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- Microalgae-associated Stenotrophomonas maltophilia enhances lutein production and biostimulant activity in Monoraphidium spPublication . Manoel, João; Štěrbová, Karolína; Saini, Mohit Kumar; Barcenas-Perez, Daniela; Cheel, José; Grivalsky, Tomás; Lakatos, Gergely Erno; Lukes, Martin; Urajová, Petra; Ferreira, Alice; Figueiredo, Daniel; Gouveia, Luisa; Masojidek, Jiri; Saurav, KumarABSTRACT: Photosynthetic organisms like microalgae can collect solar energy and transform it into biochemical compounds as other forms of energy that can be utilized in metabolic processes. In nature, microalgae coexist with bacterial communities and may maintain a symbiotic relationship. In the current study, a heterotrophic bacterium, Stenotrophomonas maltophilia was isolated from the phycosphere of a cold-adapted green microalga Monoraphidium sp. (further abbreviated as Monoraphidium). By using advanced liquid chromatography-high-resolution tandem mass spectrometry (LC-HRMS/MS), we were able to detect homoserine lactones (HSLs): 3OHC12-HSL, 3OHC10-HSL, 3OHC14-HSL, C10-HSL, C8-HSL, and OC14-HSL, produced by S. maltophilia. Further, the role of this bacterium in establishing intricate relationships and its implication on biotechnological potential was evaluated. Significant improvements were found in the lutein production of the Monoraphidium culture with bacterial supplements, achieving about 19.3 +/- 0.88 mg g-1 DW of this carotenoid compared to 13.7 +/- 1.87 mg g-1 DW in the control, which represents an increase of about 40 %. Furthermore, the biostimulant potential of Monoraphidium was evaluated using the germination tests with tomato and barley seeds. A higher germination index was observed with improvements of 55 % in tomato and 110 % in barley, respectively, as compared to the control culture, which was related to the microalgae's growth stage. The role of the bacterium was evaluated in how the intricate relationships with the microalgal culture can affect its biotechnological potential (e.g., biostimulant activity and lutein production). The current work expands our knowledge towards designing an efficient polyculture based on complementary traits and metabolic potential to maximize the yield and bioactivity in algal biotechnology.
- Editorial: The biorefineries and application of green technologies for recovering bioactive compounds from microalgaePublication . Vladic, Jelena; Gouveia, LuisaABSTRACT: Microalgae are attracting growing scientific and industrial interest as a renewable and versatile source of high-value bioactive compounds, including pigments, fatty acids, proteins, and antioxidants. Their rapid growth, ability to adapt to extreme conditions, and rich biochemical composition make them a promising resource for sustainable development across a range of applications. From cosmetics to food supplements, their potential spans multiple industries. Still, the considerable gap between encouraging laboratory research and commercially viable production remains. One of the major challenges lies in developing methods for extracting and preserving these compounds in ways that are both efficient and environmentally responsible.
- Consortium of microalgae/nitrogen-fixing bacteria as a next-generation biofertilizer, biostimulant and biopesticidePublication . Gouveia, Luisa; Ferreira, Alice; Bastos, Carolina R. V.; Avetisova, Gayane; Karapetyan, Zh.; Toplaghaltsyan, A.; Melkonyan, LusineABSTRACT: This study aims to provide insights into a new consortium of a microalga, twelve nitrogen-fixing (N-fixing) bacteria and a cyanobacterium. The microalga Tetradesmus obliquus (T), in conjunction with various N-fixing bacteria and the cyanobacterium Synechocystis sp. PCC 6803 (S), the N-fixing bacterium Sphingobacterium sp. L13G8 (5), cultivated in complete Bristol medium and in Bristol-NaNO3 free conditions, were examined. The study encompassed the analysis of their consortia, including evaluation of their growth, and potential as a biostimulant, biofertilizer and biopesticide, and assessed for sedimentation performance for targeted applications. The T and N-fixing bacteria consortia had higher growth in Bristol NaNO3-free media. The triple culture TS5 had the highest growth parameter (2.4 OD540) in the same medium, followed by T5. The consortia were employed to ascertain the efficacy of their biostimulants and biofertilizers on watercress (Lepidium sativum) and to determine the potency of their biopesticides against the fungal pathogens Fusarium oxysporum and Rhizoctonia solani (in certain consortia). Consortium T5 demonstrated the most significant impact on the seeds germination index (212.7%) and root length (6.0 cm) of L. sativum. The same consortium had a significant impact on the shoot length (4.4 cm) of L. sativum. Among all consortia that were examined, T5 exhibited significant inhibitory effects on the growth of F. oxysporum (60.6%) and R. solani (69.2%). In the same consortium, the rate of microalgal biomass sedimentation was enhanced by the N-fixing bacterium (0.4 cm h-1). Consortium T5 was the most effective in relation to growth and biomass sedimentation efficiency, in addition to its use as a biostimulant, biofertilizer and biopesticide. The created combination of microalga and N-fixing bacterium represents significant progress in the field of microalga cultivation, with notable benefits including improved biomass sedimentation and enhanced agricultural practices, as well as environmental friendliness and safety.
