玉米秸秆酶法降解洗脱解吸工艺优化研究

作为最具代表性的木质纤维素材料,玉米秸秆是生产低聚木糖的理想选择。由于以玉米秸秆直接水解产生的低聚木糖杂质含量太高,导致后期精制步骤变得非常难以进行。试验旨在研究玉米秸秆酶法降解工艺的优化,以提高玉米秸秆中低聚木糖的质量和抗氧化活性。研究结果表明,玉米秸秆酶法降解生产的低聚木糖在乙醇梯度洗脱过程中具有明显的解吸优先性; 30%乙醇洗脱液中低聚木糖的纯度最高为98.12%。玉米秸秆酶法降解生产的低聚木糖抗氧化活性测定表明,当70%乙醇洗脱液质量浓度为3 mg/mL时,低聚木糖的自由基清除率达到89.89%,可直接在抗氧化食品及饲料添加剂中应用。     

大豆秸秆木聚糖碱法提取及酶解工艺研究

大豆秸秆是木聚糖含量较高的一种农业纤维废弃物,采用碱法考察了不同前处理条件(包括预煮温度,氢氧化钠浓度,料液比和浸提时间等)对大豆秸秆中不溶性木聚糖的提取率的影响。正交实验结果表明:当预煮温度80℃,碱液浓度15%,料液比1∶20,浸提时间4 h时,大豆秸杆中不溶性木聚糖的提取率最高,为34.3%。低聚木糖是一种重要的功能性低聚糖,嗜热踝节菌木聚糖酶为低聚木糖水解用酶,通过单因素实验探究了在嗜热踝节菌木聚糖酶的催化作用下,大豆秸秆木聚糖水解的最佳条件,大豆秸秆在此方面研究尚属空白。实验结果显示:底物浓度为1.0%,酶添加量为30 U/m L,温度设定为60℃,水解时间为240 min的条件下低聚木糖的得率最高,为24.2%,在工业上具有较高的应用价值。     

超声波-复合酶法制备玉米秸秆低聚木糖工艺优化

以玉米秸秆为原料,利用超声波-复合酶法制备低聚木糖,研究超声波温度、超声处理时间、复合酶比例、复合酶添加量、酶解时间对低聚木糖制备的影响。在单因素试验的基础上,采用Box-Behnken试验设计方案对制备条件进行优化,得出制备玉米秸秆低聚木糖的最佳工艺参数为:超声温度56℃,超声处理时间40min,添加0.8%(以玉米秸秆计)复合酶(木聚糖酶和纤维素酶按照2∶1的比例组成)并酶解30 min,在此条件下,酶解液中(以玉米秸秆计)还原糖含量为36.43mg/g、可溶性总糖含量为74.32mg/g、平均聚合度为2.04。高效液相色谱法成分分析得出低聚木糖糖液的主要成分是木二糖和木三糖。     

甜高粱秸秆中半纤维素酸解液的发酵研究

高温(121℃)酸解水洗和未水洗甜高梁秸秆,测其酸解液主要成分;并利用诱变体N6和酿酒酵母TO5对其酸解液进行发酵,结果表明:水洗秸秆酸解液中木糖为主要成分,未水洗秸秆中木糖含量相对较少;发酵实验结果表明,诱变体N6能很好发酵木糖,而酿酒酵母TO5不能发酵木糖.     

玉米秸秆水解液发酵生产燃料乙醇的研究

考察水解温度、水解时间、酸浓度及固形物含量对玉米秸秆粉水解的影响,对玉米秸秆粉酸水解条件进行优化;利用高效液相色谱分析了水解液的组分及含量,其中木糖占81.15%;通过水解液乙醇发酵试验,菌株 N-1能较好地发酵木糖转化为乙醇,糖醇转化率达到0.359 g/g。     

