Metabolic pathways of the colonic metabolism of flavonoids (flavonols,flavones and flavanones) and phenolic acids

Flavonoids are metabolized by the gut microflora giving rise to a wide range of phenolic acids that may exert systemic effects in the body. Nevertheless, the colonic metabolism pathways and the function of the metabolites formed are poorly studied. In the present study, the individual colonic metabolism of three subclasses of flavonoids (flavonols, flavones and flavanones) and phenolic acids was evaluated. For this, seven standards of flavonoids (quercetin, quercetin–rhamnoside, quercetin–rutinoside, myricetin, luteolin, naringenin and kaempferol–rutinoside) and two phenolic acids (protocatechuic acid and gallic acid) were submitted to an in vitro fermentation model using rat colonic microflora. Simultaneously, a nuts–cocoa cream enriched with these standards of flavonoids was incubated and the colonic metabolism of these compounds was evaluated. The results showed that the greatest number of colonic metabolites came from the fermentation of quercetin and quercetin–rhamnoside, and the maximum concentration of fermentation products was observed after 48 h of fermentation. On the other hand, a smaller number of fermentation products were observed after the colonic fermentation of kaempferol–rutinoside, naringenin, luteolin and myricetin. The phenolic acids were slightly metabolized by the colonic microflora.     

Chemical Composition and In Vitro Polysaccharide Fermentation of Different Beans (Phaseolus vulgaris L.)

ABSTRACT:  The composition of bioactives including polysaccharide yield and resistant starch (RS) content of 4 raw and cooked bean ( Phaseolus vulgaris  L.) cultivars was evaluated. Polysaccharide was fermented  in vitro  by incubation with human gut flora under anaerobic conditions and short-chain fatty acids (SCFAs) production was compared at 6, 12, and 24 h by gas chromatography. Polysaccharide and soluble fiber contents increased upon cooking with stachyose as the major oligosaccharide. Cooked bean of cultivar Bayo Madero had the highest yield of polysaccharides (55%) and resistant starch (37%), followed by those of Negro 8025 (48% and 32%, respectively). Acetate was the most abundant SCFAs formed in all bean varieties. The concentration of SCFAs was cultivar-dependent; Bayo Madero and Negro 8025 displayed the highest concentration of butyrate (15 mmol/L), while Azufrado Higuera had the lowest and highest concentrations of acetate (39 mmol/L) and propionate (14 mmol/L), respectively. The results suggest that the common bean is an excellent source of polysaccharides that can be fermented in the colon and produce SCFAs, compounds previously reported to exert health benefits.     

Separation, identification, and quantitation of phenolic acids in Chinese waxberry (Myrica Rubra) juice by HPLC-PDA-ESI-MS

A high-performance liquid chromatography coupled with photodiode-array spectrophotometry and electrospray ionization-mass spectrometry (HPLC-PDA-ESI-MS) method for separation and determination of phenolic acids in ethyl acetate extracts from Chinese waxberry (Myrica Rubra) juice was developed. Total of 4 phenolic acids (ferulic, caffeic, sinapic, and salicylic acids) were identified by comparing their HPLC retention times, UV-Vis absorption spectra, and simultaneously recorded mass spectra with authentic standards. Quantitation was carried out by the peak area method. The calibration curves are linear over the concentration range studied with the correlation coefficients, R(2), greater than 0.99. The contents of ferulic, caffeic, sinapic, and salicylic acids in waxberry juice samples studied were 2.76, 3.58, 2.89, and 1.92 mg/L, respectively, and they occur mainly in bound forms. All relative standard deviations were less than 4%. The recoveries range from 83.6% to 95.8% for the 4 analytes. To the authors' best knowledge, this is the first report for the identification of the sinapic and salicylic acids in Chinese waxberry products.     

