金钱龟酶解多肽的分离、纯化及抗肿瘤活性研究

本研究以金钱龟肉糜为原料,通过可控的酶解技术制备抗肿瘤活性肽,研究中分别用胰蛋白酶、碱性蛋白酶、木瓜蛋白酶和复合蛋白酶protamex对金钱龟肉糜进行酶解并结合超滤初步分离得到了12种酶解产物,MTT实验表明其中胰蛋白酶酶解所得分子量小于3000 u的多肽E其抗肿瘤效果最强,当浓度为1 mg/m L时,对人肝癌细胞(Hep G-2)和人乳腺癌细胞(MCF-7)的抑制率分别为92.95%和67.08%。将E利用Sephadex G-15进一步纯化,经MTT实验筛选得到两组活性较强的多肽组份E1和E2,对Hep G-2的抗增殖效果来看E1(IC50为77.29μg/m L)强于阳性对照5-氟尿嘧啶(IC50为136.27μg/m L),对于MCF-7的抗增殖效果来看E2(IC50为214.17μg/m L)同样要强于阳性对照5-氟尿嘧啶(IC50为245.82μg/m L)。本研究中,抗肿瘤活性肽首次从金钱龟中分离得出,对于以金钱龟为原料的抗肿瘤药物和保健品的开发具有重要指导意义。     

坛紫菜木瓜蛋白酶水解肽的抗肿瘤活性研究

坛紫菜是一种重要的经济海藻,营养价值极高。为了探究坛紫菜中的抗肿瘤活性多肽,本研究以坛紫菜粉末为原料,采用反复冻融法和超声波破碎法相结合提取蛋白,使用木瓜蛋白酶对粗蛋白进行酶解。初酶解物在MTT细胞增殖检测结果的指引下,结合超滤、葡聚糖凝胶Sephadex G-15的分离纯化和MALDI-TOF质谱分析,最终筛选出具有较强抗肿瘤活性的多肽。结果显示经木瓜蛋白酶酶解物(设为P,分子量0~3 ku)分离纯化得到的多肽组份P2,包含5个多肽,对乳腺癌MCF-7和肝癌细胞Hep G-2的IC50值分别是63.64μg/m L和59.09μg/m L,其抗肿瘤活性均显著高于阳性对照5-氟尿嘧啶(对MCF-7和Hep G-2的IC50值分别是195.45μg/m L和122.72μg/m L),组份P3包含11个多肽,对肝癌细胞Hep G-2的IC50值是209.09μg/m L,也具有一定的选择性抑制作用。     

酶法制备大豆蛋白降血压肽的研究

采用5种蛋白酶对中性蛋白酶 A.S1398 酶解大豆蛋白生成的相对分子质量高的组分(HMF)进行再次水解制备血管紧张素转化酶(ACE)抑制肽.研究结果表明,Protease N 为最适用酶,优化的水解条件为:底物质量分数5%,加酶量2.0%,pH 7.0,温度55℃.在此条件下酶解120 min,水解物对ACE抑制作用最强,抑制率达到65.1%(此时水解度为11.1%);水解液经Sephadex G-15分离,得到了对 ACE 具有较好抑制作用的组分,IC50值约为40μg/mL.     

甲鱼碱溶蛋白肽的分离及抗肿瘤活性研究

为了获得具有抗肿瘤活性的肽段,研究以水解度为评价指标,评价了4种蛋白酶(酸性蛋白酶、胰蛋白酶、中性蛋白酶、木瓜蛋白酶)对甲鱼碱溶蛋白酶解效果的影响,确定中性蛋白酶为最适酶。用酶解、凝胶色谱制备甲鱼碱溶蛋白肽,用MTT肿瘤抑制实验评价抗肿瘤能力,并用HPLC测定高活性肽段的分子质量分布。结果显示:甲鱼碱溶蛋白经中性蛋白酶酶解获得较高水解度的产物,水解度为14.47%。经Sephadex G-25分离获得分子质量184~4 588 u的小分子肽,其具有较高的抗肿瘤活性,IC_(50)为32.26μg/m L。     

