木瓜蛋白酶水解鸡肉蛋白及其产物氨基酸分析研究

本文采用木瓜蛋白酶水解鸡肉蛋白研究其降解规律及酶解产物氨基酸组成。研究表明:热处理鸡肉蛋白不利于酶解;整个酶解过程中,可溶性氮、氨态氮含量呈增加趋势,但4h后增势变缓,而肽含量在水解8h达到最大值;肽分子量分布图显示大部分不溶性蛋白和分子量大于10000Da肽段在酶解4h后已经变为可溶性多肽和氨基酸,分子量在5000～10000Da间肽段酶解24h后仍大量存在,分子量在3000～5000Da间以及小于3000Da的肽占总肽量比值在整个酶解过程中呈增加趋势。与原料蛋白氨基酸组成比较表明采用木瓜蛋白酶水解鸡肉蛋白能显著提高其营养价值。     

粗酶水相提取大豆油过程中蛋白酶解与油脂释放状态

为了考察粗酶水相提取大豆油过程中蛋白酶解和油脂释放状态,监测酶解过程中总油、总蛋白提取率以及蛋白相对分子质量变化,同时采用扫描电镜、考马斯亮兰与苏丹红Ⅲ染色显微观察技术研究酶解过程中油脂和蛋白在豆粉、水解液、乳状液中的分布状态。结果表明:随酶解时间延长,蛋白酶解程度加深,油脂释放量提升,水解蛋白中相对分子质量大于33 000 Da的多肽含量逐渐降低,而相对分子质量小于5 500 Da的短肽含量增加。酶解6 h时,总蛋白和总油提取率均达到最高,分别为90. 1%和94. 2%,此时蛋白水解液中短肽(5 500 Da)含量最高,占49. 63%,相对分子质量大于33 000 Da的多肽仅占21. 15%。扫描电镜图显示由于酶解作用豆粉内的蛋白和油脂逐渐被分解和释放,原本充盈的细胞转变为空的残渣。光镜图显示,伴随豆粉酶解,水解液中蛋白含量逐渐增加,释放的油脂在水解液和乳状液中均有分布,随酶解时间延长油滴数量增加、粒径增大。     

美藤果粕多肽的提取工艺优化

以多肽的水解度为参考指标,在单因素试验基础上,利用响应面试验优化美藤果粕多肽的高温蒸煮结合酶法提取工艺,并对该工艺下制备的美藤果粕多肽进行氨基酸组成和分子量分析。结果表明,其最佳工艺参数为液料比为20∶1（mL/g）、蒸煮时间6 h、酶解温度55 ℃、酶解时间6 h、酶添加量1.16%,该条件下美藤果粕多肽的水解度为32.5%。制得的多肽分子量小于1 000 Da 的肽段占94.5%,属于小分子多肽。     

大豆球蛋白酶解物的分离及其抗氧化活性研究

以7S和11S大豆球蛋白为原料,选用Alcalase碱性蛋白酶在其最佳酶解条件下进行酶解,对酶解物进行超滤分离纯化抗氧化肽,并对其各组分进行保护系数和对·DPPH（1,1-二苯基苦酰基苯肼）自由基清除率的研究。结果显示：7S和11S大豆球蛋白Alcalase碱性蛋白酶酶解物经超滤后所得分子量小于5kDa,组分保护系数分别为2.38和2.21,其·DPPH自由基清除能力分别为75.63%和73.56%。并经高效液相色谱分析该组分的分子量分布在1000Da以下的含量最多,7S和11S酶解物超滤后组分分子量小于1000Da组分分别占81.13%占87.84%。     

亚麻多肽制备及其抗氧化活性分析

该研究以脱脂亚麻籽粕为原料,比较不同酸性蛋白酶和中性蛋白酶的酶解多肽的得率,评价了酶解多肽的抗氧化活性。结果表明:3.350黑曲酸性蛋白酶的酶解效果最好,多肽得率可达28.52%。通过蛋白酶的单因素和正交试验,确定亚麻多肽制备的最佳工艺条件为:酶解温度60℃、pH2.7、加酶量6 500 U/g,酶解时间2.5 h,此条件下亚麻多肽得率为38.80%。凝胶色谱分析表明,该酶多肽的分子量主要分布在1 000 Da～7 600 Da范围内。酶解后的亚麻多肽具有良好的还原力,当酶解液质量浓度为1.5 mg/mL时,DPPH自由基清除率为89.99%,当酶解液质量浓度为0.6 mg/mL时,超氧阴离子的清除率为85.57%。     

鸡内金酶解物制备工艺优化及抗氧化活性研究

目的：研究鸡内金的最佳酶解工艺条件,并分析其酶解物的氨基酸组成、分子量分布及抗氧化活性。方法：以鸡内金多肽得率、酶解液水解度为指标,采用正交试验优化鸡内金的酶解工艺条件;采用氨基酸分析仪、高效凝胶渗透色谱法测定鸡内金酶解物的氨基酸组成和分子量分布;并考察鸡内金酶解物对DPPH自由基和ABTS+自由基的清除能力。结果：鸡内金最佳酶解工艺条件为以中性蛋白酶和碱性蛋白酶共同酶解,酶解温度60 ℃,酶解时间9 h,酶解pH值11,蒸煮时间6 h。该条件下鸡内金多肽得率为75.68%,水解度为14.43%;鸡内金酶解物的必需氨基酸与非必需氨基酸的比值为58.56%,具有较高的营养价值;鸡内金酶解物相对分子质量＜1 000 Da所占比例为90.2%,易于消化吸收;当鸡内金酶解物质量浓度为1.5 mg/mL时,其对ABTS+自由基的清除率达到94.19%。结论：最佳酶解工艺下,鸡内金多肽得率高达75.68%,具有较好的抗氧化活性。     

