Effect of pH on retention of aroma compounds by beta-lactoglobulin

Interactions between volatile compounds and BLG in aqueous solution were studied using static and dynamic headspace techniques (exponential dilution). The intensity of interactions between methyl ketones (C7-C9), ethyl esters (C6-C9), limonene, myrcene, and beta-lactoglobulin (BLG) were estimated by determination of the relative infinite dilution activity coefficients (gamma(r)). For a constant pH value, the methyl ketones retention by BLG increased significantly with the hydrophobicity of the volatiles, whereas the retention reached a maximum for ethyl octanoate in the ester series, indicating a possible steric hindrance. For limonene and myrcene an unexpected increase in headspace concentration or "salting out" effect was noticed for acid pH. The variations of the retention according to the pH increase of the medium from pH 3 to pH 11 could be related to structural modifications of the BLG. The retention increase observed between pH 3 and pH 9 resulted from the flexibility modification of the protein, allowing better accessibility to the primary or the secondary hydrophobic sites, whereas the dramatic decrease observed at pH 11 was the consequence of the alkaline denaturation of BLG. Electrostatic interactions occurring at pH 7.5 could also explain the observed retention increase.     

Transfer of aroma compounds in water-lipid systems: binding tendency of beta-lactoglobulin

Interactions of volatile aroma compounds with protein in aqueous solutions, especially whey proteins, have received significant attention in recent years. This work attempts to improve our understanding of the mass transfer in multiphasic systems, such as emulsions at the lipid-water interface, and to reveal the role of beta-lactoglobulin in the release rate of solutes. For this purpose the rotating diffusion cell has been used. From a practical point of view it enables evaluation of the transfer through the aqueous phase, through the oil and the interfacial transfer. The effect of beta-lactoglobulin, medium pH, and solute concentration has been investigated. Benzaldehyde and 2-nonanone have been studied, and miglyol has been chosen as an oil phase. It has been demonstrated that mass transfer has a rate-limiting step, which depends on physicochemical parameters such as hydrophobicity of the volatile, diffusion and partition coefficients, and rheological properties of the aqueous phase.     

Determination of apparent binding constants for aroma compounds with beta-lactoglobulin by dynamic coupled column liquid chromatography

Apparent binding constants of aroma compounds limonene, alpha- and beta-ionone, and terpenyl acetate, with beta-lactoglobulin (BLG), were determined, using dynamic coupled column liquid chromatography, for pH values varying from 3 to 11. K(a) values varied from 2.61 to 3.21 x 10(3) M(-1) for limonene, indicating a strong interaction with BLG. Similarly, significant and close apparent binding constants were obtained for alpha- and beta-ionone, 1.7 x 10(2) and 4.5 to 5.4 x 10(2) M(-1), respectively. These data indicated that a similar mechanism is involved for the binding of these two molecules. The weaker values obtained at low pH, for alpha-ionone relative to beta-ionone, can be explained by the existence of steric hindrance. An increase of the apparent binding constant was observed, for all the compounds studied, when the pH was increased from 3 to 9. At this pH, an apparent binding constant was obtained for terpenyl acetate (1.04 x 10(2) M(-1)), whereas this determination was not possible at pH 3 and 6. The apparent binding constant increase was in agreement with the decrease of aroma compound relative activity coefficient in the presence of BLG, previously observed at this pH. It indicated a best accessibility to the same binding site. The binding constants of all the aroma compounds studied decreased at pH 11 as a result of the important release of the BLG structure previously reported.     

Interactions between beta-lactoglobulin and aroma compounds: different binding behaviors as a function of ligand structure

Interactions between beta-lactoglobulin (BLG) in its monomeric form and a wide range of aroma compounds were investigated by Fourier transform infrared (FT-IR) and 2D nuclear magnetic resonance (NMR) spectroscopies. A screening of the ligands was carried out by FT-IR through the amide I region changes of BLG upon binding. The location of two binding sites was determined by 2D NMR from the study of 10 selected ligands with different structures. All of the data suggest at least two binding behaviors as a function of the chemical class, the hydrophobicity, or the structure of the ligands. The binding of the elongated aroma compounds, such as 2-nonanone or ethyl pentanoate, within the central cavity involves residues located at the entrance of the calyx and Trp19. The binding onto the protein surface of aroma compounds that have or adopt a compact structure occurs in a site located between strand beta-G, alpha helix, and strand beta-I.     

