Bioconversion Of Isoflavones Research Articles

β-Glucosidase activity and bioconversion of isoflavones during fermentation of soymilk.

Lactobacillus rhamnosus C6 strain showed higher β-glucosidase activity as well as biotransformation of isoflavones from glycones (daidzin and genistin) to aglycones (daidzein and genistein) in soymilk. However, L. rhamnosus C2 and Lactobacillus casei NCDC297 also exhibited similar activity during soymilk fermentation. These three strains can be selected for the development of functional fermented soy foods enriched with aglycone forms of isoflavones, such as soy yoghurt, soy cheese, soy beverages and soy dahi. The study determined β-glucosidase activity of probiotic Lactobacillus cultures for bioconversion of isoflavones to aglycones in fermenting soymilk medium. Soymilk was fermented with six strains (L. rhamnosus C6 and C2, L. rhamnosus NCDC19 and NCDC24 and L. casei NCDC17 and NCDC297) at 37 °C for 12 h. The highest β-glucosidase activity and isoflavone bioconversion after 12 h occurred by L. rhamnosus C6 culture during fermentation in soymilk. Increased isoflavone aglycone content in fermented soymilk is likely to improve the biological functionality of soymilk (e.g. antioxidant activity, alleviation of hormonal disorders in postmenopausal women, etc.). Lactobacillus rhamnosus C6 culture can be used for the development of functional fermented soy-based products.

Effect of tea extract on lactic acid bacterial growth, their cell surface characteristics and isoflavone bioconversion during soymilk fermentation

In this study, the influences of tea extract (TE) supplementation on selected lactic acid bacteria (LAB) growth, their cell surface characteristics and soy isoflavone bioconversion were investigated. Soymilk-tea (SMT) was prepared by combining TE (green, oolong or black tea) with soymilk containing lactose (SML). Bacterial growth was studied by inoculating four LAB into soymilk or SMT individually. FT-IR spectroscopy and cell surface hydrophobicity (SH) assay were performed to monitor the changes in bacterial cell surface characteristics due to TE supplementation. HPLC analysis was used to evaluate isoflavone bioconversion in fermented soymilk and SMT. Results showed that TE promoted the growth of Lactobacillus delbrueckii ssp. bulgaricus (Lactobacillus bulgaricus) and L. paracasei but inhibited the growth of L. acidophilus and Streptococcus thermophilus at a concentration of 2% (weight of TE powder/volume of soymilk). Viabilities of all LAB in fermented SMT were maintained at above 7logCFU/mL during a four-week refrigeration (4°C). FT-IR spectra indicated major changes in the membrane fatty acids, proteins and cell wall polysaccharides after TE treatment. Changes in SH provided further information on the modification of bacterial cell surface by TE. Inhibition of deglycosylation was observed in fermented SMT for L. acidophilus and L. paracasei, both of which exhibited strong ability to transform isoflavone glycoside in SML, while isoflavone bioconversion was enhanced for L. bulgaricus. Our study provides some practical significance for developing novel functional soyfood by combining gut health-promoting bacteria, soy isoflavones and tea polyphenols.

Effect of ultrasound on bioconversion of isoflavones and probiotic properties of parent organisms and subsequent passages of Lactobacillus

This study evaluated the effects of ultrasound (60 W; 3 min) on growth, bioconversion of isoflavones, and probiotic properties of parent organisms and subsequent passages of Lactobacillus casei FTDC 2113. Ultrasound significantly enhanced (P < 0.05) the cell growth of the parent organisms in mannitol–soymilk fermented at 37 °C over 24 h. Such treatment also enhanced the intracellular and extracellular β-glucosidase activity, leading to increased bioconversion of isoflavones in mannitol–soymilk (P < 0.05). In addition, treated cells also exhibited better tolerance toward acidic conditions (pH 2 and pH 3) and intestinal bile salts compared to the control (P < 0.05). Ultrasound also promoted cell adhesion ability and antimicrobial activity (P < 0.05). All these positive effects were only prevalent in the parent cells without inheritance by first, second and third passage of cells. Our results suggest that ultrasound could enhance the bioactive and probiotic potentials of parent cells of L. casei FTDC 2113, and could be used in the production of probiotic foods with enhanced bioactivity.

