence of TEMPO (2,2,6,6-tetramethylpiperidine-1-oxyl radical) oxidized cellulose nanofibrils (CNF) and cellulose nanocrystals (CNC). The growth of fungi and algae was studied in liquid, agar medium, and 3D-printed nanocellulose hydrogels. The results showed that cocultures grew faster under nutrient-rich conditions than in nutrient-depleted conditions. Key cellulose-degrading enzymes, including endoglucanase and laccase activities, were higher in liquid cocultures of T. versicolor and S. vacuolatus in the presence of cellulose compared to single cultures of fungi or algae. Although similar results were observed for cocultures of T. pubescens and C. vulgaris, laccase production diminished over time in these cultures. Fungi and algae were capable of growth in 3D-printed cellulose hydrogels. These results showed that cellulase enzyme production could be enhanced by cocultivating white-rot fungi with freshwater algae under nutrient-rich conditions with TEMPO-CNF and CNC. Additionally, the growth of white-rot fungi and freshwater algae in printed cellulose hydrogels demonstrates the potential use of fungi and algae in hydrogel systems for biotechnological applications, including biofuel production and bio-based fuel cell components. IMPORTANCE Depending on the conditions used to grow fungi and algae in the lab, they can interact in a mutually beneficial or negative way. These interactions could stimulate the organisms to produce enzymes in response to the interaction. We studied how wood decay fungi and freshwater algae grew in the presence and absence of cellulose, one of the basic building blocks of wood. How fungi and algae grew in 3D-printed cellulose hydrogels was also tested. Our results showed that fungi and algae partners produced significantly larger amounts of enzymes that degraded cellulose when grown with cellulose than when grown alone. In addition, fungi and algae were shown to grow in dense nanocellulose hydrogels and could survive the shear conditions during gel structuring while 3D-printing. These cultures could potentially be applied in the biotech industry for applications like energy production from cellulose, biofuel production, and bioremediation of cellulose material.
Keywords: 3D-printing; Chlorella vulgaris; Scenedesmus vacuolatus; TEMPO; algae; cellulose; nanofibril; white-rot fungi.