A metabolomics approach to evaluate post-fermentation enhancement of daidzein and genistein in a green okara extract.

BACKGROUND: Okara is a major agri-industrial by-product of the tofu and soymilk industries. Employing food-wastes as substrates for the green production of natural functional compounds is a recent trend that addresses the dual concepts of sustainable production and a zero-waste ecosystem. RESULTS: Extracts of unfermented okara and okara fermented with Rhizopus oligosporus were obtained using ethanol as extraction solvent, coupled with ultrasound sonication for enhanced extraction. Fermented extracts yielded significantly better results for total phenolic content (TPC) and total flavonoid content (TFC) than unfermented extracts. A qualitative liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QTOF-MS) analysis revealed a shift from glucoside forms to respective aglycone forms of the detected isoflavones, post-fermentation. Since the aglycone forms have been associated with numerous health benefits, a quantitative high-performance liquid chromatography (HPLC) analysis was performed. Fermented okara extracts had daidzein and genistein concentrations of 11.782 ± 0.325 µg mL-1 and 10.125 ± 1.028 µg mL-1 , as opposed to that of 6.7 ± 2.42 µg mL-1 and 4.55 ± 0.316 µg mL-1 in raw okara extracts, respectively. Lastly, the detected isoflavones were mapped to their metabolic pathways, to understand the biochemical reactions triggered during the fermentation process. CONCLUSION: Fermented okara may be implemented as a sustainable solution for production of natural bioactive isoflavonoids genistein and daidzein. © 2021 Society of Chemical Industry.

Effects of the extract from peanut meal fermented with Bacillus natto and Monascus on lipid metabolism and intestinal barrier function of hyperlipidemic mice.

BACKGROUND: Hyperlipidemia is one of the metabolic disorders that poses a great threat to human health. This study is aimed at investigating the potential hypolipidemic properties of extract from peanut meal fermented with Bacillus natto and Monascus in mice fed with a high-fat diet. Herein, 60 male C57BL/6J mice were randomly divided into six groups: four control groups, comprised of a normal group, a model (M) group, a positive control group (atorvastatin 10 mg kg-1 ), and a nonfermented peanut meal extract group (150 mg kg-1 ), and two experimental groups, comprised of a fermented peanut meal extract low-dose group (50 mg kg-1 ) and a fermented peanut meal extract high-dose group (FH, 150 mg kg-1 ). RESULTS: Body weight (P = 0.001) and levels of serum total cholesterol (P = 0.007), triacylglycerol (P = 0.040), low-density lipoprotein cholesterol (P < 0.001), and leptin (P < 0.001) were remarkably decreased in the FH group, whereas the serum high-density lipoprotein cholesterol levels were increased (P < 0.001) by 78.3% compared with the M group. Ileum tissue stained with hematoxylin and eosin showed that the ileal villus detachments in mice were improved, and the villus height was increased by supplementation with extract from fermented peanut meal. Moreover, the expressions of intestinal ZO-1 (P = 0.003) and occludin (P = 0.013) were elevated in the FH group, compared with the M group. CONCLUSION: Extract of peanut meal fermented by B. natto and Monascus can effectively improve hyperlipidemia caused by a high-fat diet in mice, via regulating leptin and blood lipid levels, and protect the intestinal mucosal barrier, which provides evidence for its anti-hyperlipidemia effects and is a research basis for potential industrial development. © 2020 Society of Chemical Industry.

Toxicological evaluation of lotus, ginkgo, and garlic tailored fermented Korean soybean paste (Doenjang) for biogenic amines, aflatoxins, and microbial hazards.

The present study aimed to develop a consortium of nutritive fermented food products, supplemented with phytochemicals, with reduced toxicological contents. We developed new flavored Doenjang products (protein rich) fermented with lotus, ginkgo, and garlic plant extract-based Meju (termed as EMD) as the starter culture and by using traditional Meju (termed as TMD), where these plant extracts were added later during the fermentation process. Fermented Doenjang samples were analyzed for reduced levels of biogenic amines (BAs), aflatoxins, and microbial hazards, (including Bacillus cereus) as well as for their nutritive contents and antioxidant potential, after varying periods of fermentation (0, 3, 6, 9 and 12 months). All Doenjang samples prepared using plant extracts and their mixtures (1% and 10%) showed desired reduction in B. cereus counts, BAs, aflatoxins, and other foodborne pathogens as well as showed potent antioxidant abilities, including phenolic/flavonoid contents. Based on the higher efficiency in reducing various toxicants, Ginkgo biloba leaf extract added TMD samples were selected for the development of Doenjang products as an innovative approach, with great potential to improve the quality and safety of soybean fermented products in the Korean market, offering enhanced health benefits and reduced risks of toxicity.

