In the study, the researchers mimicked human gut microbiota in vitro to understand how soy isoflavones are metabolized by these microorganisms. According to Macedo, soy isoflavones can be obtained from okara (also called soy pulp or tofu dregs), an insoluble residue remaining from the production of soybean oil.
"Both protein and phenolic compounds can be extracted from okara. As a food engineer, I'm always interested in ways of using such residues. In Brazil, I know of no more promising source than soybeans to obtain the extract on an industrial scale, although in principle, equol can be obtained from all isoflavone-rich plants," she said.
Methods
The researchers produced a soy milk extract with the correct initial concentration of isoflavones for their experiments. "The industrial process used to obtain the extract for soy-based drinks sold in supermarkets doesn't have sufficient phenolic content because they're designed to serve as a source of non-animal protein," Macedo explained.
The group used different processes to obtain equol by mimicking the metabolization of the isoflavones in the extract. "In the first, we deployed enzymes to remove glucose from glycosylated isoflavones. In this case, we performed measurements to find out whether a metabolite of interest could be obtained using enzymes and to quantify the glycosylated and aglycone isoflavones at the end of the process," Macedo said.
The second strategy entailed fermenting the extract with a mixture of lactobacilli. The researchers inoculated the extract with lactic bacteria in the absence of oxygen (anaerobiosis) and analyzed the isoflavones before and after fermentation.
In the third process, they combined enzymes with probiotics. "After the enzymatic action, I inoculated the mix of probiotics. The aim was to facilitate the work of the lactobacilli by meeting them halfway with the enzymes in an attempt to speed up the process and obtain more metabolites at the end."
"We found that this combination did indeed work better. It enhanced anti-oxidant capacity, production of metabolites, and conversion of glycosylated into aglycone isoflavones. Both treatments combined had synergistic effects on soy-based products," Macedo said.
The group made a point of testing processes that can be replicated on an industrial scale, working with commercial enzymes and probiotics. "We have to develop solutions that make sense from a technological standpoint," she said.
Effects and contraindications
Because equol is very similar to estrogen, receptors of this hormone in the ovaries, womb, and breasts do not detect any difference, and in menopausal women, the organism does not react to lack of estrogen, which appears to be present, minimizing the symptoms.
"This is the point of obtaining compounds that mimic estrogen. They're also found in blackberry leaves, yams, and other plants. I don't know if they're more bioavailable or also require some kind of transformation to be absorbable by the organism," Macedo said.
According to existing knowledge, she added, phytoestrogen acts similarly to estrogen taken for hormone replacement therapy, albeit in far smaller doses.
Next steps
According to Macedo, equol can exist in two forms: R-equol and S-equol. "Only the latter is highly absorbed, but in the process used to obtain the metabolite, we couldn't find a way to separate them, so we opted for two routes. One was identification by chromatography, which distinguishes between equol and other metabolites of interest.
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