Bioconversion of isoflavones during the fermentation of Samso-Eum with Lactobacillus strains

In this study, the possible application of Lactobacillus strains as a functional starter culture to ferment Samso-Eum (SE), an oriental herbal medicine formula, and the production of bioactive isoflavones (daidzein, genistein) were investigated. Four strains of Lactobacillus (Lactobacillus plantarum KFRI 144, L. amylophilus KFRI 161, L. curvatus KFRI 166, and L. bulgaricus KFRI 344) were used for SE fermentation. Declines in pH and in viable cell counts during fermentation were investigated and the quantification of isoflavones using HPLC were performed after fermentation at 37°C for 48 h. All the tested Lactobacillus strains lowered the pH level to approximately 3.6 after 48 h and showed the highest level of growth at 24 h during SE fermentation. During the SE fermentation of the four Lactobacillus strains, the conversion of isoflavone glycosides (daidzin, genistin) into bioactive aglycones (daidzein, genistein) was observed in all of the fermentations, but with different rates depending on the strains. L. plantarum KFRI 144 and L. amylophilus KFRI 161 exhibited the highest bioconversion rate of isoflavone glycosides into their bioactive aglycones. These results demonstrate that L. plantarum KFRI 144 and L. amylophilus KFRI 161 have potentials as functional starter cultures for manufacturing fermented SE with higher isoflavone bioavailability.

Ultraviolet radiation enhanced growth of lactobacilli and their bioconversion of isoflavones in biotin-supplemented soymilk

The aim of this study was to evaluate the effect of ultraviolet (UV) treatment on growth of lactobacilli and their bioconversion of isoflavones in biotin–soymilk. Strains of lactobacilli were treated with different types of UV radiation at various doses prior to inoculation in biotin–soymilk. The impact imposed by UV treatment on lactobacilli was species/strain dependent as observed in the various parameters measured. Generally, the treatment induced oxidative damage on lactobacilli which decreased cell viability immediately after treatment. This was attributed to the increased oxidation of acyl chains and proteins in the membrane lipid bilayer, leading to increased lipid peroxidation and membrane permeability (P < 0.05). The alteration in phospholipid structures also reduced cellular membrane fluidity (P < 0.05). After removal of UV radiation, the cellularly injured bacteria cells were able to adapt, recover and resume growth during fermentation in biotin-soymilk (P < 0.05). UVB treatment of lactobacilli at 60 J/m2 showed higher intracellular and extracellular β-glucosidase activities (P < 0.05), leading to increased bioconversion of isoflavones glucosides to aglycones in biotin–soymilk. Results from this study showed that UV radiation could be applied to promote β-glucosidase activity of lactobacilli cells for the development of aglycones-enriched biotin–soymilk.

Effect of electroporation on viability and bioconversion of isoflavones in mannitol‐soymilk fermented by lactobacilli and bifidobacteria

The aim of this study was to evaluate the effect of electroporation (2.5-7.5 kV cm⁻¹ for 3.0-4.0 ms) on the growth of lactobacilli and bifidobacteria, membrane properties and bioconversion of isoflavones in mannitol-soymilk. The viability of lactobacilli and bifidobacteria decreased immediately after electroporation. This was attributed to lipid peroxidation, which led to alterations in the membrane phospholipid bilayer, specifically at the polar head, interface and apolar tail regions. Such alterations also resulted in decreased membrane fluidity and increased membrane permeability upon electroporation (P < 0.05). However, the effect was reversible and treated cells showed better growth than the control upon fermentation for 24 h at 37 °C (P < 0.05). Additionally, electroporation increased the bioconversion of glucosides to bioactive aglycones in mannitol-soymilk, which was attributed to increased intracellular and extracellular β-glucosidase activities of cells upon treatment (P < 0.05). Application of electroporation on lactobacilli and bifidobacteria could be beneficial for the development of fermented soymilk with enhanced bioactivity. Considering the enhanced bioactive aglycones, this soymilk could be useful for the prevention of hormone-dependent disorders.