Effects of high-pressure processing at low temperature on the molecular structure and surface properties of beta-lactoglobulin

High-pressure processing (HPP) was utilized to induce unfolding of beta-lactoglobulin (beta-LG). beta-Lactoglobulin solutions at concentrations of 0.5 mg/mL, in pH 7.5 phosphate buffer, were pressure treated at 510 MPa for 10 min at either 8 or 24 degrees C. The secondary structure, as determined by circular dichroism (CD), of beta-LG processed at 8 degrees C appeared to be unchanged, whereas beta-LG processed at 24 degrees C lost alpha-helix structure. Tertiary structures for beta-LG, as determined by near-UV CD, intrinsic protein fluorescence spectroscopy, hydrophobic fluorescent probe binding, and thiol group reactivity, were changed following processing at either temperature. The largest changes to tertiary structure were observed for the samples processed at 24 degrees C. Model solutions containing the pressure-treated beta-LG showed significant decreases in surface tension at liquid-air interfaces with values of 54.00 and 51.69 mN/m for the samples treated at 24 and 8 degrees C, respectively. In comparison, the surface tension for model solutions containing the untreated control was 60.60 mN/m. Changes in protein structure during frozen and freeze-dried storage were also monitored, and some renaturation was observed for both storage conditions. Significantly, the sample pressure-treated at 8 degrees C continued to display the lowest surface tension.     

Competitive binding of aroma compounds by beta-cyclodextrin.

Retention of six aroma compounds has been studied after dehydration of ternary mixtures of aroma water and beta-cyclodextrin. A maximal retention of a mole of aroma per mole of beta-cyclodextrin has been observed for five of the aroma compounds, whereas retention of benzyl alcohol can be twice as high. Retention of a mixture of aroma compounds has also been studied. It has been noted that when volatile compounds compete for the same binding sites on beta-cyclodextrin, ethyl hexanoate, 2-methylbutyric acid, and benzyl alcohol are, respectively, better retained than ethyl propionate, hexanoic acid, and hexanol. Preferential retention observed with esters can be simply explained by their difference of physicochemical properties, but for the acids and alcohols a study at the molecular scale has been necessary. The better retention of 2-methylbutyric acid can be explained by differences in the nature of interaction between the acids and their carrier. At least selectivity of retention noted for the alcohol could be due to a difference in the location of the guest and also a difference in the number of aroma molecules that can be bound per polysaccharide molecule.     

Ethanol effect on the structure of beta-lactoglobulin b and its ligand binding

The changes of structure and ligand binding properties of beta-LG B have been studied by fluorescence and circular dichroism spectroscopy in ethanolic solutions. Fluorescence measurements of retinol/beta-LG interactions at 480 nm in various ethanol concentrations show that the maximal fluorescence intensity induced by this interaction between retinol and beta-LG is observed around 20% v/v of ethanol. It is reduced to zero at 40% and 50% of ethanol. These results suggest that there are two distinct structural changes in beta-LG occurring between 20% and 30% and around 40% of ethanol. The first transition, which increases affinity and the apparent number of binding sites for retinol, may be related or similar to the Tanford transition. The strong quenching of retinol emission at 480 nm in 40% of ethanol indicates the radical transformation of beta-LG tertiary structure and the release of retinol. CD spectra at the aromatic region show that secondary and tertiary structures of beta-LG are not significantly affected between 0% and 20% of ethanol. In 30% of ethanol, beta-sheet percentage of beta-LG decreases with respect to native beta-LG (from 55% to 46%). beta-Sheet percentage in beta-LG increases in 40% and 50% alcohol (51% and 53%) relative to 30% of ethanol, which also indicates the strong rearrangement of the secondary structure of beta-LG, while its tertiary structure and beta-LG interactions are radically changed.     

精准温度控制对枸杞鲜果贮藏品质和香气成分的影响

为研究精准温控技术在枸杞鲜果保鲜中的应用效果,将枸杞分别装入泡沫箱（Control Check,CK）、泡沫箱+蓄冷剂（冰温）和精准温控箱+蓄冷剂（相温）中,记录箱内温度,结合贮藏期间枸杞的品质变化,通过主成分分析法对3组枸杞进行综合评价,并对不同处理香气成分进行分析。结果表明,精准温度控制后箱体内部温度更低,且温度波动保持在0.1 ℃以内,CK组、冰温组、相温组枸杞出现发霉腐烂现象的时间分别为10、20、40 d;贮藏40 d后,相温组枸杞的腐烂率为4.11%,远低于CK组的14.85%,色差低于3,亮度达34.12,可溶性固形物含量、可滴定酸含量、维生素C含量、类胡萝卜素含量分别较CK组高0.65个百分点、0.03个百分点、4.34 mg/100g、2.90 mg/g,相温组枸杞有效抑制腐烂率和失重率的增加,延缓色差的升高和亮度的降低,保持较高的可溶性固形物、可滴定酸、维生素C和类胡萝卜素含量,经主成分得分分析后得到3组枸杞的品质排序为相温组、冰温组、CK组,且精准温度控制贮藏的枸杞醛类、萜类等有利香气成分相对含量更高。因此,精准温度控制技术有利于枸杞鲜果的冷藏保鲜,其中相温贮藏效果更好。研究结果为枸杞鲜果精准温控保鲜提供参考。     

