金属离子对红发夫酵母的生长、细胞形态及虾青素合成的影响

主要研究了Mn2+、Mg2+和Ca2+三种金属离子对红发夫酵母(Phaffiarhodozyma)的生长、细胞形态及虾青素生物合成的影响。结果表明,在种子培养基中添加Mn2+能明显提高红发夫酵母的β-D-葡萄糖苷酶的活性并改变细胞的形态;在发酵培养基中添加0.1g/LMn2+,能明显促进细胞生长及虾青素合成;用正交试验法优化得到发酵培养基中的金属离子浓度为:0.1g/LMn2+、0.7g/LMg2+、0.2g/LCa2+,在优化后的培养基中,细胞生长速度快,摇瓶发酵57h的虾青素产量达2.95mg/L,是不加金属离子情况下发酵产虾青素最大量(发酵72h)的1.31倍。     

KNO_3对红发夫酵母生物合成虾青素的影响

比较了红发夫酵母(Phaffiarhodozyma)以KNO3、NaNO3、(NH4)2SO4及蛋白胨为氮源的细胞生长及虾青素合成过程,重点研究了KNO3对红发夫酵母生物合成虾青素的影响。结果表明:低浓度KNO3(0·1~0·9g/L)有利于细胞生长及虾青素合成,而高浓度KNO3(3·0~9·0g/L)抑制细胞生长及虾青素合成。当培养基中含有0·3g/LKNO3时,红发夫酵母的发酵过程不仅表现出细胞二次生长及虾青素二次合成现象,而且发酵周期较长,可获得较高的生物量和干细胞虾青素含量,分别为8·13g/L和0·993mg/g。     

豆腐黄浆水中补加乙醇和葡萄糖发酵虾青素的研究

在豆腐黄浆水中补加乙醇和葡萄糖发酵虾青素进行了研究,结果表明,黄浆水中添加6g/L乙醇,可提高生物量、虾青素含量以及虾青素产量,但发酵周期有所延长;黄浆水中添加10g/L葡萄糖,有利于酵母菌的生长和提高虾青素产量。     

两阶段法培养红法夫酵母生产虾青素

在法夫酵母生长过程中,培养基组成是影响细胞生长和虾青素产量的重要因素。文中通过正交设计和响应面优化方法分别对红法夫酵母生长阶段培养基和虾青素合成阶段的培养基进行了优化研究。结果表明,碳源和氟源浓度对红法夫细胞的生长及虾青素的合成有明显影响,浓度为20 g/L的葡萄糖有利于细胞的生长;而50 g/L左右的葡萄糖和高C/N有利于虾青素的合成。在此基础上提出了两阶段发酵方案。经过两阶段发酵,红法夫酵母生物量和虾青素产量达到16.8 g/L和15.015 mg/L,分别比分批培养提高了56.3%和28.7%。     

不同时间流加番茄汁对红发夫酵母生产虾青素的影响

番茄红素作为虾青素合成的前体物质,适量添加对虾青素合成起到促进作用.由于成熟番茄汁中含有大量的番茄红素,可以作为番茄红素的替代品,而且番茄汁含有的其它营养物质对红发夫酵母的生长也有一定的积极作用.这样既促进了虾青素的合成,又节约了成本,故选择番茄汁作为流加物质.结果表明,培养36h时添加番茄汁,红发夫酵母72～120h生长状态平稳,且对提高虾青素含量和产量效果显著,培养96h虾青素含量高达1318 μg/g,虾青素产量也达到最高,为17.87mg/L.     

