金龟子绿僵菌(Metarhizium anisopliae Ma83)几丁质酶的纯化及性质

由金龟子绿僵茵Ma83菌株产生的几丁质酶经硫酸铵盐盐析,Sephadex G-100柱层析,DEAE-纤维素柱层析分离纯化后,得到SDS-PAGE均一样品.酶的最适反应温度为50℃,半失活温度为65℃;酶的最适反应pH值为5.0,酶在pH4.0～7.0范围内较稳定.Ag+、Co2+、K+、Mg2+对Ma83几丁质酶有激活作用,而Hg2+、Zn2+、Pb2+对几丁质酶活力有抑制作用.经计算Ma83菌株几丁质酶对胶体几丁质的Km值为1.05 mg/mL.     

酸橙内生菌Bacillus thuringiensis Bt028几丁质酶的分离纯化及其酶学性质

为揭示酸橙内生菌Bacillus thuringiensis Bt028几丁质酶的基本酶学性质,采用离心、硫酸铵盐析、葡聚糖凝胶G-100层析及SDS-聚丙烯酰胺凝胶电泳等方法对菌株Bt028发酵液中的几丁质酶进行分离纯化鉴定,并考察该几丁质酶的最适温度、最适pH、催化动力学参数等性质。结果表明,Bt028菌株发酵液经离心、硫酸铵盐析、葡聚糖凝胶G-100层析分离纯化后获得电泳纯几丁质酶比活力为681.78 U/mg,纯化倍数为3.21,回收率15.52%。SDS-聚丙烯酰胺凝胶电泳结果表明,几丁质酶的分子质量为65 kDa。酶学性质研究结果表明,该酶的最适反应温度为60 ℃,最适反应pH为6.5,在温度低于60 ℃、pH5.5~7.5时有较好的稳定性;Mg2+、Ca2+、Hg2+、Co2+对该酶活力有抑制作用,而Cu2+和Fe3+有一定的促进作用;低浓度的甲醇、乙醇、正丙醇和二甲亚砜使酶的活性增加,但当浓度继续增大,反而会抑制酶的活性;丙酮对几丁质酶有激活作用,而甲醛对几丁质酶有抑制作用。在最适催化条件下,几丁质酶催化反应的米氏常数Km、最大反应速率Vmax、酶的转换数Kcat分别为29.533 mg/mL、108.696 μmoL/(L·min)和0.527/min。研究结果可为该酶的实际应用提供必要的工艺参数。     

SDS-Chitin-PAGE电泳法分析大葱叶几丁质酶特性

使用Sodium dodecyl sulfate-几丁质-聚丙烯胺酰胺凝胶电泳(SDS-Chitin-PAGE)方法分析了大葱葱叶几丁质酶的特性.在大葱叶组织中共检测到三种几丁质酶同工酶,分别命名为CH-A、CH-B、CH-C.分析结果表明,这三种几丁质酶具有SDS抗性,β-巯基乙醇对其活性也无显著影响.在SDS-Chitin-PAGE前,样品缓冲溶液与样品蛋白粗提液孵化处理的最适温度为24℃、时间为10～20min.凝胶电泳酶活分析表明,大葱葱叶中CH-A、CH-B的耐热温度可达60℃,CH-C的耐热温度达到70℃.葱叶中CH-A的最适pH为5.2～5.6,CH-B的最适pH为4.0～4.4,CH-C的最适pH为6.6～8.0.     