Modification of IgE binding to beta-lactoglobulin by fermentation and proteolysis of cow's milk

The effect of fermentation by Lactobacilli and of proteolytic hydrolysis of whole milk on the IgE binding ability of beta-lactoglobulin was studied using an ELISA inhibition assay. Sera from nine adult milk allergic patients were tested. The individual sera showed a similar inhibition pattern in the changes during fermentation and proteolysis. The degradation of beta-lactoglobulin was studied with liquid chromatography. In general, fermentation with Lactobacilli gave little effect on IgE binding, even though chromatography data showed a gradual degradation of beta-lactoglobulin. Proteolysis with trypsin, however, gave extensive degradation of beta-lactoglobulin and strongly decreased IgE binding. In addition, we measured the inhibition pattern of beta-lactoglobulin in various selected commercially available fermented milk products. These showed an IgE binding capacity similar to that of nonfermented high pasteurized milk.     

Modeling the partition of volatile aroma compounds from a cloud emulsion

Parameters determining the partitioning behavior of volatile compounds between a cloud emulsion and the gas phase were measured under static equilibrium headspace conditions, using volatiles (e.g., ethyl hexanoate, cymene, and octanol) representing different volatilities and different degrees of hydrophobicity. The significant factors were the molecular characteristics of the volatile and the concentration of the oil phase. The nature of the lipid (C8 and C12 triglycerides), particle size, and emulsifier type (modified starch and gum arabic) did not significantly alter volatile partitioning. An empirical model based on the partition behavior and physicochemical parameters of 67 volatile compounds was produced. This predicted the partition of volatiles (R(2) = 0.83) in cloud emulsions as a function of lipid content. The significant terms (P < 0.05) in the empirical model were Log P, Log solubility, the dipole vector, and the oil fraction.     

Effect of storage on some volatile aroma compounds in fresh-cut cantaloupe melon

Changes in volatile aroma constituents of fresh-cut cantaloupe melon with storage were determined by headspace solid-phase microextraction gas chromatography-mass spectrometry. The compounds isolated from the fruit immediately after cutting were predominantly aliphatic and aromatic esters. Storage of fruit at 4 degrees C caused a considerable decrease in concentration of esters and synthesis of the terpenoid compounds beta-ionone and geranylacetone over a period of 24 h. This change in the volatile profile with storage is consistent with that of a stress-induced defense response in the cut fruit as an adaptation process to tissue exposure and cell disruption. The same effect occurred in fruit stored at 22 degrees C and in those treated with sodium azide and ascorbic acid prior to storage. Fruit treated with ascorbic acid and sodium azide had higher concentrations of beta-ionone and geranylacetone and retained these compounds better with storage time. The reduction of esters appears to be an important early reaction step in the loss of freshness during storage of fresh-cut cantaloupe.     

Influence of temperature, modified atmosphere packaging, and heat treatment on aroma compounds in broccoli

The aroma compounds in broccoli stored in different modified atmospheres were studied. The packaging materials used were oriented polypropylene (OPP), poly(vinyl chloride) (PVC), and low-density polyethylene (LDPE) containing an ethylene-absorbing sachet. All samples were stored for either 1 week at a constant temperature of 10 degrees C or for 3 days at 4 degrees C, followed by 4 days at 10 degrees C. The atmospheres that developed inside the packaging materials differed significantly. The broccoli samples were analyzed raw and after cooking, with regard to volatile compounds, using gas-phase (headspace) extraction followed by gas chromatography-mass spectrometry (GC-MS) and GC-olfactometry. Dimethyl sulfide (DMS), dimethyl disulfide (DMDS), dimethyl trisulfide (DMTS), hexanal, 3-cis-hexen-1-ol, nonanal, ethanol, and a group of thiocyanates were selected for a detailed study because these compounds cause off-odor and can be used as indicators of stress. Significant differences were found in the aroma profiles of the broccoli samples relative to the packaging materials used for storage. Storage in OPP (14% O(2), 10.5% CO(2)) resulted in most of the off-odors, while storage in LDPE (6% O(2), 7% CO(2)) and PVC (17.9% O(2), 4% CO(2)) was found to maintain the concentration of DMS, DMDS, and DMTS during storage. Heat treatment of the broccoli increased the content of aroma compounds as well as the number of compounds containing sulfur.     