In vitro antioxidant activity of different cultivars of banana flower (Musa paradicicus L.) extracts available in India

Abstract: Six different cultivars of banana flowers (Musa paradicicus) (Kathali, Bichi, Shingapuri, Kacha, Champa, and Kalabou) were analyzed for the content of polyphenol expressed as gallic acid equivalent and flavonoid expressed as quercetein equivalent, and the in vitro total antioxidative activities of the flower extracts were compared with standard and expressed as trolox equivalent. The reducing power, 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH) and 2,2‐azinobis(3‐ethylbenzothiazoline‐6‐sulfonic acid) radical cation (ABTS?⁺) scavenging activities, inhibition of lipid peroxidation in a linoleic acid emulsion system, and liposome peroxidation system were measured and compared with respective standard antioxidants. Iron‐mediated Fenton reaction was carried out to evaluate the protective effect of the extract of banana flower (Kacha cultivar) against H₂O₂‐induced DNA damage. The Kacha variety contains the maximum amount of polyphenol (11.94 ± 0.03 mg of gallic acid equivalent/g of dry weight) and flavonoid (0.174 ± 0.001 g of quercetin equivalent/g of polyphenol). It also has the highest total antioxidant capacity, DPPH radical scavenging activity, and ABTS?⁺ radical scavenging activity with a least EC₅₀ value of 0.051 mg/mL. Hepatic cell damage in iron‐mediated Fenton reaction caused by free radicals is reduced by the banana flower extract. On the basis of the results obtained, the banana flowers are found to be a potential source of natural antioxidants. This is the first report on the antioxidant properties of the extracts from banana flowers. The study suggests that the flowers of M. paradicicus that are found in India and consumed as vegetable can provide valuable functional ingredients that help in the prevention of oxidative stress.     

Macroporous resin purification of grass carp fish (Ctenopharyngodonidella) scale peptides with in vitro angiotensin-I converting enzyme (ACE) inhibitory ability

The adsorption dynamics and thermodynamics of grass carp fish scale peptides (FSPs) onto non-polar macroporous resins (MARs), DA201-C, have been investigated. The adsorption of FSPs was affected by time, pH and peptide concentration. The adsorption process followed the Langmuir adsorption isotherm, and was endothermic (ΔH < 43 kJ/mol). The predominant force in adsorption of FSPs onto DA201-C was hydrophobic. Depending on this force, the dynamic adsorption and gradient desorption results showed that DA201-C resins were good at desalting and enriching peptides with higher contents of hydrophobic amino acids, and these peptides had higher ACE inhibitory capabilities in vitro. The lowest concentration at which the eluted fraction possessed half of its original ACE activity (IC₅₀) was 0.13 mg/ml. The results indicated that fish scale peptides produced showed good ACE-inhibitory effect in vitro and fish scales are a good source of peptides with in vitro ACE inhibitory activity.     

Taxonomic characterization and metabolic analysis of the Halomonas sp. KM-1, a highly bioplastic poly(3-hydroxybutyrate)-producing bacterium

In a brief previous report, the gram-negative moderately halophilic bacterium, Halomonas sp. KM-1, that was isolated in our laboratory was shown to produce the bioplastic, poly(3-hydroxybutyrate) (PHB), using biodiesel waste glycerol (Kawata and Aiba, Biosci. Biotechnol. Biochem., 74, 175-177, 2010). Here, we further characterized this KM-1 strain and compared it to other Halomonas strains. Strain KM-1 was subjected to a polyphasic taxonomic study. Strain KM-1 was rod-shaped and formed colonies on a plate that were cream-beige in color, smooth, opaque, and circular with entire edges. KM-1 grew under environmental conditions of 0.1%-10% (w/v) NaCl, pH 6.5-10.5 and at temperatures between 10°C and 45°C. The G+C content of strain KM-1 was 63.9 mol%. Of the 16 Halomonas strains examined in this study, the strain KM-1 exhibited the highest production of PHB (63.6%, w/v) in SOT medium supplemented with 10% glycerol, 10.0 g/L sodium nitrate and 2.0 g/L dipotassium hydrogen phosphate. The intracellular structures within which PHB accumulated had the appearance of intracellular granules with a diameter of approximately 0.5 μm, as assessed by electron microscopy. The intra- and extra-cellular metabolites of strain KM-1 were analyzed by capillary electrophoresis mass spectrometry. In spite of the high amount of PHB stored intra-cellularly, as possible precursors for PHB only a small quantity of 3-hydroxybutyric acid and acetyl CoA, and no quantity of 3-hydroxybutyl CoA, acetoacetyl CoA and acetoacetate were detected either intra- or extra-cellularly, suggesting highly efficient conversion of these precursors to PHB.     

