Development and characterization of functional ingredients from olive pomace : bioactivity and potential application

Atualmente, grande volume dos resíduos gerados ao longo da cadeia alimentar Europeia é produzido durante o processamento alimentar (19-39%). A indústria de extração de azeite não é exceção, gerando anualmente grandes quantidades de subprodutos e resíduos com elevado impacto fitotóxico, mas que são também ricos em diversos compostos bioativos e nutrientes com potencial aplicação como ingredientes alimentares. Assim sendo, é imprescindível a adoção de estratégias de redução do desperdício e de valorização dos subprodutos, no sentido de melhorar a sustentabilidade económica e ambiental da indústria alimentar. Atualmente, o bagaço de azeitona é o subproduto de maior relevo da indústria de azeite e a sua valorização adequada é também o maior desafio do sector para alcançar uma cadeia produtiva sustentável alinhada com os princípios da Bioeconomia Circular. Esta tese visou alcançar uma valorização “zero desperdício” para o bagaço de azeitona, priorizando os produtos de alto valor acrescentado sobre os de baixo valor através da produção e caracterização de ingredientes alimentares, nomeadamente ingredientes em pó ricos em vários compostos bioativos, seguida da validação in vitro das suas atividades biológicas ligadas a benefícios para a saúde, como a prevenção e redução de doenças cardiovasculares, diabetes, problemas intestinais e posterior aplicação no desenvolvimento de alimentos funcionais. Uma estratégia de “desperdício zero” foi alcançada para o bagaço de azeitona. Esta estratégia consiste num procedimento simples de fracionamento, em que se obteve uma fração líquida que revelou um alto teor de hidroxitirosol (513,61-625,76 mg/100 g de peso seco) e uma fração de polpa caracterizada por ser uma boa fonte de fibra alimentar antioxidante. Adicionalmente, obteve-se ainda uma fração de caroços que exibiu valores caloríficos significativamente elevados (18,65-18,94 MJ/kg). Estes resultados validaram o potencial valor acrescentado das frações líquidas e de polpa como ingredientes alimentares e como biocombustível no caso da fração de caroços. Após validação do potencial da fração líquida e de polpa do bagaço de azeitona como ingredientes alimentares, dois novos ingredientes em pó, estáveis e seguros, foram desenvolvidos sem qualquer etapa de extração, nomeadamente um pó enriquecido na fração líquida (LOPP) e um pó enriquecido na fração de polpa (POPP). O LOPP exibiu uma quantidade significativa de manitol (141 g/kg), potássio (54 g/kg) e hidroxitirosol/derivados (5 mg/g). O POPP exibiu uma grande quantidade de fibra alimentar (620 g/kg) associada a um teor significativo de fenólicos (7,41 mg equivalentes de ácido gálico/g fibra) com atividade antioxidante substancial. O POPP também revelou uma composição de ácidos gordos insaturados semelhante à do azeite (76% do total de ácidos gordos), sendo ainda uma fonte razoável de proteína (12%). As suas propriedades funcionais (solubilidade, capacidade de retenção de água e de óleo), capacidade antioxidante e atividade antimicrobiana foram também avaliadas, assim como a sua segurança biológica. Em síntese, a multifuncionalidade e aplicabilidade dos pós de bagaço de azeitona como fortificantes nutricionais, promotores de benefícios para a saúde e conservantes foram atestadas. A digestão simulada in vitro permitiu concluir que os fenólicos e minerais do LOPP foram afetados negativamente pela digestão, no entanto, uma bioacessibilidade significativa de potássio e hidroxitirosol foi verificada ( 57%) e as suas bioatividades foram apenas moderadamente afetadas. Em relação às suas bioatividades, pelo menos metade da atividade antioxidante (capacidade de Resumo x absorção do radical de oxigénio), de inibição da -glucosidase e de inibição da enzima conversora de angiotensina (91,98%) foram retidas e bioacessíveis após a digestão. Por sua vez, a digestão do POPP revelou uma perda significativa de fenólicos na digestão oral (62,48%), mas o papel da fibra como veículo de fenólicos permitiu recuperar uma quantidade significativa de fenólicos no estômago (77,11%) e um índice de bioacessibilidade de pelo menos 50% (tirosol e o seu glucosídeo). A fibra do POPP também interagiu positivamente com os ácidos gordos, diminuindo e facilitando bioacessibilidade dos saturados (5-6%) e dos insaturados (4-11%), respetivamente. Após realização da digestão in vitro dos pós de bagaço de azeitona, as frações disponíveis no cólon de ambos os pós exibiram uma composição abundante em compostos associados a potenciais benefícios para a saúde intestinal, nomeadamente fibra e compostos fenólicos e