Physicochemical characteristics and production of whole soymilk from Monascus fermented soybeans

Monascus fermented soybeans (MFS) were prepared by solid state fermentation. MFS showed higher solubility than the unfermented control within the acidic pH region but showed the opposite trend at pH 7 and 9. Although the emulsifying activity index of MFS was lower than that of the control, MFS had significantly higher emulsion stability at pH 7 and 9. Significant protein hydrolysis took place during fermentation, and the proportion of peak area less than 1.35 kDa was greatly increased in MFS. Whole soymilk prepared from MFS contained monacolin K (475 μg/g), and was enriched in isoflavone aglycones. Neither homogenisation nor pasteurisation caused significant changes in the isoflavone and monacolin K contents of the Monascus fermented soymilk. Total phenol content and ABTS radical scavenging activity were significantly higher in the Monascus fermented soymilk than in the control soymilk.     

Production and optimization of monacolin K by citrinin-free Monascus pilosus MS-1 in solid-state fermentation using non-glutinous rice and soybean flours as substrate

Monacolin K (MK) acts as a natural inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A reductase. Red fermented rice by Monascus spp. which is rich in MK can be used as dietary supplement to reduce the serum triglyceride and total cholesterol. A Monascus pilosus MS-1 with high-producing MK but free of citrinin had been screened in our previous investigation. Here, its solid-state fermentation parameters were optimized using statistical methods to maximize MK yield when non-glutinous rice and soybean flours were used as substrate. Maximum MK yield (17.662 mg/g) was predicted when 55 g dry substrate (20 sieve mesh), including 60 % non-glutinous rice flour and 40 % soybean flour, was put into a 250-mL conical flask, and the contents of water, acetic acid and MgSO₄·7H₂O were adjusted to 35 %, 0.6 % (v/w) and 0.004 mol/kg, respectively. Then, the substrate was mixed and sterilized, and scattered before cooling. Later, 13 % (v/w) inoculum pre-cultured for 36 h at 30 °C was used, then incubated at 30 °C for 60 h and followed by a further incubation at 24 °C up to 14 days. The verified MK yield was 18.733 mg/g, which was 106.06 % to the predicted MK yield. The MK yield was enhanced to 33 times after optimization. No citrinin was detected in Monascus pilosus MS-1 fermented products. These results indicate that citrinin-free MK can be produced under the optimized conditions.     

Characteristics of Monascus sp. isolated from Monascus fermentation products

To screen for the strains suitable for industrial applications, 15 Monacus sp. were isolated from various Monascus fermentation products. The isolated strains were belonging to Monascus pilosus, Monascus purpureus, Monascus albidulus, and Monascus femeus based on the ITS sequence and morphological characteristics. They produced globular and hyaline conidia, and red, orange, or hyaline ascomata except for M. fumeus, in which ascomata was not observed in malt extract media. Each strain produced monacolin K and citrinin at different levels on different media: 15.79–20.05 mg/L of monacolin K and 0–255 μg/L of citrinin in submerged culture of monacolin K producing media, 1.27–2.69 mg/L and about 507.4 μg/L in YES medium, 0–0.78 g/kg and 0–9.7 mg/kg in Monascus red rice products, suggesting that it is necessary to adopt the similar conditions to a specific application for screening as well. These various isolated strains would be great resources for industrial application to develop Monascus fermented functional products.     

The potential antioxidant capacity and angiotensin I-converting enzyme inhibitory activity of Monascus-fermented soybean extracts: evaluation of Monascus-fermented soybean extracts as multifunctional food additives

ABSTRACT:  The potential antioxidant capacity and angiotensin I-converting enzyme (ACE) inhibitory activity of Monascus -fermented soybean extracts (MFSE) were investigated. The average antioxidant capacities of 70% ethanol extracts from soybean after fermenting for 15 d at 30 °C were increased by a 5.2 to 7.4-fold (0.26 mM trolox equivalent/g dry weight, 91.7% 1,1-diphenyl-2-picrylhydrazyl [DPPH] radical scavenging effect at 3 mg/mL) when compared with those of the unfermented soybean extracts ( P < 0.01). The potentially significant antioxidant properties of MFSE are associated with its content of bioactive mevinolins ( r = 0.85) and isoflavone aglycones ( r = 0.98), which were derived from the soybean during Monascus -fermentation. It was also found that the water extract having a molecular mass 1 to 3 kDa showed the highest ACE inhibitory activity (65.3%), which was remarkably greater (6.5 times) than the control.     

