拟穴青蟹新型过敏原肌质钙结合蛋白的纯化、鉴定及分子克隆

为进一步认识蟹类的过敏原,采用免疫印迹方法,分析发现甲壳类过敏患者血清能与拟穴青蟹肌浆蛋白中分子质量约为21 kD的蛋白质产生特异的IgE结合反应,结果显示该蛋白可能是蟹类新型过敏原。通过硫酸铵盐析、阴离子交换和凝胶过滤柱层析等方法对21 kD-蛋白进行分离纯化,采用Western blotting和基质辅助激光解析电离飞行时间质谱（matrix assisted laser desorption ionization-time of flight-mass spectrometry,MALDI-TOF-MS）确认纯化的21 kD-蛋白为肌质钙结合蛋白（sarcoplasmic calcium binding protein,SCP）。采用SMART-RACE（SwitchingMechanism At RNA Termini-Rapid Amplification of cDNA Ends）的方法获得SCP的cDNA序列,该序列全长986 bp,开放阅读框为579 bp,编码193 个氨基酸,其理论分子质量21.94 kD,等电点4.44。拟穴青蟹SCP与甲壳类动物SCP具有较高的同源性,与昆虫SCP的同源性较差;三级结构模拟分析显示,SCP含有5 个螺旋-转角-螺旋结构区,即EF-手型结构区,并在其中两个手型结构区形成2 个钙离子结合位点;进一步预测得到SCP的4 个线性抗原表位和3 个构象性抗原表位。     

离子色谱法同时测定蛋白胨中葡萄糖、果糖、蔗糖、麦芽糖、乳糖含量

目的建立离子色谱法同时检测蛋白胨中葡萄糖、果糖、蔗糖、麦芽糖、乳糖的分析方法。方法样品用水作为提取剂,用OnGuard Ⅱ RP柱和OnGuard Ⅱ H柱净化,50mmol/LNaOH溶液作为流动相,用Carbopac PA20柱进行离子色谱分析。结果在该条件下5种糖都能得到较好分离,线性良好,葡萄糖、果糖、蔗糖、乳糖和麦芽糖的检出限为0.002、0.004、0.011、0.007、0.018 mg/L,平均回收率为90.59%～97.68%(n=9)。结论该方法操作简便、灵敏度高,可用于作为蛋白胨中葡萄糖、果糖、蔗糖、麦芽糖、乳糖的检测。     

Impact of transglutaminase on the textural, physicochemical, and structural properties of chicken skeletal, smooth, and cardiac muscles

This study examined the effects of microbial transglutaminase (MTG; 3.1 mg/ml) on chicken skeletal, smooth, and cardiac muscles; the meat containing the different muscle types was shaped into sausages and treated at 40 °C and/or 78 °C for 30 min. Although the three muscle types were obtained from the same bird, the effects of MTG addition were not uniform. All the muscle types showed a significant increase in the breaking strength (P < 0.01), but skeletal muscle exhibited the maximum increase. All samples showed a decrease in the fluorescence intensity and a significant reduction in the concentration of proteins that were extracted in a high ionic strength solution (P < 0.05). Scanning electron microscopy images and histological studies revealed that different muscle types had different physical structures and frameworks after MTG treatment, which is a reflection of the differences in the reaction specificity of MTG with different muscle proteins. Histological studies revealed that the reactions of MTG with meat proteins are both exogenous and endogenous. Cooking loss data suggested that MTG did not have any negative effect on water retention during cooking. MTG appears to be a functional and contributive substance since the results suggest that MTG can function on all muscle types that are mechanically processed for different industrial applications. MTG aggregates muscle proteins in different ways that improve their organoleptic properties such as texture, appearance, and water retention.     

