酶联适体分析法检测葡萄酒中的赭曲霉素A

建立了竞争取代酶联适体分析方法,检测葡萄酒中的赭曲霉素A(OTA)。核酸适体和OTA特异性结合导致与核酸适体杂交的短链DNA解链,解离的DNA作为捕获元素,进一步特异性结合辣根过氧化物酶(HRP),HRP催化四甲基二苯胺(TMB)底物显色,测定A450nm与浓度的线性关系,确定OTA的检出限。考察了DNA包被浓度、杂交温度和封闭液等因素对检测的影响。结果表明,在优化的条件下,所建立的竞争取代酶联适体分析法对OTA检测有高灵敏度,检测限0.88μg/L,线性范围1~100μg/L在葡萄酒中添加时,加标回收率为92.03%~106.6%,相对标准偏差RSD(n=5)小于2.1%,此方法可用于葡萄酒中OTA的快速测定。     

表面等离子共振技术快速检测食品中的 玉米赤霉烯酮毒素

目的建立基于表面等离子共振(surface plasmon resonance,SPR)技术快速检测食品中玉米赤霉烯酮毒素(zearalenone,ZEN)的方法。方法采用表面自组装技术(self-assembled monolayer,SAM)在金膜的表面修饰羧基基团,将ZEN抗原与牛血清白蛋白(albumin from bovine serum,BSA)偶联物(ZEN-BSA)通过共价键固定在芯片的表面,采用竞争法检测样品中的玉米赤霉烯酮毒素。结果该方法的检测限为8.2 ng/m L,ZEN单克隆抗体与呕吐毒素、黄曲霉毒素B_1、赭曲霉毒素、伏马毒素等没有交叉反应,与α-玉米赤霉烯醇和β-玉米赤霉烯醇交叉反应率分别为15.3%和11.5%。结论本方法具有简便、快速和高灵敏度等优势,在食品中真菌毒素的快速检测方面具有潜在的应用价值。     

Protocols for the isolation and detection of lactic acid bacteria with bacteriocinogenic potential

The objective of this study was to evaluate culture media and methodologies for isolation and detection of lactic acid bacteria (LAB) capable to produce bacteriocin-like substances. Samples of milk and cheese were pour plated on de Mann-Rogosa-Sharpe agar (MRS) and Kang-Fung-Sol agar (KFS) (both at 35 °C/48 h, under anaerobiosis), from which 389 and 256 LAB cultures were selected. The antagonistic activity of them was evaluated using the spot-on-the-lawn and two culture media: brain-heart infusion agar with catalase (BHI + C) and M17 (both at 35 °C/24 h). The proteinaceous nature of the antagonistic cultures was verified using: spot-on-the-lawn (MRS, 25 °C/24 h, under anaerobiosis) and well-diffusion (cultures amplified on modified MRS broth at 25 °C/24 h, and then neutralized using NaOH). Listeria monocytogenes ATCC 7644 was used as indicator. A larger number of antagonist cultures were isolated from MRS (83 by M17 and 65 by BHI + C) in comparison to KFS (24 by M17 and 15 by BHI + C). The spot-on-the-lawn identified a higher frequency of LAB capable of producing bacteriocin-like substances. MRS was considered to be the best culture media for the isolation of LAB capable to produce bacteriocin-like substances, activity that was better identified using the spot-on-the-lawn methodology.     

胶体金免疫层析技术在真菌毒素快速检测中的应用

胶体金免疫层析技术具有快速、简便、特异、敏感等特点, 弥补了真菌毒素传统检测方法的不足, 适用于真菌毒素快速现场筛选。本文主要介绍了胶体金免疫层析技术的原理及其在真菌毒素快速检测中的应用及发展前景。     

双抗夹心ELISA检测食品中大肠杆菌O157:H7方法研究

研究获得纯化抗大肠杆菌O157:H7IgY抗体,经检测10mg/ml纯化IgY抗体的效价为1:320;以大肠杆菌O157:H7免疫新西兰大耳白兔,获得兔抗大肠杆菌O157:H7IgG抗体,效价达1:25600。以兔抗大肠杆菌O157:H7IgG抗体稀释3200倍作为捕获抗体,抗大肠杆菌O157:H7IgY抗体为检测抗体建立双抗夹心ELISA方法检测大肠杆菌O157:H7,正交试验分析表明,捕获抗体于37℃包被2h、不封闭、抗原与捕获抗体于37℃结合2h、检测抗体浓度为0.25mg/ml、与抗原于37℃结合1h为最优反应条件。该方法对纯培养菌液检出限为105CFU/ml,具有良好的敏感性及特异性。染菌样品经在EC增菌液中选择性培养后进行双抗夹心ELISA检测,接种量为0.1～1CFU/g(ml)的样品在培养12h后可检出阳性反应,1～10CFU/g(ml)的样品在培养8h后可检出阳性反应。     

