Effects of enzyme-assisted extraction on the profile and bioaccessibility of isoflavones from soybean flour.

The effects of enzymatic extraction strategies on extractability, bioconversion, and bioaccessibility of biologically active isoflavone aglycones, total phenolic content, and antioxidant activity of aqueous extracts from full-fat soy flour were evaluated. Protease, tannase, and cellulase enzymes were used individually or in combination. Except for the protease treatment, all enzymatic treatments increased the extraction of biologically active isoflavones (daidzein and genistein) compared with the control. The use of a mixture of protease, tannase, and cellulase resulted in increased extractability and/or bioconversion of aglycones from soy flour, indicating a synergistic effect amongst the enzymes. Daidzein and genistein concentrations increased from 29.0 to 158.2 µg/g and from 27.0 to 156.5 µg/g (compared to the control), respectively. Furthermore, enzymatic extraction followed by in vitro gastrointestinal digestion significantly increased the bioaccessibility of isoflavone aglycones, total phenolic content (by 22-45%), and antioxidant activity (by 15-22%) of the extracts. These results demonstrate that enzyme selection is an efficient strategy to maximize the extraction, bioconversion, and bioaccessibility of bioactive isoflavones from soy flour, which could contribute to health benefits associated with the consumption of soy-rich products.

Combined isoflavones biotransformation increases the bioactive and antioxidant capacity of soymilk.

Isoflavones are phenolic secondary metabolites mainly occurring in soy and soybean products. Compared to glycoside forms, isoflavone aglycones present higher biological activities. This study evaluated the potential of microbial and enzymatic treatments in biotransformed isoflavones in their biologically active forms in soymilk. Seven different cultures of lactic acid bacteria and bifidobacteria associated with the action of immobilized tannase enzyme were screened for isoflavone glycoside biotransformation ability. The biotransformed soymilk samples were characterized regarding isoflavone profile, total phenolic content, and in vitro antioxidant activities. All bacterial strains showed a good growth capacity in soymilk matrix and produced ß-glucosidase enzyme, which hydrolyzed isoflavone glycosides into aglycones in soymilk after 24 h of fermentation. The microbial fermentation followed by tannase reaction (FT processes) resulted in the highest increase of bioactive aglycones (10.3- to 13.1-fold for daidzein, 10.4- to 12.3-fold for genistein, and 3.8- to 4.7-fold for glycitein), compared to control soymilk. Further, FT processes enhanced the total phenolic content (53-70%) and antioxidant activity by ORAC (69-102%) and FRAP (49-71%) assays of the soymilk matrix. Therefore, the combination of microbial fermentation and tannase treatment is a promising strategy to obtain a fermented soy product rich in bioactive isoflavones with greater health-promoting potential. KEY POINTS: • Bacterial cultures and tannase enzyme displayed isoflavone deglycosylation activity. • The addition of tannase following the fermentation maximized the isoflavone conversion. • Increased isoflavone aglycones contributed to the improved antioxidant activity of soymilk.

Biotransformation processes in soymilk isoflavones to enhance anti-inflammatory potential in intestinal cellular model.

The present study investigated, in in vitro cellular model, the modulation of intestinal inflammation by biotransformed soymilk with tannase and probiotic strains. The ability to reduce the generation of intracellular reactive oxygen species (ROS) and the antioxidant power of soy extracts were also evaluated. The results showed changes in isoflavones profile after biotransformation processes, with a significative enhancement in aglycones content. Reduction in intracellular ROS production and improvement in antioxidant capacity were observed. Anti-inflammatory responses in Caco-2 cells were also expressive. A significative decrease in interleukin 8 (IL-8) level was detected for all biotransformed samples, especially for extracts with tannase. The biotransformed soy extracts by tannase have a great potential to improve health conditions, defending the intestinal cells of oxidative damage, and acting as a possible adjuvant in inflammatory process. PRACTICAL APPLICATIONS: Soy isoflavones have been explored owing to health benefits. Only glycosylated forms are found in high concentrations in soybeans. So, microbial and enzymatic biotransformation processes aiming to increase aglycones and metabolites appear as an attractive option to enlarge the bioactivity of soy products. The present study showed a positive impact of biotransformed soymilk on antioxidant defenses systems and modulation of intestinal inflammation and could act as a nutraceutical agent.

Exploring in vitro effects of biotransformed isoflavones extracts: Antioxidant, antiinflammatory, and antilipogenic.