碱性蛋白酶酶解魔芋飞粉制备抗氧化多肽

为优化碱性蛋白酶酶解魔芋飞粉蛋白制备抗氧化多肽的条件,采用响应面分析法,以·OH清除率为响应值,研究酶解温度、酶用量、酶解p H值对制备抗氧化肽的影响。此外,还研究了不同分子质量魔芋多肽的抗氧化活性,结果表明:最佳酶解工艺参数:底物质量分数2.25%、温度55℃、酶用量3 228 U/g底物、p H 7.84、水解时间270 min。该条件下抗氧化多肽(18.35 mg/m L)的·OH清除率为73.41%,多肽得率为75.37%。通过Sephadex G-25和Sephadex G-15串联柱分离得到5个多肽组分,其中分子质量为1 500 u和1 000 u的组分抗氧化活性较高,其清除DPPH·的IC50分别为2.82 mg/m L和3.65 mg/m L;清除·OH的IC50分别为9.03mg/m L和14.16 mg/m L;抑制大鼠肝脏自发性脂质过氧化的IC50分别为0.21 mg/m L和0.66 mg/m L;抑制大鼠红细胞H2O2诱导氧化溶血的IC50分别为0.11 mg/m L和0.22 mg/m L。     

青鱼肉活性肽的制备及其抗肿瘤活性研究

以青鱼肉为原料,经二步酶解、凝胶色谱分离纯化制备抗氧化和抗肿瘤活性肽。首先以抗氧化活性和水解度为评价指标,采用单因素和正交试验优化青鱼肉活性肽的制备工艺;然后采用凝胶色谱进行分离纯化,得到不同分子质量多肽组分并分析其氨基酸组成、1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl,DPPH)自由基清除能力和抗肿瘤活性。结果表明,二步酶解青鱼肉的最优酶解组合为碱性蛋白酶-复合蛋白酶,最佳制备工艺为:pH 8.00、酶解温度50℃、料液比2∶10(g∶mL)、酶添加量6000 U/g、酶解时间3 h,在此条件的二步酶解物清除DPPH自由基的半抑制浓度(half inhibiting concentration,IC 50)为9.52 mg/mL;经Sephadex G-15分离得到5个肽组分,其分子质量范围为130~1397 Da;抗氧化实验表明,组分IV具有最强的DPPH自由基清除能力,其IC ₅₀为3.17 mg/mL,为二步水解物的3倍。细胞增殖抑制实验发现,10 mg/mL组分IV对HepG 2细胞抑制率为92.54%,表明青鱼肉活性肽同时具有抗氧化和抗肿癌活性。     

酪蛋白双酶水解物ACE抑制肽的分离纯化

采用胃蛋白酶和胰蛋白酶依次对酪蛋白进行双酶水解,制备ACE抑制肽。水解物经截留分子质量6ku的超滤膜初步分离,再通过Sephadex G-15进一步纯化,体外测定各分离产物ACE活性的半数抑制质量浓度(IC50值)。纯化得到的各组分经毛细管电泳分析肽谱、Q-TOF LC/MS检测分子质量范围。结果显示：双酶水解产物IC50值为560μg/mL,超滤流出物IC50值为250μg/mL;Sephadex G-15分离得到3个组分,组分Ⅰ的IC50值为123.41μg/mL,含有19个肽段;组分Ⅱ的IC50值为66.67μg/mL,含有14个肽段;组分Ⅲ的IC50值为64.29μg/mL,含有5个肽段。Q-TOF LC/MS测得纯化组分的分子质量范围为400～800u。     