Enhancements of isoflavone aglycones, total phenolic content, and antioxidant activity of black soybean by solid-state fermentation with Rhizopus spp.

In the present study, a solid-state fermentation process of black soybean was developed. Three filamentous fungi including Rhizopus oligosporus (BCRC 31996), R. oligosporus (NTU-5), and Rhizopus oryzae (BCRC 30894) were cultivated with steam-cooked intact and dehulled black soybeans. It was found that fermentation enhanced the total phenolic and isoflavone aglycone content as well as antioxidant activity of the black soybean methanol [80 % (v/v)] extracts. Among the three candidates, R. oligosporus BCRC 31996 was chosen as the working strain, and dehulled Tainan NO.3 black soybeans were used as the substrate based on the relatively high total phenolic content, isoflavone aglycone accumulation, and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging effect of the fermented extracts. In a 6-day fermentation, extracts of the fermented dehulled black soybeans yielded higher total phenolic content (2,876.2 μg/g) which was 1.16 times of the result from the intact soybean extracts. The DPPH scavenging effect of the fermented dehulled black soybean extract reached its maximum on the third day. The activity of β-glucosidase of dehulled black soybean samples increased with time and resulted in accumulation of isoflavone aglycones. The product of solid-state black soybean fermentation serves as a functional ingredient in the products of nutritional supplements and health foods.     

Enhancement of antioxidant activity,total phenolic and flavonoid content of black soybeans by solid state fermentation with Bacillus subtilis BCRC 14715

In the present study, a solid state fermentation of black soybeans with Bacillus subtilis BCRC 14715 was performed. The effect of fermentation on the changes of total phenolic and flavonoid content and antioxidant activities including 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging effect, and Fe²⁺-chelating ability exerted by various solvent (water, 80% methanol, 80% ethanol, 80% acetone) extracts of black soybeans was examined. It was found that fermentation enhanced the total phenolic and flavonoid content as well as antioxidant activity of the black soybean extract. Among the various extracts examined, the acetone extract of fermented black soybeans showed the highest total phenolic and flavonoid content. The acetone extract and the methanol extract of fermented black soybeans showed the highest DPPH free radical-scavenging effect and Fe²⁺-chelating ability, respectively. Analysis of extraction yields showed that the active principle associated with the DPPH radical-scavenging effect was most efficiently extracted from black soybeans using water, regardless of fermentation. Water and methanol effectively extract the Fe²⁺-chelating principles from non-fermented and fermented black soybeans, respectively.     

Antioxidant properties of water extracts from Monascus fermented soybeans

Solid-state bioprocessing of soybean by Monascus is a biotechnological strategy to produce Monascus-fermented soybeans (MFS) with more beneficial components. The objective of this study was to evaluate the antioxidant properties of cold and hot water extracts from MFS as compared to uninoculated soybeans. With regard to the EC₅₀ values of antioxidant activities, the effectiveness of cold water extracts was in a descending order: soybeans > MFS-31527 > MFS-31499 whereas that of the hot water extracts: soybeans ∼ MFS-31499 > MFS31527. Cold water extracts showed higher reducing power and lower scavenging ability on 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals than hot water extracts. For both extracts, MFS and soybeans showed comparable effective chelating abilities on ferrous ions. Total phenols were the major naturally occurring antioxidant components found. For both extracts, soybeans were more effective in antioxidant activity and scavenging ability on DPPH radicals whereas MFS-31499 and MFS-31527 were more effective in reducing power and scavenging ability on hydroxyl radicals. Based on the results obtained, MFS-31499, MFS-31527 and soybeans were relatively effective in the antioxidant properties assayed and might be potential antioxidants for application in food products.     