Detection of biogenic amines and microbial safety assessment of novel Meju fermented with addition of Nelumbo nucifera, Ginkgo biloba, and Allium sativum.

Meju, a cooked and fermented soy bean based food product, is used as a major ingredient in Korean traditional fermented foods such as Doenjang. We developed a novel type of Meju using single and combined extracts of Allium sativum (garlic clove), Nelumbo nucifera (lotus leaves), and Ginkgo biloba (ginkgo leaves) at 1% and 10% concentrations to improve the safety of Meju-based fermented products. Biogenic amines (BAs) in protein-rich fermented food products pose considerable toxical risks. The objective of this study was to investigate the effects of adding selected plant extracts in Meju samples during fermentation. Nine BAs, including tryptamine, 2-phenylethylamine, putrescine, cadaverine, agmatine, histamine, tyramine, spermidine and spermine, were isolated from Meju samples after sample derivatization with dansyl chloride and analyzed by high performance liquid chromatography. As a result, all tested Meju samples with added plant extracts showed total BAs levels in the range of 20.12 ±â€¯2.03 to 118.42 ±â€¯10.68 mg/100 g, which were below the safety limit set by various regulatory authorities (USFDA/KFDA/EFSA). However, among all tested Meju samples, LOM10 (Meju fermented with Nelumbo nucifera at 10% concentration) showed higher levels of BAs content than others either due to batch-to-batch variability or reduced beneficial microorganisms and/or due to increase in BA forming microorganisms. Also, none of the samples showed the aflatoxin level above the detection limit. Furthermore, all the tested Meju samples improved microbial safety as confirmed by the complete absence of Salmonella species and Staphylococcus aureus. However, some of the Meju samples showed the presence of coliforms (in range of 1.6 × 100-1.1 × 103 CFU/g), which is under regulatory limits. These results suggested that the use of plant extracts in Meju during fermentation have potential to improve microbial and toxicological safety of Meju products.

Preservative effect of Camellia sinensis (L.) Kuntze seed extract in soy sauce and its mutagenicity.

The purpose of this study was to investigate the applicability of green tea seed (GTS) extract as a natural preservative in food. Food preservative ability and mutagenicity studies of GTS extract and identification of antimicrobial compounds from GTS extract were carried out. The GTS extract showed only anti-yeast activity against Candida albicans with MIC value of 938µg/mL and Zygosaccharomyces rouxii with a MIC of 469µg/mL. The active compounds were identified as theasaponin E1 (1), assamsaponin A (2), and assamsaponin B (3). And GTS extracts didn't show mutagenicity because there were no dose-dependent changes in colonies of Salmonella typhimurium TA98, TA100, TA1535, TA1537, and Escherichia coli WP2uvrA regardless of the metabolic activation system. And GTS extract also showed a potent food preservation affect which eliminated all yeast below the MIC value in application test at soy sauce. Overall, these results indicate that GTS extract could be a safe and effective food preservative with anti-yeast activity.

Comparison of Antiobesity Effects Between Gochujangs Produced Using Different Koji Products and Tabasco Hot Sauce in Rats Fed a High-Fat Diet.

