野生大型真菌抑菌菌株的筛选及培养基优化

研究了9种野生食药用大型真菌次生代谢产物的抑菌活性,并优化了高抑菌活性菌株的发酵培养条件,旨在筛选具有良好抗菌活性的野生大型真菌菌株,以寻找新型抗菌物质和为其工业化生产提供参考资料。利用大肠杆菌Escherichia coli和枯草芽孢杆菌Bacillus subtilis为指示菌,采用滤纸片法测定大型真菌的抑菌活性,以摇瓶发酵法对高抑菌活性菌株的培养基配方进行优化,并验证。结果表明:绒柄小皮伞Marasmius confluens对大肠杆菌和枯草芽胞杆菌的抑菌率最高,分别达到56%和52%。最优培养基配方为:蔗糖20 g/L,酵母膏5 g/L,KH_2PO_4 3.0 g/L,MgSO_4·7H_2O 2.0 g/L,维生素B_1 10.0 mg/L,pH 7.0。在优化的条件下,绒柄小皮伞M.confluens发酵液对大肠杆菌和枯草芽孢杆菌的抑菌率分别达85%和88%,比优化前的抑菌率分别提高了0.52倍和0.69倍。野生食药用大型真菌发酵产物中存在丰富的抑菌物质,优化培养有助于提高大型真菌的抑菌活性。     

海洋细菌GM-1-1菌株摇瓶发酵培养基和发酵条件优化

GM-1-1菌株是从连云港海域分离到的一株对多种病原真菌具有较好抑制作用的海洋解淀粉芽孢杆菌(Bacillus amyloliquefaciens),为了提高该菌株发酵液的抑菌活性,以发酵液的菌体密度及对白色念珠菌(Candida albicain)的抑制作用为检测指标,采用单因素试验和正交试验对GM-1-1菌株的摇瓶发酵培养基成分及发酵条件进行优化。研究结果表明,GM-1-1菌株生长及产生抑菌物质的最佳培养基配方为:糊精20 g/L,牛肉膏15 g/L,MgSO_4 1.4 g/L,pH7.0;最佳发酵条件为:温度28℃,摇床转速220 r/min,培养时间28 h,种子液浓度10~8 cfu/mL时接种量为10%,装液量为50 mL(250 mL三角瓶)。     

培养条件对八角内生真菌发酵液抗氧化活性的 影响

对八角内生真菌Chaetomium globosum BJEF13产抗氧化活性物质的培养条件进行了研究。在筛选确定基础培养基的基础上,采用单因素试验方法考察了培养基组成及摇瓶发酵条件对菌株发酵液抗氧化活性、菌株生长的影响。基础培养基的筛选结果表明,马铃薯葡萄糖培养基、察氏培养基、马丁培养基、豆芽汁蔗糖培养基等4种常用真菌培养基中,察氏培养基优于其他3种。培养基组成和发酵条件的研究结果表明,当以4%葡萄糖、0.4%的蛋白胨、0.1%K_2HPO_4、0.05%KCl、0.05%MgSO_4和0.001%FeSO_4作为摇瓶发酵培养基时,接种量为10%,装液量为100 mL/250 mL,在初始pH为7.5、28℃、150 r/min条件下振荡培养7 d,该菌株的生长和培养液的抗氧化活性可以达到较好的水平,其生物量平均干重达0.682 6 g/100 mL,5倍稀释的乙酸乙酯萃取液对DPPH自由基的清除率可达57.51%。     

紫红曲霉与银杏叶双向发酵研究

为探讨药食用真菌与药性基质进行双向发酵的药性机理,以药食两用真菌紫红曲霉作为发酵菌株,以富含总黄酮的银杏叶药汁为药性基质进行双向发酵试验,采用单因素试验探究不同碳源、氮源、无机盐、生长因子、装液量、接种量、发酵时间、初始pH值对紫红曲霉生物量、银杏叶药汁总黄酮含量的影响。并采用正交试验优化紫红曲霉与银杏叶双向发酵条件,结果表明,在银杏叶药汁培养基装液量85 mL,接种量10%,初始pH值为5,无机盐FeSO4的条件下,其最优发酵组合为：葡萄糖浓度4%、花生粉浓度1.5%、KH2PO4浓度0.1%、MgSO4浓度0.12%,150 r/min,（28±1）℃培养11 d。在此组合下双向发酵体系中的总黄酮含量达到0.074 mg/mL,比未利用紫红曲霉发酵的对照组提高了100%;紫红曲霉生物量为13.26 g/L,比未优化前的对照组提高了76.47%。因此,紫红曲霉与银杏叶进行双向发酵可以提高银杏叶药汁总黄酮含量,并促进紫红曲霉生物量的生长。     