fibra. Assim sendo, procedeu-se a fermentação fecal in vitro das frações não digeridas retidas no cólon após a digestão gastrointestinal simulada in vitro do LOPP e POPP. Os resultados da análise do gene 16S rRNA e dos ácidos gordos de cadeia curta mostraram que estes pós não tiveram um impacto negativo na diversidade da microbiota intestinal e que promoveram uma maior produção de ácidos de cadeia curta (acetato>butirato>propionato) do que os frutooligossacarídeos. A identificação dos metabólitos fenólicos dos pós de bagaço de azeitona por LC-ESI-UHR-QqTOF-MS seguida das análises de clustering supervisionada e clustering hierárquico permitiram avaliar os padrões destes metabólitos ao longo da fermentação fecal in vitro, bem como explicar as potenciais atividades biológicas destes pós no intestino. Ambos os pós apresentaram um conteúdo fenólico e uma capacidade de absorção de radical de oxigénio significativas e capacidade de inibição da adesão de patógenos como o Bacillus cereus (22,03%) e Listeria monocytogenes (20,01%), sobretudo pelo POPP. Por fim, para validar o desempenho dos pós de bagaço de azeitona como ingredientes funcionais, LOPP e POPP foram incorporados em iogurte para aumentar o seu conteúdo em fibra alimentar, hidroxitirosol e ácidos gordos insaturados. A adição de POPP (2%) ao iogurte permitiu cumprir a condição de “fonte de fibra”, por sua vez LOPP (1%) forneceu 5 mg de hidroxitirosol/ derivados num iogurte (120 g). A adição de POPP e azeite juntamente com LOPP ou POPP permitiu uma melhoria do perfil de ácidos gordos insaturados dos iogurtes. Além disso, todos os iogurtes fortificados apresentaram um maior teor de fenólicos e atividade antioxidante. A digestão in vitro reduziu a atividade antioxidante de todas as formulações e mostrou que a incorporação de azeite juntamente com o LOPP e POPP aumentou a bioacessibilidade de hidroxitirosol e do hidroxitirosol glucosídeo, respetivamente. Contudo, todas as formulações com os pós de bagaço de azeitona podem ser consideradas boas fontes de hidroxitirosol, lípidos insaturados e fibras para os consumidores. Em suma, os resultados deste trabalho irão contribuir para a sustentabilidade da indústria do azeite no contexto da bioeconomia circular, através do desenvolvimento de ingredientes de valor acrescentado destinados à indústria alimentar com potencial impacto positivo na saúde, proporcionando ainda um biocombustível sólido.Currently, a large amount of the total European food supply chain waste (between 19 and 39%) is produced during food processing. The olive oil industry is no exception, generating large quantities of by-products and wastes annually with high phytotoxic impact, but also rich in several bioactive compounds and nutrients with potential application as food ingredients. Thus, it is imperative to implement strategies to reduce waste and valorise by-products to improve the food industry's economic and environmental sustainability. Olive pomace is the most prominent by-product of the modern olive oil industry, and its proper valorisation is one of the major obstacles to olive oil industry achieve a sustainable production chain aligned with the principles of the Circular Bioeconomy concept. This project aimed at achieving a “zero waste” valorising approach to olive pomace that prioritised the high over low added-value products, through the production and characterisation of food ingredients, namely high-value powdered ingredients rich in several bioactive compounds, followed by the validation of their biological activities related with health benefits as prevention and reduction of the prevalence of cardiovascular diseases, diabetes and gut disorders and, their application in the development of functional foods. A “zero waste” strategy was achieved for olive pomace. This strategy is based on a feasible fraction approach, and it was obtained a liquid fraction that revealed a high amount of hydroxytyrosol (513.61- 625.76 mg/100 g dry weight) and pulp fraction that was characterised to be a good source of antioxidant dietary fibre. Additionally, it was achieved a stones fraction that exhibited substantial higher heating values (18.65-18.94 MJ/kg). These results supported the potential value-added of the liquid and pulp fractions from olive pomace as functional food ingredients and as biofuel to stones. After validating the liquid and pulp fraction potential as food ingredients, new two stable and safe, powdered ingredients without any extraction step were developed, namely a liquid-enriched powder (LOPP) and a pulp-enriched powder (POPP). LOPP exhibited a significant amount of mannitol (141 g/kg), potassium (54 g/kg) and hydroxytyrosol/derivatives (5 mg/g). POPP exhibited a high amount of dietary fibre (620 g/kg) associated with a significant amount of bound phenolics (7.41 mg gallic acid equivalents/g fibre dry weight) with substantial antioxidant activity. POPP also revealed an unsaturated fatty acid composition similar to that of olive oil (76% of total fatty acids) and a reasonable source of protein (12%). Their functional properties (solubility, water‐holding and oil‐ holding capacity), antioxidant capacity and antimicrobial activity were also assessed, and their biological safety was validated. The multifunctionality and applicability as nutritional enhancers, health-benefits promoters and preservatives of the olive pomace powders were recognised. The in vitro simulated digestion allowed concluding that phenolics and minerals from LOPP were negatively affected by digestion. However, a significant bioaccessibility of potassium and hydroxytyrosol was verified ( 57%) and its bioactivities were only moderately affected. Regarding its bioactivities, at least half of the antioxidant activity (measured by Oxygen Radical Absorbance Capacity assay), -glucosidase inhibition activity and angiotensin-converting enzyme inhibitory activity (91.98%) were retained and bioaccessible after in vitro digestion. In its turn, POPP digestion Abstract xii revealed a significant loss of phenolics in the oral step (62.48%), but the dietary fibre role as phenolics’ carrier allowed recovering a significant amount of phenolics in the stomach (77.11%) and a bioaccessibility index of at least 50% (mainly for tyrosol and its glucoside). POPP dietary fibre also positively interacts with lipids decreasing the saturated fatty acids bioaccessibility (5-6%) and facilitating the unsaturated fatty acid bioaccessibility (4-11%). After the in vitro digestion system of both olive pomace powders, the colon-available fractions LOPP and POPP exhibited an abundant composition in phenolics, but also dietary fibre with potential gut health benefits as antioxidants, antimicrobial and prebiotic agents. So, the gut's potential beneficial effects were analysed by in vitro faecal fermentation of the undigested fractions retained in the colon after in vitro simulated gastrointestinal digestion. The 16S rRNA gene analysis results showed that LOPP and POPP did not negatively impact gut microbiota diversity. The short-chain fatty acids analysis showed a higher production of these fatty acids (acetate > butyrate > propionate) by LOPP and POPP than by fructooligosaccharides. The identification of phenolic metabolites by LC-ESIUHR-QqTOF-MS followed by supervised cluster and hierarchical clustering analysis allowed to evaluate the metabolite patterns of olive pomace powders phenolics throughout faecal fermentation, explaining the potential biological activities exhibited in the gut by these powders. Both powders showed significant total phenolic content and oxygen radical absorbance capacity during faecal fermentation. But also, mucin‐adhesion inhibition ability against pathogens as Bacillus cereus (22.03%) and Listeria monocytogenes (20.01%), specially POPP. Finally, to validate olive pomace powders' performance as functional ingredients, LOPP and POPP were incorporated into yoghurt to increase its content in dietary fibre, hydroxytyrosol, and unsaturated fatty acids. POPP (2%) and LOPP (1%) addition to yoghurt allowed fulfilling the condition on being a “source of fibre” and provides 5 mg of hydroxytyrosol and derivatives in a standard yoghurt (120 g), respectively. The addition of olive oil, together with olive pomace powders, was also investigated in order to understand the lipids-phenolics interaction, and not only POPP addition improved the yoghurts’ unsaturated fatty acids profile, but the olive oil addition together both olive pomace powders also improved too. Besides that, all yoghurts fortified with olive pomace powders exhibited higher total phenolic content and antioxidant activity. The in vitro digestion of yoghurts showed that olive oil incorporation together with olive pomace powders increased the bioaccessibility of hydroxytyrosol (LOPP + olive oil) and hydroxytyrosol glucoside (POPP + olive oil). Despite the reduction of antioxidant activity after digestion, the yoghurts formulations with olive pomace powders could be considered suitable carriers to deliver hydroxytyrosol, unsaturated fatty acids and fibre to the consumers. In short, the results from this work will contribute to the sustainability of the olive oil industry in circular bioeconomy context, through the development of value-added ingredients to the food industry with a positive impact in health, offering also a solid biofuel

