Effect of selected phenolic compounds on the membrane-bound adenosine triphosphatase of Staphylococcus aureus

The effect of selected phenolic compounds on the membrane-bound ATPase activity of two strains of Staphylococcus aureus was studied. These phenolic compounds were: butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), tertiary butylhydroquinone (TBHQ), propyl gallate (PG), p-coumaric, ferulic, caffeic acids, methyl paraben and propyl paraben. Cytoplasmic membrane-bound ATPase activity was measured in the presence of 0, 150, 300, 600 and 1200 μg ml⁻¹ of each phenolic compound. Among all the compounds studied, only BHA was found to significantly stimulate the activity of the enzyme. In contrast, some of the compounds were found to significantly inhibit the activity of the enzyme as was the case of TBHQ, PG, p-coumaric acid, ferulic acid and caffeic acid. The remaining compounds did not influence the activity of the ATPase at any of the concentrations tested. Strain LP ATPase as stimulated less by BHA and inhibited to a greater extent by TBHQ and PG than was the enzyme from strain A100. In contrast, LP ATPase was less inhibited by p-coumaric acid and not affected by either ferulic or caffeic acid.     

Effect of phenolic acids on glucose and organic acid metabolism by lactic acid bacteria from wine

The influence of phenolic (p-coumaric, caffeic, ferulic, gallic and protocatechuic) acids on glucose and organic acid metabolism by two strains of wine lactic acid bacteria (Oenococcus oeni VF and Lactobacillus hilgardii 5) was investigated. Cultures were grown in modified MRS medium supplemented with different phenolic acids. Cellular growth was monitored and metabolite concentrations were determined by HPLC-RI. Despite the strong inhibitory effect of most tested phenolic acids on the growth of O. oeni VF, the malolactic activity of this strain was not considerably affected by these compounds. While less affected in its growth, the capacity of L. hilgardii 5 to degrade malic acid was clearly diminished. Except for gallic acid, the addition of phenolic acids delayed the metabolism of glucose and citric acid in both strains tested. It was also found that the presence of hydroxycinnamic acids (p-coumaric, caffeic and ferulic) increased the yield of lactic and acetic acid production from glucose by O. oeni VF and not by L. hilgardii 5. The results show that important oenological characteristics of wine lactic acid bacteria, such as the malolactic activity and the production of volatile organic acids, may be differently affected by the presence of phenolic acids, depending on the bacterial species or strain.     

Addition of phenolic acids on the reduction of methanol content in wine

ABSTRACT:  Utilization of phenolic acids, including gallic acid, coumaric acid, caffic acid, cinnamic acid, and ferulic acid, for methanol reduction in wine was investigated. Enzyme activities of pectinesterase and pectin lyase decreased significantly when 0.1 mg/L of gallic acid, coumaric acid, caffic acid, cinnamic acid, or ferulic acid was added. However, no inhibition on polygalacturonase activity was observed when 0.5 mg/L of phenolic acid was added. Methanol content in commercial pectic enzyme (CPE) group increased from 11.53 ± 1.34 to 56.67 ± 3.75 ppm in the final products. Adding gallic acid or coumaric acid with CPE inhibited the increase of methanol production. In addition, when 0.2 mg/L of phenolic acid (gallic acid or coumaric acid) was added, the amount of total phenolic acid released from CPE + gallic acid or CPE + coumaric acid groups became higher than CPE group by approximately 466 and 539 mg/L, respectively. In conclusion, the values of lightness, red content, yellow content, total pigment, and total phenolic acid increased in the presence of gallic acid or coumaric acid with CPE, suggesting that adding gallic acid or coumaric acid into winemaking process is a potential method for reducing methanol content, improving wine quality, as well as increasing healthy compounds in wine production.     