壳聚糖对牡蛎ACE抑制肽的脱腥工艺研究

运用单因素试验优化牡蛎肽的酶解制备工艺,添加0.6%中性蛋白酶在p H7.0、50℃条件下酶解3 h。应用1.0%壳聚糖溶液对优化后的牡蛎酶解液进行絮凝脱腥,4 800 r/min离心15 s即可达到显著的脱腥脱脂和杂质去除效果,固形物回收率80.5%,蛋白回收率79.5%,脂肪去除率达92.4%。得到透明无腥味的牡蛎多肽后,对多肽分子量进行分析,1 000 Da以下的占93.1%,500 Da以下的约占49.7%,对牡蛎多肽的血管紧张素转换酶(angiotensinconverting enzyme,ACE)抑制活性进行研究,IC_(50)为0.475 mg/mL,为开发制备低分子量、高ACE抑制活性的脱腥牡蛎肽提供理论支持。     

高压处理-酶解大米蛋白抗氧化肽的制备及其分子量分布

本文以酶解物的抗氧化活性为指标,通过正交试验方法优化了3种高压(0.1 MPa、300 MPa、500 MPa)预处理大米蛋白的酶解条件,用高效液相色谱分析了主要酶解产物的分子量分布。结果表明,三种预处理蛋白在各自优化条件下制备的酶解物在1.0mg/mL时的最大还原力分别为0.125、0.149、0.158。常压对照组(即0.1MPa)中15-10 ku、10-1 ku和 1 ku以下等三段分子量组分所占比例分别为26.6%、46.9%、26.5%;300MPa组中的含量分别为16.5%、39.3%和44.2%;500MPa组中的含量分别为11.8%、33.1%和55.1%;可见,酶解物还原力大小与其分子量的分布之间有明显的关联性。     

红松仁多肽酶解制备工艺优化及降压降脂活性分析

为了对红松仁多肽的开发及应用提供参考，以红松仁粕为原料，采用酸浸提法提取红松仁蛋白，然后采用酶解法制备红松仁多肽。以水解度为评价指标，通过单因素实验和响应面实验对红松仁多肽酶解制备工艺条件进行了优化。同时，对最佳工艺条件下制备的多肽的分子质量分布和氨基酸组成进行分析，并测定了多肽的降压降脂活性。结果表明：红松仁多肽的最佳酶解制备工艺条件为采用分步酶解法，在酶解温度50℃、底物质量浓度6.0 g/100 mL、木瓜蛋白酶添加量10 500 U/g、酶解pH 6.0条件下酶解2.5 h,然后在碱性蛋白酶添加量10 970 U/g、酶解pH 9.0条件下酶解2.3 h,在此工艺条件下红松仁蛋白的水解度达25.94%;红松仁多肽的数均分子质量为2 272 Da,重均分子质量为4 080 Da;红松仁多肽中谷氨酸、精氨酸和天冬氨酸含量较高，必需氨基酸含量占氨基酸总量的30.23%;红松仁多肽中分子质量小于10 kDa的多肽占92%,分子质量0.5～1.0 kDa的多肽具有最高的ACE抑制活性，IC₅₀为66.05μg/mL;分子质量1.0～5.0 kDa的多肽具有最高的胰脂...     

超声辅助核桃饼脱脂和多肽制备工艺的优化

采用超声辅助核桃饼脱脂,并以脱脂核桃粉为原料制备核桃蛋白,采用碱性蛋白酶酶解核桃蛋白制备多肽。通过单因素实验和正交实验对超声辅助核桃饼脱脂和核桃多肽制备工艺条件进行优化,并对最优条件下制备的核桃多肽的特性进行分析。结果表明:超声辅助核桃饼脱脂最优条件为料液比1∶20、超声功率500 W、超声时间140 min,在最优条件下脱脂率为91.23%,蛋白损失率为11.32%;酶解制备核桃多肽的最优工艺条件为酶解温度50℃、酶解pH 9、加酶量3.0%、酶解时间5.0 h,在最优条件下水解度达到22.63%,多肽得率为88.24%。核桃多肽粗蛋白质含量约为95%,相对分子质量小于1 000 Da的多肽占比达91.61%。     

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.     

An internet compendium of analytical methods and spectroscopic information for monomers and additives used in food packaging plastics

An internet website (http://cpf.jrc.it/smt/) has been produced as a means of dissemination of methods of analysis and supporting spectroscopic information on monomers and additives used for food contact materials (principally packaging). The site which is aimed primarily at assisting food control laboratories in the European Union contains analytical information on monomers, starting substances and additives used in the manufacture of plastics materials. A searchable index is provided giving PM and CAS numbers for each of 255 substances. For each substance a data sheet gives regulatory information, chemical structures, physico-chemical information and background information on the use of the substance in particular plastics, and the food packaging applications. For monomers and starting substances (155 compounds) the infra-red and mass spectra are provided, and for additives (100 compounds); additionally proton NMR are available for about 50% of the entries. Where analytical methods have been developed for determining these substances as residual amounts in plastics or as trace amounts in food simulants these methods are also on the website. All information is provided in portable document file (PDF) format which means that high quality copies can be readily printed, using freely available Adobe Acrobat Reader software. The website will in future be maintained and up-dated by the European Commission's Joint Research Centre (JRC) as new substances are authorized for use by the European Commission (DG-ENTR formerly DGIII). Where analytical laboratories (food control or other) require reference substances these can be obtained free-ofcharge from a reference collection housed at the JRC and maintained in conjunction with this website compendium.     

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.     

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.     

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.     

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.