In vitro caecum fermentation and in vivo (Gallus gallus) of calcium delivery systems fabricated by desalted duck egg white peptides and chitosan oligosaccharide on gut health

As calcium supplements, two novel calcium delivery systems were prepared with desalted duck egg white peptides (DPs) and chitosan oligosaccharide (COS) by Amadori-linkage and transglutaminase (TGase)-induced reaction, with high calcium-binding ability of promoting calcium absorption in vivo and in vitro. However, there are relatively few studies on the gut health of calcium delivery systems. In this experiment, in vitro caecum fermentation experiment and Gallus gallus intra amniotic feeding model were used to assess the gut health of calcium delivery systems and phytic acid. Results showed that two calcium delivery systems improved the microbial community structure by inhibiting the proliferation of Escherichia coli, and promoting the proliferation of Bifidobacterium and Lactobacillus. The addition of calcium delivery systems significantly increased short chain fatty acids (SCFAs) and improved the boundary membrane function of brush nutrients by significantly increasing villi structure, surface area, cup cell expansion and production of associated genes (P < 0.05). Moreover, calcium delivery systems present significant reverse of the inhibitory effect of phytic acid on gut health through in vitro and in vivo experiments. This study suggested that DPs-COS-Ca systems were superior to the commercially available calcium supplement CaCO₃ and had excellent calcium absorption and gut health regulating effects.     

利用3-噻吩丙二酸修饰玻碳电极检测粮油中残留苯嗪草酮

利用电化学聚合法制备3–噻吩丙二酸修饰玻碳电极,应用循环伏安法研究苯嗪草酮在3–TPA修饰玻碳电极上电化学行为,考察pH值、富集时间、扫描速率及扫描范围等条件对该修饰电极检测苯嗪草酮影响。在最佳条件下,利用差分脉冲伏安法得出苯嗪草酮浓度在9.77×10–7~6.25×10–5mol/L范围内呈良好线性关系,相关系数r=0.9998,检测限为9.77×10–7mol/L。面粉、米粉和花生油样品加标回收率分别为99.2%、100.1%、93.9%,相对标准偏差为0.17%、0.25%、0.49%。     

In Silico Analysis of Selected Honey Constituents as Human Neutrophil Elastase (HNE) and Matrix Metalloproteinases (MMP 2 and 9) Inhibitors

In the present study 12 constituents of honey were evaluated on the docking behavior of human neutrophil elastase and matrix metalloproteinases (MMP 2 and MMP 9) using Discovery Studio. In addition molecular physicochemical, bioactivity, absorption, distribution, metabolism, excretion, and toxicity, and toxicity prediction by komputer assisted technology analyses were done. The molecular physicochemical analysis revealed thatall of the ligands comply with Lipinski’s rule of five. Docking studies revealed that 3-phenyllacticacid exhibited the maximum interaction energy irrespective of its target proteins.The present study provided new insight in understanding these honey constituents as potential enzyme inhibitors.     

Investigating the in vitro bioaccessibility of polyphenols in fresh and sun‐dried figs (Ficus carica L.)

