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.

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.

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."