- Reducing nutrient requirement using nitrogen-fixing bacteria for microalgae cultivationPublication . Melkonyan, Lusine; Ferreira, Alice; Bastos, Carolina R. V.; Figueiredo, Daniel; Lopes da Silva, Teresa; Avetisova, Gayane; Karapetyan, Zh.; Toplaghaltsyan, A.; Gouveia, LuisaABSTRACT: In environments, microalgae have been observed to coexist with bacteria. Different nitrogen-fixing bacteria (NFB) were isolated from Armenian soils and their growth was evaluated in co-cultivation with the microalga Tetradesmus obliquus and cyanobacteria Synechocystis sp. PCC 6803. The most effective mutualistic consortium was T. obliquus-NFB5 (Sphingobacterium sp. L13G8). This resulted an increase in both populations, chlorophyll fluorescence, biomass protein, carbohydrate content, an effect on lipid metabolism, without the need for external nitrogen. The findings demonstrated the significance of employing NFB for microalga growth, as they facilitate the essential nitrogen provision in N-free Bristol medium. Moreover, in mutualistic consortia, microalgae facilitate the exudation of dissolved organic carbon and O2 to bacteria, which, in turn, become available for bacteria, thereby reducing the necessity for energy-consuming aeration processes in co-cultivation. In return, the bacteria provide the microalgae with CO2, B vitamins and demineralize N2, P, S, thereby further supporting the growth of microalgae.
- Industrial Off-Gas Fermentation for Acetic Acid Production: A Carbon Footprint Assessment in the Context of Energy TransitionPublication . Pacheco, Marta; Brac de la Perrière, Adrien; Moura, Patrícia; silva, carlaABSTRACT: Most industrial processes depend on heat, electricity, demineralized water, and chemical inputs, which themselves are produced through energy- and resource-intensive industrial activities. In this work, acetic acid (AA) production from syngas (CO, CO2, and H2) fermentation is explored and compared against a thermochemical fossil benchmark and other thermochemical/biological processes across four main Key Performance Indicators (KPI)—electricity use, heat use, water consumption, and carbon footprint (CF)—for the years 2023 and 2050 in Portugal and France. CF was evaluated through transparent and public inventories for all the processes involved in chemical production and utilities. Spreadsheet-traceable matrices for hotspot identification were also developed. The fossil benchmark, with all the necessary cascade processes, was 0.64 kg CO2-eq/kg AA, 1.53 kWh/kg AA, 22.02 MJ/kg AA, and 1.62 L water/kg AA for the Portuguese 2023 energy mix, with a reduction of 162% of the CO2-eq in the 2050 energy transition context. The results demonstrated that industrial practices would benefit greatly from the transition from fossil to renewable energy and from more sustainable chemical sources. For carbon-intensive sectors like steel or cement, the acetogenic syngas fermentation appears as a scalable bridge technology, converting the flue gas waste stream into marketable products and accelerating the transition towards a circular economy.