Desenvolvimento de um novo conceito de refrigerante

Dissertação para obtenção de grau de mestre em Tecnologia e Segurança Alimentar – Qualidade AlimentarA indústria de refrigerantes e sumos é um subsector da indústria de bebidas não-alcoólicas, de grande impacte na economia e elevada competitividade. O desenvolvimento de novos produtos e a garantia de qualidade dos produtos são de extrema importância nesta indústria. Actualmente, o consumidor é bastante preocupado com a saúde, procurando produtos que possam não comprometer e/ou impulsionar o seu ―bem-estar‖. Surge assim, a tendência das bebidas funcionais. Contudo, o consumidor não está disposto a comprometer o sabor da bebida, pelo que a formulação deve ter em conta os ingredientes funcionais e quantidades a utilizar destes, de modo a garantir os benefícios da bebida, sem prejudicar as características sensoriais. As bebidas desportivas são um segmento de bebidas funcionais que tem pouca visibilidade no mercado português. Mas as crescentes preocupações de saúde e maior prática de exercício, têm levado a que sejam consumidas não só por atletas, mas também pelo consumidor comum, prevendo-se um crescimento deste segmento. Tal como em todas as bebidas, a funcionalidade das bebidas desportivas tem vindo a aumentar. As proteínas de soro de leite (PSL‘s) são um óptima estratégia para obter uma bebida desportiva, que para além de fornecer hidratação e melhor desempenho físico, permitem reduzir o tempo para a fadiga muscular e as dores associadas a esta. A bebida formuladano presente estudo apresenta na sua composição PSL‘s (2%), frutose (2,5%) e xarope de sacarose (5,65%). Esta bebida apresenta benefícios como melhorar o processo de recuperação e a performance. Efectuou – se ainda a formulação de bebida com 5% (m/v) de PSL‘s e de um suplemento efervescente que permite obter a bebida de 5% (m/v) a partir da bebida com 2% (m/v) de PSL‘s. A bebida enriquecida com 2% PSL‘s foi considerada satisfatória nos aromas de laranja e ananás – coco

Influence of temperature and pH on antioxidant and antihypertensive activities of a fish protein hydrolysate

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Incorporation of olive pomace ingredients in yoghurt as source of fibre and hydroxytyrosol: bioactivity and stability of throughout gastrointestinal digestion

Tânia I. B. Ribeiro thanks the Fundação para a Ciência e Tecnologia (FCT), Portugal for the PhD Grant SFRH/BDE/108271/2015, the financial support of BLC3 Association – Technology and Innovation Campus and the mentoring of Frulact and Comendador Arménio Miranda.info:eu-repo/semantics/publishedVersio

Tomato processing by-products valorisation through ohmic heating approach

Tomato by-products from processing industries have a higher potential to be reused as a source of bioactive compounds. Reliable national data on tomato by-products and physicochemical characterisation that will inform and find effective planning on tomato waste management in Portugal is absent. To help obtain this knowledge, selected Portugal companies were recruited to obtain representative samples of by-products generation, and physicochemical composition was evaluated. Furthermore, an environmental-friendly method (the ohmic heating (OH) method, which allows the recovery of bioactive compounds in absence of hazardous reagents) was also used and compared with conventional methods to explore new safe value-added ingredients. Total antioxidant capacity and total and individual phenolic compounds were also evaluated by spectrophotometric and high-performance liquid chromatography (HPLC), respectively. Tomato processing by-products have revealed a higher potential since both collected samples from companies were rich in protein (between 16.3 to 19.4 g/100 g DW, with fibre content ranging from 57.8 to 59.0 g/100 g DW). In addition, these samples contain 17.0 g/100 g of fatty acids (mainly polyunsaturated, monounsaturated and saturated, such as linoleic, oleic, and palmitic acid, respectively). Also, they present mainly chlorogenic acid and rutin as phenolic compounds. After understanding its composition, the OH was applied to determine added-value solutions to tomato by-products. With extractions, two types of fractions were obtained, namely liquid fraction rich in phenols, free sugars, and carotenoids and a solid fraction rich in fibre bound to phenols and carotenoids. This treatment has been shown to have the ability to preserve carotenoids, such as lycopene relative to conventional methods. Nevertheless, new molecules were identified by LC-ESI-UHR-OqTOF-MS analysis, such as phene-di-hexane and N-acethyl-D-tryptophan. According to the results, the OH boosts the potential of tomato by-products and can be directly introduced into the process, contributing to the circular economy and zero by-products.info:eu-repo/semantics/publishedVersio

