保加利亚乳杆菌菊芋复合汁增菌培养基的优化筛选

以菊芋为主要原料,以自行分离选育的高活力保加利亚乳杆菌(Lactobacillus bulgaricus)L.b-DR为试验菌株,研究了在菊芋汁培养基中添加单一营养因子对保加利亚乳杆菌L.b-DR细胞生长量的影响;利用正交试验优化筛选出保加利亚乳杆菌L.b-DR的菊芋复合汁增菌培养基。试验结果表明:在菊芋汁基础培养基中添加番茄汁、乳糖、蛋白胨、碳酸钙,可显著促进试验菌株的细胞生长(P<0.01);利用L934正交试验筛选出菊芋复合汁增菌培养基的最佳配比是:在菊芋汁培养基中添加7.5%番茄汁,1.5%乳糖,1%蛋白胨,0.3%碳酸钙。保加利亚乳杆菌L.b-DR经过复原脱脂乳培养基和液体MRS培养基活化后,以1%(约1×106cfu/mL)接入菊芋汁复合增菌培养基中,37℃培养16h,活菌数可达1.50×109cfu/mL,较对照菊芋汁培养基的活菌数提高23.96倍,与实验室常用的MRS培养基的活菌数相当,而其成本较实验室常用的液体MRS培养基的成本降低了2400元/t。     

乳酸菌冷冻保护剂配方设计

乳酸菌浓缩发酵剂的制备过程中,冷冻对菌体的损伤比较明显。通过在保加利亚乳杆菌和嗜热链球菌混菌菌悬液中添加不同的保护剂并测定冷冻前后乳酸菌菌数,筛选出适用的保加利亚乳杆菌和嗜热链球菌混菌的保护剂。最终确定的保加利亚乳杆菌和嗜热链球菌混菌保护剂配方为:蔗糖浓度为10%,脱脂乳浓度为10%,谷氨酸钠浓度为5%。菌体经冷冻处理后,冷冻存活率可达到95.0%,比不添加保护剂的菌体存活率大大提高。此配方也简单易得,完全可以满足生产需要。     

影响乳酸菌冷冻干燥抗性因素的研究

研究了生长培养基、菌体收获菌龄、预培养条件、保护剂等因素对保加利亚乳杆菌(Lactobacillus bul- garicus)冷冻干燥存活率和活力的影响。确定了加有1%生长因子和1%CaCO_3的NFM培养基(10%的脱脂乳),可使保加利亚乳杆菌的对数生长期延长。菌体的最佳收获菌龄为4.5 h;保护剂组成为:3%NFM+3%乳糖+3%甘油;40℃预培养1 h,其存活率可达75.5%。具有较为理想的效果。此外,在优化的试验条件下研制的冻干酸奶发酵剂制备的酸奶,达到了预定的标准。     

乳酸菌摇瓶增殖培养工艺研究

采用自行筛选的适用于作为高效冻干酸奶发酵剂的嗜热链球菌S.t-SY和保加利亚乳杆菌L.b-DR为出发菌株,在优化的麦芽复合汁增菌培养基上进行摇瓶增殖培养。通过研究培养温度、培养方式、基质起始pH值、接种量、中间补料方式等因素对乳酸菌摇瓶增殖培养的影响,优化乳酸菌摇瓶增殖培养的工艺条件。试验结果表明:在麦芽复合汁增菌培养基中乳酸菌增殖培养的最适条件是:培养温度37℃,基质起始pH值6.5～7.5,接种量0.2%～0.3%,静置培养16h,收获期S.t-SY和L.b-DR活菌数分别达到2.93×109、2.74×109cfu/mL。本试验结果为乳酸菌进一步应用麦芽复合汁增菌培养基生产高效浓缩直投式发酵剂提供了技术参数。     

嗜热链球菌与保加利亚乳杆菌麦芽复合汁增菌培养基的优化筛选

研究了在麦芽汁中添加不同的营养因子对嗜热链球菌S.t-3的细胞生长量的影响,进一步采用正交试验优化筛选出S.t-3的麦芽复合汁增菌培养基,并对保加利亚乳杆菌L.b-DR和L.b-S1进行了验证性增菌试验。结果表明,S.t-3、L.b-DR和L.b-S1在麦芽汁中能进行正常的产酸代谢,大豆蛋白胨、牛肉膏、酵母膏可显著促进S.t-3的细胞生长(P<0.05);K2HPO4不仅可以促进菌体增殖,而且还可缓冲基质pH值的变化,避免了低pH值对乳酸菌的伤害;利用L9(33)正交试验,筛选出S.t-3的麦芽复合汁增菌培养基最佳配比为:在10%的麦芽汁中添加0.5%大豆蛋白胨、0.5%酵母膏、1%牛肉膏、0.2%K2HPO4;在麦芽复合汁增菌培养基中,S.t-3、L.b-DR和L.b-S1,37℃恒温培养16 h,活菌数分别为2.42×109cfu/mL、2.85×109cfu/mL和4.81×109cfu/mL,与液体MRS培养基的活菌数相当,成本较MRS培养基降低1 000元人民币/t。     