In this study, in order to evaluate the in vitro bioaccessibility of fresh and sun‐dried figs, total antioxidant capacity (TAC), total proanthocyanidin content (TPA) and the major phenolic compounds were determined at different phases of simulated gastrointestinal (GI) tract digestion for Sarilop and Bursa siyahi fig varieties. Four major phenolic compounds (chlorogenic acid, rutin, cyanidin‐3‐glucoside (C3G) and cyanidin‐3‐rutinoside (C3R)) were investigated for GI tract digestion. The results of in vitro GI tract digestion revealed that the dialysed fraction (IN) represented 9–26% and 1–22% of the initial TAC of the whole‐fresh yellow and purple figs, respectively. Moreover, in case of 2,2‐azinobis(3‐ethylbenzothiazoline)‐6‐sulphonic acid (ABTS), TPA and chlorogenic acid contents, drying caused an increase in the IN fraction of yellow figs (38, 140, 50%, respectively). The bioaccessibility of C3G and C3R were quite low for fresh figs (0–5% of the initial values), whereas for dried figs, anthocyanins were not detected at all in the IN fraction.     

The hydroxycinnamic acid content of barley and brewers’ spent grain (BSG) and the potential to incorporate phenolic extracts of BSG as antioxidants into fruit beverages

The hydroxycinnamic acid (HA) content of starting barley for brewers’ spent grains (BSG), whole BSG and phenolic extracts from BSG was measured using high performance liquid chromatography (HPLC) and correlated with antioxidant potential. The effect of BSG phenolic extracts on antioxidant activity of fruit beverages was also assessed (using the total phenolic content (TPC), 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP) assays). The concentration of HA present in barley extract and BSG was in the order of ferulic acid (FA), p-coumaric acid (p-CA) derivatives, FA derivatives, p-CA, caffeic acid (CA) and CA derivatives. Results suggested that brewing and roasting decreased the HA content. Antioxidant activity was significantly (P < 0.05) correlated with caffeic acid (R² = 0.8309) and total HA (R² = 0.3942) concentrations. Addition of extracts to fruit beverages resulted in a significant (P < 0.05) increase in antioxidant activity of cranberry juice, measured by the FRAP assay. In vitro digestion significantly (P < 0.05) reduced TPC, DPPH and FRAP activity of the fruit beverages.     

3‐Chloropropane‐1,2‐diol (3‐MCPD) in Soy Sauce: A Review on the Formation,Reduction, and Detection of This Potential Carcinogen

Soy sauce, a dark‐colored seasoning, is added to enhance the sensory properties of foods. Soy sauce can be consumed as a condiment or added during the preparation of food. There are 3 types of soy sauce: fermented, acid‐hydrolyzed vegetable protein (acid‐ HVP), and mixtures of these. 3‐Chloropropane‐1,2‐diol (3‐MCPD) is a heat‐produced contaminants formed during the preparation of soy sauce and was found to be a by‐product of acid‐HVP‐produced soy sauce in 1978. 3‐MCPD has been reported to be carcinogenic, nephrotoxic, and reproductively toxic in laboratory animal testing and has been registered as a chemosterilant for rodent control. 3‐MCPD is classified as a possible carcinogenic compound, and the maximum tolerated limit in food has been established at both national and international levels. The purpose of this review is to provide an overview on the detection of 3‐MCPD in soy sauce, its toxic effects, and the potential methods to reduce its concentration, especially during the production of acid‐HVP soy sauce. The methods of quantification are also critically reviewed with a focus on efficiency, suitability, and challenges encountered in analysis.     

Production of triacylglycerol and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) by the toluene-degrading bacterium Rhodococcus aetherivorans IAR1

Poly (3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) produced by various bacteria has been intensively investigated as a promising biodegradable plastic, but required a supply of an expensive precursor as a secondary carbon source for its production. In a previous study, we identified a new bacterial strain, Rhodococcus aetherivorans IAR1, which synthesizes PHBV from toluene without the supply of a precursor. Toluene is the volatile organic compound most abundantly emitted to the environment. In the present paper, we show that R. aetherivorans IAR1 produces triacylglycerols (TAGs) simultaneously with PHBV. Both PHBV and TAGs were synthesized before the nitrogen source is completely exhausted. The cellular content of PHBV reached 10% of cell dry weight (CDW) and its synthesis ceased even during intermittent supply of toluene. However, accumulation of TAGs continued during cultivation and their cellular content reached 24% of CDW at the end of cultivation. Cerulenin inhibited TAG production and increased PHBV cellular content up to 30% of CDW. The mole fraction of 3-hydroxyvalerate (3HV) in PHBV produced from toluene increased from 60% to 80% during its accumulation. Fatty acid compositions of TAGs produced from acetate and toluene were different. At the end of cultivation, the mole fraction of C17:0, one of odd-carbon number fatty acids, was 5% on toluene or 10% on acetate while the mole fraction of 3HV in PHBV from toluene was as high as that in PHBV from acetate, suggesting that a C5 intermediate of toluene degradation might directly become a precursor of 3HV whereas propionyl-CoA is required for the incorporation of C17:0 into TAGs.     

