Centro de Biotecnologia e Química Fina (CBQF)
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Browsing Centro de Biotecnologia e Química Fina (CBQF) by Sustainable Development Goals (SDG) "09:Indústria, Inovação e Infraestruturas"
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- Aerobic granules synthesized with EPS and degrading strain Rhodococcus sp. FP1 for industrial wastewater treatmentPublication . Oliveira, Ana; Amorim, Catarina L.; Zlopassa, Jure; Lin, Yuemei; van Loosdrecht, Mark; Castro, Paula M. L.Background: Aerobic granular sludge-sequencing batch reactors (AGS-SBR) have been introduced as a promising and innovative wastewater treatment system economically outcompeting the conventional activated sludge system. AGS is considered a special case of suspended biofilms, composed of self-immobilized microorganisms that form spherical sludge aggregates. Microorganisms are embedded in a self-produced extracellular polymeric substances matrix thus avoiding the need of any carrier. Many synthetic organic chemicals or metabolites are being released directly into the environment after wastewater treatment processes, which are not conceived to remove them from the effluents. In spite of the AGS tolerance to toxicity, the indigenous microbial communities in biotreatment processes may not be effective in removing recalcitrant pollutants. Bioaugmentation strategies, which consists of adding specific microorganisms to the system can be a solution to overcome the difficulty to eliminate certain compounds in wastewaters. However, it is still not a well-established strategy and deserves attention to become a promising alternative. This work aimed to synthesize strong aerobic granules using EPS (extracted from AGS) and a specialized bacterial strain capable of degrading 2-fluorophenol, with the aid of specific substances to increase the cross-linking potential of EPS. Conclusions: Substance 1 was not efficient in improving the strength of the granules, due to concentration or inability to promote the crosslinking of EPS; Granules from mixture 6 were the most similar in terms of strength to the Utrecht granules. The source of EPS affected the strength of the granules, even with conditions similar to mixture 6 Control beads from mixture 7 showed to be weaker than the ones from mixture 6, reinforcing that EPS was a key component to increase the strength of the produced granules. EPS composition and concentration are important to produce strong granules that can be further used in bioaugmentation.
- Bioactive potential of olive leaf by-product throughout in vitro gastrointestinal digestionPublication . Sánchez-Gutiérrez, Mónica; Gómez-García, Ricardo; Carrasco, Elena; Rodríguez, Alejandro; Pintado, ManuelaOlive leaf, an abundant and underutilized byproduct of the olive industry, has gained attention as a potential functional ingredient due to its high content of dietary fiber and phenolic compounds. However, little is known about its bioaccessibility and transformation throughout the digestive process, limiting its application in food formulations. This study provides a comprehensive and quantitative assessment of how ground olive leaf bioactive compounds behave during gastrointestinal digestion, offering new insights into their stability and potential health benefits. The total phenolics content and antioxidant activity of ground olive leaf increased in the oral and gastric phases, decreasing slightly in the intestinal phase, with a bioaccessibility of 46% and up to 70% for the total phenolic content and antioxidant activity, respectively. The principal individual phenolic compounds identified in the intestinal phase were oleuropein, luteolin-7-glycoside, luteolin-6-glycoside and ferulic acid, with bioaccessibilities of up to 97%. The main soluble sugars (fructose, glucose, and sucrose) and organic acids (succinic, citric, and acetic acids) detected in the olive leaf samples showed different behaviors during gastrointestinal digestion: sugars increased in the oral and gastric phases but decreased in the intestinal phase, with high bioaccessibility despite reduced recovery, while organic acids remained mostly stable, except for citric acid, which decreased significantly in the intestinal phase, all showing close to 100% bioaccessibility. These results provide the first detailed evidence of the digestive fate of ground olive leaf bioactive compounds, reinforcing its potential as a functional ingredient. Its natural availability, without requiring pre-treatment, combined with its high antioxidant potential and bioaccessibility, highlights its relevance for the development of innovative food ingredients, aligning with circular economy principles and sustainable food strategies.