Fermentation with Bacillus spp. as a bioprocess to enhance anthocyanin content,the angiotensin converting enzyme inhibitory effect,and the reducing activity of black soybeans

Food possessing anthocyanins, angiotensin converting enzyme (ACE) inhibitory activity or reducing activity show beneficial effect on human health. To develop healthy food, black soybeans were fermented with either Bacillus subtilis BCRC 14715 or Bacillus sp. CN11, or a mixture of both Bacillus spp. in the present study. The anthocyanin content, the ACE inhibitory activity and the reducing power of the fermented black soybean were then examined. It was found that the ACE inhibitory activity of the extracts of bean and viscous material from the fermented black soybeans varied with extraction solvents and starter organism, yet increased as the fermentation period was extended, regardless of starter organism. After 18 h of fermentation, the water extract of bean showed less ACE inhibitory activity than did the respective 80% ethanol extract. While the water extract of viscous material showed a higher ACE inhibitory activity than the respective ethanol extract. With respect to extraction yield, it was found that the ACE inhibitor in the fermented black soybean could be extracted more efficiently with water than 80% ethanol. Fermentation with B. subtilis BCRC 14715 was also found to increase the anthocyanin content of black soybean and the reducing activity of the extracts. Finally, the 80% ethanol extract showed a higher reducing activity than the water extract.     

丛毛红曲菌固态发酵山药产莫纳可林K的影响因素研究

该研究以新鲜山药为发酵基质,采用丛毛红曲菌（Monascus pilosus）MS-1固态发酵山药,探究新鲜山药、干山药、山药品种、蔗糖、山药块茎大小、乳酸等因素分别对发酵山药中莫纳可林K（MK）产量的影响。结果表明,新鲜山药更有利产生MK,当发酵基质为新鲜铁棍山药,蔗糖添加量为3%,山药块茎大小为（0.5 cm×0.5 cm×0.5 cm）,乳酸添加量为0.3%时,MK产量可达8.10 mg/g。该研究可为山药的精深加工提供新思路,对新型红曲产品的开发也具有一定的借鉴意义。     

Optimization of ethanol extraction and further purification of isoflavones from soybean sprout cotyledon

Isoflavones from cotyledons of soybean sprouts were extracted with aqueous ethanol and further concentrated to obtain a product with a high concentration of isoflavone. The ethanol concentration, extraction time and reaction temperature were optimized by using response surface methodology (RSM). Isoflavones in aqueous ethanol were concentrated by a three-step procedure comprised of solid phase extraction (SPE) with Diaion HP-20 and Amberlite-XAD-2 adsorption columns, acid hydrolysis, and liquid–liquid extraction. The maximum amount of isoflavone in aqueous ethanol extracts (11.6 mg/g solid) was obtained when isoflavones in cotyledons (2.18 mg/ g solid) were extracted with 80–90% (v/v) aqueous ethanol above 90 °C for more than 100 min. The isoflavone extracts, obtained by SPE with a Diaion HP-20 column contained 100 mg/g solid. The liquid–liquid extraction (LLE) with ethyl ether further concentrated the extracts up to 229 mg/g solid, retaining 63% of the initial isoflavones.     

紫苏叶对红曲菌产色素和莫纳可林K及抗氧化物的影响

以高产莫纳可林K（MK）及红曲色素（MPs）、不产桔霉素的丛毛红曲菌（Monascus pilosus）MS-1为实验菌株,优化已有的固态发酵工艺,考察添加紫苏叶对红曲菌产MPs、MK及抗氧化物的影响。结果表明,添加10%紫苏叶时MPs和MK的产量最高（P＜0.05）。添加紫苏叶及紫苏叶提取液均可显著促进菌株MS-1产MPs和MK（P＜0.05）,但两种添加方式对菌株MS-1产MPs及MK的影响无显著差异（P＞0.05）。添加紫苏叶后色价和MK产量可分别达211.73 U/g和8.15 mg/g,分别为对照的1.35和1.36倍。紫苏红曲粗提物的DPPH清除率显著高于对照（P＜0.05）,可达48.71%。然而,紫苏红曲中迷迭香酸含量仅为0.032 mg/g,而紫苏叶中迷迭香酸含量高达14.71 mg/g。添加紫苏叶不仅可促使红曲菌产MPs及MK,还可能促使红曲菌产抗氧化物质,从而提高其发酵产物的抗氧化能力。     