Tomato processing by-products valorisation through ohmic heating approach

Tomato by-products from processing industries have a higher potential to be reused as a source of bioactive compounds. Reliable national data on tomato by-products and physicochemical characterisation that will inform and find effective planning on tomato waste management in Portugal is absent. To help obtain this knowledge, selected Portugal companies were recruited to obtain representative samples of by-products generation, and physicochemical composition was evaluated. Furthermore, an environmental-friendly method (the ohmic heating (OH) method, which allows the recovery of bioactive compounds in absence of hazardous reagents) was also used and compared with conventional methods to explore new safe value-added ingredients. Total antioxidant capacity and total and individual phenolic compounds were also evaluated by spectrophotometric and high-performance liquid chromatography (HPLC), respectively. Tomato processing by-products have revealed a higher potential since both collected samples from companies were rich in protein (between 16.3 to 19.4 g/100 g DW, with fibre content ranging from 57.8 to 59.0 g/100 g DW). In addition, these samples contain 17.0 g/100 g of fatty acids (mainly polyunsaturated, monounsaturated and saturated, such as linoleic, oleic, and palmitic acid, respectively). Also, they present mainly chlorogenic acid and rutin as phenolic compounds. After understanding its composition, the OH was applied to determine added-value solutions to tomato by-products. With extractions, two types of fractions were obtained, namely liquid fraction rich in phenols, free sugars, and carotenoids and a solid fraction rich in fibre bound to phenols and carotenoids. This treatment has been shown to have the ability to preserve carotenoids, such as lycopene relative to conventional methods. Nevertheless, new molecules were identified by LC-ESI-UHR-OqTOF-MS analysis, such as phene-di-hexane and N-acethyl-D-tryptophan. According to the results, the OH boosts the potential of tomato by-products and can be directly introduced into the process, contributing to the circular economy and zero by-products.This research was funded by “MultiBiorefinery: Estratégias multiuso para a valorização de uma gama alargada de subprodutos agroflorestais e das pescas: Um passo em frente na criação de uma biorrefinaria” financiado pelo Programa Operacional Competitividade e Internacionalização (POCI-01-0145-FEDER-016403) e pelo Programa Operacional Regional de Lisboa (LISBOA-01-0145- FEDER-016403), na sua componente FEDER e pela Fundação para a Ciência e Tecnologia, I.P. na componente nacional (SAICTPAC/0040/2015).info:eu-repo/semantics/publishedVersio

Are olive pomace powders a safe source of bioactives and nutrients?

"First published: 10 September 2020"BACKGROUND Olive oil industry generates significant amounts of semi-solid wastes, namely the olive pomace. Olive pomace is a by-product rich in high-value compounds (e.g. dietary fibre, unsaturated fatty acids, polyphenols) widely explored to obtain new food ingredients. However, conventional extraction methods frequently use organic solvents, while novel eco-friendly techniques have high operational costs. The development of powdered products without any extraction step has been proposed as a more feasible and sustainable approach. RESULTS The present study fractionated and valorised the liquid and pulp fraction of olive pomace obtaining two stable and safe powdered ingredients, namely a liquid-enriched powder (LOPP) and a pulp-enriched powder (POPP). These powders were characterized chemically, and their bioactivity was assessed. LOPP exhibited a significant amount of mannitol (141 g/ kg), potassium (54 g/ kg) and hydroxytyrosol/ derivatives (5 mg/g). POPP exhibited high amount of dietary fibre (620 g/ kg) associated to significant amount of bound phenolics (7.41 mg GAE/ g fibre DW) with substantial antioxidant activity. POPP also contained an unsaturated fatty acids composition similar to olive oil (76\% of total fatty acids) and showed potential as a reasonable source of protein (12 \%). Their functional properties (solubility, water-holding and oil-holding capacity), antioxidant capacity and antimicrobial activity were also assessed, and their biological safety was verified. CONCLUSION The development of olive pomace powders to apply in the food industry could be a suitable strategy to add-value to olive pomace and obtain safe multifunctional ingredients with higher health-promoting effects than dietary fibre and polyphenols itself. This article is protected by copyright. All rights reserved.TBR thanks the Fundação para a Ciência e Tecnologia (FCT), Portugal for PhD grant SFRH/BDE/108271/2015 and the financial support of Association BLC3 – Technology and Innovation Campus. This work was supported by National Funds from FCT – Fundação para a Ciência e a Tecnologia through the project MULTIBIOREFINERY – SAICTPAC/0040/2015 (POCI-01-0145-FEDER-016403). We are also grateful for the scientific collaboration under the FCT project UID/Multi/50016/2019.info:eu-repo/semantics/publishedVersio

