Browsing by Author "Pereira, Carlos D."
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- Inhibition of fish cooking wastewater oxidation with acorn extractPublication . Araújo-Rodrigues, Helena; Machado, Manuela; Pereira, Carlos D.; Pintado, Manuela E.
- Manufacture of bioactive peptide-rich concentrates from whey: characterization of pilot processPublication . Tavares, Tânia G.; Amorim, Manuela; Gomes, David; Pintado, Manuela E.; Pereira, Carlos D.; Malcata, F. XavierThis work was focused on the manufacture, at pilot scale, of cow whey protein and peptide concentrates, using selective filtration techniques—associated with hydrolysis brought about by proteolytic enzymes from Cynara cardunculus aqueous extracts, using as (optimal) conditions an enzyme/substrate ratio of 1.6% v/v, a pH of 5.2, a temperature of 55 C and an incubation time of 7 h. The profiles of proteins and peptides were assessed by liquid chromatography and electrophoresis; ca. 87% of a-lactalbumin was hydrolyzed, but essentially no degradation of b-lactoglobulin (b-Lg) was observed. A bioactive peptide concentrate, its fraction below 3 kDa and a b-Lg-rich fraction were obtained as final products, containing ca. 73, 43 and 91% w/w protein (on a total solid mass basis). All these fractions were low in lactose and salt, and their microbial loads were reduced. Said fractions are high added-value products, so they can be used as nutritional and functional ingredients—thus yielding an economically viable alternative for upgrade of whey.
- Membrane fractionation of Cynara cardunculus swine blood hydrolysate: ingredients of high nutritional and nutraceutical valuePublication . Araújo-Rodrigues, Helena; Coscueta, Ezequiel R.; Pereira, Miguel F.; Cunha, Sara A.; Almeida, André; Rosa, Ana; Martins, Rui; Pereira, Carlos D.; Pintado, Manuela E.This work proposes an innovative approach to valorise swine blood based on enzymatic hydrolysis and membrane fractionations. Hydrolysis with Cynara cardunculus enzymes, followed by microfiltration and double nanofiltration generated three high protein fractions, retentate of microfiltration (RMF; >0.5 µm) and retentate of nanofiltration (RNF; >3 kDa) with approximately 90% of protein on a dry basis and filtrate of nanofiltrate (FNF; <3 kDa) with 65%. FNF, rich in low molecular weight peptides, showed excellent antioxidants (ABTS and ORAC of 911.81 and 532.82 µmol TE g−1 DB, respectively) and antihypertensive (IC50 of 28.51 µg mL−1) potential. By peptidomics and in silico analysis, 43 unique sequences of interest were found, among which LVV-Hemorphin-7 was identified. This hemorphin was demonstrated as the main responsible for the observed bioactivity. Complementary results showed a prebiotic effect mainly for the growth of Bifidobacterium animalis Bo, as well as interesting free amino acids (mainly glutamic acid, leucine, alanine, phenylalanine and aspartic acid) and mineral (e.g., Ca, Mg, P, K and Na) profiles. No antibacterial effect was verified for the seven pathogenic bacteria tested. This study allowed obtaining new ingredient of high nutritional and nutraceutical value for human consumption, with a perspective of sustainability and industrial viability.
- A new valorisation approach of porcine blood based on enzymatic hydrolysis and membrane technology: the biological potential of resultant hydrolysate as a food ingredientPublication . Araújo-Rodrigues, Helena; Pereira, Miguel; Pereira, Carlos D.; Pintado, ManuelaBlood is one of the main meat industry by-products, which possess an interesting nutritional value and components with potential as a functionalingredient1. In this study, a new valorisation approach based on enzymatic hydrolysis and membrane technology was developed. Swine blood was cooked and enzymatically hydrolysed with Cynara cardunculusL. After hydrolysis, the product was filtered and the liquid fraction was then submitted to microfiltration (MF),using a 0.2 μm pore size membrane. MF retentate (MFR) was subject to reverse osmosis (RO) and freezedried. Subsequently, MF filtrate (MFF) was submitted to a nanofiltration (NF) 3 kDa cut-off membrane. Nanofiltration retentate fraction (NFR) was subjected to RO and freezedried. NF filtrate was resubmitted to a sequential step of NF (120 g mol-1cut-off membrane). The resultant retentate, NF filtrate (NFF) was also submitted to RO and lyophilized. The biological activities of MFR, NFR and NFF were investigated. The results suggested that MFR and NFR had approximately 90% of total protein, while NFF about 65%on a dry basis. Regarding the protein and peptide profile, all hydrolysates showed the peaks between 1200 and 14000 Da but NFF had a higher content of lower molecular size peptides. Additionally, the antioxidant capacity was evaluated by 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and oxygen-radical absorbance-capacity (ORAC) assays. NFF showed higher antioxidant activity, which was aligned with its peptide profile. The anti-hypertensive activity was also tested, and the results indicated that NFF showed a very good anti-hypertensive potential. Due to its biological properties, the NFF potential as a food functional ingredient was raised. NFF antimicrobial potential was also investigated however, no antimicrobial effect was registered for the pathogenic strains tested. Regarding the free amino acids and mineral profile, a high content of glutamic acid, leucine, alanine, phenylalanine and aspartic acid as well as phosphorus, magnesium, calcium, sodium and potassium was registered.