- Breaking the virus: yeast glucans as an effective alternative to acyclovir in HSVI treatmentPublication . Tavares-Valente, Diana; Moreira, Helena; Sousa, Pedro; Amorim, Manuela; Conde, António; Pintado, Manuela; Fernandes, João; Azevedo-Silva, JoãoGlucans, structural polysaccharides in the yeast cell wall, are known for their biological and immunomodulatory capacities, helping in prevention and management of infections. Herpes simplex virus type 1 (HSVI) is a prevalent infection that causes great comorbidity and is challenging to treat due to the adverse effects of standard antiviral drugs like acyclovir. This study assessed the potential of yeast glucans extracted from two different origins − a steviol-glycoside producing strain and a wild-type strain- to circumvent HSVI infection, either in vitro and ex vivo. Treatment with glucans in keratinocytes and macrophages in vitro reduced cell infection similarly to acyclovir. However, unlike acyclovir, glucans demonstrated an immunostimulatory effect, increasing the production of IL-1β, TNF-α and IL-6. Additionally, both glucans were formulated with squalane for skin application. This formulation improved glucans penetration in the skin, restored skin structure and reduced the cytopathic effect of HSVI infection. In summary, this study highlights yeast glucans as a natural therapeutic alternative for HSVI treatment, offering an option with an excellent safety profile. Moreover, using glucans from industrial side-streams promotes a sustainable approach, contributing to the circular economy.
- Carnosic acid production from sugarcane syrup by engineered yeast in fed-batch fermentationPublication . Carsanba, Erdem; Fernandes, Sara; Beato, Felipe; Carvalho, Luís Carlos; Pintado, Ana; Lopes, Ana; Ribeiro, Mónica; Leal, Tânia; Pintado, Manuela; Oliveira, CarlaPhenolic diterpene carnosic acid (CA) is widely used in the food, nutritional health, and cosmetic industries due to its antioxidative and antimicrobial properties. This work aimed to overproduce CA in Saccharomyces cerevisiae from sugarcane syrup in fed-batch 2 L bioreactor fermentation. A geranylgeranyl diphosphate (GGPP)-producing strain modified with genes encoding the enzymes copalyl diphosphate synthase (Pv.CPS), miltiradiene synthase (Ro.KSL2), hydroxy ferruginol synthase (Ro.HFS), CA synthase (Ro.CYP76AK8), CYP reductase (At.ATR1), and transketolase (TKL1) was used. Lowering the feed rate from 12–26 g/L/h to 7–8 g/L/h, and the use of a dynamic dissolved oxygen (DO) trigger (min. 10%, max. 40%, threshold 70%) instead of a DO trigger of 30%, enhanced CA production by 27%. As a result, the highest CA titer ever reported to date, 191.4 mg/L, was obtained in 4-day fermentation. This study shows the feasibility of engineered yeast to produce CA from the sustainable feedstock sugarcane syrup.
- Dry-aged beef: a global review of meat quality traits, microbiome dynamics, safety, and sustainable strategiesPublication . Ribeiro, Ana J.; Silva, Filipe; Teixeira, Paula; Saraiva, Cristina M.Dry-aged beef is valued for its tenderness, complex aroma, and concentrated flavor. However, variability in aging protocols and limited understanding of underlying biological and technological processes can compromise consistent quality and safety. This review examines factors influencing meat characteristics, including genetics, breed, sex, age, diet, intramuscular fat deposition, antioxidant reserves, and endogenous enzyme pools. Postmortem biochemical pathways, such as proteolysis, lipid oxidation, and nucleotide degradation, are discussed in relation to flavor and texture development. The dynamics of surface microbiota are analyzed, highlighting the succession from psychrotrophic spoilage bacteria to molds and yeasts, which collectively form an enzymatic crust that contributes umami and nutty notes while providing antimicrobial barriers. Regulatory frameworks in major markets are reviewed, alongside valorization strategies that convert crust trimmings into umami-rich powders, bioactive peptides, starter cultures, or industrial enzymes. Despite advances, critical knowledge gaps remain, including the functional roles of minor crust taxa, the efficacy of defined starter cultures or bacteriophage blends, and standardized methods for texture and flavor measurement. By focusing on these biochemical and microbiological mechanisms and their applications, this review provides a roadmap for transforming dry aging into a reproducible, safe, and high-quality process in modern meat science.