Determination of Zn,Cu, Cd,Pb, Ni and Cr in some Slovenian foodstuffs

An acid-assisted microwave digestion procedure was optimised for the determination of trace elements in foodstuffs by the use of flame and electrothermal atomic absorption spectrometric (FAAS, ETAAS) techniques. Zn, Cu, Cd, Pb, Ni and Cr were determined in cabbage, wheat, potato, instant milk, fish, eggs and baby foodstuffs. The repeatability of measurements was tested in the cabbage sample and was found to be ±3.3% for Zn, 4.6% for Cd, ±6.8% for Cu and ±15.5% for Cr. The worse value obtained for Cr was probably due to its inhomogeneous distribution and very low concentration in the cabbage. Concentrations of Ni and Pb in the cabbage sample were below the limit of detection (0.2 mg kg^–1). The accuracy of the analytical procedure was checked by the analysis of the standard reference material CRM 1570 Trace and minor elements in spinach leaves and IAEA 336 Trace and minor elements in lichen. Good agreement between the certified or indicative and determined values was obtained. The results indicate that the concentrations of Zn, Cu, Cd, Pb, Ni and Cr found in cabbage, wheat and potato are comparable to the literature data. Cd, Pb and Ni were not found to be the contaminants in the foodstuffs analysed. Higher concentrations of Ni were found only in baby food "okolenik", arising from the cocoa that "okolenik" contained. It was demonstrated that the Cr content is very low in baby food, yolk and white of egg and in fish (<0.05 mg kg^–1 Cr). Elevated concentrations of trace elements were found only in mussels that accumulate heavy metals from the marine environment.     

Pilot survey of commercial pasteurized milk consumed in the metropolitan area of Rio de Janeiro,Brazil, for tetracyclines residues,including the 4-epimers of oxytetracycline,tetracycline and chlortetracycline

This pilot survey aimed to assess the occurrence of tetracyclines and the 4-epimers of oxytetracycline, tetracycline and chlortetracycline in commercial pasteurized milks sold in the metropolitan area of Rio de Janeiro, Brazil, between October 2009 and March 2010. A liquid chromatography–tandem mass spectrometry (LC–MS/MS) method, developed and validated in our laboratory, was used. All 100 analyzed samples were compliant, but 14 contained oxytetracycline in concentrations ranging from the method limit of detection (3.7?µg?l^–1) to the method limit of quantification (12.2?µg?l^–1). One sample contained oxytetracycline and tetracycline simultaneously (at a concentration slightly higher than method limit of quantification, 7.0?µg?l^–1). The presence of 4-epioxytetracycline and 4-epitetracycline in contaminated samples with the parent drugs could not be confirmed as traces were detected only in the quantification MRM transition. No other tetracyclines were detected.     

超声波对低嘌呤脱脂豆浆溶解性、保水性及粘度的影响

为了研究超声波处理对低嘌呤脱脂豆浆溶解性、保水性及黏度的影响,以脱脂豆粕为原料,采用超声波法,通过单因素及正交试验,研究最佳超声波处理条件.研究结果表明,最佳超声波处理条件为超声功率315 W、超声时间40 min、超声温度60℃,此条件下的黏度为1.4 mPa·s、保水性为81.83％、溶解性为58.15％.超声波处理的溶解性比未经超声波处理提高了22.35％.因此,超声波处理能显著提高低嘌呤脱脂豆浆的溶解性及低嘌呤脱脂豆腐的保水性,降低低嘌呤脱脂豆浆的黏度.     

Bioactive compound content,antioxidant activity,deoxynivalenol and heavy metal contamination of pearled wheat fractions

Wheat kernels are rich in antioxidant compounds, that are mainly present in the outer bran layers and which are removed during milling. Unfortunately, several contaminants, e.g., mycotoxins and heavy metals, are also concentrated in the external layers. Pearling of 3 wheat varieties gave five fractions (each 5% of the original grain weight), starting from the outer layer until the inner kernel, designated as 0–5%, 5–10%, 10–15%, 15–20%, 20–25%, respectively. The remaining 75% of the inner kernel was also collected. Dietary fibre, free phenolic acids and total antioxidant activity decreased progressively from the external to the internal layers. However, the 5–10% fraction was richer in β-glucan content than the external one (0–5%). Heavy metals were only found in the most external fraction. Deoxynivalenol contamination decreased from the external to the internal layers: 64% of total contamination of kernel was found in the 0–5% and 5–10% fractions. The 10–15% kernel fraction offered the best compromise between high nutritional value and low contamination risk.     