乙醇对法夫酵母发酵合成虾青素的影响

利用摇瓶培养法夫酵母JMU-MVP14菌株,通过乙醇传感器检测乙醇含量,研究乙醇对法夫酵母细胞生长及虾青素合成的影响。结果表明,在10 g/L和20 g/L葡萄糖初始培养基中分别添加1～2 g/L乙醇时,均可提高虾青素的产量;而添加高浓度乙醇,则会抑制细胞生长和虾青素的合成。以10 g/L葡萄糖为初始培养基,控制发酵中恒定乙醇浓度为2 g/L,虾青素质量浓度可达到50.1 mg/L,比只添加2 g/L乙醇提高了20.3%,且虾青素细胞产率达到13.1 mg/g,葡萄糖的利用率能达到77.2%以上,说明恒定乙醇控制策略更有利于法夫酵母虾青素的合成。在10 g/L初始葡萄糖浓度,恒定乙醇浓度2 g/L,并每24 h补加5 g/L葡萄糖的优化条件下,虾青素产量最大值达到55.3 mg/L,虾青素细胞产率为19.7 mg/g,比对照组(14.1 mg/g)提高了39.7%。     

氮饥饿/过氧化氢协同胁迫促进法夫酵母合成虾青素

虾青素是一种强抗氧化剂,在细胞抵御外界环境胁迫方面起到重要作用。研究了氮饥饿/过氧化氢协同胁迫对法夫酵母合成虾青素的影响,结果表明:低含氮量有利于虾青素的合成,随着碳氮比的降低,虾青素的合成受到抑制,最佳的碳氮比为3∶0.6,每克干细胞虾青素的含量最高达0.32mg/g;在此基础上添加5mmol/L过氧化氢协同胁迫法夫酵母可进一步提高虾青素的产量,在发酵24h(对数生长期)加入3mmol/L的H2O2,36h补加2mmol/L,虾青素产量达到0.59mg/g,比空白对照提高63.9%。     

低氮促进红发夫酵母合成虾青素机理的研究

培养基中初始氮(硫酸铵)浓度对红发夫酵母虾青素合成有明显的影响,低氮能够促进红发夫酵母合成虾青素.运用代谢通量分析的方法,定量分析不同氮浓度下红发夫酵母的中心碳代谢表明:生成乙酰辅酶A的反应的通量与合成虾青素反应的通量正相关,并且这两个通量都随培养基中初始氮浓度的降低而升高.这说明乙酰辅酶A是红发夫酵母合成虾青素的一个限制因素.进一步测定红发夫酵母丙酮酸代谢相关酶的活性表明:参与丙酮酸脱氢酶旁路的两种酶--乙醛脱氢酶和乙酰辅酶A合成酶在红发夫酵母中的活性极低;然而,柠檬酸裂解酶的活性较高,并且与红发夫酵母虾青素的合成明显呈正相关.该结果说明,柠檬酸裂解反应决定着红发夫酵母细胞质中乙酰辅酶A的供给,从而显著影响着其虾青素的合成.低氮条件能够提高柠檬酸裂解酶的活性和限制蛋白质的合成,从而增加乙酰辅酶A的供给,促进红发夫酵母合成虾青素.     

不同培养方式对红发夫酵母产虾青素的影响

通过间歇流加培养、指数流加培养、混合碳源-番茄汁流加培养和恒速流加培养等4种培养方式的实验,说明进行流加培养有利于红发夫酵母细胞生长及虾青素的合成。与分批培养相比,所有流加模式下生物量及虾青素产量都有不同程度地提高。其中流加混合碳源-番茄汁获得了最好的实验结果,最终的生物量、虾青素含量和产量分别达到33.71g/L、1106μg/g和36.26mg/L。     

几种物质对红发夫酵母生长和虾青素合成的影响

研究2-脱氧葡萄糖、胡萝卜素、番茄红素等物质对红发夫酵母生长及虾青素合成的影响。番茄红素的添加可以明显提高红发夫酵母细胞内虾青素含量,番茄红素的添加量为700μg/L时,虾青素含量与产量均达到最高,分别为1468μg/g、17.59 mg/L,对照空白分别提高了72%、69%。结果表明番茄红素适量添加对虾青素合成起到促进作用,但成本较高,由于番茄汁富含番茄红素并且成本较低,可以作为替代品进行流加,节约成本。     

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.