Bioconversion of Isoflavones and the Probiotic Properties of the Electroporated Parent and Subsequent Three Subcultures of Lactobacillus fermentum BT 8219 in Biotin-Soymilk

This study was aimed at an evaluation of the potential inheritance of electroporation effects on Lactobacillus fermentum BT 8219 through to three subsequent subcultures, based on their growth, isoflavone bioconversion activities, and probiotic properties, in biotin-supplemented soymilk. Electroporation was seen to cause cell death immediately after treatment, followed by higher growth than the control during fermentation in biotin-soymilk (P<0.05). This was associated with enhanced intracellular and extracellular beta-glucosidase specific activity, leading to increased bioconversion of isoflavone glucosides to aglycones (P<0.05). The growing characteristics, enzyme, and isoflavone bioconversion activities of the first, second, and third subcultures of treated cells in biotin-soymilk were similar to the control (P>0.05). Electroporation affected the probiotic properties of parent L. fermentum BT 8219, by reducing its tolerance towards acid (pH 2) and bile, lowering its inhibitory activities against selected pathogens, and reducing its ability for adhesion, when compared with the control (P<0.05). The first, second, and third subcultures of the treated cells showed comparable traits with that of the control (P>0.05), with the exception of their bile tolerance ability, which was inherited to the treated cells of the first and second subcultures (P<0.05). Our results suggest that electroporation could be used to increase the bioactivity of biotin-soymilk via fermentation with probiotic L. fermentum BT 8219, with a view towards the development of functional foods.

Growth, bioconversion of isoflavones and probiotic properties of parent and subsequent passages of Lactobacillus upon ultraviolet radiation

The objective of this study was to evaluate the effects of ultraviolet (UV) radiation (UVB; 90 J/m2) on growth, bioconversion of isoflavones and probiotic properties of parent and subsequent passages of L. casei FTDC 2113. UV radiation significantly enhanced (P < 0.05) the growth of parent cells in mannitol-soymilk fermented at 37°C for 24 h. This had led to an enhanced intracellular and extracellular β-glucosidase activity with a subsequent increase in bioconversion of isoflavones in mannitol-soymilk (P < 0.05). UV radiation also promoted (P < 0.05) the tolerance of parent cells towards acidic condition (pH 2 and 3) and intestinal bile salts (oxgall, taurocholic and cholic acid). In addition, parent treated cells also exhibited better (P < 0.05) adhesion ability to mucin and antimicrobial activity compared to that of the control. All these positive effects of UV radiation were only prevalent in the parent cells without inheritance by first, second and third passage of cells. Although temporary, our results suggested that UV radiation could enhance the bioactive and probiotic potentials of L. casei FTDC 2113, and thus could be applied for the production of probiotic products with enhanced bioactivity.

Effects of ultrasound on growth, bioconversion of isoflavones and probiotic properties of parent and subsequent passages of Lactobacillus fermentum BT 8633 in biotin-supplemented soymilk

This study aimed to evaluate the effects of ultrasound on Lactobacillus fermentum BT 8633 in parent and subsequent passages based on their growth and isoflavone bioconversion activities in biotin-supplemented soymilk. The treated cells were also assessed for impact of ultrasound on probiotic properties. The growth of ultrasonicated parent cells increased (P<0.05) by 3.23–9.14% compared to that of the control during fermentation in biotin-soymilk. This was also associated with enhanced intracellular and extracellular (8.4–17.0% and 16.7–49.2%, respectively; P<0.05) β-glucosidase specific activity, leading to increased bioconversion of isoflavones glucosides to aglycones during fermentation in biotin-soymilk compared to that of the control (P<0.05). Such traits may be credited to the reversible permeabilized membrane of ultrasonicated parent cells that have facilitated the transport of molecules across the membrane. The growing characteristics of first, second and third passage of treated cells in biotin-soymilk were similar (P>0.05) to that of the control, where their growth, enzyme and isoflavone bioconversion activities (P>0.05) were comparable. This may be attributed to the temporary permeabilization in the membrane of treated cells. Ultrasound affected probiotic properties of parent L. fermentum, by reducing tolerance ability towards acid (pH 2) and bile; lowering inhibitory activities against selected pathogens and reducing adhesion ability compared to that of the control (P<0.05). The first, second and third passage of treated cells did not exhibit such traits, with the exception of their bile tolerance ability which was inherited to the first passage (P<0.05). Our results suggested that ultrasound could be used to increase bioactivity of biotin-soymilk via fermentation by probiotic L. fermentum FTDC 8633 for the development of functional food.

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 β-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.