Distribution of phenolic yeasts and production of phenolic off-flavors in wine fermentation

The activity of wine yeasts to decarboxylate ferulic and p-coumaric acids is one of their biological properties related to the production of phenolic off-flavors (POF) in wine-making. We examined POF productivity in 116 strains of wine yeast, 74 strains of wild yeast (Saccharomyces cerevisiae) and 23 strains of non-Saccharomyces yeast, and found that a majority of these yeasts were POF-producing strains. The frequency distribution of POF-producing strains was 81 to 95% in wine yeasts, 85 to 97% in wild yeasts and 78 to 83% in non-Saccharomyces yeasts based on the POF test with addition of ferulic and p-coumaric acids to grape juice medium. The Rhodotorula, Candida, Cryptococcus, Pichia, Hansenula, and Brettanomyces strains had high or moderate POF productivity among the 20 non-Saccharomyces species. The decomposition rate of ferulic acid correlated with POF production and the critical concentration of phenolic acid (free form) in grape must was estimated to be more than 10 mg/l. Segregation of POF phenotype and Southern blot analysis of phenolic wine yeasts suggest that POF production is controlled by the POF gene (PAD1). The results showed the frequent distribution of phenolic yeasts in the wine-making environment. These suggest the importance of controlling POF production by using wine yeast strains of low POF productivity. The grapes must be prepared by a suitable process to prevent the increase in phenolic acid content.     

Metabolism of food phenolic acids by Lactobacillus plantarum CECT 748

Phenolic acids account for almost one third of the dietary phenols and are associated with organoleptic, nutritional and antioxidant properties of foods. This study was undertaken to assess the ability of Lactobacillus plantarum CECT 748^T to metabolize 19 food phenolic acids. Among the hydroxycinnamic acids studied, only p-coumaric, caffeic, ferulic and m-coumaric acids were metabolized by L. plantarum. Cultures of L. plantarum produced ethyl and vinyl derivatives from p-coumaric and caffeic acids, 4-vinyl guaiacol from ferulic acid, and 3-(3-hydroxyphenyl) propionic acid from m-coumaric acid. Among the hydroxybenzoic acids analysed, gallic acid and protocatechuic acid were decarboxylated to pyrogallol and catechol, respectively. Inducible enzymes seem to be involved, at least in m-coumaric and ferulic acid metabolism, since cell-free extracts from cultures grown in the absence of these phenolic acids were unable to metabolize them. Further work is needed for the identification of the enzymes involved, since the knowledge of the metabolism of phenolic compounds is an important issue for the food industry.     

Ability of Lactobacillus brevis strains to degrade food phenolic acids

The potential to degrade 15 food phenolic acids was investigated for several Lactobacillus brevis strains isolated from different sources. All the strains analysed in this study showed a similar metabolism on phenolic acids. Among the cinnamic acids assayed, only p-coumaric, ferulic and caffeic acids were metabolized by the L. brevis strains. These acids were decarboxylated to produce their corresponding vinyl derivatives. Contrarily to the results previously reported on Lactobacillus plantarum, the L. brevis strains analysed in this study were unable to subsequently reduce or metabolize these vinyl derivatives. In L. brevis, vinyl phenol, vinyl catechol, and vinyl guaiacol were the final metabolic products from p-coumaric, caffeic or ferulic acids, respectively. From the benzoic acids analysed, and similarly to L. plantarum strains, only gallic and protocatechuic acids were modified by L. brevis strains. Both acids were decarboxylated to pyrogallol and catechol, respectively. Currently, the enzymes involved in the metabolism of phenolic acids in L. brevis remain uncharacterized.     

荞麦蜜酚酸含量的高效液相色谱测定及其抗氧化作用的研究

以中国产荞麦蜜为原料,采用高效液相色谱-电化学检测法(HPLC-ECD)测定其中的原儿茶酸、没食子酸、咖啡酸、丁香酸、p-香豆酸、绿原酸、阿魏酸和鞣花酸等8种酚酸的含量。结果表明,中国荞麦蜜中含有原儿茶酸、没食子酸、咖啡酸、p-香豆酸、阿魏酸等五种酚酸,其含量分别为没食子酸为461μg/100g蜂蜜,p-香豆酸为150μg/100g蜂蜜,咖啡酸为79μg/100g蜂蜜,原儿茶酸为30μg/100g蜂蜜,阿魏酸为39μg/100g蜂蜜。本文还对荞麦蜜酚酸提取物的抗氧化作用进行了研究,结果显示荞麦蜜酚酸提取物对超氧阴离子自由基、羟基自由基、DPPH自由基和脂质过氧化均有抑制作用。     

