类柠檬苦素生物转化与脱苦研究进展

类柠檬苦素中柠檬苦素的苦味是世界柑橘属果汁加工业的一大障碍,类柠檬苦素生物脱苦法是目前改善苦味的首选方法。本文主要概述类柠檬苦素的结构与分类、自然生物转化与“后苦”现象、酶法脱苦、固定化细胞脱苦和基因工程脱苦等方面的国内外研究进展。     

采收期、贮藏时间及加工单元操作对赣南脐橙汁苦味物质含量的影响

探讨赣南脐橙采收期、贮藏时间及加工单元操作对橙汁苦味物质(柠檬苦素、诺米林)含量的影响,为降低橙汁苦味提供参考。测定不同采收期与贮藏时间及不同加工单元操作下的橙汁中的苦味物质含量,重点研究橙汁热处理过程中苦味物质的变化并进行动力学模型拟合。结果表明,随着采收期的延长,橙汁中柠檬苦素与诺米林的含量逐渐降低,但在贮藏期间,橙汁中的柠檬苦素与诺米林含量先升高后降低;破碎汁中柠檬苦素与诺米林含量均高于挤压汁,酶解工艺后破碎汁和挤压汁中柠檬苦素含量均大幅升高,而诺米林未被检出;热处理温度越高、pH值越低,橙汁中苦味物质含量越高,当pH值>6.0时,柠檬苦素和诺米林均未在橙汁中检出,同时可用联合反应模型拟合热处理过程中橙汁柠檬苦素和诺米林含量变化。脐橙采收期与贮藏期和加工单元操作特别是热处理条件均对橙汁苦味物质的含量影响较大,后续可通过延迟采收期与贮藏时间、优化取汁方法及控制热处理参数等措施降低橙汁的苦味物质含量。     

柑桔发酵酒脱苦技术研究

柑枯发酵酒中苦味物质主要由柠檬苦素,柚皮苷等组成,本研究采用吸附法,酶制剂等脱苦方法,确定最佳酶制剂脱苦工艺参数。     

柠檬苦素高效降解菌的分离筛选与鉴定

筛选出在酸性环境下高效降解柠檬苦素的菌株,扩展了生物法消除柑橘果汁中的苦味物质的应用领域。从新鲜醋醅中粗筛并分离出以柠檬苦素为唯一碳源的8株菌,其相应的柠檬苦素降解率均达70%以上,其中6、7、8号菌的柠檬苦素降解率较高,分别为95.14%、98.21%和94.31%。比较3株高效降解菌的胞外蛋白、胞内蛋白和菌液的柠檬苦素降解能力,3株菌中具有降解柠檬苦素的活性蛋白酶主要分布在细胞外,属于胞外蛋白酶,胞内蛋白对柠檬苦素几乎无降解作用,其中6号菌株胞外酶的柠檬苦素降解活性最强,柠檬苦素降解率为45.57%。形态学观察及分子生物学鉴定6、7、8号菌均属于拉乌尔菌属(Raoultella sp.)。菌株6具有高效降解柠檬苦素能力,能够成为今后降解柠檬苦素微生物源的优势菌种。     

不同甜橙品种果汁中柠檬苦素含量的变化

为了减少柠檬苦素对橙汁品质的影响,为橙汁加工的原料品种和加工时间选择提供参考,对20个甜橙品种的鲜榨果汁与巴氏灭菌果汁,在7个不同成熟期,采用高效液相色谱法检测了柠檬苦素和诺米林含量。结果表明:各成熟阶段的所有测试甜橙品种的鲜榨果汁与巴氏灭菌果汁均能检测到柠檬苦素,巴氏灭菌果汁中柠檬苦素含量更高,平均是鲜汁的约2倍;随着果实的成熟,柠檬苦素含量总体逐步降低,到11月中旬时,试验甜橙品种的鲜榨果汁与巴氏灭菌果汁的柠檬苦素含量已低于人的苦味阈值,适于橙汁加工;不同甜橙品种甚至不同脐橙品种之间,柠檬苦素的含量存在明显差异;诺米林含量与柠檬苦素含量呈现类似变化,但含量更低,不是导致橙汁苦味的主要因素。此外,短期的冷冻贮藏对柠檬苦素的含量影响不大。     