保加利亚乳杆菌浓缩培养的研究

对培养保加利亚乳杆菌的基础培养基进行筛选,确定为MRS培养基。然后优化培养条件:发酵时间为12h,起始pH值为5.80,发酵温度为40℃。最佳培养基:MRS培养基 7.5%番茄汁 12%麦芽汁 0.009mol/LCaCl2 2%乳清。通过中和试验,保加利亚乳杆菌菌体浓度可达到9.55×109mL-1。     

保加利亚乳杆菌高密度培养的初步研究

对培养保加利亚乳杆菌的基础培养基进行优选,确定为基础MRS培养基.然后优化培养条件发酵时间为12h,起始pH为6.4,发酵温度为37℃.培养基MRS培养基 7.5%番茄汁 12%麦芽汁 5%海带汁 2%乳清.通过正交试验证明,保加利亚乳杆菌菌体浓度可达到1.0×1012 cfu/mL.     

保加利亚乳杆菌冷冻干燥保护剂的筛选

采用响应面分析法筛选并优化了保加利亚乳杆菌冻干保护剂,研究发现在众多物质中脱脂乳、海藻糖、菊粉、麦芽糊精对菌种保护作用突出,以冻干后细胞存活率为响应值,当脱脂乳5.94%,海藻糖7.75%,菊粉8.39%,麦芽糊精9.04%作为混合保护剂使用时,细胞存活率达到90.46%。使用液氮进行预处理并且保护剂的体积为菌体两倍时可以使菌体存活率达到91.93%.     

冷冻保护剂对直投式木葡萄酸醋杆菌发酵剂菌体细胞特性的影响

本文研究了1%谷氨酸、5%海藻糖、10%脱脂乳及复合0.575%谷氨酸+0.075%海藻糖+6.4%脱脂乳作为冷冻保护剂对直投式木葡萄酸醋杆菌(GBX)发酵剂菌体细胞特性的影响,通过测定GBX发酵剂菌体细胞的DNA泄露、细胞液总抗氧化能力、SOD活性、纤维素合成酶活性和ATP酶活性变化等情况来衡量GBX菌体细胞的生理活性变化。结果发现:添加单一1%谷氨酸、5%海藻糖、10%脱脂乳以及复合0.575%谷氨酸+0.075%海藻糖+6.4%脱脂乳作为保护剂均能明显抑制冷冻干燥过程中菌体细胞的DNA泄露,提高细胞总抗氧化能力、SOD活性、纤维素合成酶和ATP酶活性,表明添加冷冻保护剂能够一定程度上保护菌体细胞,其中复合保护剂对菌体细胞的保护效果最佳,电子扫描显微镜也验证了冷冻保护剂能确实降低菌体细胞在冷冻干燥过程中损害程度,从而使菌体细胞保持良好的生理活性。     

酸奶冻干发酵剂制备中保加利亚乳杆菌不同菌株特性的研究

研究了不同来源的7株保加利亚乳杆菌在乳中的生长与发酵特性、后酸化活性以及抗冷冻干燥特性。结果表明,7菌株在乳中42℃发酵,L.b-S1和L.b-DR凝乳时间最短,为3 h,凝乳后的活菌数、pH值、滴定酸度均无显著差别,活菌数均达1×108 mL-1以上,pH值均达4.5～5.0,滴定酸度均达90～100°T;7菌株在发酵后的酸乳中冷藏期间,L.b-S1和L.b-DR的后酸化活性最低,4℃冷藏21 d,酸度上浮不足10°T,pH值下降0.2～0.4,活菌数下降1个log数量级左右;7菌株在以脱脂乳为保护剂的冷冻干燥试验中,L.b-S1抗冻干性最强,其次为L.b-DR,其冻干存活率分别达31.46%和20.39%。     

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.     

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.     

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.     

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.     

Influence des ions sur le pouvoir hydratant de l'ure e: e tude sur peau de porc ex vivo

This study deals with the influence of ions (NaCl and MgSO4) in a W/O emulsion containing 10% urea. Moisturization kinetics are assessed by corneometry on pig skin ex vivo. The formula's influence on urea penetration is measured by infrared spectrometry with an ATR device and the stripping method. Corneometry and spectroscopy were chosen to record simultaneously the hydratation levels and urea localization into superficial cell layers. Urea crystallization after evaporation of emulsions and aqueous solutions is described. Results show that urea does not hydrate nor penetrate when applied to the skin through an aqueous gel. In a W/O emulsion, sodium chloride increases the ability of urea to moisturize without improving penetration. In vitro urea crystallization is disturbed by sodium chloride or magnesium sulphate for solutions and emulsions. This stabilization by ions is correlated with good moisturization values. The stabilization of urea in the solute state provided by ions increases its water epidermal binding capacity without enhancing penetration.     

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.