Reaction of apple pectin with ammonia

Amidated apple pectins were prepared in aqueous and aqueous-alcoholic media, and the factors influencing both ammonolysis and hydrolysis of the ester groups of apple pectin by ammonia were studied.
In aqueous solution, the extent of ammonolysis and hydrolysis was very dependent upon ammonia concentration. In aqueous alcohol, the products were also dependent upon the particular alcohol and its concentration, increasing concentrations suppressing hydrolysis; the overall rate of reaction is much slower than in the absence of alcohol. The product balance between amonolysis and hydrolysis of the ester groups is influenced by the polarity of the medium, and by the concentration of NH⁺₄ and OH⁻.
It was shown that the extents of ammonolysis and hydrolysis of the ester groups are significantly affected by the polarity of the medium and the acid-base equilibrium of the interaction of ammonia with water, i.e. NH₃+H₂O ↔ NH⁺₄+OH⁻. The content of free ester and amidated carboxyl groups in the pectin can be regulated by varying the concentration of ammonia, temperature, reaction time and the polarity of the medium. The rate of enzymic hydrolysis of amidated pectins decreases with increasing amide content. A mechanism for the reaction is proposed.     

Antihypertensive peptides from whey proteins fermented by lactic acid bacteria

Food Science and Biotechnology - In this study, whey proteins were fermented with 34 lactic acid bacteria for 48 h at 37 °C and their ability to inhibit angiotensin...     

苹果汁中与混浊有关的活性蛋白质的分离纯化

采用疏水作用色谱PhenylSepharoseCL-4B和固相萃取C18柱分离纯化了浓缩苹果汁和鲜榨苹果汁中与混浊有关的活性蛋白质。经过SDS-PAGE,结果表明浓缩苹果汁和鲜榨苹果汁中的活性蛋白质为同一类蛋白,均为疏水性蛋白质,其分子量为36kDa。     

豆腐酸浆老汤中产酸菌的分离筛选与鉴定

利用MRS选择性培养基,从分别采自山东邹平、陕西榆林、河北涞源、云南石屏、云南建水及云南元阳的6份酸浆老汤中分离纯化出12种菌落形态不一样的乳酸菌,分别采用MC改良培养基和发酵黄浆水p H及产酸量的监测对12种菌进行初筛及复筛,筛选出一株产酸较快、较高的乳酸菌,并对该菌株进行了菌落形态、生理生化及16S r DNA序列分析的鉴定实验,结果表明:该菌株为革兰氏阳性菌,菌落特征为白色,圆形,表面突起,不透明,边缘整齐,镜检发现该菌为球菌、直径为0.8μm,四联或成串排列,经生理生化及16S r DNA序列鉴定确定该菌为肠球菌属中的屎肠球菌(Enterococcus faecium),遗传稳定性较好。     

苹果果胶研究进展

苹果(Malus pumila Mill.)是我国北方广泛种植的经济作物。近年来,很多学者从苹果渣中提取分离得到苹果果胶。现代医学和生物学研究表明,苹果果胶具有抗氧化、降血脂、抗菌及抗癌的功效。本文介绍了苹果果胶的结构、组分分级与特性研究,着重阐述了目前国内外对于苹果果胶的提取、分离的最新进展及其生物活性,讨论了目前研究中存在的问题,并对其发展前景进行了展望。     

Effects of fermentation by lactic acid bacteria on the antigenicity of bovine whey proteins

BACKGROUND: The main whey proteins α‐lactalbumin (α‐LA) and β‐lactoglobulin (β‐LG) are considered as the major allergens in cow's milk. Microbial fermentation can produce some proteolytic enzymes, which can induce the degradation of milk protein allergens. In this study, the effects of fermentation by lactic acid bacteria on the antigenicity of α‐LA and β‐LG were investigated using indirect competitive ELISA. Meanwhile, the proteolysis of milk proteins was detected by TNBS assay and SDS‐PAGE electrophoresis. RESULTS: Fermentation by lactic acid bacteria could significantly reduce the antigenicity of α‐LA and β‐LG in skim milk. Combined strains of Lactobacillus helveticus and Streptococcus thermophilus were the most effective in reducing the antigenicity of both whey proteins. In addition, α‐LA and β‐LG antigenicity decreased to a lower value at 6 h of fermentation and at 0.5 d of cold storage by fermentation with the combined strains. The results of TNBS assay and SDS‐PAGE electrophoresis showed that lactic acid bacteria strains used in this study hydrolysed whey proteins only to a limited extent. CONCLUSION: The fermentation with lactic acid bacteria is an effective way to reduce whey proteins antigenicity. Copyright © 2010 Society of Chemical Industry     