Enhanced growth and bioconversion of isoflavones in prebiotic-soymilk fermented by UV-treated lactobacilli and bifidobacteria

The aim of this study was to evaluate the effects of ultraviolet (UV) radiation (ultraviolet A (UVA), ultraviolet B (UVB) and ultraviolet C (UVC) at 30–90 J/m2) on the membrane properties of lactobacilli and bifidobacteria, and their bioconversion of isoflavones in prebiotic-soymilk. UV treatment caused membrane permeabilization and alteration at the acyl chain, polar head and interface region of membrane bilayers via lipid peroxidation. Such alteration subsequently led to decreased (p < 0.05) viability of lactobacilli and bifidobacteria immediately after the treatment. However, the effect was transient where cells treated with UV, particularly UVA, grew better in prebiotic-soymilk than the control upon fermentation at 37°C for 24 h (p < 0.05). In addition, UV treatment also increased (p < 0.05) the intracellular and extracellular β-glucosidase activity of lactobacilli and bifidobacteria. This was accompanied by an increased (p < 0.05) bioconversion of glucosides to bioactive aglycones in prebiotic-soymilk. Our present study illustrated that treatment of lactobacilli and bifidobacteria with UV could develop a fermented prebiotic-soymilk with enhanced bioactivity.

ACE inhibitory activity and bioconversion of isoflavones by Lactobacillus in soymilk supplemented with B‐vitamins

PurposeThe aim of this study is to examine the bioactive properties of lactobacilli‐fermented B‐vitamin soymilk, namely the in‐vitro antihypertensive property and bioconversion of isoflavone glucosides to aglycones.Design/methodology/approachLactobacillus acidophilus BT 1088, L. fermentum BT 8219, L. acidophilus FTDC 8633 and L. gasseri FTDC 8131 were investigated for their bioactive potential and enhanced bioconversion of isoflavones in soymilk supplemented with individual B‐vitamins at a concentration of 1 mg/L.FindingsThe supplementation of thiamine, riboflavin, niacinamide, calcium pantothenate, biotin and folic acid enhanced the ACE‐inhibitory activity of lactobacilli in soymilk accompanied by a lower IC50 value compared to the control (P&lt;0.05). The β‐glucosidase specific activity of lactobacilli was also enhanced on supplementation of B‐vitamins, leading to increased bioconversion of isoflavones in soymilk. The concentration of genistein was decreased, accompanied by an increased concentration of genistein on fermentation in the presence of thiamine, niacinamide, biotin, calcium pantothenate and folic acid. Additionally, the supplementation of niacinamide, calcium pantothenate, biotin and folic acid also led to lower concentrations of malonyl daidzin, indicating increased hydrolysis of malonyl daidzin to daidzin. Results from the present study indicated that the supplementation of B‐vitamins could enhance the bioactive potential and bioconversion of isoflavones in lactobacilli‐fermented soymilk.Originality/valueThis work has shown that the supplementation of B‐vitamins in lactobacilli fermented soymilk has exerted in vitro ACE‐inhibitory activity and increased the accumulation of bioactive isoflavone aglycones. To the authors' knowledge, this is the first evaluation reporting on such aspects.

Enhanced growth of lactobacilli and bioconversion of isoflavones in biotin-supplemented soymilk upon ultrasound-treatment

This study aimed at utilizing ultrasound treatment to further enhance the growth of lactobacilli and their isoflavone bioconversion activities in biotin-supplemented soymilk. Strains of lactobacilli ( Lactobacillus acidophilus BT 1088, L. fermentum BT 8219, L. acidophilus FTDC 8633, L. gasseri FTDC 8131) were treated with ultrasound (30 kHz, 100 W) at different amplitudes (20%, 60% and 100%) for 60, 120 and 180 s prior to inoculation and fermentation in biotin-soymilk. The treatment affected the fatty acids chain of the cellular membrane lipid bilayer, as shown by an increased lipid peroxidation ( P < 0.05). This led to increased membrane fluidity and subsequently, membrane permeability ( P < 0.05). The permeabilized cellular membranes had facilitated nutrient internalization and subsequent growth enhancement ( P < 0.05). Higher amplitudes and longer durations of the treatment promoted growth of lactobacilli in soymilk, with viable counts exceeding 9 log CFU/mL. The intracellular and extracellular β-glucosidase specific activities of lactobacilli were also enhanced ( P < 0.05) upon ultrasound treatment, leading to increased bioconversion of isoflavones in soymilk, particularly genistin and malonyl genistin to genistein. Results from this study show that ultrasound treatment on lactobacilli cells promotes ( P < 0.05) the β-glucosidase activity of cells for the benefit of enhanced ( P < 0.05) isoflavone glucosides bioconversion to bioactive aglycones in soymilk.

