菌株F21来源α-淀粉酶的酶学性质研究

该研究对菌株F21所产的α-淀粉酶进行纯化和酶学性质研究。 通过硫酸铵盐析、疏水层析等方法纯化后,α-淀粉酶酶活达到 4 616.3 U/mL。 酶学性质研究表明,该酶最适pH值为4.8,最适温度为55 ℃,且酶在pH 4.0～9.0及低于45 ℃的条件下稳定性较高;Ca2+ 对酶活有较强激活作用,Fe2+及Fe3+对酶活有较强的抑制作用。     

Bacillus subtilis AS35发酵麦糟产β-葡聚糖酶粗酶性质的研究

研究了由Bacillus subtilis AS35发酵麦糟制备的β-葡聚糖酶的性质,研究表明,该酶在55℃~65℃时酶活力较高,最适反应温度为60℃;最适反应pH值为5.5,在pH值为4.0~7.0时有较高的稳定性,在4℃保存24h后,残余酶活超过85%;在1mmol/L的浓度条件下,Fe2+、Co2+、Ca2+,尤其是Co2+对酶活性有明显的激活作用;Pb2+、Fe3+、A13+对酶活性有明显的抑制作用。     

纤维素酶高产担子菌株的筛选鉴定及酶学性质初探

从河北省棉田土壤样中分离出一株纤维素酶高产菌株,经中科院微生物所鉴定,初步确定为一株担子菌(a basidiomycetous specie),将其命名为担子菌LKY01.对该菌株酶学性质进行了初步探索.研究表明,本酶反应最适pH值为5.0左右;最适反应温度约60℃;Co2+、Mn2+对本酶酶活有明显的抑制作用,Cu2+、Fe2+、Ca2+、Na+、K+、Sr2+、Mg2+对本酶有一定的促进作用;在50℃下本酶有较好的稳定性,其酸碱稳定范围在pH 4.0～8.0.     

郫县豆瓣曲曲霉的筛选、分子鉴定及其酶学性质的研究

以自然接种的瓣曲为研究对象,应用示踪和基于基质特性组合的菌种筛选技术,筛选出适合其酿造特性的分离株No17.通过形态学观察和26S rDNA、ITS序列的分子生物学鉴定,该分离株为Aspergillus oryzae,命名为A.oryzae SZP17.该菌株的中性蛋白酶和α-淀粉酶的酶学性质研究表明,中性蛋白酶的最适反应温度为40℃,20℃～40℃时比较稳定,最适反应pH值为7.5,耐酸性较强,Cu2+和Mn2+对其具有激活作用,Fe3+和Fe2+对其具有抑制作用.α-淀粉酶的最适反应温度为60℃,低于50℃比较稳定,最适反应pH值为6.0,pH值>7.5时易失活,各种金属离子对其酶活影响较小.     

三七渣固态发酵所产淀粉酶的酶学特性研究

考察了黑曲霉固态发酵三七渣所产淀粉酶的酶学特性。研究结果表明,该淀粉酶的最适温度为60℃;当温度为30~50℃时,具有较好的热稳定性,保温处理2 h后其酶活仍可保持在90%以上;其最适pH为5.0;当pH为3.0~5.0时,具有较好的pH稳定性,在4℃冷藏保存2 h后其酶活仍可保持在80%以上;Mn2+对该淀粉酶有轻微的激活作用,K+、Na+、Mg2+、Fe3+、Zn2+和Ni2+对该淀粉酶有弱抑制作用,Cu2+则对其有强抑制作用。该淀粉酶米氏常数Km值为0.64 mg·mL-1,最大反应速率Vmax为0.45 mg/min。     