UV-C辐照对凡纳滨对虾虾头主要内源酶酶学性质的影响

以凡纳滨对虾虾头为研究对象,探究UV-C辐照对其主要内源酶的影响。用功率为30 W的UV-C紫外灯以20 cm高度、辐照20 min,处理虾头均浆液,检测内源酶酶活,分析比较辐照前后温度、pH对其内源酶酶活的影响规律。从虾头中检测出酸性及碱性蛋白酶、脂肪酶、几丁质酶和多酚氧化酶等酶活性; UV-C辐照前后,脂肪酶、几丁质酶的最适pH有所变化,脂肪酶最适pH由10变为6～10,几丁质酶最适pH由5变为6左右;各主要内源酶最适温度无明显变化; UV-C辐照后,在最适条件下酸性蛋白酶酶活升高23. 99%,碱性蛋白酶升高37. 49%,脂肪酶升高21. 37%,几丁质酶升高37. 70%,多酚氧化酶升高46. 52%。结果显示,UV-C辐照对凡纳滨对虾虾头酸性及碱性蛋白酶、脂肪酶、几丁质酶和多酚氧化酶等主要内源酶均有激活作用。     

绿僵菌Ma83固态发酵几丁质酶和部分酶学性质的初步研究

研究了金龟子绿僵菌MetarhiziumanisopliaeMa83菌株产几丁质酶的固态发酵条件和粗酶液的酶学性质。研究结果显示 ,当以麸皮∶蚕蛹粉为 3∶1作为培养基 ,最适氮源为1%NaNO3,起始pH为 6 5 ,发酵温度为 31℃ ,接种量为 2mL液态种子时酶活力最高。此外 ,添加稻壳对Ma83产几丁质酶有促进作用。该菌株在生长 2d时 ,酶活力达 33 9U /g干培养基。酶的最适反应温度为 5 0℃ ,最适反应 pH为 6 0 ;不同温度保温 1h后 ,酶的半失活温度为 42℃。不同 pH值的缓冲溶液中 (2 5℃ )放置 1h后 ,酶在pH 5 0～ 6 0条件下稳定性最高     

Chitinolyticbacter meiyuanensis SYBC-H1几丁质酶的性质及在虾皮水解中的作用

以一株产几丁质酶细菌Chitinolyticbacter meiyuanensisSYBC-H1为材料,对几丁质酶进行了分离纯化及酶学性质研究。通过硫酸铵沉淀、DEAE-cellulose阴离子交换介质和SephadexG-100分子筛层析柱处理其所产几丁质酶,获取纯度为95%以上的几丁质酶。酶学性质表明,该酶的最适温度为40℃,最适pH值为6.5,在50℃以下较为稳定,在pH值为5.0时最稳定,β-巯基乙醇有利于防止半胱氨酸氧化而增加几丁质酶稳定性,EDTA可增加该酶稳定性,Na+,K+对几丁质酶有明显的激活作用,Zn2+,Mn2+,Fe2+,Fe3+对几丁质酶有较大的抑制作用。SYBC-H1几丁质酶可把虾皮水解为N-乙酰氨基葡萄糖,水解率为21.8%。     

一种海洋几丁质酶产生菌的筛选及酶学性质研究

海洋是自然界几丁质主要来源地，滋养出种类繁多可降解几丁质的微生物，因此从海洋中筛选产高活性几丁质酶的细菌，是获得高产几丁质酶微生物的有效途径。该文以胶体几丁质为唯一碳源，从渤海滩涂筛选到一株具有降解几丁质特性的细菌Y-8,对其进行鉴定并研究其几丁质酶酶学性质。Y-8菌株经分子鉴定为副溶血性弧菌(Vibrio parahaemolyticus),发酵上清液经SDS-PAGE及蛋白质谱分析，发现其存在2种几丁质酶，分别为Chi1几丁质外切酶，氨基酸残基数为848,理论分子质量为87.6 kDa; Chi2几丁质内切酶，氨基酸残基数为1 054,理论分子质量为112.9 kDa。Y-8所产生的几丁质酶能够高效降解胶体几丁质，获得单一产物N-乙酰氨基葡萄糖，其最适反应温度为55℃,45℃保温1 h,仍能保持70%以上活性；最适pH为6.0,在pH 4.0～9.0、37℃保温1 h后相对酶活性保留55%以上，具有较好的稳定性。10 mmol/L的Mn²⁺能使几丁质酶活性提高362%,而EDTA以及SDS、吐温-20、吐温-80对酶活力具有抑制作用。     