Changes of Phenolic Acids During Aging of Organic Wines

Grapes (Vitis vinifera L. origin var: Carignane, Cabernet Sauvignon, Merlot, Grenache, Columbard, and Semillon) were cultivated and processed according to accepted organic agriculture and organic wine techniques. Aged wines (5 years) were evaluated for changes of their phenolic acids. The highest reduction of gallic acid concentrations were determined in Cabernet Sauvignon (24.36 mg/L) and Carignane (16.00 mg/L) wines. The quantities of p-hydroxybenzoic acid decreased mostly in Carignane (22.47 mg/L) and Columbard (20.84 mg/L) wines. The decreases of syringic acid were dominant in Cabernet Sauvignon (2.34 mg/L) and Carignane (1.69 mg/L) wines. In the case of ferulic acid, the highest reduction was determined in Cabernet Sauvignon (3.97 mg/L) wines. The decreases of p-coumaric acid contents were the same and mostly in Merlot (1.06 mg/L) and Grenache (1.035 mg/L) wines. The principal component analyses results demonstrated the relations among aged wines produced from different grape varieties and their phenolic acids. The distribution of data was accumulated into two groups. The first group included total phenols, gallic acid, p-hydroxybenzoic acid, syringic acid, p-coumaric acid, and Merlot/Carignane/Grenache wines. The second one included ferulic acid and Cabernet Sauvignon wine.     

Antimicrobial activity of binary combinations of natural and synthetic phenolic antioxidants against Enterococcus faecalis

This study investigated the antimicrobial activity of 3 natural (thymol, carvacrol, and gallic acid) and 2 synthetic [butylated hydroxyanisole (BHA) and octyl gallate] phenolic compounds, individually and in binary combinations, on 4 dairy isolates of Enterococcus faecalis with different virulence factors (β-hemolytic, gelatinase, or trypsin activities; acquired resistance to erythromycin or tetracycline; and natural resistance to gentamicin). A checkerboard technique and a microdilution standardized method were used. All compounds individually tested exhibited antimicrobial activity against E. faecalis, with minimal inhibitory concentrations (MIC) ranging from 30 μg/mL (octyl gallate) to 3,150 μg/mL (gallic acid), although no significant differences were detected among strains to each phenolic compound. Carvacrol in combination with thymol or gallic acid, and gallic acid combined with octyl gallate showed partial synergistic inhibition of all E. faecalis strains. The most effective combinations were thymol + carvacrol and gallic acid + octyl gallate, as the MIC for each of these compounds was reduced by 67 to 75% compared with their respective individual MIC. These results highlight the possibility of using combinations of these phenolic compounds to inhibit the growth of potential virulent or spoilage E. faecalis strains by reducing the total amount of additives used in dairy foods.     

On the phenolic acids of vegetables. IV. Hydroxycinnamic acids and hydroxybenzoic acids of vegetables and potatoes (author's transl)]

Lettuce, endive and chicory exclusively, cornsalad and sweet fennel almost exclusively contain caffeic acid derivatives beside traces of ferulic acid. Parsley exclusively and spinach almost exclusively show p-coumaric acid derivatives. Compared to root, fruit and seed vegetables the contents of phenolic acids in green leaves are considerably high. Rhubarb is the only vegetable, which contains gallic acid (chief phenolic acid) beside hydroxycinnamic, protocatechuic and vanillic acid derivatives. Furthermore hydroxybenzoic acid derivatives (salicylic, gentisic and vanillic acid) occur in cornsalad, sweet fennel, parsley and spinach in small concentrations; cornsalad shows p-hydroxybenzoic acid (ca. 20 mg/kg). Onions (Allium cepa) contain almost only protocatechuic acid beside small amounts of p-hydroxybenzoic and vanillic acid. In the outer dry coloured skins protocatechuic acid reaches concentrations up to 2% of plant material; the internal pulpy tissues show lower concentrations (ca. 20 mg/kg). On the contrary to the bulbs the green leaves of onions like chive and leek contain almost exclusively compounds of ferulic and p-coumaric acid. Garlic even shows a different phenolic acid pattern of skins and internal tissues. The caffeic acid derivatives of potatoes are mainly localized to a 1--2 mm thick outer layer. The different localization of phenolic acids in the different parts of vegetable plants is discussed.     

