罗非鱼酶解多肽的乳化性及乳化稳定性研究

考察罗非鱼蛋白酶解蛋白酶种类及其水解度(DH)对罗非鱼蛋白酶解多肽的乳化性及乳化稳定性的影响,并探讨多肽的疏水性与其乳化性及乳化稳定性之间的关系。研究发现,蛋白酶种类及罗非鱼蛋白水解度对酶解多肽乳化性及乳化稳定性有不同的影响,中性蛋白酶和复合风味蛋白酶酶解罗非鱼可以得到乳化性和乳化稳定性均较好的多肽。低水解度的多肽乳化性及乳化稳定性高,DH=5%时,经中性蛋白酶酶解的多肽乳化性及乳化稳定性分别为62.05%和28.90%,经复合风味蛋白酶酶解的多肽乳化性及乳化稳定性分别为67.46%和44.89%。不同DH酶解多肽疏水性与乳化性及乳化稳定性趋势大致相同。     

复合蛋白酶水解棉粕制备多肽的工艺优化

为提高棉粕蛋白资源的利用率,考察了复合蛋白酶水解脱酚棉粕制备多肽的工艺。以多肽得率和水解度为评价指标,比较了不同蛋白酶对棉粕蛋白水解能力的差异,确定了以胰蛋白酶与米曲霉蛋白酶按照酶活比3∶1构建的复合蛋白酶作为水解棉粕蛋白的最佳蛋白酶。采用单因素试验及响应面试验对影响棉粕蛋白水解的因素底物蛋白质量浓度、加酶量、p H、酶解温度以及酶解时间进行了分析,并确定了棉粕蛋白水解的最佳工艺条件为:棉粕蛋白质量浓度25 g/L,加酶量5 000 U/g,p H 8. 91,酶解温度50～55℃,酶解时间5. 92 h。在最佳工艺条件下,水解棉粕蛋白的多肽得率可以达到71. 85%。多肽的氨基酸分析结果表明,酶水解制备的多肽能基本保持原棉粕蛋白的氨基酸组成,其氨基酸种类较齐全,8种必需氨基酸含量丰富,营养价值较高。     

复合蛋白酶水解螺旋藻制备多肽的工艺优化

螺旋藻富含优质蛋白质,为有效利用这一蛋白质资源,试验探究了复合蛋白酶水解螺旋藻制备多肽的工艺路线。通过单一酶解及复合酶解试验考察了不同蛋白酶对螺旋藻水解效果的差异,以及不同蛋白酶对螺旋藻水解作用的协同效果,由此筛选出适合螺旋藻水解的最佳蛋白酶为:由碱性蛋白酶、胰蛋白酶、木瓜蛋白酶按酶活比2∶1∶1的比例构成的复合蛋白酶。采用单因素及响应面试验对复合蛋白酶水解螺旋藻的工艺条件进行了优化,得到螺旋藻酶解的最适工艺条件为:螺旋藻浓度50 g/L,pH 9.0,加酶量6 000 U/g蛋白,温度52℃,酶解时间6.2 h。在优化的工艺条件下,水解螺旋藻的多肽得率可以达到(77.82±1.21)%。多肽的氨基酸组成分析结果显示,酶解制备得到的多肽基本上保持了原螺旋藻蛋白的氨基酸组成,氨基酸组成齐全,必需氨基酸占到氨基酸总量的43%以上,具有很高的营养价值。研究结果表明,复合蛋白酶水解螺旋藻制备多肽的工艺路线具有可行性。     

酶法水解卵黄蛋白制备多肽的工艺优化

利用酶法制备卵黄蛋白多肽。比较复合风味蛋白酶、中性蛋白酶、碱性蛋白酶、复合蛋白酶水解卵黄蛋白的效果,确定碱性蛋白酶与复合风味蛋白酶为复合酶解的工艺用酶。采用响应面分析法,以水解度、多肽含量为响应值,研究加酶量、酶用量比、复合酶解时间比、pH值对制备多肽工艺的影响。结果表明：酶法水解卵黄蛋白制备多肽的最佳工艺条件为：卵黄蛋白质量浓度10g/100mL、温度55℃、pH7.2,按0.8g/100mL添加碱性蛋白酶水解2h后,再按0.4g/100mL添加复合风味蛋白酶水解2h,在该条件下水解度和多肽含量分别为(13.31±0.5)%和(1.85±0.5)mg/mL。     