- Enhancing carob flour (Ceratonia siliqua L.) for by-product utilization in food industries: carob syrup production, functional profiling and applicationPublication . Vilas-Boas, Ana Martins; Brassesco, María Emília; Quintino, Andreia C.; Medronho, Bruno; Vieira, Margarida C.; Brandão, Teresa R. S.; Silva, Cristina L. M.; Silva, Beatriz; Azevedo, Miguel; Pintado, ManuelaThe focus on by-product valorization in the food industry, particularly from the carob pod, underscores a commitment to sustainability and resource efficiency. This fruit, sourced from the leguminous evergreen carob tree (Ceratonia siliqua L.), is renowned for its adaptable flavour and nutritional value, in Mediterranean regions such as Portugal. Its production yields significant by-products, presenting environmental challenges when not managed efficiently. Innovative approaches, including integral carob flour production, aim to optimize utilization while minimizing waste and energy consumption. This study repurposed carob waste to produce novel, value-added ingredients like carob syrup, by thermal hydrolysis of integral carob flour using water at 1:3 solid-to-liquid ratio - obtaining up to 50 % solubility yield. The resulting syrup exhibited 72 % °Brix, a melting temperature (Tm) of approximately 130 °C and predominantly viscous behavior with minimal elastic (solid-like) response. Lastly, the syrup was incorporated into a carob-based brigadeiro, replacing conventional glucose-fructose syrup. Simulated gastrointestinal digestion revealed enhanced bioaccessibility of sugars and phenolics, and increased antioxidant activity during the intestinal phase. Despite sugar availability, the prebiotic activity of the syrup decreased when embedded in the brigadeiro matrix, potentially due to interactions with polyphenols or organic acids. Cytotoxicity and permeability assays confirmed safety at ?0.5 % (w/v) and supported intestinal barrier integrity. These findings support the use of integral carob flour for producing multifunctional ingredients, contributing to circular economy models while meeting consumer demands for healthier, sustainable food products.
- A flow-based method as a more sustainable tool for total protein monitoring in industrial processesPublication . Teixeira, Raquel; Ribas, Tânia C. F.; Almeida, André; Pintado, Manuela; Rangel, António O. S. S.The growing interest in protein hydrolysates highlights the need for fast and reliable analytical methods to assess their nutritional and functional properties. Total protein quantification is a key parameter for characterizing by-products, monitoring hydrolysis processes, and evaluating hydrolysates. In this work, a sustainable flow-based spectrophotometric method was developed and optimized using the Biuret method [1]. The flow injection analysis (FIA) system employs a double injection strategy in a continuous stream, significantly reducing reagent consumption compared to the conventional procedure, with a 75% decrease in Biuret reagent and an 83% reduction in sodium hydroxide, while maintaining analytical accuracy (Figure 1). Optimization of flow and chemical parameters enabled robust performance across different matrices used during enzymatic hydrolysis. The dynamic range of the method was 0.100 results were in agreement with those obtained with the reference method (<10% deviation). Additionally, the herein proposed method reduced the analysis time from 30 to 3 minutes per sample (triplicate). The proposed protocol offers a faster, greener, and reliable tool for the food industry, supporting sustainability in quality control and nutritional evaluation.