菌株SFGi-1产赤霉素发酵条件的研究

通过摇瓶培养对菌株SFGi-1产赤霉素的培养基和培养条件进行了优化,确定最佳发酵培养基为液化淀粉100g/L,花生饼粉15g/L,豆粕粉3g/L,硫酸镁0.75g/L,磷酸二氢钾1.0g/L:培养条件为培养温度30℃,接种量5%,250mL摇瓶装液量为35mL,起始DH值为5.5,发酵时间204h.在此条件下,菌株SFGi-1赤霉素发酵最高效价可达到1424mg/L.     

米根霉液态发酵产糖化酶工艺条件研究

米根霉具有较强的产淀粉糖化酶能力。本研究通过对米根霉液态发酵工艺条件的研究来提高其产糖化酶能力,为红曲黄酒纯种酿造技术奠定基础。以米根霉为出发菌株,大米液化液作为碳源进行摇瓶发酵,通过正交试验优化液态培养基配方,并通过单因素试验得出米根霉的培养条件。优化的摇瓶发酵最佳培养基为:液化液浓度50%,NH4Cl 5 g/L,KH2PO41 g/L,MgSO4·5H2O 0.3 g/L,FeSO4·7H2O 0.4 g/L,CaCO38 g/L;发酵条件为:初始pH6.0,装液量50 mL,摇床转速150r/min,接种量6%,发酵温度为32℃。该发酵条件下,菌株摇瓶发酵液的糖化酶活力达到196.6 U/mL±8.1 U/mL。     

益生菌的筛选及其发酵特性研究

为获得性能优良的益生菌生产菌株,通过乳酸菌的溶钙圈筛选,耐酸、耐胆盐检测及体外抗氧化和分解胆固醇能力测试进行了菌株的选育;对筛选菌株进行16S rRNA基因测序及生理生化特性鉴定;采用正交试验优化培养基配方和发酵条件。结果表明,从牛奶储罐中分离、筛选到一株各项性能优良的菌株,编号为SX68,经鉴定其为植物乳杆菌（Lactobacillus plantarum）。优化培养基配方为：蛋白胨10 g/L,牛肉膏10 g/L,酵母提取物5 g/L,磷酸氢二钾3 g/L,柠檬酸氢二胺4 g/L,乙酸钠5 g/L,葡萄糖30 g/L,吐温-80 1 mL/L,MgSO4·7H2O 0.58 g/L,MnSO4·4H2O 0.25 g/L,pH 6.4;最佳发酵条件为：起始pH 6.5,发酵温度35 ℃,接种量20 mL/L,溶氧量＜100 mL/L,搅拌转速50 r/min,罐压0.03 MPa。在此优化条件下,菌株SX68最高发酵生物量（OD600 nm值）为15.68,活菌数为9.6×1012 CFU/mL,可为其用于益生菌生产奠定基础。     

产红色素赤红球菌的筛选鉴定和产色素条件的优化

将一株土壤中分离筛选出的产红色素菌株采用生理生化及分子生物学鉴定,根据菌株的16Sr DNA序列分析结果及生理生化特性,该菌株归属为赤红球菌(Rhodococcus ruber)。以此菌株为出发菌株,通过单因素筛选及正交试验对赤红球菌发酵色素培养基以及培养条件进行优化研究。确定最优培养基配方为:果糖10 g/L、蛋白胨5 g/L、酵母粉15 g/L、氯化钠10 g/L、玉米浆15 g/L;最佳培养条件为:培养温度28℃、摇瓶装液量50 mL/250 mL、转速190 r/min、接种量4%、p H6.5,发酵液产菌体干重达到6.6 g/L,红色素粗品产量达到0.93 g/L。     

重组大肠杆菌产谷氨酰胺转氨酶培养基及发酵条件优化

对已构建好的表达谷氨酰胺转氨酶的大肠杆菌Rosetta DE3的摇瓶培养条件及发酵条件进行优化,所获优化培养基配方为葡萄糖4.0g/L,酵母膏3.0g/L,NH4Cl 4.0g/L,Na2HPO42.0g/L,K2HPO41.0g/L,MgSO4.7H2O 1.0g/L,NaCl 3.0g/L。得菌株发酵培养的最佳优化条件为装液量为25mL,接种量为5%,加IPTG浓度为0.6mmol/L,诱导时间为4h。     

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%。     

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.     