In vitro protein digestibility and mineral availability of green beans (Phaseolus vulgarisL) as influenced by variety and pod size

Three varieties of green beans (Cleo, Strike and Sentry) were harvested and sorted into four fractions according to pod size (diameter <7 mm; 7–8·5 mm; 8·6–10 mm and >10 mm). Ash content and dietary fibre increased significantly as pod size increased mainly in Cleo and Strike beans. Strike showed the highest fibre content (378·0 g kg⁻¹) but the lowest carbohydrate (364·6 g kg⁻¹) and ash (68·4 g kg⁻¹) values. Mean values for Fe and Mg content were higher in Cleo beans (70·9 and 27·1 mg kg⁻¹, respectively), Zn, Cu and Mg were higher in Strike beans (48·7 mg kg⁻¹, 22·4 mg kg⁻¹ and 3·15 g kg⁻¹, respectively) while Na and Ca values were maximum in Sentry (459·1 mg kg⁻¹ and 7·11 g kg⁻¹, respectively). Trypsin inhibitor was negatively related to in vitro protein digestibility but no relationship was found between this last parameter and phytic acid content. This antinutrient, together with dietary fibre, and a negative influence on in vitro mineral dialysability of green beans. © 1998 SCI     

Characterization of polyphenoloxidase (PPO) and total phenolic contents in medlar (Mespilus germanica L.) fruit during ripening and over ripening

Characterization of polyphenoloxidase (PPO) enzyme and determination of total phenolic concentrations during fruit ripening and over ripening in medlar (Mespilus germanica L.) were determined. During ripening, PPO substrate specificity, optimum pH and temperature, optimum enzyme and substrate concentrations were determined. Among the five mono- and di-phenolic substrates examined ((p-hydroxyphenyl) propionic acid, l-3,4-dihydroxyphenylalanine, catechol, 4-methylcatechol and tyrosine), 4-methylcatechol was selected as the best substrate for all ripening stages. A range of pH 3.0–9.0 was also tested and the highest enzyme activity was at pH 7.0 throughout ripening. The optimum temperature for each ripening stage was determined by measuring the enzyme activity at various temperatures over the range of 10–70 °C with 10 °C increments. The optimum temperatures were found to be 30, 20 and 30 °C, respectively, for each ripening stage. Optimum enzyme and substrate concentrations were found to be 0.1 mg/ml and 40 mM, respectively. The V_max and K_m value of the reaction were determined during ripening and found to be 476 U/mg protein and 26 mM at 193 DAFB (days after full bloom) – stage 1, 256 U/mg protein and 12 mM at 207 DAFB – stage 2, 222 U/mg protein and 8 mM at 214 DAFB – stage 3. For all ripening stages sodium metabisulfite markedly inhibited PPO activity. For stage 1 of ripening, Cu²⁺, Hg²⁺ and Al³⁺, for stage 2, Cu²⁺ and Hg²⁺, and for stage 3, Cu²⁺, Hg²⁺, Al³⁺ and Ca²⁺ strongly inhibited diphenolase activity. Accordingly, it can be concluded that as medlar fruit ripen there is no significant changes in the optimum values of polyphenoloxidases, although their kinetic parametres change. As the fruit ripening progressed through ripe to over-ripe, in contrary to polyphenoloxidase activity, there was an apparent gradual decrease in total fruit phenolic concentrations, as determined by using the aqueous solvents and water extractions.