“黑美人”马铃薯多酚氧化酶的特性研究

以邻苯二酚为底物,在413nm处测定黑美人马铃薯多酚氧化酶(PPO)的活性,研究了温度、pH、底物浓度对其活性的影响,并建立了酶促褐变反应动力学方程,探讨了L-半胱氨酸、抗坏血酸、柠檬酸、EDTA及亚硫酸氢钠五种抑制剂对酶促褐变的抑制效果。结果表明:黑美人马铃薯多酚氧化酶最适反应pH为6.3;最适反应温度为35℃;酶促褐变反应动力学符合米氏方程描述的单底物酶促反应动力学,以邻苯二酚为底物,Km=0.0011mol/L,Vmax=142.23U/min.g;动力学方程为V=142.23[S]/(0.0011+[S]);五种抑制剂均对PPO酶促褐变具有抑制作用,其强弱依次为亚硫酸氢钠>L-半胱氨酸>抗坏血酸>EDTA>柠檬酸。     

冬瓜多酚氧化酶的特性及抑制剂研究

以邻苯二酚为底物,采用'分光光度法在420 nm 处测定冬瓜多酚氧化酶(PPO)的活性,研究了不同温度和 pH 值对其活性的影响,并建立了酶促褐变反应动力学方程,探讨了抗坏血酸、亚硫酸氢钠、柠檬酸和 EDTA-2Na 4 种抑制剂对冬瓜酶促褐变的抑制效果.结果表明,冬瓜多酚氧化酶的最适温度为20℃,最适pH值为6.0;冬瓜多酚氧化酶的米氏方程符合单底物酶促反应动力学,相应的动力学参数为 Kin=0.0118 mol/L,Vm=0.0586 U/min;化学抑制剂抗坏血酸、亚硫酸氢钠、柠檬酸和EDTA-2Na 对多酚氧化酶都有抑制作用,其活性分别为对照的25%、32%、62%和57%.     

芒果多酚氧化酶的特性及抑制研究

以邻苯二酚为底物,采用分光光度法在420nm处测定芒果多酚氧化酶(PPO)的活性,研究了温度、pH值、底物浓度对其活性的影响,并建立了酶促褐变反应动力学方程,探讨了L-半胱氨酸、抗坏血酸、亚硫酸氢钠、柠檬酸和醋酸五种抑制剂对芒果酶促褐变的抑制效果。结果表明,芒果多酚氧化酶的最适温度为50℃,最适pH值为6.86;酶促褐变反应动力学符合米氏方程所描述的单底物酶促反应动力学,以邻苯二酚为底物的最大反应速度Vmax=192(U/min·g),Km=0.0173mol/L,相应的动力学方程为v=192[S]/(0.0173+[S]);五种物质对该酶均表现出不同的抑制作用,抑制效果为:抗坏血酸>L-半胱氨酸>亚硫酸氢钠>柠檬酸>醋酸。     

茄子多酚氧化酶特性及控制研究

茄子多酚氧化酶(polyphenol oxidase,PPO)是引起酶促褐变,导致果实褐变的主要原因.采用匀浆法从茄子中提取多酚氧化酶粗液,以邻苯二酚为底物,在λ420nm处研究温度、pH值、底物浓度对茄子果实中PPO活性的影响,建立相应的酶促褐变反应动力学方程.探讨L-半胱氨酸、抗坏血酸、亚硫酸氢钠、柠檬酸4种抑制剂对茄子酶促褐变的抑制效果.结果表明,茄子多酚氧化酶的最适温度为30℃,最适pH值为6.0及8.5(可能存在同工酶),酶促褐变反应动力学符合米氏方程所描述的单底物酶促反应动力学,相应的动力学参数Km=0.0215 U/(min·g),Vmax=0.539U/(min·g),相应的动力学方程为v=0.539[S]/(0.0215+[S]);供试4种抑制剂防止果实褐变的效果由强到弱依次是:抗坏血酸、亚硫酸氢钠、L-半胱氨酸、柠檬酸.另外,茄子PPO对高温较为敏感,在100℃时加热105 s可钝化酶活性.     