发酵大豆食品中染料木素含量的ELISA测定

染料木素是大豆发酵食品中的主要异黄酮成分。制备了牛血清白蛋白-染料木素免疫抗原和卵白蛋白-染料木素包被抗原,并对它们进行紫外和SDS-聚丙稀酰胺凝胶电泳鉴定。抗染料木素抗体的效价为1:1600。建立竞争ELISA测定发酵大豆制品中染料木素含量方法,染料木素测定浓度范围为6.25～150μg/L;回收率为87%～102%;亲和常数为1.62×108;与大豆甙元、芒柄花素的交叉反应率分别为9.8%、4.57%;与槲皮素、儿茶素、单宁和核黄素等结构类似物无交叉反应。同时将ELISA法与薄层层析法进行比较。     

无酵母生淀粉酒精发酵的研究

利用根霉—3(Rhizopus—3)麸曲进行无酵母生淀粉酒精发酵研究。以生玉米淀粉为原料,无酵母酒精发酵醪液酒度8.5％(v/v,20℃),淀粉利用率84.87％。研究了该工艺过程中的一些条件。发现发酵终产物乙醇对生淀粉糖化酶的活性有抑制作用。生淀粉的糖化是该过程的限速步骤。15gRhizopus—3鲜曲的酒化力与10ml酒母相当。发酵时添加一定量的α-淀粉酶、纤维素酶或果胶酶有协同作用,可提高淀粉利用率。同时探讨了无酵母生淀粉酒精发酵的机制,认为选育具有高活性生淀粉糖化酶和酒化酶等复合酶系的菌种是关键问题。     

Mechanism of action of Melaleuca armillaris (Sol. Ex Gaertu) Sm. essential oil on six LAB strains as assessed by multiparametric flow cytometry and automated microtiter-based assay

Little is known concerning the effects of essential oils (EOs) against lactic acid bacteria (LAB), either of the human gastrointestinal microflora or those involved in industrial processes. GC and GC/MS analysis of Melaleuca armillaris EO resulted in the identification of 68 compounds comprising 99.6% of the oil. Eucalyptol (1,8-cineole) was the major compound (68.9%) and the composition was largely dominated by the oxygenated monoterpenes fraction (77.3%). The anti-LAB activities of the EO were first checked by the disc diffusion assay. In a second phase, time-survival kinetics of each strain incubated with increasing concentrations of the EO (0.25, 2.5, 5 and 25 μg/ml) were established using an automated microtiter assay (Bioscreen C). Bacteriostatic or bactericidal effects were noticed depending on the studied strain and on the applied concentration of the EO. The mathematical modelling of the kinetics showed that in presence of increasing concentrations of M. armillaris EO, the lag phases of growth were extended (0.69%–97.5%) and both the growth rate and final cell density were reduced. Variations depending on the strain were noticed.     

Antioxidant activities of aqueous extracts of selected plants

The antioxidant properties of 25 edible tropical plants, expressed as Trolox equivalent antioxidant capacity (TEAC), were studied using DPPH (1,1-diphenyl-2-picrylhydrazyl free radical) scavenging and reducing ferric ion antioxidant potential (FRAP) assays. Their cupric ion chelating activities (CCA) and total polyphenol contents (TPC) were also determined. A strong correlation between TEAC values obtained for the DPPH assay (TEAC_DPPH) and those for the FRAP assay (TEAC_FRAP) implied that compounds in the extracts were capable of scavenging the DPPH free radical and reducing ferric ions. A satisfactory correlation of TPC with TEAC_DPPH and TEAC_FRAP suggested that polyphenols in the extracts were partly responsible for the antioxidant activities while its correlation with CCA was poor, indicating that polyphenols might not be the main cupric ion chelators. Principal component analysis (PCA) indicated that TEAC_DPPH, TEAC_FRAP and TPC contributed to the total variation in the antioxidant activities of the plants.