In this study, we compared the antiobesity effects between gochujangs prepared using different koji products and Tabasco hot sauce in rats fed a high-fat diet (HFD). Male Sprague-Dawley rats were fed HFD containing four different types of 10% gochujang powder or 0.25% commercial Tabasco sauce powder for 8 weeks. The body weight gain, liver and epididymal and mesenteric fat pad weights, serum leptin levels, and lipogenesis-related mRNA levels of HFD-gochujang supplementation groups were significantly decreased compared with those of the HFD group. In addition, gochujang supplement significantly reduced adipocyte size; hepatic triglyceride and total cholesterol levels; the occurrence of fatty liver deposits and steatosis by inhibiting lipogenesis through downregulation of fatty acid synthase, acetly-CoA carboxylase, and glucose-6-phosphate-dehydrogenase. These effects were greater in the gochujang-supplemented groups than the Tabasco hot sauce-supplemented group. The gochujang prepared by nutritious giant embryo rice koji and soybean koji was most effective in terms of antiobesity effects, compared with the other tested gochujangs. In gochujangs, the antiobesity effects are mediated by high levels of secondary metabolites such as isoflavone, soyasaponin, capsaicin, and lysophosphatidylcholine. The current results indicated that the gochujang products have the potential to reduce fat accumulation and obesity.

Reduction of Aflatoxin B1 Toxicity by Lactobacillus plantarum C88: A Potential Probiotic Strain Isolated from Chinese Traditional Fermented Food "Tofu".

In this study, we investigated the potential of Lactobacillus plantarum isolated from Chinese traditional fermented foods to reduce the toxicity of aflatoxin B1 (AFB1), and its subsequent detoxification mechanism. Among all the investigated L. plantarum strains, L. plantarum C88 showed the strongest AFB1 binding capacity in vitro, and was orally administered to mice with liver oxidative damage induced by AFB1. In the therapy groups, the mice that received L. plantarum C88, especially heat-killed L. plantarum C88, after a single dose of AFB1 exposure, showed an increase in unabsorbed AFB1 in the feces. Moreover, the effects of L. plantarum C88 on the enzymes and non-enzymes antioxidant abilities in serum and liver, histological alterations of liver were assayed. The results indicated that compared to the control group, L. plantarum C88 alone administration induced significant increase of antioxidant capacity, but did not induce any significant changes in the histological picture. Compared to the mice that received AFB1 only, L. plantarum C88 treatment could weaken oxidative stress by enhancing the activity of antioxidant enzymes and elevating the expression of Glutathione S-transferase (GST) A3 through Nuclear factor erythroid (derived factor 2) related factor 2 (Nrf2) pathway. Furthermore, cytochrome P450 (CYP 450) 1A2 and CYP 3A4 expression was inhibited by L. plantarum C88, and urinary aflatoxin B1-N7-guanine (AFB-N7-guanine), a AFB1 metabolite formed by CYP 1A2 and CYP 3A4, was significantly reduced by the presence of viable L. plantarum C88. Meanwhile, the significant improvements were showed in histological pictures of the liver tissues in mice orally administered with viable L. plantarum C88. Collectively, L. plantarum C88 may alleviate AFB1 toxicity by increasing fecal AFB1 excretion, reversing deficits in antioxidant defense systems and regulating the metabolism of AFB1.

Probiotic Soy Product Supplemented with Isoflavones Improves the Lipid Profile of Moderately Hypercholesterolemic Men: A Randomized Controlled Trial.

BACKGROUND: Cardiovascular disease is the leading cause of worldwide morbidity and mortality. Several studies have demonstrated that specific probiotics affect the host's metabolism and may influence the cardiovascular disease risk. OBJECTIVES: The aim of this study was to investigate the influence of an isoflavone-supplemented soy product fermented with Enterococcus faecium CRL 183 and Lactobacillus helveticus 416 on cardiovascular risk markers in moderately hypercholesterolemic subjects. DESIGN: Randomized placebo-controlled double-blind trial Setting: São Paulo State University in Araraquara, SP, Brazil. PARTICIPANTS: 49 male healthy men with total cholesterol (TC) >5.17 mmol/L and <6.21 mmol/L Intervention: The volunteers have consumed 200 mL of the probiotic soy product (group SP-10(10) CFU/day), isoflavone-supplemented probiotic soy product (group ISP-probiotic plus 50 mg of total isoflavones/100 g) or unfermented soy product (group USP-placebo) for 42 days in a randomized, double-blind study. MAIN OUTCOME MEASURES: Lipid profile and additional cardiovascular biomarkers were analyzed on days 0, 30 and 42. Urine samples (24 h) were collected at baseline and at the end of the experiment so as to determine the isoflavones profile. RESULTS: After 42 days, the ISP consumption led to improved total cholesterol, non-HDL-C (LDL + IDL + VLDL cholesterol fractions) and electronegative LDL concentrations (reduction of 13.8%, 14.7% and 24.2%, respectively, p < 0.05). The ISP and SP have prevented the reduction of HDL-C level after 42 days. The C-reactive protein and fibrinogen levels were not improved. The equol production by the ISP group subjects was inversely correlated with electronegative LDL concentration. CONCLUSIONS: The results suggest that a regular consumption of this probiotic soy product, supplemented with isoflavones, could contribute to reducing the risk of cardiovascular diseases in moderately hypercholesterolemic men, through the an improvement in lipid profile and antioxidant properties.