Development of a reversed-phase high performance liquid chromatography (RP-HPLC) procedure for the simultaneous determination of phenolic compounds in peanut skin extracts

A reversed-phase high performance liquid chromatography (RP-HPLC) procedure was developed for simultaneous determination of five phenolic acids, two stilbenes and eight flavonoids in peanut skin extract. A C₁₈ column fitted with diode array detection at 250 and 320, 280 and 370, and 306 nm for phenolic acids, flavonoids and stilbenes, respectively, with mobile phase consisting of 0.1% formic acid in water and 0.1% formic acid in 100% acetonitrile. Phenolic compounds were eluted with good resolution (Rs > 1.5) within 95 min as follows: gallic, protocatechuic, epigallocatechin, catechin, β-resorcylic (internal standard), caffeic, procyanidin B₂, epicatechin, epigallocatechin gallate, p-coumaric, ferulic, piceid, epicatechin gallate, catechin gallate, resveratrol and quercetin. The variation in recovery and reproducibility in peak area was <11 and <2.5%, respectively. The correlation coefficients, r, of calibration curves of the 15 compounds were >0.999. The method was used to quantify phenolic compounds in peanut skin extracts from Runner, Virginia and Spanish peanuts.     

Study of the inhibitory activity of phenolic compounds found in olive products and their degradation by Lactobacillus plantarum strains

Lactobacillus plantarum is the main species responsible for the spontaneous fermentation of Spanish-style green olives. Olives and virgin oil provide a rich source of phenolic compounds. This study was designed to evaluate inhibitory growth activities of nine olive phenolic compounds against four L. plantarum strains isolated from different sources, and to explore the L. plantarum metabolic activities against these phenolic compounds. None of the nine compounds assayed (oleuropein, hydroxytyrosol, tyrosol, as well as vanillic, p-hydroxybenzoic, sinapic, syringic, protocatechuic and cinnamic acids) inhibited L. plantarum growth at the concentration found in olive products. Oleuropein and tyrosol concentrations higher than 100 mM were needed to inhibit L. plantarum growth. On the other hand, sinapic and syringic acid showed the highest inhibitory activity since concentrations ranging from 12.5 to 50 mM inhibited L. plantarum growth in all the strains analyzed. Among the nine compounds assayed, only oleuropein and protocatechuic acid were metabolized by L. plantarum strains grown in the presence of these compounds. Oleuropein was metabolized mainly to hydroxytyrosol, while protocatechuic acid was decarboxylated to catechol. Metabolism of oleuropein was carried out by inducible enzymes since a cell-free extract from a culture grown in the absence of oleuropein was unable to metabolize it. Independent of their isolation source, the four L. plantarum strains analysed showed similar behaviour in relation to the inhibitory activity of phenolic compounds, as well as their ability to metabolize these compounds.     

益生菌发酵蓝莓花色苷的代谢规律

选用嗜热链球菌、两岐双歧杆菌、植物乳杆菌进行纯种发酵含有蓝莓花色苷提取液的培养液,考察了益生菌发酵过程中活菌数、体外抗氧化性、有机酸、单体花色苷和酚酸等组分的变化情况。结果表明,发酵48 h后,三种益生菌的活菌数均增至8.0 lg CFU/mL左右,两岐双歧杆菌的活菌数最高(P<0.05),发酵后不同菌株发酵样的ABTS+·清除能力显著提升(P<0.05),其中两岐双歧杆菌发酵样品抗氧化能力最强(P<0.05);但植物乳杆菌发酵样品中乳酸含量最高(P<0.05);发酵过程中单体花色苷含量呈下降趋势,两岐双歧杆菌发酵样品花色苷组分与其它两株菌发酵的样品差异较大;绿原酸、对香豆酸和咖啡酸的含量总体呈现下降趋势,而没食子酸、丁香酸和阿魏酸含量呈上升趋势,主成分分析图中三株益生菌发酵样的分布差异较大。以上结果为进一步解释益生菌发酵代谢花色苷的机理提供了参考。     