红发夫酵母中3R,3’R-虾青素的分离纯化和结构鉴定

研究红发夫酵母中3R,3’R-虾青素的分离纯化工艺条件,并利用高效液相色谱串联大气压化学电离源质谱(high performance liquid chromatography-atmospheric pressure chemical ionization-mass spectrometry,HPLCAPCI-MS)、核磁共振(nuclear magnetic resonance,NMR)和高效液相色谱-紫外光谱(high performance liquid chromatography-ultra violet,HPLC-UV)对纯化得到的虾青素结构进行鉴定。结果表明:利用细胞玻璃研磨器进行研磨提取、硅胶柱层析分离和结晶提纯相结合的方法,能够有效减少反式虾青素的异构化,得到的虾青素纯度高于95%。采用高效液相色谱-质谱仪(high performance liquid chromatography-mass spectrometry,HPLC-MS)、超导核磁共振谱仪(1H和13C NMR)以及Chiralpak IC手性固定化纤维柱分析,显示制备的反式虾青素几乎都以3R,3’R结构组成。     

高效液相色谱法同时测定可乐中苯甲酸、糖精钠、安赛蜜和咖啡因

目的建立高效液相色谱同时测定可乐中苯甲酸、糖精钠、安赛蜜和咖啡因的方法,并应用于国际FAPAS能力验证。方法样品经超声脱气用水稀释,采用C_(18)色谱柱分离,以乙酸铵缓冲液(0.02 mol/L)/甲醇为流动相,采用梯度洗脱分离,在230 nm、272 nm波长下进行检测。结果苯甲酸、糖精钠、安赛蜜和咖啡因线性范围为2.0~200μg/m L,线性关系良好,相关系数大于0.999,回收率在97%~103%之间,RSD为0.5%~2.8%之间,检出限在0.005~0.02μg/m L之间。结论该法灵敏度高、重现性好,用于FAPAS能力验证时结果接近中值。     

Amino acids,fatty acids,and dietary fibre in edible seaweed products

The nutritional compositions of 34 edible seaweed products of the Laminaria sp., Undaria pinnatifida, Hizikia fusiforme and Porphyra sp. varieties were analyzed.     

Effects of brown rice, rice bran, and polished rice on colon carcinogenesis in rats

Rice is a nutritious staple food with health-promoting activity. This study investigated the effects of brown rice, rice bran, and polished rice on 1,2-dimethylhydrazine (DMH)-induced colon carcinogenesis in rats. The colons were examined for preneoplastic lesions and the expression of pro-inflammatory proteins. Lipid peroxidation was determined in livers and plasma; antioxidant-associated parameters were determined in livers. The results showed that consumption of medium-level of rice bran significantly reduced the number of aberrant crypt foci (ACF) and altered their distribution. Brown rice and rice bran significantly reduced cyclooxygenase-2 (COX-2) expression of the middle colon. Brown rice, rice bran, and polished rice had no significant effect on plasma and hepatic lipid peroxides and hepatic antioxidant-associated parameters. Thus, rice bran may be one candidate of the active rice fraction that protects the colon against DMH-induced early carcinogenesis in rats and may be a novel dietary supplement for chemoprevention of colon cancer.     