柑橘类果汁脱苦技术研究进展

柚皮苷和类柠檬苦素是柑橘类果汁的主要苦味物质,其存在明显影响了柑橘类果汁的品质。介绍了脱除柑橘类果汁苦味的一些技术：代谢脱苦、吸附脱苦、酶法脱苦、屏蔽法脱苦、分离技术脱苦和基因脱苦等。     

柑橘类的功能性成分研究概况

概述了柑橘类的功能性成分,包括类黄酮类、类柠檬苦素类、胡萝卜素类以及其它一些生理活性物质。其中,类黄酮类和类柠檬苦素类是柑橘中的最主要生理活性成分。     

柑橘中苦味物质对HeLa细胞增殖与凋亡的作用机制

目的：以‘高台’蜜柚种子和白色海绵层为原料,提取柠檬苦素、诺米林和柚皮苷3 种苦味物质,分别探究它们对宫颈癌HeLa细胞增殖和凋亡的作用机制。方法：选择超声波辅助有机溶剂萃取和重结晶法,采用Acquity UPLC BEH C18色谱柱（2.1 mm×50 mm,1.7 μm）,对3 种苦味物质提取、分离纯化和检测;在50 μmol/L的工作浓度下,以3 种苦味物质对HeLa细胞进行处理,利用细胞增殖检测、细胞划痕和逆转录-聚合酶链式反应实验进行抑癌作用的验证。结果：柠檬苦素类似物和柚皮苷的提取率分别为5.21 mg/g和7.72 mg/g;柠檬苦素、诺米林和柚皮苷的纯度分别为94.08%、93.91%和93.39%;抑癌活性实验证实了3 种苦味物质能抑制HeLa细胞增殖和迁移。     

柑桔类果汁脱苦法

柑桔类水果包括柠檬、脐橙、酸橙、甜橙、蜜柑、柚、葡萄柚、红桔、金桔等,一般中晚熟柑桔在鲜食时无苦味,但当它们经过榨汁、过滤以及杀菌处理等加工成果汁后,都带有一种令人讨厌的苦味,从而限制了果汁在食品工业中的普遍应用。柑桔类果汁之所以有苦味,主要是由于外皮、内膜、筋络、种籽中含有的两类苦味化合物而造成。一类是黄酮类化合物,其苦味主要是柚苷;另一类是柠檬苦素类化合物,其苦味代表物是柠碱和诺米林,因此,为了去除苦味,扩大果汁的应用,我们有必要对柑桔类果汁的脱苦法作一介绍,以供生产厂家和同行参考。     

几种柑橘类果汁中主要苦味物在加工过程中含量变化的研究

目的:研究榨汁温度、热处理、过滤等加工技术对几种柑橘类果汁主要苦味成分的影响, 为脱苦工艺提供试验依据.方法:采用高效液相色谱法检测3种柑橘类果汁中柚皮苷和柠檬苦素类化合物的含量, 比较不同的预处理方式、加热温度、加热时间对其含量的影响.结果:不同品种的柑橘果汁的主要苦味物质含量差别较大, 果实预冷至4℃后榨汁可减少苦味,果汁苦味物随热处理温度的升高而增加, 同时柚皮苷和柠檬苦素类化合物的含量分别在加热至20 min和15 min时达到最大值.结论:榨汁温度、热处理、过滤等加工技术对柑橘类水果的苦味物质含量具有重要的影响,控制适宜的工艺参数有利于减少果汁中苦味物质的含量.该研究对于柑橘果汁的脱苦工艺具有重要的参考价值.     

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.     

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.     

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.     

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.     

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.     

Food-borne Listeria monocytogenes risk assessment

Listeria monocytogenes is ubiquitous in the environment and in food processing plants. Consequently, foods are frequently contaminated. However, the occurrence rate of listeriosis is only about five cases per million people per year. Listeriosis primarily strikes immunocompromised individuals, pregnant women and the elderly with a fatality rate of 20-25%. The FDA is in the process of finishing a risk assessment that is being conducted as an initial step in reviewing its approach to maximizing the public protection from foodborne L. monocytogenes . The risk assessment evaluated the presence and quantitative levels of L. monocytogenes in 21 groups of ready-to-eat foods. The potential growth of L. monocytogenes between retail point-of-sale, where contamination data originated, and consumption was modelled. The frequency and amount of consumption of these foods completed the data for the exposure assessment. For the hazard characterization or dose response part of the risk assessment, data from animal studies, virulence assays and epidemiological investigations were used to estimate the likelihood of illness for different human groups from consuming different numbers of L. monocytogenes . This risk assessment is a virtual review of current scientific knowledge. Quantitative modelling provides greater insight than a qualitative review and also indicates the uncertainty about our knowledge. The risk assessment does not attempt to define an acceptable or tolerable level of L. monocytogenes consumption or propose changes in regulations. These decisions are the responsibility of risk managers who consider additional factors such as food preferences, technical feasibility and societal values when evaluating regulatory policies.     

