不同发酵条件对Fusarium. solani ZH0101产木聚糖酶的影响

以本实验室筛选到的一株产木聚糖酶但不产纤维素酶的真菌Fusarium solani ZH0101为供试菌株,利用麦秸为诱导物,对产木聚糖酶的液体发酵培养基进行优化。对发酵时间、麦秸添加量、培养基起始pH值、磷酸盐、金属离子、无机氮源和有机氮源对产酶的影响进行研究,优化最佳的培养条件。优化后的液体发酵培养基条件为：麦秸20g/L、KH2PO4 4g/L、CH3COONH4 1g/L、酵母膏2g/L和起始pH值为6.0,发酵时间为14d。优化条件下所产无纤维素酶活力的木聚糖酶酶活力为26.85U/mL,能比未优化前的20.82U/mL提高近30%。     

纳豆激酶液态发酵工艺优化

在单因素实验的基础上,用响应面法和正交实验对Bacillus subtilis液态发酵生产纳豆激酶的培养基和发酵条件进行了优化。实验结果表明,最佳产酶培养基组成为麦芽糖2%,酵母膏4%,CaCl20.03%,pH7.0;最佳发酵条件为接种量1%,培养温度34℃,摇床转速200r/min,装液量100mL/500mL(挡板瓶),培养48h达到产酶高峰,产酶活力可达4300U/mL。     

黑曲霉ATCC16404固体发酵豆皮产生木聚糖酶的研究

以豆皮为原料,用黑曲霉ATCC16404固体发酵产木聚糖酶,采用DNS测定酶活,研究了最佳产酶培养基配比,优化产酶最佳条件。单因素实验结果表明,最佳培养基配方为：以硫酸铵为氮源,矿质元素添加硫酸镁,添加量均为2%（w/w以豆皮干重计）,豆皮颗粒大小为25～50目,不外加碳源,不加表面活性剂。选择料液比、初始pH、培养时间、接重量优化产酶条件,正交实验结果表明,最佳产酶条件为：料液比1∶1.00（w/v）、初始pH5.5,培养时间3d,1.0×107/mL浓度孢子悬浮液接种0.3mL/10g,在此条件下酶活为366.29U/g。     

几丁质酶菌株L012的产酶条件研究

在前期实验的基础上,进一步对优良菌株L012的产酶条件进行了研究.结合单因素实验和正交实验结果,确定了该菌株的最佳产酶培养基组成(g/L):细粉几丁质10,淀粉3,玉米浆3,NaNO3 1.0,K2HPO4 1.05,KH2PO4 0.45,NaCl 0.1,MgSO4·7H2O 0.3,FeSO4·7 H2O 0.03,pH 7.0;最佳培养条件:起始pH 7.0,培养温度30 ℃,摇瓶装液量30 mL,摇床转速180 r/min,接种菌龄24h,接种量10％,发酵时程48 h.条件优化后,菌株L012产酶水平从原来的0.75 IU/mL提高到2.31 IU/mL.     

响应面法优化产β-内酰胺酶菌Bacillus cereus B03的发酵条件

为快速高效地提高菌株Bacillus cereus B03的产酶能力,采用响应面法对Bacillus cereus B03产β-内酰胺酶的发酵培养基进行优化。首先通过单因素实验研究了不同碳源及浓度、不同氮源及浓度、不同金属离子及浓度、氯化钠、磷酸氢二钾以及温度、pH、接种量、装液量对菌株产酶活力的影响,然后设计Plackett-Burman试验筛选出影响产酶的3个显著性因素:温度、pH、接种量。在此基础上,最后设计Box-Behnken中心组合试验确定最优产酶发酵条件。结果表明,最佳发酵培养基成分为葡萄糖20 g/L、酵母浸粉20 g/L、NaCl 2 g/L、MgSO₄·7H₂O 0.2 g/L,K₂HPO₄·3H₂O 4 g/L,最佳产酶发酵条件为发酵温度37 ℃,pH为7.3,接种量3%,装液量50 mL/250 mL。在此优化条件下,该菌株产β-内酰胺酶的酶活力为113278.7 U/mL,为优化之前酶活(88792.7 U/mL)的1.28倍。本研究为进一步工业化开发利用性状稳定且高产β-内酰胺酶的菌株提供借鉴。     

重组大肠杆菌摇瓶发酵产环糊精葡基转移酶条件的优化

对一株表达环糊精葡基转移酶基因工程菌E.coli BL21(DE3)产环糊精葡基转移酶的发酵条件进行了研究.利用单因素实验和正交实验获得该菌株产环糊精葡基转移酶的最佳条件为:种子培养时间14h,接种量3%,培养温度37℃,培养基初始pH8.0,发酵培养基的组成为玉米粉1.5%、玉米浆6%、蛋白胨1.0%,500mL三角瓶装液量为50mL.210r/min 振荡培养,其发酵液酶活可达5010U/mL.     