黏质沙雷氏菌胞外几丁质酶的纯化及特性

黏质沙雷氏菌几丁质酶发酵上清液经硫酸铵沉淀、透析、DEAE-琼脂糖凝胶阴离子交换层析和苯基-琼脂糖凝胶疏水层析,得到电泳纯的几丁质酶和几丁质结合蛋白CBP21。该几丁质酶和CBP21分子质量分别约为58 ku和21 ku,CBP21对该几丁质酶水解几丁质增效明显。几丁质酶反应最适温度为50℃,最适pH约为6.5～7.0。该酶在55℃以下、pH 4.5～8.0范围内稳定。酶的Km值为0.22 mg/mL,Vm为1.26μmol/(min.mg)。金属离子K+、Sn2+、Mn2+对酶有一定激活作用,而Pb2+、Hg2+和Cu2+则强烈抑制其活性。该几丁质酶的糖基含量约为3.3%。EDTA和2-ME可分别提高酶活力65%和105%。H2O2强烈抑制酶活力,提示其活性中心可能存在硫氢基。     

短短芽孢杆菌几丁质酶的分离纯化及酶学性质

探讨来源于短短芽孢杆菌FM4B的几丁质酶（chitinase）分离纯化过程及其性质。菌株FM4B摇瓶发酵液经过离心、乙醇分级沉淀及葡聚糖凝胶G-100层析纯化,用变性聚丙烯酰胺凝胶电泳确定其分子质量。结果获得几丁质酶纯品,酶的纯化倍数为7.19,酶活回收率为30.6%,分子质量为66 kD;酶活力在pH 5.0～9.0和80 ℃以下稳定,最适pH值为6.0,最适温度为50 ℃;Cu2+、Hg2+、Pb2+、Co2+以及Zn2+对该酶有强烈的抑制作用,Mg2+ 和Ca2+对该酶的活力具有一定的促进作用;该几丁质酶对多种霉菌有显著的抑菌效果。菌株FM4B分泌的几丁质酶稳定性好,且对多种霉菌抑制效果明显,具有较高的潜在利用价值。     

大肠杆菌表达几丁质酶对不同底物的酶学性质及其降解产物分析

采用重组大肠杆菌大量表达几丁质酶LICHI18A,经Ni-NTA柱纯化后,对不同底物的酶学性质进行了研究,以Lineweaver-Burk双倒数作图法测定米氏常数,采用薄层色谱(TLC)和高效液相色谱(HPLC)对降解产物进行分析。结果表明,该几丁质酶分子量为54 k Da,对水溶性壳聚糖的Km和Vmax分别为4.04 mg/m L和222.2μmol/(min·mg);对水溶性壳聚糖的反应最适温度和p H分别为60℃和7.0;对胶体几丁质的Km和Vmax分别为2.913 mg/m L和2.836μmol/(min·mg),反应最适温度和p H分别为40℃和5.0。Ni2+、Mg2+、Ca2+、Zn2+对胶体几丁质的降解有明显的促进作用,但对水溶性壳聚糖的降解没有促进作用,且Ni2+和Zn2+有一定的抑制作用。Tween-80对降解胶体几丁质起一定的促进作用,而不同浓度的SDS有抑制酶活作用,且浓度越高抑制作用越强。TLC和HPLC分析结果表明,该酶降解胶体几丁质的产物主要为几丁二糖和单糖,而水溶性壳聚糖的降解产物较为复杂,主要有单糖、几丁二糖和甲壳寡糖。     

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.     

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.     

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.     

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.     

Announcing a new international peer-reviewed journal:Food Science and Human Wellness now accepting manuscript submission in English

<正>We are pleased to announce the launch of a new international peer-reviewed journal-Food Science and Human Wellness,ISSN 2213-4530,which is an open access journal,produced and hosted by Elsevier B.V.on behalf of Beijing Academy of Food Sciences.Food Science and Human Wellness is an international peer-reviewed English journal that provides a forum for the dissemination of the