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.     

Food allergy—towards predictive testing for novel foods

The risks associated with IgE-mediated food allergy highlight the need for methods to screen for potential food allergens. Clinical and immunological tests are available for the diagnosis of food allergy to known food allergens, but this does not extend to the evaluation, or prediction of allergenicity in novel foods. This category includes foods produced using novel processes, genetically modified (GM) foods, and foods that might be used as alternatives to traditional foods. Through the collation and analysis of the protein sequences of known allergens and their epitopes, it is possible to identify related groups which correlate with observed clinical cross-reactivities. 3-D modelling extends the use of sequence data and can be used to display eptiopes on the surface of a molecule. Experimental models support sequence analysis and 3-D modelling. Observed crossreactivities can be examined by Western blots prepared from native 2-D gels of a whole food preparation (e.g. hazelnut, peanut), and common proteins identified. IgEs to novel proteins can be raised in Brown Norway rat (a high IgE responder strain), and the proteins tested in simulated digest to determine epitope stability. Using the CSL serum bank, epitope binding can be examined through the ability of an allergen to cross-link the high affinity IgE receptor and thereby release mediators using in vitro cell-based models. This range of methods, in combination with data mining, provides a variety of screening options for testing the potential of a novel food to be allergenic, which does not involve prior exposure to the consumer.     

Peptides from milk proteins and their properties

This review has attempted to study the literature pertaining to peptides derived from milk proteins. Hydrolysis of milk proteins to generate peptides has been practiced for a long time and it was recognized early on in this process that the taste of hydrolyzates might hinder use of these products in food formulations. Modification of protein is necessary to form a more acceptable or utilizable product, to form a product that is less susceptible to deteriorative reactions and to form a product that is of higher nutritionall quality. Modifications may be achieved by a number of chemical and enzymatic means. This review has considered only enzymatic modification of dairy proteins. Modified proteins contain peptides and some of these peptides have been purified and their functionalities have been compared with unmodified proteins. This paper has examined the literature pertaining to improvement in functionality of enzyme-modified proteins. Improvements in solubility, emulsification, foaming and gelation were examined. There is limited information available on the sequence of the peptides necessary to improve the functional characteristics of proteins. Knowing the sequences of desirable functional peptides can lead to genetic alteration of proteins to improve functionality. Addition of synthetic peptides to intact proteins may be another way in which the functionality of proteins can be augmented. Some of the peptides in milk proteins are capable of affecting biological functions of an organism. These effects can be antimicrobial and probiotic, i.e., prevent the growth and proliferation of undesirable and pathogenic organisms, or they may promote the growth of desirable bacteria in the digestive tract of humans and animals. Peptides derived from milk protein have been shown to exert digestive and metabolic effects as well. They may also influence the immune system. These biological effects may play an important role in the development of medical foods that treat or mitigate the effects of diseases. Proteins are allergens and therefore it is possible that products derived from modification of proteins may also be allergens. The known literature about the allergenicity of peptides derived from milk proteins has been examined in this article. Last, but not the least, the taste attributes of peptides is also considered. Bitterness of hydrolyzates is a common occurrence and the origins of these bitter peptides and possible ways of mitigating this sensory defect has been discussed. Many of the peptides that enhance functionality and exert biological activity are likely to be bitter. Therefore, the bitter taste of hydrolysis products has to be dealt with in boosting the functional or nutraceutical aspects of foods containing these peptides. Analytical techniques for sequencing peptides have become more accessible and purification of peptides is commercially feasible. Computer based modeling techniques have aided the prediction of structures in these peptides. These advances, coupled with the advances in biotechnology, promise to revolutionize the future of nutraceutical and functional foods.