β-甘露聚糖酶高产菌株发酵条件优化

从土壤中分离出1株产β-甘露聚糖酶的优良菌株Bacillus sp.QYW-1,具有发酵周期短且产酶活力高等特性,初始酶活力21.85 U/mL。在单因素实验对培养基及培养条件优化的基础上利用Plackett-Burman实验设计对影响产酶的重要因素进行筛选。实验发现,影响该菌株产酶的重要因素是魔芋粉、蛋白胨及硫酸镁。最陡爬坡实验和Box-Behnken实验得到响应面(RSM)优化的最佳培养基为:魔芋粉26 g/L,蛋白胨10 g/L,MgSO43.8 g/L,NaCl 10 g/L,KCl 6 g/L,NaNO36 g/L,K2HPO43 g/L,初始pH6.5。在此条件下菌株发酵产β-甘露聚糖酶酶活力为233.86 U/mL,与模型预测值相符,与单因素优化后的酶活力115.62 U/mL相比,提高了102%。     

Pseudomonas sp. W7产低温蛋白酶培养基及培养条件的优化

采用Plackett-Burman法对影响假单胞菌（Pseudomonas sp.）W7产低温蛋白酶的因素进行显著性分析,筛选出对产酶影响显著的3 个因素：葡萄糖、蛋白胨和MgSO4;然后用最陡爬坡试验逼近最大产酶区域;应用Box-Behnken原理设计和响应面分析确定产酶培养基的最佳组合为：葡萄糖4.25 g/L、蛋白胨7.00 g/L、干酪素4.00 g/L、K2HPO4 5.00 g/L、KH2PO4 1.00 g/L、CaCl2 0.26 g/L、MgSO4 0.14 g/L,低温蛋白酶的酶活力达到了183.9 U/mL,比优化前提高了21%。另外,对Pseudomonas sp. W7的产酶条件进行了优化,在单因素的基础上,利用正交试验设计,最终确定产酶的最优培养条件为：培养温度为20 ℃、初始pH值为7.0、接种量为3%、装液量为20%、摇床转速为100 r/min,在最佳条件时测得的酶活力为193.2 U/mL。通过生长曲线和产酶曲线的测定,确定产酶的培养时间为48 h。     

酸橙内生菌Bacillus thuringiensis Bt028产几丁质酶的发酵条件优化

为了提高分离自酸橙果实中的一株内生细菌Bacillus thuringiensis Bt028产几丁质酶的性能。采用单因素实验方法考察了培养基组成和发酵条件对该菌株产几丁质酶的影响,并在此基础上采用响应面方法对该菌株产几丁质酶的摇瓶发酵条件进行了优化。单因素实验结果表明胶体几丁质浓度、培养温度和初始pH对该菌株产几丁质酶影响明显;响应面试验结果表明,当胶体几丁质浓度为0.8%、酵母粉浓度为1%、初始pH为6.9、接种量3%、装液量为100 mL/250 mL、30 ℃培养48 h时,该菌株产几丁质酶性能最佳,发酵液中几丁质酶酶活可达10.48 U/mL。研究结果为该菌株几丁质酶的进一步开发与应用奠定了基础。     

高去酰胺蛋白酶产生菌的诱变选育及产酶条件优化

以地衣芽孢杆菌2709为出发菌株,采用氮离子注入技术,在最佳注入剂量为1.5×1015ions/cm2的条件下进行诱变处理,筛选得到一株去酰胺度高达48.99%的突变株SDL-9,去酰胺度提高了85.57%,而肽键水解度相对较弱。并对SDL-9的产酶条件进行优化,结果表明：最佳碳源为质量浓度为2.0g/100mL的玉米粉,最佳氮源为质量浓度3.0g/100mL的大豆蛋白,质量浓度为0.04g/100mL Fe3+对产酶具有明显的促进作用,质量浓度为0.9g/100mL谷氨酰胺对产酶有明显的诱导作用,最佳起始pH值为7.0,最佳发酵温度35℃,在此条件下进行发酵产酶,测得去酰胺度为57.1%,肽键水解度为13.2%。     

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.