Degradability of phenolic acid-hemicellulose esters: A model system

A synthetic model system containing p-coumaric and ferulic acids esterified to hemicellulose was used to study polysaccharide degradability. Oatspelts xylan was fractionated into A_linear, B_linear and B_ranched fractions prior to synthetic esterification with phenolic acids at treatment concentrations of 0, 25, 50, 75 and 100 g phenolic acid per kg hemicellulose. Concentrations of phenolic acids esterified to the hemicellulose fractions were determined by alkaline hydrolysis. In-vitro dry matter disappearance (IVDMD) and degradability of hemicellulose neutral sugars were measured after 48 h ruminal fermentations. Esterification efficiency of the phenolic acids to the hemicellulose fractions was low (0.3 to 13.9%) and greater for p-coumaric than ferulic acid (4.7 vs 3.1%, respectively). Concentration of esterified phenolic acids was negatively correlated with IVDMD for the A_linear and B_branched fractions. Ferulic acid appeared to be more inhibitory to IVDMD than p-coumaric acid. Generally the degradability of the side chain sugars of the hemicellulose fractions was negatively correlated with esterified phenolic acid concentrations. Xylose degradation was only correlated with esterified ferulic acid level in the A_linear fraction. The in-vitro ruminal fermentations resulted in the degradation of the majority of the phenolic acid esters. Analysis of the synthetic phenolic acid-hemicellulose esters by ¹³C NMR and FTIR was unable to prove the presence of monomeric phenolic acid esters. The presence of phenolic acid polyesters was unlikely because of the solubility of the synthetic phenolic acid-hemicellulose esters. The neutral sugar degradation data suggest that esterification of the phenolic acids was limited to sugars with primary hydroxyl groups. While this model system was useful for studying cell wall degradation, future studies must employ model systems in which the chemical constituents being tested accurately model those found in nature.     

Differentiation of Romanian Wines on Geographical Origin and Wine Variety by Elemental Composition and Phenolic Components

Several wines were analyzed by high-performance liquid chromatography (HPLC-PDA), inductively coupled plasma mass spectrometry (ICP-MS), and flame atomic absorption spectroscopy (FAAS) in order to associate the concentration levels of the chemical parameters with the geographical origin of wines and wine varieties. Thus, the concentrations of 22 elements (Li, Be, Co, Ni, Cs, U, Pb, V, As, Ba, Cr, Cu, Zn, Al, Mn, Rb, Sr, Fe, Ca, Mg, Na, and K) and seven phenolic compounds (gallic acid, (+)-catechin, (?)-epicatechin, p-coumaric acid, ferulic acid, trans-cinnamic acid, and resveratrol) were analyzed in 22 wines from two different wine-producing areas of Romania. Among the phenolic compounds, (+)-catechin, (?)-epicatechin, p-coumaric acid, ferulic acid, and resveratrol concentrations were the most useful markers for wine differentiation by geographical origin and wine variety, whereas Ba, Be, Cr, Cs, Li, Mg, Na, Ni, Sr, U, and Zn were the main inorganic parameters selected. To explore the distribution pattern of the samples, aimed at differentiating the wine samples by geographical region and wine variety, principal component analysis (PCA) was applied to the obtained data.     

Phenolic acids and flavonoids profiles of extracts from edible wild fruits and their antioxidant properties

In this study, the total phenolic, total flavonoids, phenolic compounds, the mineral content, and antioxidant activity of fruit extracts of seven wild species (Crataegus monogyna Jacq., Prunus spinosa L., Rosa canina L., Hippophaë rhamnoides L., Rubus fruticosus L., Prunus padus, Cornus mas L.) were investigated. The results indicated significant differences (p < 0.05) in the total phenolics and total flavonoids content, between the seven analyzed species. These ranged from 184.69 to 727.29 mg GAE/100 g FW and 17.27 to –165.55 mg QE/100 g FW, respectively. The antioxidant activity found in fruits was not directly affected by the total phenolic content (TPC). This activity was linked to a larger extent to the type of individual phenolic compounds and to a lesser extent to the TPC, because fruits with higher TPC have not always presented the highest values of antioxidant activity. HPLC analysis of methanolic extract showed the presence of phenolic acids (i.e. gallic, vanillic, chlorogenic, caffeic, syringic, p-coumaric, ferulic, sinapic, salycilic, elagic, and trans-cinnamic) and flavonoids (i.e. catechin, epicatechin, rutin, myricetin, and quercetin). Significant differences (p < 0.05) were observed in each individual mineral between fruits from wild flora. The fruits tissues of wild species turned out to be a good source of calcium (Ca), magnesium (Mg), sodium (Na), iron (Fe), manganese (Mn), copper (Cu), chromium (Cr), zinc (Zn), and boron (B). The results demonstrated that wild species possessed great potential for food production as sources of bioactive compounds such as phenolic compounds and minerals, for food supplements or functional foods.     