棉籽多肽的制备及其自由基清除能力研究

以碱溶酸沉法提取的棉籽蛋白为原料,分别用碱性蛋白酶、中性蛋白酶、胃蛋白酶、木瓜蛋白酶对其进行水解,比较4种蛋白酶水解棉籽蛋白的水解度及棉籽多肽自由基清除能力。结果表明:中性蛋白酶水解所得棉籽多肽自由基清除能力较强。在单因素试验基础上,采用正交优化试验,得到中性蛋白酶水解制备棉籽多肽最佳条件为:水解时间5 h,水解温度45℃,pH 7.5,加酶量8 000U(100 mL 5%棉籽蛋白溶液)。在最佳条件下棉籽蛋白的水解度为11.23%,制备的棉籽多肽对超氧阴离子自由基清除率为72.48%,对羟自由基清除率为89.01%,其10倍稀释液对DPPH自由基清除率为84.67%,与0.5 mg/mL VC的自由基清除能力相当。     

响应面法优化元宝枫籽粕酶解工艺及多肽功能特性研究

本文以元宝枫籽粕为原料,采用碱性蛋白酶法对元宝枫籽粕进行酶解,以酶解时间、加酶量、pH、酶解温度、料液比为考察因素,酶解多肽得率为评价指标,在单因素实验的基础上,根据Box-Behnken中心组合实验设计对元宝枫籽粕碱性蛋白酶酶解多肽制备工艺进行优化,并对优化工艺获得的酶解多肽进行了氨基酸组成、吸水性、吸油性、起泡性质、乳化性质和表面疏水性等功能特性表征。结果表明:最优的酶解制备工艺为:酶解时间3.3 h,pH为10,加酶量为3%,酶解温度为55 ℃。在最优制备工艺条件下元宝枫籽粕碱性蛋白酶酶解多肽得率为40.13%±0.15%。氨基酸组成分析表明酶解多肽所含八种必需氨基酸量高达20.3%,远高于国际粮农组织所建议成人所需必需氨基酸量。此外,酶解多肽的吸油性（4.553 g/g）高于大豆蛋白（2.61 g/g）,其表面疏水性（1365.3）与大豆7S球蛋白的表面疏水性相似,乳化性和乳化稳定性略低于大豆分离蛋白。本研究所获得的元宝枫籽粕碱性蛋白酶酶解多肽具有较好的功能特性,这也表明它可作为一种潜在的功能成分应用于食品中,为元宝枫籽粕的新应用开发提供数据和理论支撑。     

酶法制取棉籽水解蛋白研究

棉籽蛋白是我国一种重要的优质植物蛋白,对棉籽蛋白采用酶法水解,能显著改善功能特性,通过研究得出:酶处理的最适参数范围为温度40～45℃,时间5～7小时,PH7～8,加酶量2～3％;棉籽水解蛋白DH为17～18时乳化性较好,DH为13～14时起泡能力较强。     

微波辅助酶解棉籽粕的条件优化及酶解产物功能性质研究

研究了微波辅助碱性蛋白酶和风味蛋白酶双酶酶解棉籽粕的工艺条件.通过单因素实验确定了碱性蛋白酶酶解的最佳工艺条件为:微波温度60℃,微波功率500 W,酶加量5％(以底物质量计),酶解时间15 min;风味蛋白酶酶解的最佳工艺条件为:微波温度60℃,微波功率600W,酶加量5％(以底物质量计),酶解时间15 min.参照单因素优化条件,对棉籽粕进行连续酶解,酶解液多肽含量为13.32 mg/mL.棉籽粕经过微波连续双酶酶解后,吸油性、起泡性、乳化性等功能性质得到改善.     