The present study aimed to investigate, in in vitro assays, the antilipogenic and antiinflammatory potential as well as the antioxidant capacity of biotransformed soymilk by tannase and ß-glycosidase enzymes. The results showed a significant enhancement of the antioxidant capacity, especially by biotransformed soymilk with free tannase (SFT), corresponding to an increase of 2.3 and 1.25 times by oxygen radical absorbance capacity (ORAC) and ferric reducing antioxidant power (FRAP) assays, respectively. The lipid accumulation reduction by 3T3-L1 adipocytes assay was not significant. However, the antiinflammatory responses were expressive. In lipopolysaccharide-stimulated RAW 264.7 macrophages, SFT reduced around 37 times TNF-α expression at the highest tested concentration of the sample. Other inflammatory parameters, as IL-6 and nitric oxide, were no longer detected when the cells were treated with SFT and soymilk with immobilized enzymes, respectively. The biotransformed soy extracts with tannase have great potential to act as a nutraceutical, protecting the cells against oxidative damage and helping maintain health under inflammatory stress. PRACTICAL APPLICATIONS: Soy isoflavones have been associated with several beneficial effects on human health, including inhibition capacity of lipid accumulation in adipocytes, antiinflammatory properties, and antioxidant potential. However, the isoflavones bioavailability differs among their chemical forms, and studies have shown that the higher health benefits are conferred by aglycones and their metabolites, such as equol, compared to the other forms. For this reason, the enrichment of isoflavone aglycones and metabolites in soy-based products has attracted growing attention. The present study was focused on developing a bioprocess able to produce a rich extract with soy isoflavones metabolites, with increased bioactive potential for application as a functional ingredient or a nutraceutical.

A new biotechnological process to enhance the soymilk bioactivity.

Equol, a daidzein metabolite produced exclusively by intestinal bacteria in some, but not all, humans, exhibits a wide range of beneficial health effects owing to its superior nutraceutical effect compared with isoflavones of soy. The aim of this work was to develop bioprocesses capable of increasing the bioactive properties of soymilk and, most importantly, increase the equol content by a biotechnological process in vitro. Biotransformation processes based on soymilk fermentation by probiotic lactic bacteria and application of the enzyme tannase caused an increase in the bioactive isoflavones and antioxidant capacity of soymilk. Furthermore, these processes approximately resulted in a 10-fold increase in the equol content of the soymilk. This is the first study to produce a significant equol concentration in soymilk using enzymatic processing only. The results suggest a new and effective biotechnological process, with major commercial potential, capable of producing a bioactive soy extract that intends to be "functional for everyone."

Caracterização química da castanha de sapucaia (Lecythis pisonis Cambess. ) da região da zona da mata mineira / Chemical characterization of sapucaia nuts (Lecythis pisonis Cambess. ) from zona da mata mineira region

Atualmente as castanhas têm recebido atenção especial, pois são fontes naturais de vitaminas, minerais, proteínas e ácidos graxos essenciais, podendo assim contribuir para a dieta humana e de animais. Pesquisas recentes confirmam que esses alimentos são fontes ainda de compostos bioativos, os quais podem trazer benefícios significativos à saúde humana. Este trabalho teve como objetivo avaliar a composição química de castanhas da espécie sapucaia (Lecythis pisonis Cambess.) localizadas na zona da mata mineira. Foram analisadas a composição centesimal (lipídios, proteínas, carboidratos, cinzas e umidade), teor de minerais por espectrometria de plasma e perfil lipídico por cromatografia gasosa. A composição centesimal revelou 54,8% de lipídios; 26,82% de proteínas; 5,01% de carboidratos; 3,17% de cinzas e 10,2% de umidade. Quanto ao perfil lipídico, 43,1% eram ácidos graxos polinsaturados, 41,7% ácidos graxos monoinsaturados e 15,2% ácidos graxos saturados. Os minerais fósforo, magnésio e manganês se destacaram pelos elevados teores, 941; 343 e 4,8 mg.100-1, respectivamente. A castanha de sapucaia é uma potencial fonte energéticoprotéica e de minerais importantes para a saúde, mas deve ter sua toxicidade avaliada.

Currently the nuts have received special attention of researchers because they are natural sources of vitamins, minerals, protein and essential fatty acids, could also contribute to the diet of humans and animals. Recent research confirms that these foods are also sources of bioactive compounds, which can bring significant benefits to human health through regular consumption. This study evaluated the chemical composition of sapucaia nut (Lecythis pisonis Cambess.) of the state of Minas Gerais. We analyzed the chemical composition (lipids, proteins, carbohydrates, ash and moisture), and the mineral content by mass spectrometry, and the lipid profile by gas chromatography. The chemical composition showed 54,8% lipids; 26,82% protein; 5,01% carbohydrates; 3,17% ash and 10,2% moisture. The lipid, 43,1% were polyunsaturated fatty acids, monounsaturated fatty acids 41,7% and 15,2% saturated fatty acids. The minerals phosphorus, magnesium and manganese are highlighted by high levels, 941; 343 and 4,8 mg.100-1, respectively. Sapucaia nut is a potential protein-energy and minerals source, but its toxicity must be evaluated.