- A flow-based method as a more sustainable tool for total protein monitoring in industrial processesPublication . Teixeira, Raquel; Ribas, Tânia C. F.; Almeida, André; Pintado, Manuela; Rangel, António O. S. S.Introduction: This proposal aims to promote the circular economy, minimizing waste and enhancing environmental sustainability. In collaboration with several companies, the project uses by-products, subjecting them to hydrolysis processes to obtain hydrolysates. Objective: The main objective is to develop a flow spectrophotometric method to quantify the total soluble protein of these by-products, valuing them in the production of new products, such as animal feed, among others. Conclusion: This method is a robust, versatile, and accurate tool for quality control in the food industry, supporting a circular economy.
- Innovative processing and sterilization techniques to unlock the potential of silk sericin for biomedical applicationsPublication . Veiga, Anabela; Ramírez-Jiménez, Rosa Ana; Santos-Rosales, Víctor; García-González, Carlos A.; Aguilar, Maria Rosa; Rojo, Luis; Oliveira, Ana L.Silk sericin (SS), a by-product of the textile industry, has gained significant attention for its biomedical potential due to its biocompatibility and regenerative potential. However, the literature lacks information on SS processing methods and the resulting physicochemical properties. This study represents the first step in protocol optimization and standardization. In the present work, different processing techniques were studied and compared on SS extracted from boiling water: evaporation, rotary evaporation, lyophilization, and dialysis, which presented a recovery yield of approximately 27–32%. The goal was to find the most promising process to concentrate extracted SS solutions, and to ensure that the SS structure was highly preserved. As a result, a new cryo-lyophilization methodology was proposed. The proposed method allows for the preservation of the amorphous structure, which offers significant advantages including complete dissolution in water and PBS, an increase in storage stability, and the possibility of scaling-up, making it highly suitable for industrial and biomedical applications. The second part of the work focused on addressing another challenge in SS processing: efficient and non-destructive sterilization. Supercritical CO2 (scCO2) has been gaining momentum in the last years for sterilizing sensitive biopolymers and biological materials due to its non-toxicity and mild processing conditions. Thus, scCO2 technology was validated as a mild technique for the terminal sterilization of SS. In this way, it was possible to engineer a sequential cryo-lyophilization/scCO2 sterilization process which was able to preserve the original properties of this natural silk protein. Overall, we have valorized SS into a sterile, off-theshelf, bioactive, and water-soluble material, with the potential to be used in the biomedical, pharmaceutical, or cosmetic industries.
- Life cycle assessment of floating wetland islands for crop productionPublication . Carrillo, Valentina; Pereira, Sofia; Afonso, Carlos A.; Calheiros, CristinaBackground & Aim: Floating wetland islands (FWI) are innovative, nature-based solutions for water management and ecological restoration, rooted in circular economy and sustainable development [1,2]. They consist of floating media planted with macrophytes that act as biofilters by absorbing pollutants like nitrogen and phosphorus, thereby enhancing crop production [3]. FWI in hydroponic crops can be a viable alternative for agricultural production, allowing a yield per square meter by taking advantage of the nutrients present in the water and optimizing the use of space. This study aims to evaluate the feasibility of crop production in FWI using a hydroponic system, estimating the potential yield per square meter. Additionally, a life cycle assessment (LCA) will be performed to quantify the environmental impact and ensure resource efficiency in this innovative agricultural approach. Methods: Rainwater runoff will be used and characterized to evaluate parameters such as nutrients, organic matter, dissolved oxygen, salinity, pH, electrical conductivity and temperature. The study will consist of a prototype FWI with three replicates and a control, using expanded clay on the floating platform. Plants will be selected for humans and animals. Biomass production per m², foliar analysis and chlorophyll will be measured. Finally, LCA will be carried out from “cradle to grave”, considering the limits and scope of the system in three types of scenarios. Expected results: The feasibility of hydroponic crop production in FWI is expected, analyzing yield per square meter, associated to the plants health and establishment. The LCA will allow us to assess the environmental impact and identify opportunities to optimize the sustainability and efficiency of the system according to the planned approach. system according to the intended approach.