茶渣多肽制备及其对羟自由基的清除能力

分别以酸性蛋白酶、中性蛋白酶、碱性蛋白酶以及复合蛋白酶水解茶渣粗蛋白制备茶渣多肽,并对多肽的羟自由基清除能力进行考察。通过单因素试验证明:复合蛋白酶的效果最好,其最佳水解工艺条件为粗蛋白液质量分数0.5%、加酶量600U/g、温度55℃、pH8.0、水解5h,在此条件下,多肽对羟自由基的清除能力达27.3%;碱性蛋白酶的效果也较好,其最佳工艺为蛋白液质量分数0.5%、加酶量400DU/g、pH8.5、温度50℃、时间1.0h,在此条件下,多肽对羟自由基的清除能力达25.3%,与前者的清除能力没有显著差异,但用时少4h;利用碱性蛋白酶作为酶源制得茶渣多肽粗品中含有蛋白质、多肽、游离氨基酸分别为28.4%、11.0%、1.5%,其对羟自由基的半清除率IC50为8.432mg/mL,相同条件下VC的IC50为0.897mg/mL。     

复合蛋白酶水解黑豆粕制备多肽的工艺优化

为充分利用黑豆粕中的蛋白质资源,研究了复合蛋白酶水解黑豆粕制备多肽的工艺路线。从蛋白酶的筛选入手,考察了不同蛋白酶对黑豆粕水解效果的差异,并通过复配比例优化了最佳复合蛋白酶的组成。采用单因素试验及响应面试验确定了复合蛋白酶水解黑豆粕的最佳工艺条件。结果表明:最佳复合蛋白酶为胰蛋白酶、碱性蛋白酶与桔青霉蛋白酶按酶活比1∶3∶2复配;最佳工艺条件为加酶量5 670 U/g,底物蛋白质量浓度40 g/L,pH 9.55,水解温度50℃,水解时间6 h。在优化的工艺条件下水解黑豆粕,多肽得率可以达到82.44%。水解液中多肽的相对分子质量均在7.8 kDa以下,大部分多肽的相对分子质量小于3.3 kDa;多肽基本上保持了黑豆粕蛋白的氨基酸组成,具有较高的营养价值。     

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.     

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.     

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.     

Analysis of benzo[a]pyrene in spiked fatty foods by second derivative synchronous spectrofluorimetry after microwave-assisted treatment of samples

A simple, rapid and inexpensive method has been developed for the determination of benzo[a     

The development of reference materials for paralytic shellfish poisoning toxins in lyophilized mussel. II: Certification study

This paper describes the second part of a project undertaken to develop certified mussel reference materials for paralytic shellfish poisoning toxins. In the first part two interlaboratory studies were undertaken to investigate the performance of the analytical methodology for several PSP toxins, in particular saxitoxin and decarbamoyl-saxitoxin in lyophilized mussels, and to set criteria for the acceptance of results to be applied during the certification exercise. Fifteen laboratories participated in this certification study and were asked to measure saxitoxin and decarbamoyl-saxitoxin in rehydrated lyophilized mussel material and in a saxitoxin-enriched mussel material. The participants were allowed to use a method of their choice but with an extraction procedure to be strictly followed. The study included extra experiments to verify the detection limits for both saxitoxin and decarbamoyl-saxitoxin. Most participants (13 of 15) were able to meet all the criteria set for the certification study. Results for saxitoxin.2HCl yielded a certified mass fraction of <0.07 mg/kg in the rehydrated lyophilized mussels. Results obtained for decarbamoyl-saxitoxin.2HCl yielded a certified mass fraction of 1.59+/-0.20 mg/kg. The results for saxitoxin.2HCl in enriched blank mussel yielded a certified mass fraction of 0.48 +/- 0.06 mg/kg. These certified reference materials for paralytic shellfish poisoning toxins in lyophilized mussel material are the first available for laboratories to test their method for accuracy and performance.     

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.