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- Optimizing bacterial nanocellulose production from eucalyptus bark: A circular approach to wastewater management and resource recoveryPublication . Rodrigues, Ana Cristina; Martins, Daniela; Duarte, Maria Salomé; Marques, Susana; Gama, Miguel; Dourado, Fernando; Carvalho, Ricardo; Cavaleiro, AnaABSTRACT: The production cost of bacterial nanocellulose (BNC) is a major limitation to its widespread use. However, this limitation can be addressed by using alternative low-cost substrates and high-yield strains. Agro-industrial wastederived substrates offer a cost-effective and sustainable solution, but their high organic load often requires additional downstream wastewater treatments. Here, we optimized static BNC production using eucalyptus bark hydrolysate (EBH) as a low-cost carbon source and proposed a circular approach for wastewater management. Optimization was performed using response surface methodology - central composite design. The optimized EBH medium yielded a 39.7-fold increase compared to standard medium, with a maximum BNC production of 8.29 f 0.21 g/L. Fermentation wastewater only (WaF) and combined with BNC washing streams (WaW) revealed high levels of organic matter, namely chemical oxygen demand (COD) of 159.0 f 2.0 and 41.1 f 0.3 g/L, and volatile solids (VS) of 99.5 f 0.9 and 26.3 f 0.2 g/L, respectively, requiring treatment before disposal. A sequential anaerobic-aerobic digestion was investigated for wastewater treatment and valorisation. Anaerobic digestion proved to be effective in treating the wastewater: methanization percentages over 87 % were achieved, and methane productions of 486 f 2 and 544 f 30 L/kg VS were obtained from WaF and WaW, respectively. Subsequent aerobic treatment was unsuccessful in further reducing COD levels (approximately 1.5 g/L). Notably, treated wastewater was recycled into the production process up to 45 % without affecting the BNC yield. This study provides valuable insights into the optimization of BNC production from lignocellulosic biomass and the management of wastewater streams, contributing to the development of a more sustainable and economically viable process.
- Nanobubble-enhanced oxygen transfer in bacterial nanocellulose production: Comparative evaluation with static and airlift systemsPublication . Rodrigues, Ana Cristina; Martins, Daniela; Carvalho, Ricardo; Marques, Susana; Belo, Isabel; Espina, Begona; Dourado, Fernando; Gama, MiguelABSTRACT: Despite the unique properties of bacterial nanocellulose (BNC), oxygen limitation during large-scale production impairs microbial metabolism and cellulose synthesis, leading to high production costs and limited commercial success. Static fermentation can achieve high titers, but industrially it is operationally challenging. Agitated systems like airlift (AL) bioreactors, allow faster production but typically yield lower titers. This study pioneered the use of an agitated bioreactor equipped with a nanobubble (NB) generator, and its performance was compared with that of static and AL systems, employing a newly isolated Komagataeibacter sp. strain from kombucha cultivated in Eucalyptus bark hydrolysate and corn steep liquor. Key monitored parameters included dissolved oxygen, cell density, pH, sugar and lactic acid contents, and BNC production. The obtained BNC was characterized for its crystallinity, thermal stability, degree of polymerization, morphology and fiber size. The AL-and NB-derived BNC exhibited a denser network structure, lower crystallinity index, and lower polymerization degrees than that from static culture. NB technology generated stable nanobubbles (size: 95.8 f 12.9 nm; zeta potential:-14.2 f 8.6 mV). At 1 L. min-1 airflow, compared to AL, the NB bioreactor achieved a 6-fold higher volumetric mass transfer coefficient (kLa 35.9 f 1.2 h-1) and oxygen transfer rate (OTR: 309.7 f 10.2 mg.L-1.h-1). It supported greater cell density but maintained a similar BNC volumetric productivity to that of the AL (0.023 g.L-1.h-1), and moderately higher (near 280%) than that of static culture (0.0082 g.L-1.h-1). Thus, the improved oxygen levels provided by the NB system favored biomass growth rather than BNC production, suggesting that further optimization is needed to redirect carbon flux toward BNC production.