Probiotic Bacillus spp. in Soy-Curd: Nutritional, Rheological, Sensory, and Antioxidant Properties.

UNLABELLED: The focus of this study was to coculture probiotic Bacillus spp. with dairy starter cultures namely, Streptococcus thermophilus and Lactobacillus bulgaricus for enhanced nutritional properties of soy-curd. Subsequently, rheological, sensory, and antioxidant properties of soy-curd along with mineral as well as fatty acid composition were analyzed. Data revealed an increase in the cell viability of probiotic Bacillus spp. on coculturing rather than as mono-culture. Proximate analysis showed higher nutritional value along with increased trace elements. UFA/SFA ratio, rheology, and sensory properties of probiotic soy-curd were in the acceptable range. Probiotic soy-curd showed higher antioxidant activity as measured by the ability to scavenge free radicals. No significant difference in the overall quality within the probiotic products was observed. However, B. flexus MCC2427 cocultured product displayed slightly better attributes than other samples. In general, the results suggest that soy-curd can be a suitable carrier for probiotic Bacillus spp. and the enhanced nutritional and antioxidant properties could be of additional advantage to combat malnutrition problem. PRACTICAL APPLICATION: In order to supply consumers with intriguing probiotic products for improving health benefits, several criteria including technological and functional properties should be considered as a quality control measures. Further, a meaningful level of probiotics has to be viable to exhibit beneficial effect. Hence, present work has been carried out to improve the quality of soy-curd by supplementation of probiotic Bacillus spp. These Bacillus spp. are well characterized native probiotic cultures with potential functional attributes including antimicrobial, antioxidant, anticholesterol activity (Shobharani and Halami 2014). Hence, the application of these cultures will encourage for development of food product with wider health benefits.

Characterization of a novel ß-cypermethrin-degrading Aspergillus niger YAT strain and the biochemical degradation pathway of ß-cypermethrin.

Aspergillus niger YAT strain was obtained from Chinese brick tea (Collection number: CGMCC 10,568) and identified on the basis of morphological characteristics and internal transcribed spacer (ITS) sequence. The strain could degrade 54.83 % of ß-cypermethrin (ß-CY; 50 mg L(-1)) in 7 days and 100 % of 3-phenoxybenzoic acid (3-PBA; 100 mg L(-1)) in 22 h. The half-lives of ß-CY and 3-PBA range from 3.573 to 11.748 days and from 5.635 to 12.160 h, respectively. The degradation of ß-CY and 3-PBA was further described using first-order kinetic models. The pathway and mechanism of ß-CY degraded by YAT were investigated by analyzing the degraded metabolites through high-performance liquid chromatography (HPLC) and liquid chromatography-mass spectrometry (LC-MS). Relevant enzymatic activities and substrate utilization were also investigated. ß-CY degradation products were analyzed. Results indicated that YAT strain transformed ß-CY into 3-PBA. 3-PBA was then gradually transformed into permethric acid, protocatechuic acid, 3-hydroxy-5-phenoxy benzoic acid, gallic acid, and phenol gradually. The YAT strain can also effectively degrade these metabolites. The results indicated that YAT strain has potential applications in bioremediation of pyrethroid insecticide (PI)-contaminated environments and fermented food.

[Transformation of baicalin and wogonoside through liquid fermentation with Bacillus natto].