葡萄糖接枝改性对米渣分离蛋白乳化性及结构的影响

利用湿热法合成米渣分离蛋白-葡萄糖接枝产物,以研究美拉德反应条件及反应程度对接枝产物乳化性和结构的影响。以温度、pH值、时间、米渣分离蛋白和葡萄糖质量比为试验因素,并利用响应面法优化试验。结果显示,接枝产物乳化性同实验因素之间不呈线性关系。在温度79.13℃,pH 10.6,时间16 min,质量比1∶1的反应条件下制得的接枝产物乳化性为7 778 m2/g,为米渣分离蛋白的1.46倍。在此条件下蛋白接枝产物表面疏水性降低;红外光谱结果表明米渣分离蛋白以共价键的形式引入糖分子,蛋白接枝产物的α-螺旋结构含量减少,β-折叠和不规则卷曲结构含量增加。电镜扫描结果显示蛋白改性后表面结构分散,聚集体之间的界限分明。     

重组水稻 α-半乳糖苷酶的分离纯化及酶学性质研究

对重组水稻α- 半乳糖苷酶的分离纯化及其酶学性质进行研究。结果表明：该重组酶分子质量约为59kD;最适酶反应温度为45℃,最适pH 值为5.0;酶活力在0～20℃,pH4.0～7.0 最为稳定;酶学动力学常数Km 为0.78mmol/L,Vmax 为10.16mmol/(mg·min)。多数的金属离子和有机离子对酶催化作用没有影响,Hg2+、Ag+及－ SH 基团抑制剂p-chloromercuribenzoic acid(pCMB)对酶活性有强烈抑制作用。     

BKP竹浆生产工艺的改进

介绍了雅安中竹纸业有限责任公司BKP竹浆洗、筛、漂工艺流程的改进情况。通过改进工艺和加强工艺控制，提高了生产能力，黑液提取率≥98％，碱回收率提高，实现了碱自给，降低了漂白中段废水量，污水排放达到国家标准。     

κ-Casein Macropeptides from Cheese Whey: Physicochemical, Biological, Nutritional, and Technological Features for Possible Uses

κ-Casein macropeptide (CMP), one of the components of whey, is produced during cheese making. Whey production has grown enormously in the last three decades and will continue to grow, along with cheese production. There is an increased interest in research for new applications of food industry by-products in order to avoid their extensive elimination and negative environmental implications. In this respect, CMP has been the focus of extensive scientific research that has proven its value as a functional and bioactive peptide, as well as a source of biologically active peptides. This article evaluates the possibilities and future perspectives of the use of CMP and related peptides obtained from cheese whey from different ruminant species. Physicochemical, technological, biological, and nutritional aspects are considered, and processes for analysis, fractionation, and separation are reviewed. The objective is to help to promote further exploitation of cheese industry coproducts for the preparation of high added-value ingredients to be included in the composition of nutraceuticals or functional food products.     

LC–MS/MS characterization,antidiabetic, antioxidative,and antibacterial effects of different solvent extracts of Anamur banana (Musa Cavendishii)

The main objective of this study was to examine the phenolic compounds and the antibacterial, antioxidant, anti-α-glucosidase and anti-α-amylase activities of the different extracts (methanol, ethanol and hexane) of Musa cavendishii collected from the Anamur district in Turkey. LC–MS/MS was used to identify phenolic compounds. Quinic acid, acotinic acid, hesperidin and amentoflavone were identified in methanol extract. These phenolic compounds, excluding hesperidin, were also identified in the ethanol extract. Methanolic extract appeared the most active in all enzyme inhibition, antibacterial and antioxidative activity assays which is mainly due to its rich phenolic content. The methanol extract of banana showed the highest anti-α-glucosidase and anti-α-amylase activities with IC50 values of 5.45 ± 0.39 mg/mL, 9.70 ± 0.29 mg/mL, respectively. This study showed that methanol and ethanol extract, especially the methanol extract, have potential for use in the development of functional foods for reducing the diabetes and bacterial risks.     