浓香型大曲中蛋白酶菌株的分离及酶学性质研究

从浓香型大曲中分离得到1株产蛋白酶优势菌株,并将该菌所产蛋白酶经硫酸铵分级沉淀、透析、吸附树脂处理,得到蛋白酶粗酶。对该酶的最适pH值、pH值稳定性、最适温度、温度稳定性、金属离子影响等进行研究。结果表明,该菌株产蛋白酶的最适pH值为7,pH值稳定性较差;最适温度为40℃,热稳定性随温度增加而降低;金属离子Li+和螯合剂EDTA对该酶具有抑制作用,金属离子Mn2+、Fe3+、Cu2+、K+、Zn2+、Mg2+、Ca2+等均对该酶具有激活作用,其中Zn2+对该酶的激活作用最强。     

红曲霉产生淀粉酶固态发酵条件的优化及部分酶学性质

以红曲霉M2为研究对象,在单因素试验的基础上,确定无机盐水添加量、培养时间、培养温度和培养基初始pH四个因素的取值范围,以生淀粉酶活力为指标设计正交试验优化了培养条件,并对最优条件下生淀粉酶的部分酶学性质进行了研究。结果表明:温度为32℃,pH值5.0,无机盐水量为14 mL,培养时间6 d时酶活最高可达445.37 u/g;该酶的最适反应温度为40℃,最适反应pH为5.0,在pH值为5.0⁷.0范围内较稳定,热稳定性较差,在60℃保温2 h残余酶活只有23.9%;Zn2+,Mn2+,Cu2+对该酶有促进作用,Ba2+和Fe3+对该酶有抑制作用,Na+,K+,Mg2+对该酶基本无影响。     

中国对虾蛋白酶性质的研究

采用酶学分析方法研究了中国对虾蛋白酶的性质。结果表明,该酶最适温度为60℃,最适pH为9.0,是一种碱性蛋白酶。此酶在50℃以下及pH 7.0~10.0稳定性较好。以酪蛋白为底物,蛋白酶的最适底物浓度为2%。60℃时,该酶的半衰期为22 min,70℃时则为7 min。低浓度的EDTA对酶活有抑制作用,Mn2+、Ca2+、Mg2+对蛋白酶有一定的激活作用,而Cu2+、Zn2+、Fe2+、Pb2+对酶活有不同程度的抑制作用,可推断该酶是一种金属蛋白酶。     

灰树花漆酶的双水相萃取及酶学性质

研究双水相萃取法分离灰树花漆酶。以PEG6000/(NH4)2SO4作为成相系统,通过单因素试验研究酶液添加量、PEG6000质量分数、(NH4)2SO4质量分数和体系pH对灰树花漆酶萃取效应的影响,并通过正交试验确定最佳萃取条件,同时进行纯化漆酶的酶学性质研究。结果表明,在pH为6时,PEG6000/(NH4)2SO4双水相萃取的最佳条件为:酶液添加量5%,PEG6000质量分数25%,(NH4)2SO4质量分数17%,此时漆酶的分配系数和酶活收率分别高达7.6和96.6%。纯化漆酶的最适反应温度和最适pH分别为45℃和2.97,在30~45℃和pH 3.5~5.0之间具有很好的稳定性。金属离子Cu2+、Zn2+、Mg2+、Mn2+、K+、Ca2+对漆酶酶活有促进作用,其中Cu2+和Mg2+的促进作用最显著,而Co2+、Fe2+、Na+对漆酶酶活有抑制作用,其中Fe2+的抑制作用最明显。     

粗糙脉孢菌木聚糖酶酶学性质的研究

对粗糙脉孢菌木聚糖酶粗酶的酶学性质进行了研究,结果表明:该木聚糖酶的最适反应温度50℃、pH值为5.8,温度低于50%、pH 4.6～7.4时该酶稳定性较好;金属离子Ca2+、Mg2+、Mn2+、Fe2+对该酶的活性有促进作用,K+、Na+、Pb2+、Cu2+、Fe3+、Al3+对酶的活性有抑制作用;EDTA、SDS、DMSO对该酶活性的抑制作用较小;以木聚糖为底物的酶动力学常数Km和Vmax值分别为1.30g/L和1.35μmol/(min·mL).     

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.