产角蛋白酶菌株的筛选及发酵条件优化

采用以羽毛为唯一碳氮源的无机盐培养基,通过考察HC值(水解圈直径与菌落直径的比值)、羽毛的降解程度,筛选得到10株产角蛋白酶的菌株,经测定10株菌株摇瓶发酵液的蛋白酶活力,确定了一株产酶能力较强的菌株CJPE209,并对菌株进行鉴定,菌株CJPE209被鉴定为芽孢杆菌(Bacillus sp.),其发酵液酶活为267 U/mL。通过单因素实验,确定了菌株CJPE209产角蛋白酶的最适发酵条件:发酵周期48 h,培养基装液量25 mL/250 mL,发酵温度37℃,初始pH7,摇床转速220 r/min,发酵液酶活为337.6 U/mL,是优化前的1.26倍。     

产脂肪酶菌株的筛选鉴定及产酶条件优化

以橄榄油为唯一碳源,采用油脂同化平板从食堂废弃物中筛选出一株产脂肪酶菌株HFE722。通过测定与分析该菌株16S rRNA基因序列,鉴定该菌株为芽孢杆菌（Bacillus sp.）。菌株HFE722在初始条件（发酵温度30 ℃,接种量为1%,自然pH,装液量为100 mL/250 mL,摇床转速为200 r/min）下培养36 h,测得发酵液上清液脂肪酶酶活为2.17 U/mL。优化后所得菌株HFE722产酶的最适发酵条件为：发酵温度30 ℃,发酵周期为36 h,接种量为1%（V/V）,初始pH 7.0,装液量为50 mL/250 mL,摇床转速为160 r/min。在最佳发酵条件下,发酵液上清液酶活可达到5.8 U/mL,酶活较优化前提高了167.28%。     

传统发酵酸牛奶中产蛋白酶酵母菌的筛选及产酶条件优化

从新疆哈萨克族传统发酵酸牛奶中分离筛产蛋白酶酵母菌菌株,并确定了各菌株的产酶能力。试验从新疆塔城地区采集酸牛奶样品,以蛋白酶活性作为筛选指标进行初筛、复筛,得到5株高产蛋白酶菌株并对其产酶条件进行优化。经形态观察及26S rDNA序列分析鉴定菌株D1-3为Pichia kudriavzevii,E1-5为Issatchenkia orientalis,S1-7为Kluyveromyces marxianus,W1-10为Candida parapsilosis,S1-16为Saccharomyces cerevisiae。对其产酶条件优化得出:在最佳产酶发酵液条件下,菌株D1-3最佳产酶活力达78 U/m L、S1-7为79 U/m L、S1-16为55 U/mL、E1-5为82 U/mL、W1-10为64 U/mL,其最适产酶温度为28℃,W1-10为37℃;菌株最适产酶初始p H、接种量和时间分别为5.0、3%、48 h。     

多粘菌素E降解酶产生菌的筛选及其产酶条件的研究

多粘菌素类抗生素在畜牧业中广泛使用,其药物残留给环境及人类健康带来了严重影响.从实验室保藏的若干蛋白酶产生菌中筛选得到高效降解多粘菌素的菌株DC-01,经形态观察和16S rRNA序列分析鉴定为地衣芽孢杆菌(Bacillus licheniformis).采用抑菌圈法测定菌株B.licheniformis DC-01在各种产酶条件下的发酵液对多粘菌素E的降解能力,结果表明:该菌株最佳产酶条件为温度37~44℃,初始pH值7.0~8.0,接种量8%,250mL烧瓶装液量50mL,发酵培养基中碳氮源组合为5.2%蔗糖和4.0%大豆蛋白胨.在最适发酵培养基中,37℃,200r/min摇床振荡培养24h产生的粗酶液37℃下对10 000U/mL多粘菌素E作用1h的降解率达92.1%,为多粘菌素E的残留处理提供了良好的菌种资源.     