- Novel eco-friendly method to extract keratin from hairPublication . Cassoni, Ana C.; Freixo, Ricardo; Pintado, Ana I. E.; Amorim, Manuela; Pereira, Carlos D.; Madureira, Ana Raquel; Pintado, Manuela M. E.Pig slaughtering for meat production is a major worldwide industry that generates many byproducts that can cause environmental problems. Nonetheless, it is possible to have a management of these residues toward the reuse and valorization of these byproducts. One opportunity focuses on the extraction of value-added compounds such as keratin since pig hair is composed by 80% of this protein. There are some methods for keratin extraction from human hair as other sources such as wool and feathers, but they bring many problems such as pollution, time-consumption, and high costs. This work uses a commercial detergent belonging to the category of degreasers, capable of fast and efficient dissolution of pig hair, followed by a simple filtration to remove residues from skin trimmings with fat and lard. The resulting solution is submitted to an ultrafiltration process to obtain a solution with higher protein content and purity, both at laboratory scale and pilot scale-up. Use of this keratin green extraction method allows to obtain a keratin product with protein purity up to 70% with a yield extraction of about 50%, which although lower than the previous methods allow protein integrity and cleaner technology. This method allows a relevant valorization of pig hair, using for the first time a simple, cost-effective, and environmentally friendly approach.
- Peptide-rich yeast fractions from brewer’s spent yeast: a scalable fractionation approach and their functional application in bakery productsPublication . Brassesco, María Emilia; Paupério, Ana; Pereira, Carlos D.; Ferreira, João Paulo; Pintado, ManuelaBrewer’s spent yeast (BSY), a significant brewing by-product, poses environmental challenges and opportunities for valorization as a sustainable protein source. This study focuses on transforming BSY into high-value functional ingredients for food applications. A green, sustainable, and scalable process was developed to extract bioactive compounds from BSY at both laboratory and pilot scales, yielding peptide-rich fractions with robust antioxidant properties. These extracts were incorporated into prototype formulations, including protein-enriched crackers, demonstrating their potential as natural, nutritious ingredients. Physicochemical, compositional, and functional characterizations validated their application viability. The antioxidant potential of BSY fractions was confirmed through total phenolic compounds and ABTS and oxygen radical absorbance capacity assays, where the retentate from the 10 kDa ultrafiltration fraction on the pilot scale exhibited superior bioactivity, supporting its selection as the most suitable fraction for food formulations. Additionally, the transition from laboratory to pilot scale revealed slight variations in protein retention and bioactive compound recovery, emphasizing the need for process optimization. These findings highlight BSY’s potential to support circular economy practices by reducing waste while enhancing the nutritional and functional value of food products.
- Revealing the potential brewer’s spent yeast: sustainable innovations for alternative protein development and waste reduction in the agri-food sectorPublication . Brassesco, María Emilia; Paupério, Ana Isabel; Coscueta, Ezequiel; Pereira, Carlos D.; Ferreira, João Paulo; Pintado, ManuelaIntroduction: Brewer’s spent yeast (BSY), a major brewing by-product, accumulates annually, posing environmental challenges [1]. Research highlights BSY’s potential as a sustainable alternative protein source in animal feed and human food. With the market expected to reach $2.17 billion by 2030, BSY offers a promising solution for waste reduction and protein replacement. However, more research is needed to optimize its use in human food products, particularly regarding flavor, texture, and processing. Valorizing BSY supports circular economy principles, transforming it into functional, high-protein ingredients for various applications. Objectives: This study aims to valorize BSY by producing differentiated functional extracts. These extracts, along with other relevant by-products (such as tomato flour and wheat germ), will be used to formulate protein-rich snacks. An innovative green process was employed to promote sustainable practices in the agri-food sector and introduce novel, natural ingredients for developing alternative protein food products. Conclusions: The RUF10 fraction showed a significant increase in antioxidant activity compared to RUF40 and a better production yield than PUF10. Crackers formulated with varying RUF10 concentrations exhibited notable differences in physicochemical properties. The protein content in crackers P6 was notably, but this resulted in reduced water activity and increased firmness, as indicated by lower fracturability. Future work should focus on understanding the peptide profile of yeast fractions and analyzing the antioxidant activity within the food matrix, providing deeper insights into the functional properties of BSY-derived products in food applications. In conclusion, the incorporation of BSY fractions successfully enhanced the nutritional profile, aligning with sustainability goals and offering a viable alternative for protein-rich food products.