Effect of Ultrasound on the Growth of Probiotics and Bioconversion of Isoflavones in Prebiotic-Supplemented Soymilk

The objective of the present study was to evaluate the effects of ultrasound on the growth of probiotics and bioconversion of isoflavones in prebiotic-soymilk. Previous studies have shown that ultrasound elevated microbial enzymatic activity and growth by altering cellular membranes. The growth of probiotics was significantly decreased (P < 0.05) immediately after ultrasound treatment, attributed to membrane permeabilization, cell lysis, and membrane lipid peroxidation upon ultrasound treatment. The ultrasound treatment also caused alteration at the acyl chain, polar head, and interface region of the probiotic membrane phospholipid bilayers. The cells treated with ultrasound showed recovery from injury with subsequent increase in growth upon fermentation in soymilk (P < 0.05). Ultrasound treatment at 100 W for 2 and 3 min also enhanced (P < 0.05) the intracellular and extracellular β-glucosidase activity of probiotics, leading to increased (P < 0.05) bioconversion of glucosides to aglycones in the prebiotic-soymilk. Our present study illustrated that ultrasound treatment could produce bioactive synbiotic-soymilk with increased concentrations of bioactive aglycones.

Effects of the β‐glycosidase reaction on bio‐conversion of isoflavones and quality during tofu processing

Isoflavones are the most common group of phytoestrogens which are present in significantly large amounts in soybean and soy products such as tofu. Isoflavones occur naturally in glycoside forms having lower bioavailability than their aglycone forms. beta-Glycosidase acts as a bio-catalyst for the conversion of isoflavone glycosides to isoflavone aglycones, raising the bioavailability of isoflavones; therefore, it can be used to improve the quality of tofu. We need to establish process conditions for the optimal outcome of the enzyme reaction in tofu. By using the beta-glycosidase (0.02% w/v) reaction at 55 degrees C for 30 min, a maximum 84.5% conversion of isoflavone glycoside to isoflavone aglycone was obtained. The enzyme reaction caused no significant effects on the sensory acceptability of soft tofu. The hardness of enzyme-treated hard tofu increased with the coagulant amount whereas prolonged heating resulted in decrease of hardness. Incorporation of enzyme reaction before the coagulation process during soft tofu processing provided a sufficient bio-conversion of isoflavones at optimal conditions. beta-Glycosidase can be effectively used for the bioconversion of isoflavones in soft tofu manufacturing process at optimal reaction conditions before the onset of coagulation process.

Angiotensin I-converting enzyme inhibitory activity and bioconversion of isoflavones by probiotics in soymilk supplemented with prebiotics

Lactobacillus sp. FTDC 2113, L. acidophilus FTDC 8033, L. acidophilus ATCC 4356, L. casei ATCC 393, Bifidobacterium FTDC 8943 and B. longum FTDC 8643 were incorporated into soymilk supplemented with fructooligosaccharides (FOS), inulin, mannitol, maltodextrin and pectin. The objective of the present study was to evaluate the effects of prebiotics on the bioactivity of probiotic-fermented soymilk. Proteolytic activity was increased in the presence of FOS, while the supplementation of inulin and pectin increased the angiotensin I-converting enzyme inhibitory activity accompanied by lower IC50 values. The β-glucosidase activity was also enhanced in the presence of pectin. This led to higher bioconversion of glucosides to aglycones by probiotics, especially genistin and malonyl genistin to genistein. Results from this study indicated that the supplementation of prebiotics enhanced the in-vitro antihypertensive effect and production of bioactive aglycones in probiotic-fermented soymilk. Therefore, this soymilk could potentially be used as a dietary therapy to reduce the risks of hypertension and hormone-dependent diseases such as breast cancer, prostate cancer and osteoporosis.

Production of a High Value-Added Soybean Containing Bioactive Mevinolins and Isoflavones

The production of mevinolin, a potent hypocholesterolemic drug, and the bioconversion of isoflavones were investigated in soybeans fermented with Monascus pilosus KFRI-1140. The highest yields of 2.94 mg mevinolins and 1.13 mg isoflavone aglycones per g dry weight of soybean were obtained after 20 days of fermentation. Mevinolin was present in the fermentation substrate predominantly in the hydroxycarboxylate form (open lactone, 94.8～96.7%), which is currently being used as an hypocholesterolemic agent. The significant (p＜0.01) bioconversion (96.6%) of the glucoside isoflavones (daidzin, glycitin, genistin) present in the soybean to the bioactive aglycones (daidzein, glycitein, genistein), with a 15.8-fold increase of aglycones was observed. The results suggest that Monascus-fermented soybean has potential as a novel medicinal food or multifunctional food supplement.