具ACE抑制活性的油茶饼粕酶解物的制备及分离表征

以油茶饼粕为原料,研究通过5种蛋白酶水解制备的茶粕酶解物ACE抑制率随酶解时间的变化规律,结果表明除蛋白酶C外,其余4种蛋白酶水解的茶粕酶解物ACE抑制率随酶解时间呈现先增加后减小的趋势。比较不同蛋白酶水解的茶粕酶解物ACE抑制活性得出经蛋白酶A水解后的茶粕酶解物具有较高的ACE抑制率(94.4%);为获得高纯度并具有高ACE抑制活性的酶解组分,文章采用CM-Sephadex C-25阳离子交换色谱和RP-HPLC纯化经蛋白酶A酶解的茶粕酶解物。由CM-Sephadex C-25阳离子交换色谱分离得到一个组分A,通过RP-HPLC进一步分离组分A得到6个组分。对ACE抑制率最高的组分A-2进行RP-HPLC二次纯化,得出该分离组分达到色谱纯,其半数抑制浓度(IC_(50))为98μg.m L~(-1)。     

黄鳍金枪鱼皮胶原肽酶解工艺及抗氧化活性研究

以金枪鱼皮为原料,以肽得率、水解度和感官评价为指标,确定金枪鱼皮胶原肽酶解的最优工艺条件是:中性蛋白酶,酶解时间4 h,酶添加量1 000 U/g,此时肽得率29.03%。体外抗氧化能力测定结果:金枪鱼皮胶原蛋白酶解物具有一定的抗氧化活性,总还原力是VC总还原力的60%;与VC相比,DPPH·清除率(IC50=0.43 mg/m L)、O22-清除率(IC50=3.67 mg/m L)、卵磷脂过氧化物清除率(IC50=14.67 mg/m L)较低,·OH清除率(IC50=0.39 mg/m L)、H2O2清除率(IC50=0.05 g/m L)较高。该酶解物能显著降低油脂的过氧化值,然而降低幅度略低于VC。     

海蜇生殖腺酶解肽的抗氧化活性和ACE抑制活性研究

为实现海蜇加工副产物生殖腺的高值化利用,以海蜇性腺脱脂粉为原料,选用中性蛋白酶、木瓜蛋白酶、胰蛋白酶和碱性蛋白酶分别对海蜇生殖腺进行水解,以酶解产物(JGHP)的水解度、DPPH清除活性和ACE抑制活性为指标,选择制备海蜇生殖腺活性肽的工具酶。经超滤膜将酶解产物分成JGPH-P1(大于3000 u)、P2(1000~3000 u)、P3(小于1000 u)三个部分,分别测定三部分的DPPH清除活性和ACE抑制活性。结果表明,中性蛋白酶水解所得产物的水解度、DPPH清除率和ACE抑制活性均优于其他三种蛋白酶;JGHP超滤后,其中组分JGHP-P3的活性较高。利用电子自旋共振(ESR)技术检测JGHP-P3对DPPH·、·OH、O-2·的清除活性。其ACE抑制活性IC50为1.06 mg/m L,DPPH·、·OH、O-2·的清除活性IC50分别为0.38、0.63、0.59 mg/m L。由此可见,经中性蛋白酶水解得到的JGPH中小于1000 u的组分具有较高的抗氧化和ACE抑制活性。     

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.     

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     

Chinese CTP Market

According to Printing and Printing Equipment Industries Association of China（PEIAC）＇s statistics to the plate manufucturer in China, in 2013, the actual offset plate production has reached 346 million square meters in China. Among them, the CTP production volume was 245 million square meters, up by 11% than that of last year; the total sales of the CTP plate was 239 million square meters, up by 13%.     

Preparatory Work of Print China 2015 is Going Smoothly

正Held at Guangdong Modern International Exhibition Center,Print China 2015 will cover 7exhibition halls,besides the original Hall No.3,4,5,6,7,the newly built F zone of Hall 3 will be used too.The total area will be140,000 square meters.Hall 3:Offset and large printing equipment,package printing equipment,post press