Efeito dos resíduos de café seco e fermentado por Monascus ruber no metabolismo de camundongos Apo E / Effect of dry coffee residues fermented with Monascus ruber on the metabolism of Apo E mice

FUNDAMENTO: A aterosclerose é uma doença inflamatória crônica de origem multifatorial que ocorre em resposta à agressão endotelial. O fungo Monascus ruber apresenta atividade hipocolesterolêmica e polifenóis presentes no resíduo de café apresentam atividade antioxidante, podendo auxiliar na prevenção de doenças cardiovasculares. O resíduo de café possui quantidade significativa de açúcares fermentescíveis, constituindo-se em substrato apropriado para o cultivo de fungos. OBJETIVO: O objetivo deste estudo foi avaliar o efeito dos resíduos de café seco e fermentado por Monascus ruber no metabolismo lipídico de camundongos knockout Apo E. MÉTODOS: O ensaio biológico foi realizado com 30 camundongos knockout para o gene Apo E, divididos em cinco grupos e submetidos a diferentes tratamentos. Foi realizada a prospecção fitoquímica e quantificação de compostos fenólicos dos resíduos fermentado e sem fermentar. O soro dos animais foi analisado utilizando kits enzimáticos e o tecido aórtico incluso em parafina e corado com H/E para realização da análise histopatológica. RESULTADOS: O resíduo de café sem fermentar 2%, em relação ao grupo controle, diminuiu em 42% o nível sérico de triacilgliceróis e em aproximadamente 41% a fração VLDL-c. Os grupos dos animais alimentados com 10% de resíduo não fermentado e 2% de resíduo fermentado diminuíram a área de lesão 10,5% e 15,4%, respectivamente, quando comparados ao controle. O resíduo fermentado apresentou um teor de compostos fenólicos superior ao resíduo não fermentado. CONCLUSÃO: O presente estudo mostra que a fermentação do resíduo de café apresenta potencial efeito benéfico sobre as doenças cardiovasculares, especialmente a aterosclerose.

BACKGROUND: Atherosclerosis is a chronic inflammatory disease of multifactorial origin, which occurs in response to endothelial injury. The fungus Monascus ruber has hypocholesterolemic activity, and the polyphenols present in coffee residue have an antioxidant activity and can help prevent cardiovascular diseases. Coffee residue has a significant amount of fermentable sugars, being an adequate substrate for growing fungi. OBJECTIVE: The objective of this study was to assess the effect of dry coffee residue fermented with Monascus ruber on the lipid metabolism of ApoE knockout mice. METHODS: The biological assay was performed with 30 ApoE knockout mice, divided into five groups and undergoing different treatments. The phytochemical prospection and quantification of phenolic compounds of the fermented and non-fermented coffee residues were performed. The sera of the animals were analyzed by using enzyme kits, and the aortic tissue was embedded in paraffin and stained with hematoxylin and eosin to undergo histopathological analysis. RESULTS: Comparing with the control group, the group receiving 2% non-fermented coffee residue showed a reduction of 42% in the serum levels of triacylglycerols and of approximately 41% in VLDL-c. The groups receiving 10% non-fermented coffee residue and 2% fermented coffee residue showed reductions in the lesion areas of 10.5% and 15.4%, respectively, as compared with the control group. The fermented coffee residue showed a higher content of phenolic compounds as compared with the non-fermented coffee residue. CONCLUSION: The present study showed that coffee residue fermentation has a potentially beneficial effect on cardiovascular diseases, especially atherosclerosis.

Effect of dry coffee residues fermented with Monascus ruber on the metabolism of Apo E mice.

BACKGROUND: Atherosclerosis is a chronic inflammatory disease of multifactorial origin, which occurs in response to endothelial injury. The fungus Monascus ruber has hypocholesterolemic activity, and the polyphenols present in coffee residue have an antioxidant activity and can help prevent cardiovascular diseases. Coffee residue has a significant amount of fermentable sugars, being an adequate substrate for growing fungi. OBJECTIVE: The objective of this study was to assess the effect of dry coffee residue fermented with Monascus ruber on the lipid metabolism of ApoE knockout mice. METHODS: The biological assay was performed with 30 ApoE knockout mice, divided into five groups and undergoing different treatments. The phytochemical prospection and quantification of phenolic compounds of the fermented and non-fermented coffee residues were performed. The sera of the animals were analyzed by using enzyme kits, and the aortic tissue was embedded in paraffin and stained with hematoxylin and eosin to undergo histopathological analysis. RESULTS: Comparing with the control group, the group receiving 2% non-fermented coffee residue showed a reduction of 42% in the serum levels of triacylglycerols and of approximately 41% in VLDL-c. The groups receiving 10% non-fermented coffee residue and 2% fermented coffee residue showed reductions in the lesion areas of 10.5% and 15.4%, respectively, as compared with the control group. The fermented coffee residue showed a higher content of phenolic compounds as compared with the non-fermented coffee residue. CONCLUSION: The present study showed that coffee residue fermentation has a potentially beneficial effect on cardiovascular diseases, especially atherosclerosis.