Characterization of some Allium hybrids by aroma precursors, aroma profiles, and alliinase activity

Various Allium hybrids, obtained by the crossbreeding of Allium cepa (onion) as the mother plant and six taxonomically distant wild species obtained by embryo rescue, were investigated with special respect to their individual profiles of cysteine sulfoxides as well as enzymically and nonenzymically formed aroma substances. Alliinase (EC 4.4.1.4) catalyzes the conversion of odorless (+)-S-alk(en)yl-L-cysteine sulfoxides into volatile thiosulfinates. These thiosulfinates were converted to a variety of sulfides by steam distillation. SPME-gas chromatography (GC) and high-performance liquid chromatography (HPLC) used for the analysis of aroma components and their precursors permitted a high sample throughput, so that numerous gene bank accessions and Allium breeding materials were analyzed within a comparatively short time. Cysteine sulfoxides as well as alliinase activity were found in all investigated samples at different levels, but (+)-S-methyl-L-cysteine sulfoxide (methiin) was the most abundant sulfoxide present. (+)-S-(trans-1-Propenyl)-L-cysteine sulfoxide (isoalliin) is typical for onion and was found in all investigated hybrids. The pattern of the other cysteine sulfoxides depended strongly on the parent plants used. The profile of aroma components corresponded with the related pattern of aroma precursors (cysteine sulfoxides). Successful hybridization was proven by randomly amplified polymorphic DNA analysis. Together with these established marker techniques, HPLC and SPME-GC analysis provide support to breeding projects designed to improve the health and aroma properties of Allium hybrids.     

Effects of heat treatment and pectin addition on beta-lactoglobulin allergenicity

The specific effects of heat treatment and/or addition of low/high-methylated pectin (LMP/HMP) on the allergenicity of beta-lactoglobulin (beta-Lg) and its hydrolysis products were investigated through a two-step in vitro digestion approach. beta-Lg was first hydrolyzed by pepsin and then by a trypsin/chymotrypsin (T/C) mixture done in a dialysis bag with a molecular weight cutoff of 1000. The protein digestion was followed by SDS-PAGE electrophoresis performed on each digestion product, and their in vitro allergenicity was analyzed by immunoblotting. Such procedure was applied on beta-Lg samples mixed with the two kinds of pectin before or after heating (80 degrees C, 25 min) to determine the respective impact of heat treatment and pectin addition. Heat denaturation improved significantly the susceptibility of beta-Lg against the pepsin and the T/C. This effect, which was coupled to a reduction in immunoreactivity of the digested beta-Lg, appeared to be distinctively modulated by LMP and HMP. Through nonspecific interaction with the beta-Lg, pectin could reduce the accessibility of cleavage sites and/or epitope sequences. This mechanism of action is discussed in relation to the intra- and intermolecular interactions between beta-Lg and pectin initiated under the experimental conditions.     

Effect of 1-methylcyclopropene on volatile emission and aroma in cv. Anna apples

The rapidly ripening summer apple cultivar Anna was treated with 0.1 micro L(-1) and 1 microL L(-1) 1-methylcyclopropene (MCP) at harvest and kept at 20 degrees C, or stored for 5 weeks at 0 degrees C and then transferred to 20 degrees C. Total volatiles were not reduced by treatment with 0.1 microL L(-1) MCP, but were 70% lower in fruits treated with 1 microL L(-1) MCP than in untreated fruits. Ethylene production was 50% and 95% inhibited by 0.1 microL L(-1) and 1 microL L(-1) MCP, respectively. The volatiles produced by fruit at harvest were predominantly aldehydes and alcohols, with some acetate esters as well as 2-methyl butyl acetate and beta-damascenone. During ripening, the acetate and butyrate esters increased greatly and alcohols and aldehydes decreased. MCP-treated apples retained more alcohols, aldehydes, and beta-damascenone volatiles than did untreated apples. Sensory evaluation found that control and 0.1 microL L(-1) treated apples developed more fruity, ripe, and overall aromas, but the preference was for the 1 microL L(-1) treated apples with a less ripe aroma.     