发光杆菌产几丁质酶的工艺优化

为提高海洋发光杆菌Photobacterium sp.LG-1产低温几丁质酶的酶活性,利用响应面法对其发酵产酶条件进行了优化。Plackett-Burman实验结果表明,影响菌株产酶的三个主要因素分别为发酵温度、发酵时间和酵母膏含量。菌株产酶的最适条件为:胶体几丁质浓度12.0 g/L、酵母膏浓度4.5 g/L、转速220 r/min、发酵温度20℃、装液量75 mL/250 mL、接种量1%、初始pH7.0、发酵时间120 h,几丁质酶的最大酶活为5.10 U/mL。实际平均酶活经验证与预测酶活相近,几丁质酶活比优化前提高了11.50%。为下一步低温几丁质酶的降解机制研究及在工业中的应用提供参考。     

烟曲霉菌株发酵产壳聚糖酶条件的优化

通过单因子试验和正交试验对选育的一株产壳聚糖酶菌株UC-233进行发酵条件优化,结果表明其最适氮源是黄豆粕粉,最适碳源是壳聚糖,其最适发酵条件是:黄豆粕粉3.3%,胶体壳聚糖1.5%,培养基初始pH为4.0,250mL三角瓶装液量为50mL,接种量为1mL,发酵温度是30℃,发酵时间为48h,在此条件下菌株发酵液的壳聚糖酶活力达39.5U/mL,是优化前的1.43倍.     

传统阳江豆豉发酵过程中米曲霉的筛选及发酵条件优化

对从广东阳江豆豉的曲醅中筛选的米曲霉菌株进行培养基和发酵条件优化,结果表明,该菌产酶最适培养基的组成为添加3%的酵母粉、1%的葡萄糖、3%的硝酸钠和0.50%的氯化钙于基础PDA培养基中。该菌株产酶最优发酵条件为温度30℃,pH值为7~8,250 mL三角瓶中装瓶量为120 mL。在此最优条件下进行生长曲线绘制,培养12 h时,开始出现白色菌落,到36 h时,白色菌丝体明显增多,蛋白酶活力也持续增大,菌株培养48 h时,黄绿色孢子开始产生,蛋白酶活力达到最大值,到60 h时,菌株增长明显减慢,产生大量孢子,颜色较深,蛋白酶活力开始下降。综合菌量和蛋白酶活力,培养48 h为最适发酵时间。     

产低温碱性蛋白酶菌株QD-1的鉴定及产酶条件和酶学性质

目的:对来自裙带菜的1株产低温碱性蛋白酶内生菌株QD-1进行菌种鉴定,产酶条件和粗酶性质研究。方法:选取菌株性状优良、产蛋白酶活力较高的菌株QD-1,对其进行形态学及分子生物学16S rDNA鉴定,并研究其最适发酵条件。考察了碳源、氮源、发酵时间、培养温度、起始pH值等因素对菌株产蛋白酶的影响,并对其酶学性质进行研究。结果:经鉴定该菌株为芽孢杆菌属,命名为Bacillus sp.QD-1;该菌株在以蔗糖为碳源,蛋白胨为氮源,培养基初始pH 9.0,培养温度25℃,接种量10%的条件下150 r/min振荡培养48 h,所产蛋白酶活性最高达995 U/m L。该蛋白酶的最适作用温度20℃,最适作用pH 9.0,Mn~(2+)对该酶有较强的激活作用,Zn~(2+),EDTA和PMSF对该酶有抑制作用。结论 :从裙带菜内生菌QD-1发酵液中获得的蛋白酶是一种酶活较高的低温碱性蛋白酶。     

一株产大豆蛋白酶的芽孢杆菌发酵条件的优化

对一株产大豆蛋白酶芽孢杆菌(Bacillussp.D-16-9)的发酵条件进行了优化,研究各种碳源、氮源及无机盐对产酶的影响,应用正交试验优化发酵培养基组成。结果发现菌株D-16-9可以利用无机氮源生长,但用来产酶则很差;有机氮源利于生长和产酶,适当添加诱导物更利于产酶;酶合成模式属于滞后合成型,大量收获在菌体生长的稳定期。通过试验确定发酵培养基的最佳组成:5.0%玉米粉、3.0%大豆豆粕、0.05%氯化钾0、.05%硫酸镁;确定最适发酵条件为:起始pH10,培养温度30℃、种龄24 h、接种量10%,250 mL三角瓶中装入100 mL发酵培养基,摇瓶转速170 r/min,发酵时间120 h。综合最佳的培养基组成和培养条件,最终大豆蛋白酶活达6 500 U/mL。优化后蛋白酶活性由4423.20U/mL提高到6505.60U/mL。     

降解豆粕菌株的筛选及发酵工艺优化

利用产蛋白酶能力较强的菌株将脱脂豆粕原料中的大豆蛋白降解为大豆多肽.以蛋白酶活力及水解度为指标筛选出降解豆粕能力较强菌株W-13,并以水解度为指标对菌株发酵降解豆粕的条件进行了优化,得到最适发酵条件为:11%豆粕、2%玉米粉、3%接种量、发酵时间48 h,在最适培养基上蛋白水解度为65%.     

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.