西葫芦多酚氧化酶酶学特性研究

以邻苯二酚为底物,采用分光光度法对西葫芦多酚氧化酶(PPO)的酶学特性及不同抑制剂对多酚氧化酶活性的影响进行研究。结果表明,西葫芦多酚氧化酶最适pH为6.6,最适温度为35℃,90℃处理5min可使该酶失活。多酚氧化酶催化的酶促褐变反应动力学符合米氏方程,动力学参数为Km=0.0417mol/L,Vmax=208.33U。四种抑制剂对PPO的抑制效果由强到弱依次为:亚硫酸氢钠>L-半胱氨酸>抗坏血酸>柠檬酸。     

砀山酥梨多酚氧化酶酶学特性及抑制效应的研究

多酚氧化酶是酶促褐变的关键酶,其特性与对其抑制效应研究一直是果蔬酶促褐变生理生化研究的重要内容,本试验以邻苯二酚为底物,采用分光光度法对砀山梨多酚氧化酶的酶学特性及不同抑制剂对多酚氧化酶活性的影响进行了研究。结果表明：砀山梨PPO的最适pH为4.5,最适温度为34℃;短时间高温能显著抑制PPO活性;PPO催化的酶促褐变反应动力学符合米氏方程,该酶促反应的最大速率为178.57U/min,酶反应速度为最大反应速度1/2时的底物浓度为0.125mol/L;柠檬酸、L-半胱氨酸、抗坏血酸,亚硫酸氢钠较柠檬酸能较好地抑制PPO活性,随着浓度的升高,抑制效应逐渐增强,但综合研究表明L-半胱氨酸抑制效应较好。     

毛酸浆多酚氧化酶的酶学特性研究

以毛酸浆中多酚氧化酶(PPO)为研究对象,采用分光光度法在420nm处对其酶学特性进行研究。结果表明,毛酸浆PPO的底物邻苯二酚的最适质量分数为0.04%;最适pH为7.0;最适温度为35℃;100℃处理2min,可使酶完全失活;PPO催化的酶促褐变反应动力学符合米氏方程,动力学参数为Km=0.025mol/L,Vmax=125U/min;抗坏血酸、柠檬酸、L-半胱氨酸和亚硫酸钠对PPO酶活性的抑制作用均随浓度的增大而加强,抑制能力由强到弱依次为:抗坏血酸柠檬酸L-半胱氨酸EDTA亚硫酸氢钠。     

山药多酚氧化酶的特性和抑制研究

以邻苯二酚为底物,410nm处测定山药多酚氧化酶(PPO)的活性,研究了pH、温度、底物浓度、酶浓度、抑制剂和激活剂对山药多酚氧化酶活性的影响。结果表明,山药PPO最适反应pH为6.5,最适反应温度为40℃;柠檬酸、草酸、L-抗坏血酸、亚硫酸氢钠及半胱氨酸五种抑制剂对山药PPO均有抑制作用,其强弱依次为:L-半胱氨酸>抗坏血酸>草酸>柠檬酸>亚硫酸氢钠;硫酸铜和三氯化铁均有加速酶促褐变作用,强弱为CuSO_4>FeCl_3。     

芦荟多酚氧化酶特性的研究

研究了芦荟多酚氧化酶的基本性质。以邻苯二酚为底物,采用分光光度法在4 10nm处测定芦荟多酚氧化酶的活性,研究了温度、pH值、底物浓度以及酶浓度对其活性的影响,并建立了酶促褐变反应动力学方程。实验结果表明,芦荟多酚氧化酶的最适温度为4 0℃,最适pH值为6 5 ,其酶促褐变反应动力学符合米氏方程所描述的单底物酶促反应动力学,相应的动力学参数Km=0 2 8mol/L ,Vmax=2 32×10 -2 U/min。文中还研究了利用蛋白酶抑制褐变的方法,以芦荟多酚氧化酶液(mL)∶木瓜蛋白酶(mL ,浓度0 5mg/mL) =5∶1的比例向芦荟多酚氧化酶粗酶液中添加木瓜蛋白酶,在5 5℃、pH5 7条件下,水解10min可较好地抑制多酚氧化酶的活性,从而抑制芦荟的酶促褐变。     

抑制剂对杏多酚氧化酶抑制作用

使用冷丙酮分别从黄金杏和晚红杏中提取多酚氧化酶(PPO),以邻苯二酚为底物,利用分光光度计法研究几种抑制剂在不同浓度下对2种杏PPO的抑制作用及酶促反应动力学曲线。结果表明, 2种杏最适波长均为410nm。以邻苯二酚为底物, 2种杏多酚氧化酶的酶促褐变反应均符合米氏方程,晚红杏、黄金杏PPO的动力学参数米氏常数分别为0.042 mol/L和0.114 mol/L。V_(max)分别为100 U/min和167 U/min。抑制剂对2种杏多酚氧化酶的抑制作用存在一定差异。对黄金杏PPO抑制作用:抗坏血酸(40 mmol/L)柠檬酸(10 mmol/L)L-半胱氨酸(10mmol/L)亚硫酸氢钠(10 mmol/L)。对晚红杏PPO抑制作用:亚硫酸氢钠(10 mmol/L)柠檬酸(10 mmol/L)抗坏血酸(40 mmol/L)L-半胱氨酸(10 mmol/L), EDTA-2Na对2种杏PPO抑制作用均较差。     