豆腐酸浆老汤中产酸菌的分离筛选与鉴定

利用MRS选择性培养基,从分别采自山东邹平、陕西榆林、河北涞源、云南石屏、云南建水及云南元阳的6份酸浆老汤中分离纯化出12种菌落形态不一样的乳酸菌,分别采用MC改良培养基和发酵黄浆水p H及产酸量的监测对12种菌进行初筛及复筛,筛选出一株产酸较快、较高的乳酸菌,并对该菌株进行了菌落形态、生理生化及16S r DNA序列分析的鉴定实验,结果表明:该菌株为革兰氏阳性菌,菌落特征为白色,圆形,表面突起,不透明,边缘整齐,镜检发现该菌为球菌、直径为0.8μm,四联或成串排列,经生理生化及16S r DNA序列鉴定确定该菌为肠球菌属中的屎肠球菌(Enterococcus faecium),遗传稳定性较好。      

Isolation of caseins from whey proteins by microfiltration modifying the mineral balance in skim milk

The objective of this work was to study the effect of different salts and salt concentration on the isolation of casein micelles from bovine raw skim milk by tangential flow microfiltration. Tangential flow microfiltration (0.22 μm) was conducted in a continuous process adding a modified buffer to maintain a constant initial sample volume. This buffer contained calcium chloride (CaCl₂), sodium phosphate (Na₂HPO₄), or potassium citrate (K₃C₆H₅O₇) in concentrations ranging from 0 to 100 mM. The concentrations of caseins and whey proteins retained were determined by sodium dodecyl sulfate-PAGE and analyzed using the Scion Image software (Scion Corporation, Frederick, MD). A complete isolation of caseins from whey proteins was achieved using sodium phosphate in the range of 10 to 50 mM and 20 times the initial volume of buffer added. No whey proteins were detected at 50 mM but this was at the expense of low caseins being retained. When lower sodium phosphate concentrations were used, the amount of caseins retained was higher but a small amount of whey proteins were still detected by sodium dodecyl sulfate-PAGE. Among the salts tested, calcium chloride at 50 mM and all volumes of buffer showed the higher retention of casein proteins. The highest casein:whey protein ratio was found at 30 mM CaCl₂, but no complete casein micelle isolation was achieved. Potassium citrate was the most ineffective salt because a rapid loss of caseins and whey proteins was observed at all concentrations and with low quantities of buffer added during the filtration process. Our results show the potential of altering the mineral balance in milk for isolation of casein micelles from whey proteins in a continuous tangential flow microfiltration system.     

Bioutilisation of whey for lactic acid production

The disposal of whey, the liquid remaining after the separation of milk fat and casein from whole milk, is a major problem for the dairy industry, which demands simple and economical solutions. The bioconversion of lactose present in whey to valuable products has been actively explored. Since whey and whey permeates contain significant quantities of lactose, an interesting way to upgrade this effluent could be as a substrate for fermentation. Production of lactic acid through lactic acid bacteria could be a processing route for whey lactose and various attempts have been made in this direction. Immobilised cell technology has also been applied to whey fermentation processes, to improve the economics of the process. A fermentative means of lactic acid production has advantages over chemical synthesis, as desirable optically pure lactic acid could be produced, and the demand for optically pure lactic acid has increased considerably because of its use in the production of poly(lactic acid), a biodegradable polymer, and other industrial applications. This review focuses on the various biotechnological techniques that have used whey for the production of lactic acid.     

从苹果渣中提取果胶的工艺研究

探讨了以苹果渣为原料提取果胶的最佳工艺条件,实验结果表明萃取液的pH值为2.0、料液比为1∶13、温度为85℃、水解时间为1.5h为提取果胶的最佳工艺条件,产率达14.04%。     

Isolation of bovine immunoglobulins and lactoferrin from whey proteins by gel filtration techniques

Whey is a suitable source of immunoglobulins and lactoferrin to enrich infant formulas. Gel filtration on Sephacryl S-300 and on Fractogel TSK HW-55 was used to isolate immunoglobulins from colostral whey, acid whey, and Cheddar cheese whey. The SDS-PAGE and immunoelectrophoresis techniques indicated that the purity of the fractions from fractionation on Sephacryl S-300 was better than that by fractionation on TSK HW-55 column. Biological activity of fractions from the Sephacryl S-300 column as assessed by immunochemical analysis was 99, 83.3, and 92% for colostral, acid, and sweet wheys. The well-proven antimicrobial agent, lactoferrin, was isolated from sweet whey by heparin-attached Sepharose. Lactoferrin selectively adsorbed to the column was subsequently eluted with 5 mM Veronal-HCl containing .5 M NaCl, pH 7.4. Purity of the isolated protein was confirmed by SDS-PAGE and immunoelectrophoresis.     