玉米秸秆酸催化预水解过程中纤维素和半纤维素变化规律研究

采用正交实验分析法,通过考察预水解后固相中半纤维素去除率、纤维素保留率以及液相中木糖和葡萄糖的含量,探讨了在液比为1∶5的条件下,处理温度、保温时间和冰乙酸用量对玉米秸秆酸催化预水解过程的影响,进而为玉米秸秆农业废弃物的高值化利用提供一定的理论基础。结果表明,在处理温度190℃、冰乙酸用量1%、保温时间120 min时半纤维素去除率最高,而处理温度、保温时间以及冰乙酸用量对纤维素保留率的影响均不显著。液相中的木糖含量在冰乙酸用量2%、处理温度180℃、保温时间120 min时较高;而液相中的葡萄糖含量在冰乙酸用量3%、处理温度170℃、保温时间30 min时较高。     

生物机械法处理玉米秸秆制备纸浆、葡萄糖和木糖

开发了基于机械生物处理法的玉米秸秆利用技术，该技术将玉米秸秆加工为机械浆、葡萄糖和木糖。首先对风干的玉米秸秆进行单螺旋搓丝和筛分，实现皮瓤分离，所得皮、瓤的质量比为1∶8；所得的玉米秸秆皮进行机械磨浆和0.18 m m缝筛精制，所得纸浆相对玉米秸秆的得率为75%。硫酸盐针叶木浆和玉米秸秆机械浆进行配抄，针叶浆加入量为20%时，纸浆撕裂指数为6.0 mN·m2^/g、抗张指数为22.7 N.m/g。所得的玉米秸秆瓤加入纤维素酶和木聚糖酶进行酶解糖化，50℃、48 h的葡萄糖产量为37.5 kg/t玉米秸秆瓤，木糖产量为16.28 kg/t玉米秸秆瓤。     

运动发酵单胞菌和休哈塔假丝酵母混合发酵秸秆产乙醇

首先考察了运动发酵单胞菌(Zymomonas mobilis,Z.mobilis)和休哈塔假丝酵母(Candida shehatae,C.shehatae)分别利用葡萄糖或木糖为唯一底物时的乙醇发酵特性;其次,研究了单菌及双菌发酵混合糖(葡萄糖和木糖比例为3∶1)的产乙醇性能;在此基础上,考察了单菌和双菌培养应用于玉米秸秆发酵产乙醇的可行性。结果表明,Z.mobilis和C.shehatae的接种方式显著影响混合糖发酵产乙醇的效率,当分步接种发酵48 h后,乙醇产量达到最高,为25.77 g/L。当以60 g/L Na OH预处理的玉米秸秆为底物时,分步接种可高效利用秸秆酶解产物(葡萄糖和木糖)实现乙醇产量的最大值(22.34 g/L),提示Z.mobilis和C.shehatae分步接种发酵可最大化地发酵秸秆中的葡萄糖和木糖生产乙醇,有利于乙醇的高产率生产,可为实现纤维原料生产燃料乙醇的工业化提供参考。     

代谢木糖生产乙醇的基因工程菌研究进展

木糖广泛存在于林业及农业废弃物中 ,木糖发酵生产乙醇是再生资源的有效利用。文中报道了近几年来在利用基因工程菌发酵木糖生产乙醇方面的研究进展。重点介绍了大肠杆菌、酿酒酵母、树干毕赤酵母及运动发酵单胞菌的基因改造情况。     

玉米秸秆酶解制备阿魏酰聚糖

玉米秸秆经多级旋风磨机械预处理后物理结构发生改变,更易被相应的酶水解从而释放出阿魏酰聚糖。相同酶解条件下,多级旋风磨预处理后的秸秆的阿魏酰聚糖含量是粗秸秆的3.34倍;用商品复合酶Validase TRL、HSP 6000BG和Rapidase Smart Clear,木糖苷酶及阿拉伯呋喃糖苷酶分别酶解机械预处理后的玉米秸秆,Validase TRL酶解效果最佳,阿魏酰聚糖得率为1.46μmol/g玉米秸秆,是不加酶时的3.48倍;协同酶解的最优组合为Validase TRL 100 U/g,木糖苷酶50U/g和阿拉伯呋喃糖苷酶50U/g,该条件下阿魏酰聚糖的得率为2.87μmol/g;酶解液经大孔树脂分离纯化后,阿魏酰聚糖的回收率为81.44%。     

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.     

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.     

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.     

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.     

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.     

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.     

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.