Reliable identification of grapevine rootstock varieties using RAPD PCR on woody samples

Since grapevine ( Vitis spp .) rootstock material is being traded increasingly as disbudded woody material a lack of distinctive morphological features on such material necessitates an alternative and reliable means of identification. Methods described here were developed for rapid and efficient extraction of DNA from woody samples rich in phenolic compounds and polysaccharides, and for subsequent identification of varieties by RAPD PCR. Using these methods, and with the application of only one selected RAPD primer, we were able to differentiate sixteen rootstock varieties, including the seven varieties most commonly used in Germany. Problems commonly encountered with reproducibility of RAPD patterns were avoided by choosing primers with a dinucleotide sequence and a high G/C content that allowed a rather high annealing temperature of 45°C. Methods described here should also be useful for other horticultural crops, especially those with woody tissues rich in phenolic compounds and polysaccharides.     

Chlorohydrins of bisphenol A diglycidyl ether (BADGE) and of bisphenol F diglycidyl ether (BFDGE) in canned foods and ready-to-drink coffees from the Japanese market

BADGE.2HCl and BFDGE.2HCl were determined in 28 samples of ready-to-drink canned coffee and 18 samples of canned vegetables (10 corn, 5 tomatoes and 3 others), all from the Japanese market. HPLC was used as the principal analytical method and GCMS for confirmation of relevant LC fractions. BADGE.2HCl was found to be present in one canned coffee and five samples of corn, BFDGE.2HCl in four samples of canned tomatoes and in one canned corn. No sample was found which exceeded the 1mg/kg limit of the EU for the BADGE chlorohydrins. However the highest concentration was found for the sum of BFDGE.2HCl and BFDGE.HCl.H₂O at a level of 1.5mg/kg. A Beilstein test confirmed that all cans containing foods contaminated with BADGE.2HCl or BFDGE.2HCl had at lest one part coated with a PVC organosol.     

Aroma characterization of various apricot varieties using headspace–solid phase microextraction combined with gas chromatography–mass spectrometry and gas chromatography–olfactometry

The characterization of the aromatic profile of several apricot cultivars with molecular tracers in order to obtain objective data concerning the aromatic quality of this fruit was undertaken using headspace–solid phase microextraction (HS–SPME). Six apricot cultivars were selected according to their organoleptic characteristics: Iranien, Orangered, Goldrich, Hargrand, Rouge du Roussillon and A4025. The aromatic intensity of these varieties measured by HS–SPME–Olfactometry were defined and classified according to the presence and the intensity of grassy, fruity and apricot like notes. In the six varieties, 23 common volatile compounds were identified by HS–SPME–GC–MS. Finally, 10 compounds, ethyl acetate, hexyl acetate, limonene, β-cyclocitral, γ-decalactone, 6-methyl-5-hepten-2-one, linalool, β-ionone, menthone and (E)-hexen-2-al were recognized by HS–SPME–GC–O as responsible of the aromatic notes involved in apricot aroma and considered as molecular tracers of apricot aromatic quality which could be utilized to discriminate apricot varieties.     

European priorities for research to support legislation in the area of food contact materials and articles

A strong science base is required to underpin the planning and decision-making process involved in determining future European community legislation on materials and articles in contact with food. Significant progress has been made in the past 5 years in European funded work in this area, with many developments contributing to a much better understanding of the migration process, and better and simpler approaches to food control. In this paper this progress is reviewed against previously identified work-areas (identified in 1994) and conclusions are reached about future requirements for R&D to support legislation on food contact materials and articles over the next 5 or so years.     

Textural Properties of Cold-set Gels Induced from Heat-denatured Whey Protein Isolates

A 9% whey protein (WP) isolate solution at pH 7.0 was heat-denatured at 80°C for 30 min. Size-exclusion HPLC showed that native WP formed soluble aggregates after heat-treatment. Additions of CaCl2 (10–40 mM), NaCl (50–400 mM) or glucono-delta-lactone (GDL, 0.4–2.0%, w/v) or hydrolysis by a protease from Bacillus licheniformis caused gelation of the denatured solution at 45°C. Textural parameters, hardness, adhesiveness, and cohesiveness of the gels so formed changed markedly with concentration of added salts or pH by added GDL. Maximum gel hardness occurred at 200 mM NaCl or pH 4.7. Increasing CaCl2 concentration continuously increased gel hardness. Generally, GDL-induced gels were harder than salt-induced gels, and much harder than the protease-induced gel.     