This experiment aimed to explore and research the process of preparing baicalein and wogonin through liquid fermentation with Bacillus natto. Active enzymes of produced by B. natto was used for the biological transformation of baclin and wogonoside, in order to increase the content of the haicalein and wogonin in the scutellaria. With the content of the baicalein and wogonin as evaluating indexes, the effects of carbon source, nitrogen source, the types and suitable concentration of inorganic salt, medium pH, granularities of medical materials, liquid volume in flask, shaking speed, liquid-to-solid ratio, fermentation time on the fermentation process were studied. The optimal process conditions for liquid fermentation of scutellaria were 1.0% of peptone, 0.05% of NaCl, pH at 6, the granularities of medical materials of the scutellaria screened through 40-mesh sifter, 33% of liquid, shaker incubator speed at 200 r x min(-1), liquid-to-solid ratio of 5:1, temperature at 37 degrees C, fermentation for 6 days, baclin's conversion rate at 97.6% and wogonoside's conversion rate at 97% in the scutellaria. According to the verification test, the process was stable and feasible, and could provide data reference for the industrial production.

Doenjang, a fermented Korean soybean paste, inhibits lipopolysaccharide production of gut microbiota in mice.

Doenjang has been reported to exhibit antioxidant, fibrinolytic, antimutagenic, anticancer, and antiobesity effects. In our preliminary study, doenjang decreased fecal lipopolysaccharide (LPS) levels in mice. Therefore, we investigated the effect of doenjang on the composition of gut microbiota in mice. Treatment with doenjang significantly increased the number of bifidobacteria cultured in BL media, compared with mice not treated with doenjang. However, doenjang decreased the number of Enterobacteriaceae cultured in DHL media. Doenjang significantly suppressed the ß-glucuronidase activity, but did not influence α-/ß-glucosaminidase and α-/ß-glucosidase activities. When gut microbiota in mice treated with or without doenjang was analyzed by pyrosequencing, doenjang induced a significant modulation of the populations of the dominant gut microbiota. At the phylum level, doenjang treatment resulted in a significant decrease of Firmicutes and an increase of Bacteroidetes, which led to a decrease in the Firmicutes to Bacteroidetes ratio in gut microbiota. At the family level, the number of Ruminococcaceae and Lachnospiraceae were significantly decreased, while the number of Odoribacter_f was increased in doenjang-treated mice. Of colonic tight junction proteins, occludin, ZO-1, and claudin-1 in mice, occludin alone was significantly increased by treatment with doenjang. Although treatment with doenjang seemed to suppress NF-κB activation, it was not significant. Doenjang significantly suppressed tumor necrosis factor-α expression, whereas it did not influence interleukin (IL)-1ß and IL-6 expression. However, doenjang increased IL-10 expression. Based on these findings, doenjang may promote gut health by regulating gut microbiota and its LPS concentrations and suppressing harmful enzyme production.

6-Hydroxydaidzein enhances adipocyte differentiation and glucose uptake in 3T3-L1 cells.

Fermented soybean foods have been shown to reduce incidence of diabetes and improve insulin sensitivity. 6-Hydroxydaidzein (6-HD) is a bioactive ingredient isolated from fermented soybean. In this study, we examined the effects of 6-HD on adipocyte differentiation and insulin-stimulated glucose uptake, as well as the mechanisms involved. In our experiments, 6-HD enhanced 3T3-L1 adipocyte differentiation and insulin-stimulated glucose uptake in a dosage-dependent manner. In addition, 6-HD increased peroxisome proliferator-activated receptor gamma (PPARγ) gene expression and PPARγ transcriptional activity. 6-HD increased CCAAT/enhanced binding protein alpha (C/EBPα) expression as well. Although having no effects on glucose transporter type 4 (GLUT4) gene expression, 6-HD facilitated GLUT4 protein translocation to the cell membranes. Our results indicate that 6-HD exhibited the actions of promoting adipocyte differentiation and improving insulin sensitivity by increasing the expression of C/EBPα and facilitating the translocation of GLUT4 via the activation of PPARγ, suggesting that 6-HD can be promising in diabetes management.

Hypoallergenicity of various miso pastes manufactured in Japan.