- Bio-oil from hydrothermal liquefaction of microalgae cultivated in wastewater: An economic and life cycle approachPublication . Silva, Thiago; Junior, Maurino Magno de Jesus; Magalhães, Iara; Ananias, Marina Stefany; Saleme Aona de Paula Pereira, Alexia; Rodrigues, Fábio de Ávila; Delgado dos Reis, Alberto José; Calijuri, Maria LuciaABSTRACT: Although microalgae are a promising sustainable biofuel feedstock, their energy-intensive production and most environmental assessments rarely achieve the desired trade-off between productivity and sustainability. In this context, this study aims to evaluate the economic and environmental feasibility of producing bio-oil via hydrothermal liquefaction (HTL) of wastewater-grown microalgae at an industrial scale. Four scenarios varied production scale and steam source: sugarcane bagasse (SCB) in SC1 and SC3, liquefied petroleum gas (LPG) in SC2 and SC4. Each scenario processed microalgae at 300 degrees C for 30 min. Smaller-scale feedstock (1332.9 kg/h) in SC1 and SC2 produced 34.6 kg/h of bio-oil, while the larger feedstock (85,554.4 kg/h) in SC3 and SC4 yielded 2222.2 kg/h. Microalgae biomass cultivation costs dominated overall expenses (56-75 %). Economic analyses indicated minimum selling prices of 3.82-8.52 USD/kg, exceeding the average literature figure of 1.57 USD/kg. Life Cycle Assessment (LCA) showed SCB reduced fossil resource depletion by 14.97 % compared to LPG but increased emissions of nitrogen oxides, particulates, and toxic compounds, which are manageable via selective catalytic reduction and flue gas desulphurization. Cyclohexane as a solvent elevated human carcinogenic toxicity, greener alternatives could reduce toxicity but may cost more, requiring further cost analysis. Advancing this biorefinery route requires optimization of cultivation and processing costs, adoption of environmentally benign solvents, and implementation of emission control strategies to enable economically feasible and environmentally sustainable bio-oil production.
- Strawberry Tree Fruit Residue as Carbon Source Towards Sustainable Fuel Biodesulfurization by Gordonia alkanivorans Strain 1BPublication . Paixão, Susana M.; Silva, Tiago; Salgado, Francisco; Alves, LuísABSTRACT: Biodesulfurization (BDS) is a clean technology that uses microorganisms to efficiently remove sulfur from recalcitrant organosulfur compounds present in fuels (fossil fuels or new-generation fuels resulting from pyrolysis and hydrothermal liquefaction). One of the limitations of this technology is the low desulfurization rates. These result in the need for greater amounts of biocatalyst and lead to increased production costs. To mitigate this issue, several approaches have been pursued, such as the use of alternative carbon sources (C-sources) from agro-industrial waste streams or the co-production of high-added-value products by microorganisms. The main goal of this work is to assess the potential of strawberry tree fruit residue (STFr) as an alternative C-source for a BDS biorefinery using Gordonia alkanivorans strain 1B, a well-known desulfurizing bacterium with high biotechnological potential. Hence, the first step was to produce sugar-rich liquor from the STFr and employ it in shake-flask assays to evaluate the influence of different pretreatments (treatments with 1-4% activated charcoal for prior phenolics removal) on metabolic parameters and BDS rates. Afterwards, the liquor was used as the C-source in chemostat assays, compared to commercial sugars, to develop and optimize the use of STFr-liquor as a viable C-source towards cost-effective biocatalyst production. Moreover, the high-market-value bioproducts simultaneously produced during microbial growth were also evaluated. In this context, the best results, considering both the production of biocatalysts with BDS activity and simultaneous bioproduct production (carotenoids and gordofactin biosurfactant/bioemulsifier) were achieved when strain 1B was cultivated in a chemostat with untreated STFr-liquor (5.4 g/L fructose + glucose, 6:4 ratio) as the C-source and in a sulfur-free mineral-minimized culture medium at a dilution rate of 0.04 h-1. Cells from this steady-state culture (STFr L1) achieved the highest desulfurization with 250 mM of dibenzothiophene as a reference organosulfur compound, producing a maximum of approximate to 213 mM of 2-hydroxibyphenil (2-HBP) with a corresponding specific rate (q2-HBP) of 6.50 mu mol/g(DCW)/h (where DCW = dry cell weight). This demonstrates the potential of STFr as a sustainable alternative C-source for the production of cost-effective biocatalysts without compromising BDS ability. Additionally, cells grown in STFr L1 also presented the highest production of added-value products (338 +/- 15 mu g/g(DCW) of carotenoids and 8 U/mL of gordofactin). These results open prospects for a future G. alkanivorans strain 1B biorefinery that integrates BDS, waste valorization, and the production of added-value products, contributing to the global economic viability of a BDS process and making BDS scale-up a reality in the near future.