Rheological properties of whey protein isolate stabilized emulsions with pectin and guar gum

Dynamic oscillatory and steady-shear rheological tests were carried out to evaluate the rheological properties of whey protein isolate (WPI) stabilized emulsions with and without hydrocolloids (pectin and guar gum) at pH 7.0. Viscosity and also consistency index of emulsions increased with hydrocolloid concentration. At γ = 20 s⁻¹, the value of viscosity of the emulsion with 0.5% (w/v) pectin was about fivefold higher than that of the emulsion without pectin. Flow curves were analyzed using power law model through a fitting procedure. Flow behaviour index of all emulsions except for containing 0.5% (w/v) guar gum was approximately in the range of 0.9–1.0, which corresponds to near-Newtonian behaviour. The shear thinning behaviour of emulsions containing 0.5% (w/w) guar gum was confirmed by flow behaviour index, n, of 0.396. Both storage (G′) and loss modulus (G″) increased with an increase in frequency. Emulsions behaved like a liquid with G″ > G′ at lower frequencies; and like an elastic solid with G′ > G″ at higher frequencies. Effect of guar gum was more pronounced on dynamic properties. Phase angle values decreased from 89 to <10° with increasing frequency and indicated the viscoelasticity of WPI-stabilized emulsions with and without pectin/guar gum.     

Properties of whey proteins obtained from different whey streams

The physio-chemical characteristics of whey proteins (WPs) in sweet, salty, native and acid whey were investigated and compared. WPs from acid whey were characterised by hydrophobically and covalently driven protein aggregation. Covalent aggregation in acid whey consisted of both thiol/disulphide and non-thiol/disulphide mediated reactions. Fourier transform infrared data characterised this protein aggregation as a β-sheet attraction causing subtle changes in the secondary structure. In contrast, WPs in salty whey aggregated via van der Waals, hydrogen, electrostatic interactions and covalent bonds. Both thiol/disulphide and non-thiol/disulphide interactions led to cross-linked β-sheets, disrupting the secondary protein structures. This aggregation exposed hydrophobic segments while oxidising a high number of free thiol groups. The absence of these types of WP aggregation in sweet or native whey highlighted the fact that elevated salt concentration in salty whey or heat treatment applied during production of acid whey are largely responsible for structural differences.     

Hydrolysis of apple pectin by the coordinated activity of pectic enzymes

The hydrolysis of pectin from apples, cv. Budimka fruit (Pyrus mahus L.), by individual and/or combined action of fungal polygalacturonase from Aspergillus niger (FPG), fungal pectin esterase from A. niger (FPE) and plant tomato pectin esterase (PPE) was studied. The optimum pH values for individual actions of FPG, FPE and PPE on 1% apple pectin were determined to be 4.5, 3.5 and 6.5, and the optimum temperatures were 40, 45, and in the range 45–50 °C, respectively. FPE was found to be more efficient for the hydrolysis of the apple pectin than was PPE from tomato. By measuring the initial velocities on 1% apple pectin it was confirmed that the PG expressed no effect on the PE activity. By using the combination of FPG (162 U/l) and FPE (60 U/l), e.g., in a respective ratio of 2.5, an efficient pectin degradation process, with a viscosity reduction of η/η₀ = 1.05, could be reached in less than 2 h. This process produced about 160 ppm of methanol in the pectin digest. The long term hydrolysis reaction of the apple pectin with FPG (162 U/l) and FPE (27 U/l) achieved a degree of hydrolysis of around 29% after 12 h and consisted mostly of trimers (28.4%).     

苹果果胶膜法分离工艺研究

对苹果果胶超滤法分离条件和不同分子量的果胶与其性质的关系进行研究,采用5种不同截流分子量(MWCO)的超滤膜分离苹果果胶.结果表明:在跨膜压差为0.08MPa,料液浓度为1g/L,室温下,不同分子量的果肢获得了良好的分离.分子量与其理化性质有着对应的关系,分子量越大,其半乳糖醛酸含量和酯化度均越高,透光率减小.同时.超滤时果胶液的脱色有良好作用.本实验为工业上果胶的纯化、分离提供理论依据.     