Miso paste (miso), a fermented soybean food, is popular in Japan and other Asian countries. However, the soybean is known to induce an allergenic response in some individuals. In the present study, we evaluated the allergenicity of various kinds of miso available in Japan. Total proteins were extracted from Amakuti-kome miso, Karakuti-kome miso, Mugi-miso and Mame-miso, and the protein profiles were analyzed. The major protein bands detected in the intact soybean extract were not present in any of the miso samples, which instead showed various low molecular weight protein bands of approximately 10-25 kDa. The existence levels of six major soybean allergens were determined by Western blotting using specific antibodies. We found that the allergen levels varied among miso and allergen types; however, allergen levels were consistently lower in miso than in the soybean extract. We obtained similar results for IgE-ELISA experiments using serum IgE from soybean allergy patients. Taken together, these results indicate that compared to soybean extract, various types of miso contain small quantities of intact soybean allergens. Additionally, several lines of evidence indicated that the allergen levels were exceptionally low in the dark-colored Karakuti-kome miso and Mame-miso, which are produced with relatively long fermentation periods, suggesting that the duration of fermentation might be a key factor in the hypoallergenicity of miso.

Reduction of tyramine by addition of Schizandra chinensis baillon in Cheonggukjang.

This study was performed to examine the microorganisms responsible for the high tyramine content of Cheonggukjang, a traditional Korean fermented soy food, and to establish a technology for controlling the growth of these microorganisms. The tyramine content in 13 collected Cheonggukjang samples averaged 604.9 mg/kg. Since the tyramine content measured from most samples was sufficient to cause harm to the human body, it is necessary to control its production in food. Enterococci were confirmed to be the bacterial species producing most of the tyramine through the microbial examination and were present in high numbers from not detected (<10(1)) to 7.0 × 10(10) colony-forming units (CFU)/g. To control the growth of enterococci, various plant extracts with antimicrobial activity, common salts, and variable temperature conditions were tested. It was found that 4 samples among the 159 plant extracts had a strong antimicrobial activity in Cheonggukjang, especially against Enterococcus faecium, showing viable cell counts of <10(1)-10(3) CFU/g after 24 h of ripening, which were significantly lower values compared to the control (10(9)-10(11) CFU/g). The Cheonggukjang with the addition of the four plant extracts showed ∼83%-95% lower concentrations of tyramine compared to the control. Cheonggukjang prepared with the Schizandra chinensis Baillon extract had the lowest tyramine content without sacrificing the sensory quality. Not only was the bacterial species of E. faecium reduced more remarkably, by up to 10(3) CFU/g compared to the 10(9)-10(11) CFU/g shown in the control, but it also decreased the tyramine content by up to 91%.

Discovery of an S-equol rich food stinky tofu, a traditional fermented soy product in Taiwan.

A recent epidemiological study showed that daily intake of mg quantities of S-equol is required for health-promoting effects in menopausal women. However, the maximum equol content in food was reported to be approximately 130 µg/100 g in egg yolk. The objective of this study was to find a high equol-containing food. We measured the equol content of 33 egg yolks and 21 fermented soybean foods. Equol was detected in 28 egg yolks at the maximum content of 43 µg/100 g. In the fermented soybean foods, equol was detected only in stinky tofu. We examined 16 stinky tofu samples purchased during different seasons and the average equol content was 1.39 mg/100 g, ranging from 0.34 to 2.68 mg/100 g. Equol was present in stinky tofu as the S-enantiomeric form and as an aglycon type. This is the first report demonstrating that stinky tofu contains high levels of S-equol, which may exert beneficial effects in menopausal women.

Conjugated linoleic acid conversion by six Lactobacillus plantarum strains cultured in MRS broth supplemented with sunflower oil and soymilk.

