多酚氧化酶的提取及分离纯化研究进展

多酚氧化酶(PPO)是自然界中分布极广的一种金属蛋白酶,能催化邻-苯二酚氧化成邻-苯二醌,也是许多果蔬等农产品酶促褐变的主要原因。本文综述了国内外多酚氧化酶的提取及分离纯化技术的研究进展,包括传统提取方法(如缓冲液匀浆法、丙酮提取法、丙酮粉提取法等)和新型提取方法(超声波辅助提取法和超高压提取法等),传统经典分离纯化方法(如溶剂法、沉淀法)和新型分离纯化方法(如凝胶色谱分离法和离子交换色谱分离法等),旨在为多酚氧化酶的研究和应用,特别是为抑制农产品酶促褐变提供参考。     

蘑菇中多酚氧化酶的活性及影响活性的区素

多酚氧化酶存在于蘑菇、马铃薯、苹果和桃子等果蔬中。多酚氧化酶是一种含铜氧化酶(含铜近0.17％),它催化空气中的氧去氧化二酚类物质的反应。例如邻二酚很快变成棕褐色的邻二醌%.多酚氧化酶的作用,可使蘑菇发生褐变,使感官质量下降。     

高灌蓝越橘多酚氧化酶的提取和纯化

采用丙酮粉沉淀抽滤法、硫酸铵分级沉淀法及苯基琼脂糖疏水性层析法对高灌蓝越橘多酚氧化酶进行提取和纯化。在提取和纯化中，用加入不同的酚吸收剂和抑制剂并比较酶活性的高低来确认最佳方法。结果表明在粗提中加入酚吸收剂、增溶剂和抑制剂提取效果较好，苯基琼脂糖疏水性层析法对此类酶纯化效果较佳并为后来的辅酶的分离提供了较高的纯化倍数。     

圆蘑菇多酚氧化酶的特性研究

采用0.2 mol/L磷酸盐缓冲溶液匀浆法从圆蘑菇中提取多酚氧化酶(PPO),并对该多酚氧化酶的酶学特性进行研究.实验结果表明,以邻苯二酚为底物,该酶的最适pH为6.4,最适温度为30℃,Km值为0.032 3 mol/L,Vmax值为0.0164 U/min,100 ℃热处理1.5 min可完全钝化PPO的活力,10 mmol/L的EDTA、L-半胱氨酸或柠檬酸亚锡二钠能有效抑制PPO活力.     

带肉梨汁护色及加工工艺的研究(Ⅰ)——子母梨中多酚氧化酶特性的研究

本文用丙酮粉法从子母梨组织中提取多酚氧化酶,经萃取后,分别用量压法或分光光度法测定了pH、温度,酶浓度、底物浓度对多酚氧化酶酶活力的关系,分别求出最适pH值、最适温度、V_(max)和K_m;测定了该酶体系的热稳定性情况;初步研究了SO_2和VitC对多酚氧化酶的抑制作用。     

澳洲青苹多酚氧化酶的分离纯化研究

采用丙酮粉沉淀抽滤法从澳洲青苹中提取多酚氧化酶.提取的粗酶液经30%～90%饱和度硫酸铵分级沉淀、透析除盐、交联葡聚糖G-75凝胶色谱层析、后经PEG 6000浓缩,得到纯化的多酚氧化酶,纯化倍数为7.49.纯化后的酶液经十二烷基磺酸钠-聚丙烯酰胺凝胶电泳,采用考马斯亮蓝R-250染色显示一条电泳带.通对过PPO与邻苯二酚反应产物的波谱扫描显示,其产物在λ=410nm下有最高吸收.     