Prebiotic effects of olive pomace powders in the gut: In vitro evaluation of the inhibition of adhesion of pathogens, prebiotic and antioxidant effects

Supplementary data related to this article can be found at https:// doi.org/10.1016/j.foodhyd.2020.106312Olive pomace is a biowaste rich in polyphenols and insoluble dietary fibre with high potential to develop new value chains towards a sustainable and circular bioeconomy. Regarding gut health, olive pomace phenolics and insoluble dietary fibre (after possible fermentation) could act as antioxidants, antimicrobial and prebiotic agents. These potential beneficial effects on the gut were analysed for two powders from olive pomace: liquid-enriched powder (LOPP) - mostly source of phenolics - and pulp-enriched powder (POPP) - main source of insoluble dietary fibre. LOPP and POPP were subjected to an in vitro simulated gastrointestinal digestion followed by in vitro faecal fermentation. The undigested fraction retained in the colon was analysed regarding its potential antioxidant, antimicrobial and prebiotic effects. LOPP and POPP did not impact the gut microbiota diversity negatively, showing a similar ratio of Firmicutes/Bacteroidetes compared to a positive control (FOS). LOPP exhibit a positive (similar to FOS) effect on the Prevotella spp./Bacteroides spp. ratio. Both powders also promoted more the production of short-chain fatty acids (mainly acetate?>?butyrate?>?propionate) than FOS. Both powders showed also significant total phenolic content and oxygen radical absorbance capacity during faecal fermentation until 48?h. Besides that, these powders showed mucin-adhesion inhibition ability against pathogens, principally POPP against Bacillus cereus (22.03?±?2.45%) and Listeria monocytogenes (20.01?±?1.93%). This study demonstrates that olive pomace powders have prebiotic effects on microbiota, including the stimulation of short-chain fatty acids production, potential antioxidant and antimicrobial activity which could improve human gut health.Tˆania I. B. Ribeiro thanks the FCT - Fundação para a Ciência e Tecnologia, Portugal and Association BLC3 – Technology and Innovation Campus, Centre Bio R&D Unit for the PhD Grant SFRH/BDE/108271/2015. This work was supported by National Funds from FCT - Fundação para a Ciência e a Tecnologia, Portugal through the project MULTI-BIOREFINERY - SAICTPAC/0040/2015 (POCI-01-0145-FEDER- 016403). We would also like to thank the scientific collaboration under the FCT project UID/Multi/50016/2019.info:eu-repo/semantics/publishedVersio

AgroForest biomass and circular bioeconomy: case studies

The agroforest sector plays a leading role as a biomass supplier to obtain bio-based products that allowed an acceleration in the circular bioeconomy transition. This chapter applied a mixed-methods review to identify new attractive bio-based products and to evaluate its market potential in Portugal. Forest biomass was identified as an excellent raw material for (1) low-carbon building materials, (2) biotextiles, and (3) bioplastics. The potential of agro-food waste to obtain new bio-based materials was also emphasised. The new bioproducts identified have high potential and attractive markets. It was estimated that a 5% market share of these bioproducts in the global construction, textiles, and plastics markets in 2030 corresponds to an aggregate increase in revenues of 260-579 million € per year in Portugal. The environmental sustainability implications arising from the diffusion of these new biomaterials are also highlighted, focusing on the decarbonisation of the economy.info:eu-repo/semantics/publishedVersio

Screening of enzymatic hydrolysis of mixed fish by-products

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