- Alternative feedstocks for high-quality biodiesel: Lipid production from eucalyptus bark hydrolysate by Yarrowia lipolytica W29 using different cultivation modesPublication . Dias, Bruna; Lopes, Marlene; Marques, Susana; Gírio, Francisco; Belo, IsabelABSTRACT: Microbial lipids produced by yeasts from lignocellulosic biomass are a promising feedstock for the biodiesel industry, providing a renewable energy source as an alternative to traditional fossil fuels. This study investigated the potential of Yarrowia lipolytica W29 to produce lipid-rich biomass from undetoxified sugar-concentrated eucalyptus bark hydrolysate (EBH). The lipid concentrations achieved in batch cultures (13.4 g L-1) were the highest for wild-type Y. lipolytica strains in lignocellulosic hydrolysates. Different two-stage cultivation modes (repeated batch, continuous-feeding fed-batch, and pulse fed-batch) were studied to enhance biomass and lipid production. The cell and lipid mass was higher in pulse fed-batch and continuous-feeding fed-batch cultures than batch cultures. Production of citric acid, a side product of industrial interest, was improved in the continuous-feeding fed-batch culture. Microbial lipids produced by Y. lipolytica W29 were highly unsaturated and mainly composed of oleic acid (50% to 53%). The estimated properties of the biodiesel that would be obtained from these intracellular lipids would meet the international biodiesel standards EN 14214 and ASTM D6751. This study demonstrates the feasibility of using EBH for Y. lipolytica lipid production and promotes the sustainable production of high-quality biodiesel from lignocellulosic feedstocks.
- Bridging gaps in biorefineries: The unexplored role of social dimension in life cycle assessment researchPublication . Ortigueira, Joana; Lopes, TiagoABSTRACT: This review examines the disregarded role of social dimensions in Life Cycle Assessment (LCA) within biorefinery implementation, addressing the question: "How can the inclusion of social factors in LCA improve sustainability assessments, and what are the implications of the limited Social Life Cycle Assessment (S-LCA) studies in biorefineries?" A systematic literature review was conducted using Web of ScienceTM, focusing on studies that integrate social dimensions in LCA. Bibliometric analysis using the bibliometrix R-package and VOSviewer identified key trends, influential papers, and research gaps. Results revealed a significant gap in incorporating social dimensions into biorefinery LCA, with most studies focusing primarily on environmental and economic impacts. Limited attention is given to social aspects such as community well-being, labor rights, and social equity. Case studies that included social factors demonstrated a more comprehensive sustainability assessment, emphasizing the importance of stakeholder engagement and social acceptability in biorefinery projects. This review highlights the need for standardized social indicators and methodologies to integrate social dimensions effectively. The lack of S-LCA in biorefinery implementation reflects a critical gap in sustainability assessments. Addressing this requires developing a unified S-LCA methodology, fostering interdisciplinary collaboration, and encouraging stakeholder participation to ensure diverse perspectives are considered. Ultimately, incorporating social dimensions is essential for achieving a more balanced and comprehensive evaluation of biorefinery sustainability.
- Design optimisation of five pilot-scale two-stage vertical flow-constructed wetlands for piggery wastewater treatmentPublication . Karan, N.; Gogoi, Jayanta; Ganguly, Anasuya; Brito, António; Marques dos Santos, C.; de Oliveira Corrêa, Diego; Gouveia, Luisa; Mutnuri, SrikanthABSTRACT: With growing pig farming, sustainable piggery wastewater treatment methods are essential for environmental protection. This study evaluated five pilot-scale two-stage vertical flow-constructed wetlands (VFCWs) with varying configurations of aeration, plantation, and saturation zones. Three VFCW configurations (1VFCW, 2VFCW, and 3VFCW) were unsaturated, while 4VFCW and 5VFCW were saturated in the second stage (up to 60 and 90 cm, respectively). The 5VFCW featured a stacked configuration with no space between its two stages. Passive aeration was selectively applied in 2VFCW, 3VFCW, 4VFCW, and 5VFCW, while plants were present in most configurations except the control. Saturated 4VFCW achieved the highest removal efficiency for TN (77.03 ± 16.24%) and NO3− (46.06 ± 45.96%), while the stacked 5VFCW showed the highest removal for chemical oxygen demand (COD) (94.17 ± 4.85%) and Total ammoniacal nitrogen (TOC) (86.35 ± 6.78%). Unsaturated 1VFCW excelled in TAN removal (98.89 ± 0.33%), and the control system (C) showed the highest removal efficiency for PO43− (90.38 ± 6.52%) and TOC (87.52 ± 9.83%). Overall, 4VFCW emerged as the most balanced and effective system, supported by an optimal combination of aerobic and anaerobic conditions that facilitated sequential nitrification and denitrification, along with an extended hydraulic retention time due to saturation.