- Revealing the potential brewer’s spent yeast: sustainable innovations for alternative protein development and waste reduction in the agri-food sectorPublication . Brassesco, María Emilia; Paupério, Ana Isabel; Coscueta, Ezequiel; Pereira, Carlos D.; Ferreira, João Paulo; Pintado, ManuelaBackground: Brewer’s spent yeast (BSY), the second most significant by-product of the brewing industry, represents an abundant biomass resource with considerable environmental implications due to its annual accumulation. This study aims to valorize BSY by-products producing differentiated functional extracts that can be used to formulate protein-rich snacks. By enhancing the value of this by-product, the research aims to promote sustainable practices within the agri-food sector and introduce novel natural ingredients for developing alternative protein food products. Methods: The experiments were conducted at a pilot scale, starting with an initial yeast mass of 200 L. A pre-treatment procedure involving washing and centrifugation was implemented to eliminate residual microorganisms, suspended material, and certain bitter compounds. Subsequently, BSY underwent an integrated process to produce peptide extracts. This process involved inducing autolysis through heat treatment to facilitate the hydrolysis of yeast extracts. Following centrifugation, the resulting liquid fraction was subjected to membrane filtration with different cut-off points (40 and 10 kDa) to generate distinct fractions. The extracts were then freeze-dried and underwent physicochemical characterization. The most promising extract was used to develop prototypes for savory snacks, with evaluations focusing on parameters such as color (using the CIELAB color system), texture, and water activity. Results: The derived fractions demonstrated high protein content along with significant amounts of minerals and fiber. Furthermore, the successful formulation of protein-rich savory snacks demonstrated the feasibility of repurposing these extracted fractions as functional ingredients in the production of high-protein products for human consumption. Conclusion: This research highlights the potential of using BSY by-products to create value-added ingredients and products, thereby advancing sustainability goals and aligning with the principles of the circular economy in the food industry.
- The potential of acorn extract treatment on PUFAS oxidative stability: a case study on fish cooking wastewaterPublication . Araújo-Rodrigues, Helena; Ribeiro, Tânia Bragança; Machado, Manuela; Pereira, Carlos D.; Pintado, Manuela E.Fish byproducts are valuable sources of Ω-3 polyunsaturated fatty acids (PUFAs). Their valorization potentially alleviates pressure on this sector. This study uses a circular economy approach to investigate the oil fraction from sardine cooking wastewater (SCW). Analysis of its fatty acid (FA) profile revealed promising PUFA levels. However, PUFAs are highly susceptible to oxidation, prompting the exploration of effective and natural strategies to replace synthetic antioxidants and mitigate their associated risks and concerns. An antioxidant extract from acorn shells was developed and evaluated for its efficacy in preventing oxidative degradation. The extract exhibited significant levels of total phenolic compounds (TPC: 49.94 and 22.99 mg TAE or GAE/g DW) and antioxidant activities (ABTS: 72.46; ORAC: 59.60; DPPH: 248.24 mg TE/g DW), with tannins comprising a significant portion of phenolics (20.61 mg TAE/g DW). LC-ESI-UHR-QqTOF-MS identified ellagic acid, epicatechin, procyanidin B2 and azelaic acid as the predominant phenolic compounds. The extract demonstrated the ability to significantly reduce the peroxide index and inhibit PUFA oxidation, including linoleic acid (LA), eicosapentaenoic (EPA), and docosahexaenoic acid (DHA). This approach holds promise for developing stable, functional ingredients rich in PUFAs. Future research will focus on refining oil extraction procedures and conducting stability tests towards the development of specific applications.