Formation and characterization of noncovalent ternary complexes based on whey protein concentrate,high methoxyl pectin,and phenolic acid

Protein-polysaccharide-polyphenol noncovalent ternary complexes possess unique physicochemical, structural, and functional properties. In the present study, ternary complexes based on whey protein concentrate (WPC; 2%, wt/vol) and high methoxyl pectin (HMP; 0.5%, wt/vol) complexes and 0.2 to 0.6% (wt/vol) chlorogenic acid (CA) or rosmarinic acid (RA) were formed and characterized at 3 pH values (4, 4.5, and 5). The pH conditions were decided according to phase diagram of WPC and HMP during acidification. Fluorescence quenching experiments indicated that WPC-HMP complexes bound RA stronger than CA and the binding constant increased with increasing pH for both phenolic acids. Particle size of ternary complexes decreased and absolute ζ-potential increased with pH values changing from 4 to 5, and RA influenced the particle size of WPC-HMP complexes greater than CA. The CA and RA in ternary complexes showed good stability against UV light with pH order of pH 5 > pH 4.5 > pH 4. Fourier-transform infrared spectroscopy spectra indicated the involvement of hydrogen bonding between WPC-HMP and CA or RA. Antibacterial tests showed that ternary complexes had good antibacterial activity against Staphylococcus aureus and Escherichia coli at concentrations of 6.2 mg/mL and the ability increased with decreasing pH values. All ternary complexes possessed strong scavenging radical capacities with median inhibitory concentration (IC₅₀) values ranging from 2.71 ± 0.05 to 6.20 ± 0.41 μg/mL. Antioxidative ability of ternary complexes increased as pH went up and WPC-HMP-RA showed significantly higher antioxidative property compared with WPC-HMP-CA. Data may provide useful information for rational design of ternary complexes and applications of the formed complexes in food matrices such as beverages and emulsions.     

Functionality of extrusion--texturized whey proteins

Whey, a byproduct of the cheesemaking process, is concentrated by processors to make whey protein concentrates (WPC) and isolates (WPI). Only 50% of whey proteins are used in foods. In order to increase their usage, texturizing WPC, WPI, and whey albumin is proposed to create ingredients with new functionality. Extrusion processing texturizes globular proteins by shearing and stretching them into aligned or entangled fibrous bundles. In this study, WPC, WPI, and whey albumin were extruded in a twin screw extruder at approximately 38% moisture content (15.2 ml/min, feed rate 25 g/min) and, at different extrusion cook temperatures, at the same temperature for the last four zones before the die (35, 50, 75, and 100 degrees C, respectively). Protein solubility, gelation, foaming, and digestibility were determined in extrudates. Degree of extrusion-induced insolubility (denaturation) or texturization, determined by lack of solubility at pH 7 for WPI, increased from 30 to 60, 85, and 95% for the four temperature conditions 35, 50, 75, and 100 degrees C, respectively. Gel strength of extruded isolates increased initially 115% (35 degrees C) and 145% (50 degrees C), but gel strength was lost at 75 and 100 degrees C. Denaturation at these melt temperatures had minimal effect on foaming and digestibility. Varying extrusion cook temperature allowed a new controlled rate of denaturation, indicating that a texturized ingredient with a predetermined functionality based on degree of denaturation can be created.     

The chemical characteristics of apple pectin influence its fermentability in vitro

The aim of the present study was to characterize the fermentability of different apple pectins, as soluble dietary fibres, in vitro. High- and low-methoxyl pectins were fermented completely and very fast by human colonic bacteria, whereas pectic acid and amidated pectin were degraded more slowly. The main fermentation metabolites, i.e. short-chain fatty acids (SCFA) and gases, were produced in lower amounts from pectic acid and amidated pectin. Based on the higher production of SCFA, pectins with a higher degree of methoxylation might be the best candidates for food enrichment due to a beneficial effect on colonic health. On the other hand, the longer persistence of the slowly degraded pectins in the gut may be of advantage as binding and diluting agents for carcinogens.     

乳清分离蛋白酶解物的分离条件

采用试管法研究了SP Sephadex C-25对乳源抗氧化肽的分离条件,确定SP Scphadex C-25色谱分离条件为:上样量3mg/mL IE;起始缓冲液:20mmol/L的醋酸缓冲液(pH4.0);NaCI浓度为Imol/L.