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.     

低温带皮菜籽粕微粉的不同粒级部分的功能特性

为研究低温带皮菜籽粕微粉的不同粒级部分的功能特性,以经低温脱脂的带皮菜籽粕为原料,经微粉碎后筛分成212~425μm、150~212μm和106~150μm的3个不同粒级的微粉样品,检测这些样品的吸水性、吸油性、乳化性和乳化稳定性、蛋白质体外消化率。结果表明:1 3个不同粒级的微粉样品之间的粗纤维含量存在显著差异,表明三者的结构组成成分有一定差异。23个微粉样品的乳化活性和乳化稳定性随粒度级别的减小而显著增加(P0.01)。33个微粉样品的蛋白质体外消化率随粒度级别的减小而显著增加(P0.01)。4不同粒级带皮菜籽粕微粉样品的吸水性与吸油性受其结构组成物质不同和粒度的双重影响,与粒度的相关性不明显。     

Microbiology of food taints

Fresh and processed foods are often spoilt by the presence of undesirable flavours and odours caused by microbial action. The aim of this paper is to review the current knowledge of microbiologically induced taints that occur in a wide range of foodstuffs, including meats, poultry, fish, crustaceans, milk, dairy products, fruits, vegetables, cereals and cereal products. Examples have been chosen where the compounds responsible for the taint have been identified and sufficient data obtained to demonstrate the involvement of microorganisms. However, in some cases the full identity of the causative organism may not have been elucidated. The types of microorganisms covered by this review include bacteria, fungi, yeasts, actinomycetes and cyanobacteria. Although cyanobacteria do not in general infect foods, their presence in aqueous systems and water supplies can lead to off-flavours in aquatic organisms and processed foodstuffs. Several examples of each of these processes are discussed. Wherever possible, the likely biosynthetic pathway used by the microorganism to produce the offending compound in a foodstuff is indicated.     

The development of reference materials for paralytic shellfish poisoning toxins in lyophilized mussel. I: Interlaboratory studies of methods of analysis

This paper describes the first part of a project undertaken to develop mussel reference materials for Paralytic Shellfish Poisoning (PSP) toxins. Two interlaboratory studies were undertaken to investigate the performance of the analytical methodology for several PSP toxins, in particular saxitoxin (STX) and decarbamoyl-saxitoxin (dc-STX) in lyophilized mussels, and to set criteria for the acceptance of results to be applied during the second part of the project: the certification exercise. In the first study, 18 laboratories were asked to measure STX and dc-STX in rehydrated lyophilized mussel material and to identify as many other PSP toxins as possible with a method of their choice. In the second interlaboratory study, 15 laboratories were additionally asked to determine quantitatively STX and dc-STX in rehydrated lyophilized mussel and in a saxitoxin-enriched mussel material. The first study revealed that three out of four postcolumn derivatization methods and one pre-column derivatization method sufficed in principle to determine STX and dc-STX. Most participants (13 of 18) obtained acceptable calibration curves and recoveries. Saxitoxin was hardly detected in the rehydrated lyophilized mussels and results obtained for dc-STX yielded a CV of 58% at a mass fraction of 1.86 mg/kg. Most participants (14 out of 18) identified gonyautoxin-5 (GTX-5) in a hydrolysed extract provided. The first study led to provisional criteria for linearity, recovery and separation. The second study revealed that 6 out of 15 laboratories were able to meet these criteria. Results obtained for dc-STX yielded a CV of 19% at a mass fraction of 3.49mg/kg. Results obtained for STX in the saxitoxin-enriched material yielded a CV of 19% at a mass fraction of 0.34mg/kg. Saxitoxin could not be detected in the PSP-positive material. Hydrolysis was useful to confirm the identity of GTX5 and provided indicative information about C1 and C2 toxins in the PSP-positive material. The methods used in the second interlaboratory study showed sufficiently consistent analysis results to undertake a certification exercise to assign certified values for STX and dc-STX in lyophilized mussel.