银杏外种皮多酚氧化酶的研究

本文采用不同条件抽提银杏外种皮多酚氧化酶(简称PPO),比较了抽提条件对PPO性的影响;利用乙醇分级法对银杏外种皮多酚氧化酶进行初步分离纯化,比活力从49．8μ/mg提高到292μ/mg;研究pH值、温度、金属离子对乙醇分级提纯后的外种皮多酚氧化酶的影响,发现银杏外种皮中的多酚氧化酶的最适温度为30℃,最适pH值在7．6,Mg^2 对酶有明显的激活作用,而Na^ 、K^ 、Fe^2 则对酶有抑制作用。     

小麦多酚氧化酶酶学特性的研究

选取3个小麦品种作为试验材料,以邻苯二酚为底物,采用分光光度法在410nm处测定小麦多酚氧化酶（PPO）的活性,研究温度、pH值、底物浓度对其活性的影响以及酶促动力学常数Km,结果表明：小麦多酚氧化酶的最适温度范围为60～75℃．最适pH值为4．0～4．6,描述的单底物酶促反应动力学,相应的动力学参数Km=0．19mol／L．vmax=4．04×102（mol·L^-1）·min-1。     

枇杷果实酶促褐变控制研究

枇杷果实在贮藏及加工过程中极易产生酶促褐变．本文采用分光光度法，研究了几种因素对枇杷果实多酚氧化酶活性的影响。结果表明：多酚氧化酶的最适pH为6．3，抗坏血酸、L—半胱氨酸、柠檬酸、草酸、柠檬酸．抗坏血酸混合物可明显抑制该酶活性。     

米曲霉氨基酰化酶纯化方法的研究

比较了盐析法和有机溶剂沉淀法对于米曲霉氨基酰化酶的纯化效果。分别使用硫酸铵、无水乙醇和丙酮对米曲霉氨基酰化酶进行纯化，结果发现：有机溶剂沉淀法的纯化效果优于盐析法。采用丙酮二次沉淀法纯化的氨基酰化酶比酶活为93.20U／mg，纯化倍数为3．26：通过乙醇二次沉淀，得到的酶液比酶活为93.22U／mg，纯化倍数为3．10，而采用盐析法纯化的酶液的比酶活为56.01U／mg，纯化倍数为2.06。从酶的纯化效果方面考虑，丙酮沉淀法最佳，但从工业生产的可行性方面考虑，乙醇沉淀法较好。     

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.     

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.     

低温带皮菜籽粕微粉的不同粒级部分的功能特性

为研究低温带皮菜籽粕微粉的不同粒级部分的功能特性,以经低温脱脂的带皮菜籽粕为原料,经微粉碎后筛分成212~425μm、150~212μm和106~150μm的3个不同粒级的微粉样品,检测这些样品的吸水性、吸油性、乳化性和乳化稳定性、蛋白质体外消化率。结果表明:1 3个不同粒级的微粉样品之间的粗纤维含量存在显著差异,表明三者的结构组成成分有一定差异。23个微粉样品的乳化活性和乳化稳定性随粒度级别的减小而显著增加(P0.01)。33个微粉样品的蛋白质体外消化率随粒度级别的减小而显著增加(P0.01)。4不同粒级带皮菜籽粕微粉样品的吸水性与吸油性受其结构组成物质不同和粒度的双重影响,与粒度的相关性不明显。     

Microbiology of food taints

Fresh and processed foods are often spoilt by the presence of undesirable flavours and odours caused by microbial action. The aim of this paper is to review the current knowledge of microbiologically induced taints that occur in a wide range of foodstuffs, including meats, poultry, fish, crustaceans, milk, dairy products, fruits, vegetables, cereals and cereal products. Examples have been chosen where the compounds responsible for the taint have been identified and sufficient data obtained to demonstrate the involvement of microorganisms. However, in some cases the full identity of the causative organism may not have been elucidated. The types of microorganisms covered by this review include bacteria, fungi, yeasts, actinomycetes and cyanobacteria. Although cyanobacteria do not in general infect foods, their presence in aqueous systems and water supplies can lead to off-flavours in aquatic organisms and processed foodstuffs. Several examples of each of these processes are discussed. Wherever possible, the likely biosynthetic pathway used by the microorganism to produce the offending compound in a foodstuff is indicated.     