Exploring the bioactive potential of brewers spent grain ohmic extracts

Ohmic based brewer's spent grain (BSG) hydro-ethanolic extracts were produced and their composition and biological potential through evaluation of their antibacterial, antibiofilm and antihypertensive activities were characterized. Ohmic Heating Extraction (OHE) was performed using two aqueous extractions of ethanol (60 and 80% ethanol:water (v/v)). A polyphenolic profile was performed and 4-hydroxybenzoic acid was the compound displaying the highest concentration (125.86 μg/g BSG) and the extract of 60% (v/v) hydro-ethanolic solution was the one with highest antioxidant activity (30.70 mg ascorbic acid/g BSG). Nevertheless, both extracts proved to have no mutagenic activity, and both were capable of inhibiting 50% of the angiotensin-I converting enzyme (ACE) enzyme, the most effective being the one obtained using 80% ethanol:water (v/v). BSG extracts displayed an inhibitory effect against Bacillus cereus at 2.50 and 0.625 mg/mL, for 80% (v/v) and 60% (v/v) ethanol:water, respectively and to a lesser extent against Listeria monocytogenes, at 5.00 and 2.50 mg/mL, for 80% (v/v) and 60% (v/v) ethanol:water, also the same concentrations inhibited biofilm formation.     

Comparison of extraction solvents and techniques used for the assay of isoflavones from soybean

The impact of extraction solvents and techniques on the assay of isoflavones from soybean was investigated. This systematic study was undertaken to address substantial variations in the solvents and procedures used for the extraction of isoflavones from soybeans by different research groups as described in the recent peer-reviewed published literature. Comparison of four previously optimized and commonly deployed solvent mixtures (acetonitrile:water 58:42 (v/v); ethanol:water 70:30 (v/v); methanol:water 90:10 (v/v); superheated pressurized water) was carried out for the extraction of isoflavones. In addition, we also examined the extraction efficiencies of three additional new solvent mixtures (dimethyl sulphoxide:acetonitrile:water, 5:58:37 (v/v/v); dimethyl sulphoxide:ethanol:water, 5:70:25 (v/v/v); Genapol:water 5:95 (v/v)) for the extraction of isoflavones from soybeans. Assessments of six commonly used extraction techniques (shaking, vortexing, sonication, stirring, Soxhlet, and pressurized liquid extraction (PLE)) with an optimized extraction solvent mixture was also performed. Both, the total isoflavones content and the isoflavones HPLC profile varied significantly with different extraction solvents and techniques. Optimum total isoflavones recoveries from soybean samples were obtained with dimethyl sulphoxide:ethanol:water (5:75:25, v/v/v) solvent mixture using a PLE. Intermediate extraction recoveries of total isoflavones from soybean samples were obtained with the other extraction solvent mixtures and techniques tested. The extraction efficiencies of isoflavones with shaker, vortex, stirring, and Soxhlet were between 65% and 70% as compared to PLE. Total isoflavones extracted by the sonication procedure was 93.3% as compared to PLE.     

Composition and antioxidant activity of raisin extracts obtained from various solvents

Experiments were conducted to determine if the contents of phenolics and browning reaction products and antioxidant activity of raisin extracts were closely dependent on the extraction solvent. Enhanced extraction yields were obtained from solvent containing higher water concentrations. However, total phenolic content (TPC) was highest for extracts obtained from solvent to water ratios of 60:40 (v/v), whereby the extract obtained from ethanol:water (60:40, v/v) had the highest TPC of 375 mg gallic acid equivalents (GAE)/100 g extract. HMF content was highest in extracts obtained from 60% solvent, regardless of solvent type. The extract obtained from 60% methanol had the highest HMF content at 199 μg/g extract. Although the 60% solvents provided extract with high antioxidant components, the antioxidant activity of raisin extracts obtained from 80:20 (v/v) solvent/water was significantly higher than other raisin extracts obtained from different solvent concentrations. Phenolic acids, HMF, and low-molecular-weight flavonoids were responsible for the antioxidant activity, but not the high-molecular-weight flavonoids.     

