高分子量透明质酸产生菌选育及发酵条件优化

以Streptococcus equi SH-O为出发菌,用5-溴尿嘧啶(5-BU)、紫外线和N-甲基-N-硝基-N-亚硝基胍(NTG)为诱变剂进行复合诱变,选育到1株遗传稳定性好、产高分子量透明质酸(HA)的突变菌SH-109;在500mL摇瓶和5L发酵罐中,分别对其生产高分子量HA的发酵条件进行了优化.多因素正交试验表明发酵培养基的最佳配比为:50g/L蔗糖、100g/L混合氮源、10g/L葡萄糖、2.0g/L MgSO4·7H2O、2.0g/L Na2HPO4·12H2O、1.0g/L NaHCO3、0.1g/L UTP.在pH7.6、转速550r/min条件下有利于高分子量HA的合成,突变菌SH-109发酵生产HA相对分子量达4.36×106Da.     

复合诱变选育大分子量透明质酸高产菌株

大分子量透明质酸高产菌株的选育是工业化生产透明质酸的关键,本实验以兽疫链球菌为出发菌,采用亚硝基甲基脲和15keV低能氮离子进行复合诱变,筛选出一株溶血素和透明质酸分解酶双缺陷型突变菌株SZ-36,采用摇瓶发酵培养后,透明质酸的产量最高达到3.16g/L,相对分子量达到2.42×106u,比原始菌株分别提高了267.4%和137.3%,并经10次传代以后,突变菌株SZ-36具有良好的传代稳定性,为工业化生产透明质酸提供了优良菌株。     

pH调控方式和温度对透明质酸发酵过程的影响

本文以一株产透明质酸(hyaluronic acid, HA)的兽疫链球菌为研究对象,在5.00 L发酵罐和250 mL摇瓶中考察了温度和pH对HA发酵过程中菌体生长、HA产量、HA分子量大小以及分布的影响。结果显示,当pH为7.00时适合菌体生长,而在产物合成阶段更高的pH条件8.00下HA比生成速率更高。据此,设计了分段和周期性改变发酵过程pH的调控模式,研究表明:分段pH调控方式与pH为7.00时相比HA浓度没有提高,但是pH在7.00和8.00之间进行的分段调节时分子量提高了10.60%,同时显著降低了多分散系数,另外在pH在7.00和8.00之间进行的分段调节的同时在33℃的培养温度下收获时HA分子量提高了18.70%,而为后续工业化生产分子量更为集中的HA,同时降低不同分子量HA分离成本提供了方向。     

pH值对透明质酸钠溶液溶血性测定的影响

目的探讨pH值对透明质酸钠（HA）溶液溶血性的影响。方法调整0．4％HA溶液的pH值,以1％羊血悬浮液检测溶血性。结果0．4％HA溶液的pH〈4时,溶血检测阳性;调整pH5．5—8．0时,溶血阴性。结论0．4％HA溶液在pH〈4时,其溶血性检测结果为假阳性。检测时应将HA溶液的pH调至5．5～8．0。     

pH值对透明质酸钠溶液溶血性测定的影响

目的 探讨pH值对透明质酸钠(HA)溶液溶血性的影响.方法 调整0.4%HA溶液的pH值,以1%羊血悬浮液检测溶血性.结果 0.4%HA溶液的pH<4时,溶血检测阳性;调整pH 5.5～8.0时,溶血阴性.结论 0.4%HA溶液在pH<4时,其溶血性检测结果为假阳性.检测时应将HA溶液的pH调至5.5～8.0.     

pH值对透明质酸钠溶液溶血性测定的影响

目的探讨pH值对透明质酸钠(HA)溶液溶血性的影响。方法调整0.4%HA溶液的pH值,以1%羊血悬浮液检测溶血性。结果0.4%HA溶液的pH<4时,溶血检测阳性;调整pH 5.5~8.0时,溶血阴性。结论0.4%HA溶液在pH<4时,其溶血性检测结果为假阳性。检测时应将HA溶液的pH调至5.5~8.0。     

复合诱变选育透明质酸高产菌株

以马疫链球菌AF-0为出发菌株,用LiCl结合紫外线及硫酸二乙酯(DES)进行复合诱变,涂布筛选培养基,从中筛选出溶血素和透明质酸降解酶的双缺陷型突变菌株,经摇瓶发酵考察得到一株产量较高的菌株C4,并经多次传代,其遗传性基本稳定,但随传代次数增多,HA产量有下降趋势;其透明质酸产量达3.32g/L,分子量达1.45×106u,相对于原始出发菌株,产量及分子量分别提高了78%和40%.     

透明质酸-明胶复合膜的制备及理化性质研究

为研发保鲜效果良好的生物基可食用膜,采用流延法将不同分子量的透明质酸(Hyaluronic acid,HA)与明胶制成透明质酸-明胶复合膜,测定其厚度、密度、抗拉强度、不透明度、色度、溶解性、水蒸气透过率和水分含量等理化指标,分析HA分子量对复合膜理化性质的影响,并使用红外光谱表征复合膜的结构。结果表明,随着HA分子量的增加,复合膜厚度、水分含量和不透明度呈现先增加后降低,溶解性和水蒸气透过率降低。红外光谱中酰胺A区特征峰向短波迁移,表明HA分子与明胶分子之间存在氢键作用。透明质酸分子量为(80~100)×104 Da时,复合膜的综合性能最佳。HA-明胶复合膜的开发在食品包装和保鲜方面有潜在的应用价值。     

乳酸发酵短杆菌中的赖氨酸代谢控制发酵

以本室选育的赖氨酸产生菌AL 0 39为出发菌株 ,以亚硝基胍诱变 ,筛选到一株氟丙酮酸敏感型突变株FP 0 94,其赖氨酸产量比出发菌株提高 37.5 %。进而研究了生物素、醋酸钙对该菌产赖氨酸的影响。     

体外酶法水解制备小分子透明质酸

为了制备分子量分布集中的小分子透明质酸,在6 g/L的高分子量透明质酸水溶液中加入一定梯度重组水蛭透明质酸酶,并控制水解时间。结果表明,通过控制加入重组水蛭HAase的量和水解时间,根据水解分子量下降规律,可以制备出分子量分布集中的小分子HA,包括水解终产物透明质酸四糖和六糖。同时,通过高效体积排阻色谱和琼脂糖凝胶电泳,可以证明水解产物的分子量分布集中。     

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.     

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.     

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.     

低温带皮菜籽粕微粉的不同粒级部分的功能特性

为研究低温带皮菜籽粕微粉的不同粒级部分的功能特性,以经低温脱脂的带皮菜籽粕为原料,经微粉碎后筛分成212~425μm、150~212μm和106~150μm的3个不同粒级的微粉样品,检测这些样品的吸水性、吸油性、乳化性和乳化稳定性、蛋白质体外消化率。结果表明:1 3个不同粒级的微粉样品之间的粗纤维含量存在显著差异,表明三者的结构组成成分有一定差异。23个微粉样品的乳化活性和乳化稳定性随粒度级别的减小而显著增加(P0.01)。33个微粉样品的蛋白质体外消化率随粒度级别的减小而显著增加(P0.01)。4不同粒级带皮菜籽粕微粉样品的吸水性与吸油性受其结构组成物质不同和粒度的双重影响,与粒度的相关性不明显。     

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.