Analysis for organic residues from aids to polymerization used to make plastics intended for food contact

Polymers intended for food contact use have been analysed for organic residues which could be attributed to a range of substances employed as polymerization aids (e.g. initiators and catalysts). A wide range of polymers was extracted with solvents and the extracts analysed by gas chromatography-mass spectrometry (GC-MS). The overwhelming majority of substances identified were not derived from aids to polymerization but were oligomers, additives and adventitious contaminants. However, a small number of substances were identified as initiator residues. These included tetramethylsuccinonitrile (TMSN) which was observed in two polymers and it derived from recombination of two azobisisobutyronitrile (AIBN) initiator radicals. Methyl benzoate, benzoic acid, biphenyl and phenyl benzoate were detected in one poly(methyl methacrylate) sample and in two polyvinylchlorides and they are thought to be derived from benzoyl peroxide initiator. TMSN was subsequently targeted for analysis of poly-(methyl methacrylate) plastics using proton nuclear magnetic resonance spectrometry (¹     

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

Experiments were performed to characterize the kinetics of the permeation of different medium molecular weight model permeants: bisphenol A, warfarin and anthracene, from liquid paraffin, through a surrogate potential functional barrier (25 microns-thick orientated polypropylene--OPP) into the food simulants olive oil and 3% (w/v) acetic acid. The characterization of permeation kinetics generally observed the permeation models previously reported to explain the experimental permeation results obtained for a low molecular weight group of model permeants. In general, the model permeants exhibited behaviour consistent with their relative molecular weights with respect to (a) the time taken to attain steady-state permeation into the food simulant in which they were more soluble, (b) their subsequent steady-state permeation rates, and (c) their partition between liquid paraffin and the OPP membrane.     

The development of reference materials for paralytic shellfish poisoning toxins in lyophilized mussel. I: Interlaboratory studies of methods of analysis

This paper describes the first part of a project undertaken to develop mussel reference materials for Paralytic Shellfish Poisoning (PSP) toxins. Two interlaboratory studies were undertaken to investigate the performance of the analytical methodology for several PSP toxins, in particular saxitoxin (STX) and decarbamoyl-saxitoxin (dc-STX) in lyophilized mussels, and to set criteria for the acceptance of results to be applied during the second part of the project: the certification exercise. In the first study, 18 laboratories were asked to measure STX and dc-STX in rehydrated lyophilized mussel material and to identify as many other PSP toxins as possible with a method of their choice. In the second interlaboratory study, 15 laboratories were additionally asked to determine quantitatively STX and dc-STX in rehydrated lyophilized mussel and in a saxitoxin-enriched mussel material. The first study revealed that three out of four postcolumn derivatization methods and one pre-column derivatization method sufficed in principle to determine STX and dc-STX. Most participants (13 of 18) obtained acceptable calibration curves and recoveries. Saxitoxin was hardly detected in the rehydrated lyophilized mussels and results obtained for dc-STX yielded a CV of 58% at a mass fraction of 1.86 mg/kg. Most participants (14 out of 18) identified gonyautoxin-5 (GTX-5) in a hydrolysed extract provided. The first study led to provisional criteria for linearity, recovery and separation. The second study revealed that 6 out of 15 laboratories were able to meet these criteria. Results obtained for dc-STX yielded a CV of 19% at a mass fraction of 3.49mg/kg. Results obtained for STX in the saxitoxin-enriched material yielded a CV of 19% at a mass fraction of 0.34mg/kg. Saxitoxin could not be detected in the PSP-positive material. Hydrolysis was useful to confirm the identity of GTX5 and provided indicative information about C1 and C2 toxins in the PSP-positive material. The methods used in the second interlaboratory study showed sufficiently consistent analysis results to undertake a certification exercise to assign certified values for STX and dc-STX in lyophilized mussel.     

The Ministry of Environmental Protection Issued "Feasible Technology Guidelines for Pollution Prevention and Control in Wood Pulping Process of the Paper Industry (Trial)" and Two Other Guiding Technical Documents

On December 27t＂, 2013, the Ministry of Environmenta Protection announced that, in order to implement ＂The Environmental Protection Law of the People＇ s Republic of China＂, improve the working system in environmenta protection technologies, and promote technologica advancement in pollution prevention, the Ministry of Environmental Protection sponsored the formulation of three guiding technical documents including ＂Feasible Technology Guidelines for Pollution Prevention and Contro n Wood Pulping Process of the Paper Industry （Trial）＂     

1~(st) Intelli-Tissue~ EcoEc Tissue Machine Supplied by PMP Group Successfully Put into Operation in China

正On April 29th,2014,Intelli-Tissue EcoEc tissue machine supplied by PMP Group successfully put into operation at Hebei Xuesong Paper Co.,Ltd.,this is the first such kind of paper machine of PMP Group in China.