Analysis of physiochemical composition and antioxidant properties between hulls of the genetically modified glyphosate-tolerant soybean and northeast soybean

In this study, the physiochemical and antioxidant properties of the soybean hulls from the genetically modified glyphosate-tolerant soybeans (line 40-3-2) and local cultivar northeast soybeans were investigated. The levels of fat, total phenolic, total extractable pectin and soluble dietary fiber in northeast soybeans hulls were less than that in glyphosate-tolerant soybeans hulls, respectively. The antioxidant capacity of total phenolic, water soluble pectin, and soluble dietary fiber showed that DPPH free radical scavenging activities of glyphosate-tolerant soybeans hulls were 118.23, 57.34 and 197.22 μg AAE/g, which were 2.3, 1.2 and 9.4 times of northeast soybeans hulls, respectively (p < 0.05), and FRAP of glyphosate-tolerant soybeans hulls were 401.67, 747.51 and 328.53 μg AAE/g, which were 1.8, 8.7 and 4.8 times of northeast soybeans hulls (p < 0.05). Glyphosate-tolerant soybeans hulls extract showed the stronger antioxidant activity, which was positively correlated with total phenolic content (r = 0.890, p = 0.001). It provides evidence on developing value-added utilization of hulls, soybean processing by-products, as nutraceuticals or functional food ingredients.     

Soluble phenolics and antioxidant properties of soybean (Glycine max L.) cultivars with varying seed coat colours

This research was the first to investigate nutritional components, including soluble phenolics (isoflavones and anthocyanins), protein, oil, and fatty acid as well as antioxidant activities in different coloured seed coat soybeans (yellow, black, brown, and green) for two crop years. The soluble phenolics differed significantly with cultivars, crop years, and seed coat colours, while protein, oil, and fatty acid exhibited only slight variations. Especially, malonylgenistin and cyanidin-3-O-glucoside compositions had the most remarkable variations. Green soybeans had the highest average isoflavone content (3079.42 μg/g), followed by yellow (2393.41 μg/g), and black soybeans (2373.97 μg/g), with brown soybeans showing the lowest value (1821.82 μg/g). Anthocyanins showed only in black soybeans, with the average contents of the primary anthocyanins, cyanidin-3-O-glucoside, delphinidine-3-O-glucoside, and petunidin-3-O-glucoside, quantified at 11.046, 1.971, and 0.557 mg/g, respectively. Additionally, Nogchae of green soybean and Geomjeongkong 2 of black soybean may be recommended as potential cultivars owing to the highest average isoflavone (4411.10 μg/g) and anthocyanin (21.537 mg/g) contents. The scavenging activities of 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) radicals also differed remarkably, depending upon isoflavone and anthocyanin contents, with black soybeans exhibiting the highest antioxidant effects.     

Phenolics and antioxidant activity of lentil and pea hulls

Hulls obtained by pilot-scale dehulling process from green and red lentils and yellow peas were fractionated into hulls (> 500 ??m) and residues (< 500 ??m). These fractions together with the corresponding whole seeds were extracted with four solvents, aqueous (70%) acetone, (80%) ethanol, hot water (70-80 °C) and water (22 ± 1 °C) and evaluated for antioxidant activity in relation to phenolic contents. Aqueous acetone (70%, v/v) extracted the highest level of total phenolics at about 87 mg of catechin equivalent per gram of sample from lentil hulls followed by hot water, water and aqueous ethanol (80%, v/v). Red lentil hull with maximum concentration of phenolic compounds exhibited the strongest antioxidant activity of 260 mg (1040 ??M) trolox equivalent/g hull. Hot water and water extracted significantly higher total phenolics from whole pulse and their residue than other solvents, but with weaker antioxidant activity. Aqueous ethanol was always the best extractant of phenolic antioxidant from yellow pea irrespective of its millstream. The most potent phenolic antioxidant was obtained from water extract of green and red lentil hulls, followed by those extracted with ethanol, acetone, and hot water. Total phenolic content was highly correlated with antioxidant activity of pulses and its millstreams.     