UNLABELLED: Six strains of Lactobacillus plantarum, isolated from traditional dairy products of minority nationalities, were evaluated for their ability to produce conjugated linoleic acid (CLA) from free linoleic acid in vitro. All the 6 strains were found to be capable of converting linoleic acid to CLA when using sunflower oil as substrate or during soymilk fermentation. The inhibitory effect of linoleic acid on the growth of the L. plantarum was also discussed. The production of CLA was increased with adding high concentration of substrate in sunflower oil and IMAU60042 produced the highest CLA both in sunflower oil and soymilk. The CLA was composted by 2 isomers: cis9, trans11-CLA and tran10, cis12-CLA, and cis9, tran s11-CLA covered the most part of the total CLA formed except for L. plantarum P8. The production of CLA was decreased during the storage of fermented soymilk. The CLA contents decreased significantly in the first week, also more quickly in 2 wk. Especially, tran10, cis12-CLA decreased more rapidly than cis9, tran11-CLA. No dramatic change was observed among other 8 fatty acids in soymilk. The proportion of unsaturated fatty acids varied after fermentation with different L. plantrum strains, but all decreased the during storage. The research on the ability of converting CLA of L. plantrum strains could be basis for the future research and development of fermented soymilk products. PRACTICAL APPLICATION: Desirable probiotic traits, such as acid and bile tolerance, aggregation activity, and antibacterial activity, have been proved for the 6 Lactobacillus plantarum strains. The 6 L. plantarum strains might be used in the fermentation of soymilk to produce multifunctional probiotic soymilk products, especially the rich CLA contents.

Acetylcholinesterase inhibitory activity of Chinese sufu (fermented tofu) ethanol-extract.

Acetylcholinesterase (AChE) inhibitory activities of commercial sufu and self-produced sufu were investigated in this experiment. The anti-AChE activities of commercial sufu samples of 15 brands, sourcing from various parts of China, and self-produced sufu, fermented with Actinomucor elegans 3.118, were measured. The results indicated that ethanol extract of Chinese sufu exhibited significant inhibitory activity against AChE in vitro. The inhibitory activity of No. 5 sufu was the strongest (IC50, 0.191 mg/ml), while the pre-fermented sufu showed the highest inhibitory activity during sufu manufacturing. In addition, soybean extracts and potato extracts were used to culture A. elegans 3.118 in order to estimate which culture was preferable for the production of these AChE inhibitors. The soybean extracts, after fermentation by A. elegans 3.118, showed higher anti-AChE activity than did the potato extracts. The IC50 of the soybean extracts was 1.29 µg/ml.

Enhanced growth of lactobacilli and bioconversion of isoflavones in biotin-supplemented soymilk by electroporation.

This study aimed at utilizing electroporation to further enhance the growth of lactobacilli and their isoflavone bioconversion activities in biotin-supplemented soymilk. Strains of lactobacilli were treated with different pulsed electric field strength (2.5, 5.0 and 7.5 kV/cm) for 3, 3.5 and 4 ms prior to inoculation and fermentation in biotin-soymilk at 37°C for 24 h. Electroporation triggered structural changes within the cellular membrane of lactobacilli that caused lipid peroxidation (p < 0.05) and alteration of membrane fluidity (p < 0.05). This was due to the application of electric potential difference across membrane that induced pores formation and subsequently increased membrane permeability. Reversible permeabilized cells resumed growth to >9 log CFU/ml after fermentation in biotin-soymilk (p < 0.05). Lactobacilli cells treated at electric field strength of 7.5 kV/cm for 3.5 ms also showed enhanced ß-glucosidase activity (p < 0.05) compared to lower doses and control, leading to increased bioconversion of isoflavones glucosides to aglycones in biotin-soymilk (p < 0.05). Results from this study show that electroporation could be used to produce biotin-soymilk with increased bioactive aglycones.

Antiosteoporotic effects of Lactobacillus -fermented soy skim milk on bone mineral density and the microstructure of femoral bone in ovariectomized mice.

Osteoporosis is a major skeletal disease associated with loss of estrogen in postmenopausal women. In this study, Lactobacillus paracasei subsp. paracasei NTU 101 (NTU 101F) and Lactobacillus plantarum NTU 102 (NTU 102F) were used as starters to ferment soy skim milk. This was then used as a nutritional supplement for 8 weeks to ovariectomized (OVX) mice. This study reveals that soy skim milk fermented with lactobacilli can increase the contents of aglycone isoflavones, soluble calcium, and vitamin D(3). The trabecular bone volumes and trabecular number of the distal femur in mice fed NTU 101F increased by a factor of 1.48 and 1.74 compared with the OVX group. The bone network density and thickness of the distal metaphyseal trabecular in mice fed NTU 101F and Fosamax was significantly greater than that of OVX mice. These results suggest that fermented soy skim milk can attenuate bone loss in OVX mice and lower the risk of osteoporosis.