Synthesis and evaluation of antioxidant and antifungal activities of novel ricinoleate-based lipoconjugates of phenolic acids

Syntheses of four castor oil fatty acid-based novel lipoconjugates of phenolic acids were carried out following Mitsunobu methodology. The lipid part consists of methyl ricinoleate and its saturated analogue, methyl-12-hydroxystearate and the phenolic moieties are ferulic and vanillic acid. Synthesised compounds are evaluated for antioxidant activity using three in vitro assays (DPPH radical scavenging assay, DSC studies for oxidative induction temperature of linoleic acid and autoxidation of linoleic acid in Tween 20 micellar medium) and compared with three widely used antioxidants in the food industry, BHT, α-tocopherol, and dodecyl gallate. Synthesised compounds are found to exhibit good antiradical activity. These compounds also exhibited very good antifungal activity against studied fungal strains. All these results suggested the applicability of the synthesised compounds as potent lipophilic antioxidants for combating oxidative stress.     

Analysis of non-coloured phenolics in red wine: Effect of Dekkera bruxellensis yeast

The non-coloured phenolic composition of red wine obtained from Touriga Nacional grapes growing in the Dão region (Portugal) was determined by HPLC/DAD and 16 compounds were identified and quantified. The effect of nine different Dekkera bruxellensis strains on the levels of phenolic acids and flavonoids of the wine were also evaluated. In all samples (inoculated and non inoculated), phenolic acids predominated over flavonoids. In inoculated samples a considerable rise in the amounts of gallic acid was observed (about 33%) while the amounts of t-CAFTA, t-COUTA, caffeic and p-coumaric acids suffered a statistically significant decrease.     

Isolation and structure elucidation of phenolic compounds in Chinese olive (<Emphasis Type="Italic">Canarium album</Emphasis> L.) fruit

Chinese olive (Canarium album L.), one native and well-known tropical fruit tree in the southeast of China, contain a large amount of phenolics and possess great pharmacological activities. In this study, phenolics were extracted from Chinese olive fruit using 80% (v/v) aqueous acetone, and seven phenolic compounds were isolated and purified by Polyamide column and Toyopearl HW-40 column chromatography from crude extracts. Their structures were elucidated by high performance liquid chromatography coupled to diode array detection and electrospray ionization mass spectrometry (HPLC-DAD-ESI-MS), and where possible by ¹H NMR and ¹³C NMR spectrometry. Except gallic acid and hyperin, five phenolic compounds including methyl gallate, ethyl gallate, corilagin, kaempferol-3-glucoside and amentoflavone were first identified in Chinese olive.     

ANTIOXIDANT ACTIVITY OF WHITE WINE EXTRACTS AND SOME PHENOLIC ACIDS TOWARD CORN OIL OXIDATION

The ability of white wine extracts and some phenolic acids to inhibit corn oil oxidation was evaluated by monitoring the absorbance at 234 nm and the p-anisidine value. Wine extracts, each at 120 mg/L total phenolics, were tested in corn oil stripped of tocopherols kept at 50C. Two extracts rich in phenolic acids significantly inhibited oil oxidation, the most active being equivalent to butylated hydroxyanisol (BHA) at 200 mg/L. Moreover, four phenolic acids were tested in corn oil stripped of tocopherols kept at 50C and in edible corn oil kept at 110C. Gallic acid and caffeic acid, each at 80 mg/L, appeared to be stronger antioxidants than BHA at 200 mg/L. The present results indicate that some white wine phenolics, such as phenolic acids, may be strong antioxidants in corn oil. Moreover, the antioxidant activity of gallic acid and caffeic acid indicates their potential application in corn oil.

PRACTICAL APPLICATIONS

The antioxidant activity of white wine phenolic extracts toward corn oil oxidation and the characterization of active phenolic compounds may be useful in developing nutrient antioxidants for corn oil. Moreover, the antioxidant activity of gallic acid and caffeic acid indicates their potential application in corn oil.