Tests of potential functional barriers for laminated multilayer food packages. Part I: Low molecular weight permeants

The advent of the functional barrier concept in food packaging has brought with it a requirement for fast tests of permeation through potential barrier materials. In such tests it would be convenient for both foodstuffs and materials below the functional barrier (sub-barrier materials) to be represented by standard simulants. By means of inverse gas chromatography, liquid paraffin spiked with appropriate permeants was considered as a potential simulant of sub-barrier materials based on polypropylene (PP) or similar polyolefins. Experiments were performed to characterize the kinetics of the permeation of low molecular weight model permeants (octene, toluene and isopropanol) from liquid paraffin, through a surrogate potential functional barrier (25 μm-thick oriented PP) into the food simulants olive oil and 3% (w/v) acetic acid. These permeation results were interpreted in terms of three permeation kinetic models regarding the solubility of a particular model permeant in the post-barrier medium (i.e. the food simulant). The results obtained justify the development and evaluation of liquid sub-barrier simulants that would allow flexible yet rigorous testing of new laminated multilayer packaging materials.     

Occurrence of bisphenol-F-diglycidyl ether (BFDGE) in fish canned in oil

The levels of bisphenol-F-diglycidyl ether (BFDGE) were quantified as part of a European survey on the migration of residues of epoxy resins into oil from canned fish. The contents of BFDGE in cans, lids and fish collected from all 15 Member States of the European Union and Switzerland were analysed in 382 samples. Cans and lids were separately extracted with acetonitrile. The extraction from fish was carried out with hexane followed by re-extraction with acetonitrile. The analysis was performed by reverse phase HPL C with fluorescence detection. BFDGE could be detected in 12% of the fish, 24% of the cans and 18% of the lids. Only 3% of the fish contained BFDGE in concentrations considerably above 1mg/kg. In addition to the presented data, a comparison was made with the levels of BADGE (bisphenol-A-diglycidyl ether)analysed in the same products in the context of a previous study.     

Contribution of European research to risk analysis

The European Commission's, Quality of Life Research Programme, Key Action 1—Health, Food & Nutrition is mission-oriented and aims, amongst other things, at providing a healthy, safe and high-quality food supply leading to reinforced consumer confidence in the safety of European food. Its objectives also include the enhancing of the competitiveness of the European food supply. Key Action 1 is currently supporting a number of different types of European collaborative projects in the area of risk analysis. The objectives of these projects range from the development and validation of prevention strategies including the reduction of consumers risks; development and validation of new modelling approaches; harmonization of risk assessment principles, methodologies, and terminology; standardization of methods and systems used for the safety evaluation of transgenic food; providing of tools for the evaluation of human viral contamination of shellfish and quality control; new methodologies for assessing the potential of unintended effects of genetically modified (genetically modified) foods; development of a risk assessment model for Cryptosporidium parvum related to the food and water industries; to the development of a communication platform for genetically modified organism, producers, retailers, regulatory authorities and consumer groups to improve safety assessment procedures, risk management strategies and risk communication; development and validation of new methods for safety testing of transgenic food; evaluation of the safety and efficacy of iron supplementation in pregnant women; evaluation of the potential cancer-preventing activity of pro- and pre-biotic ('synbiotic') combinations in human volunteers. An overview of these projects is presented here.     

Influence des ions sur le pouvoir hydratant de l'ure e: e tude sur peau de porc ex vivo

This study deals with the influence of ions (NaCl and MgSO4) in a W/O emulsion containing 10% urea. Moisturization kinetics are assessed by corneometry on pig skin ex vivo. The formula's influence on urea penetration is measured by infrared spectrometry with an ATR device and the stripping method. Corneometry and spectroscopy were chosen to record simultaneously the hydratation levels and urea localization into superficial cell layers. Urea crystallization after evaporation of emulsions and aqueous solutions is described. Results show that urea does not hydrate nor penetrate when applied to the skin through an aqueous gel. In a W/O emulsion, sodium chloride increases the ability of urea to moisturize without improving penetration. In vitro urea crystallization is disturbed by sodium chloride or magnesium sulphate for solutions and emulsions. This stabilization by ions is correlated with good moisturization values. The stabilization of urea in the solute state provided by ions increases its water epidermal binding capacity without enhancing penetration.