Effects of caseins on thermal stability of bovine beta-lactoglobulin

Casein fractions have been shown to act as molecular chaperones and inhibit aggregation of whey proteins in dilute solutions (< or =1% w/v). We evaluated if this approach would stabilize protein solutions at higher concentration and thermal processing temperatures desired for beverage applications. Mixtures of beta-lactoglobulin (BLG) (6% w/v) with either beta-casein (BCN) (0.01-2% w/v) or alpha s-casein (ACN) (2% w/v) were adjusted to pH 6.0 and heated (70-90 degrees C) for 20 min, cooled, and then analyzed to determine the degree of aggregation. Aggregation was determined by solution turbidity as optical density (OD) at 400 or 600 nm. The addition of 0.05% (w/v) BCN or greater caused a drop in turbidity for solutions heated at 70-90 degrees C. In contrast, inhibition was observed in BLG-ACN mixtures at 70 degrees C but not at > or =75 degrees C. Moreover, prolonged heating (90 min) of BLG with 2% (w/v) BCN (pH 6.0) at 90 degrees C produced a clear solution while BLG-ACN solutions formed translucent gels after heating for 15 min. The weight-averaged molar mass and root-mean-square (rms) radius of soluble aggregates were determined by size exclusion chromatography in conjunction with multiangle laser light scattering (SEC-MALS). SEC-MALS confirmed the turbidity results by showing that the BLG-BCN mixture (8% w/v protein) produced aggregates with lower molar mass and smaller rms radius (majority 20-40 nm). These results showed that BCN is a feasible component to stabilize higher concentrations of whey proteins in beverages.     

戴尔有孢圆酵母调控晚采小芒森葡萄酒乙酸和香气

为降低甜型葡萄酒中的挥发酸,进一步增加其香气复杂度,该研究利用本土戴尔有孢圆酵母R12与酿酒酵母NX11424同时接种和顺序接种发酵,研究其对贺兰山东麓产区晚采小芒森葡萄酒乙酸及香气成分的调控.与酿酒酵母单菌发酵相比,本土戴尔有孢圆酵母与酿酒酵母按照5:1、20:1、50:1的不同菌体数量比例同时接种和间隔5 d顺序接...     

气象因子对高香气烤烟品种主要香味前体物含量的影响

为研究气象因子对高香气烤烟品种主要香味前体物质含量的影响,以烤烟品种‘云烟87’为对照,对高香气烤烟品种‘豫烟11号’在5个不同地点的主要香味前体物质含量进行了比较,并分别对2个品种的香味前体物质与气象因子进行了关联度分析。研究结果表明:‘豫烟11号’主要香味前体物质含量在每个地点均显著高于‘云烟87’。烟叶类胡萝卜素在低纬度地区的含量显著低于高纬度地区,多酚类物质的含量随着海拔降低、成熟期光照时数增加而逐渐增加,叶片腺毛分泌物整体上高纬度地区的含量显著高于低纬度地区,以河南卢氏的含量最高。2个品种的烟叶类胡萝卜素含量受成熟期日照时数、旺长期平均气温等光温因素影响较大,而烟叶大田生长期间日照时数的长短对多酚类物质含量影响较大。2个品种的烟叶石油醚提取物和腺毛分泌物含量与气候因素的灰色关联度趋势有较大差异,‘豫烟11号’烟叶石油醚提取物含量受烟叶生长期间的日照时数影响较大,‘云烟87’则受平均气温的影响较大。‘豫烟11号’烟叶腺毛分泌物含量受成熟期日照时数和旺长期平均气温等的影响较大,‘云烟87’则受伸根期平均气温、成熟期蒸发量和成熟期平均气温等的影响较大。高香气烤烟品种‘豫烟11号’在选择种植区域时,应充分考虑成熟期日照时数和旺长期平均气温对品质的影响。     

Effect of some fat replacers on the release of volatile aroma compounds from low-fat meat products

The effect of fat content and carbohydrate fat-replacers on the release of volatile odor compounds from beefburger, salami, and frankfurter has been investigated. The reduction in fat content in any of the three meat products studied resulted in a tendency toward an increase in the quantities of volatiles released in the headspace. Tapioca starch and maltodextrin appear to delay the release of certain classes of compounds selectively; for instance, tapioca starch appears to slow the release of some Maillard products while maltodextrin has a similar effect on terpenes. In contrast, oat fiber decreases the release of most of the compounds analyzed. Thus, the addition of carbohydrate fat-replacers to low-fat meat products could assist the flavor qualities of low-fat meat products by slowing down the release of odor compounds.