Effects of selected herb extracts on iron-catalyzed lipid oxidation in soybean oil-in-water emulsion

Effect of herb extracts on the oil oxidation of an oil-in-water emulsion containing iron was studied. The emulsion comprised tocopherol-stripped soybean oil (40 g), citrate buffer (60 g, pH 4.0), xanthan gum (35mg), and FeSO₄ (0.5mg) with 80% ethanol extracts of rosemary (Rosmarinus officinalis), basil (Ocimum basilicum), peppermint (Mentha piperita), thyme (Thymus vulgaris), or oregano (Origanum vulgare). Hydroperoxide contents and p-anisidine values were significantly (p<0.05) lower in the emulsions with the added herb extracts compared with the control emulsions. The antioxidant activity of the basil and peppermint extracts containing higher proportions of rosmarinic acid was significantly higher (p<0.05) than that of the other herb extracts, but the polyphenol degradation rate was not significantly different. The antioxidant activity of the basil and peppermint extracts showed concentration-dependence within 400 mg/kg. This study suggests that the oxidative stability of oil-inwater emulsion containing iron can be improved by addition of basil or peppermint extract.     

蒸汽爆破预处理对粉葛总黄酮及抗氧化性的影响

实验选取爆破压力1.0、2.0 MPa,维持压力时间30、40、60、80 s,对粉葛进行预处理。利用紫外分光光度法表征爆破前后总黄酮的变化,旨在探讨蒸汽爆破预处理对粉葛总黄酮及抗氧化性的影响。抗氧化活性利用1,1-二苯基-2-三硝基苯肼（1,1-diphenyl-2-picrylhydrazyl,DPPH）清除率进行评价。结果表明：经过蒸汽爆破预处理后,粉葛的总黄酮提取量、甲醇提取物得率和抗氧化性得到显著提高。在最优处理条件下,总黄酮（以葛根素计）提取量达到5.43 mg/g,是未经预处理粉葛提取量的2.32 倍;甲醇提取物得率为16.92%;抗氧化活性得到显著增加,DPPH清除活性的半数抑制浓度（50% inhibitory concentration,IC50）由30.65 g/L降低至10.10 g/L。将蒸汽爆破预处理应用于粉葛活性物质的提取,可实现提取量和抗氧化能力的有效提高。     

高产开环型monacolin K红曲霉菌株的筛选及其在红曲酒酿造中的应用

该研究采用38株红曲霉菌株固态发酵制备功能红曲米,采用高效液相色谱法测定红曲米中monacolin K和桔霉素含量,筛选高产开环型莫纳可林K(monacolin K)红曲霉菌株,通过形态观察及分子生物学技术对筛选菌株进行鉴定,并将其应用于红曲酒酿造,分析红曲酒的品质。结果表明,筛选得到1株高产开环型monacolin K、不产桔霉素的菌株ABQ2,经鉴定,其为丛毛红曲霉（Monascus pilosus）。采用该菌株发酵制备功能红曲米中的monacolin K含量达到25.10 mg/g,开环率为90.5%,且桔霉素未检出。采用菌株ABQ2酿造的红曲酒酒精度为12%vol,酸度为1.91 g/100 mL,monacolin K含量为0.48 mg/mL,其理化指标均符合相关行标要求;与小曲酒相比,红曲酒中总酯含量（433.65 mg/L）极显著增加（P<0.01）,其中乳酸乙酯和乙酸乙酯含量最高,高级醇类含量（145.52 mg/L）略低,主要为正丙醇、异丁醇、正戊醇、β-苯乙醇等。