QuEChERS-超高压液相色谱-串联质谱法同时测定水产品中的6种丁香酚类麻醉剂

建立了同时测定水产品中的丁香酚、异丁香酚、甲基丁香酚、甲基异丁香酚、乙酸丁香酚酯、乙酰基异丁香酚残留量的超高压液相色谱-串联质谱法。样品用乙腈提取,经50 mg乙二胺-N-丙基硅烷、100 mg C₁₈、150 mg Mg SO₄净化,采用Kinetex C₁₈(2.1 mm×100 mm,2.6μm)色谱柱分离,以甲醇-水作为流动相进行梯度洗脱,电喷雾离子源正负快速切换,多反应监测模式检测,内标法定量。丁香酚和异丁香酚的线性范围为2～100 ng/m L,其他4种丁香酚类药物的线性范围为1～100 ng/m L,线性相关系数均不低于0.999。丁香酚类药物检出限(信噪比为3)为0.5～1.5μg/kg,定量限为2.0～5.0μg/kg。6种丁香酚类麻醉剂的方法回收率为67.0%～106.4%,相对标准偏差为1.2%～15.7%。该方法能实现对水产品中丁香酚类麻醉剂的准确、快速检测,测定结果可靠。     

动物源弯曲菌分离纯化鉴定方法优化

【目的】弯曲菌是重要的食源性人畜共患病原菌,通常难以诊断,而动物源方向缺少系统的检测分析方法及相应的优化。为了获得更适宜分离动物来源的空肠弯曲菌和结肠弯曲菌,提高分离效率,降低分离成本,特开展方法优化,填补动物源弯曲菌分离方法建设的空白,为耐药菌株的有效追踪溯源和风险评估提供依据。【方法】对已有的空肠弯曲菌和结肠弯曲菌分离纯化鉴定方法Preston肉汤和Bolton肉汤增菌;CCDA选择性培养分离、弯曲菌显色培养分离、Skirrow选择性培养分离;生化鉴定和分子生物学鉴定做了筛选优化,针对目前存在的分离纯化和鉴定方法进行了调查比较,得到了更适合于健康动物来源的粪便和盲肠中的分离方法。【结果】牛血清预增菌、CCDA选择性分离、弯曲菌显色培养基纯化、哥伦比亚血琼脂进行扩增及分子学方法鉴定,同时建议采用厌氧条件进行培养。该流程简单易行,成本较低,适合于绝大多数实验环境条件,是获取弯曲菌的首选流程,便于推广实施。【结论】通过将现研究阶段存在的大多数检测方法进行了综合分析,整理出一套针对于动物来源的粪便及肠道内容物的检测分析方法,对健康动物中弯曲菌的分离效率能够提高30%,同时空肠弯曲菌和结肠弯曲菌的分离效率无明显差异。     

韭菜β-木糖苷酶的分离纯化与部分性质研究

新鲜韭菜经匀浆、缓冲液提取、硫酸铵分级沉淀、羧甲基离子交换层析(carboxymethyl-sephar ose,CMSepharose)再通过Su perdex-200凝胶过滤层析,从而获得电泳纯的β-木糖苷酶。该酶的比活力达到18.25 U/mg,纯化倍数为12.59,回收率为1.83%,分子质量为123.02 k D,亚基分子质量为61.51 k D。通过对β-木糖苷酶的酶学性质研究得到最适温度为65℃,最适p H值为4。该酶在25~55℃及p H 3.0~5.0的范围内有较好的稳定性;在最适条件下测得其米氏常数(K_m)值为0.28 mmol/L;甲醇、乙醇、异丙醇及十二烷基硫酸钠(sodium dodecyl sulfate,SDS)和Ag+对该酶具有抑制作用,Mn~(2+)和Co~(2+)对该酶具有一定的激活作用。     

燕麦中β-葡聚糖的分离纯化及结构表征

以我国山西产燕麦(晋燕8号)为原料提取并分离纯化,得到Oglu-1和Oglu-2两个燕麦β-葡聚糖级分,经凝胶渗透色谱层析和Sephacryl-400SF柱层析,证实Oglu-1为单一组分,纸层析和HPLC分析表明该级分多糖由葡萄糖为单一糖基组成,红外光谱和核磁共振分析证明该多糖分子链中存在β-(1→4)和β-(1→3)两种糖苷键,其比例为2.1:1,该结果与国外学者的研究基本一致。采用凝胶渗透色谱和激光光散射联用仪(GPC-LLS)测定Oglu-1的重均相对分子质量为4.49×105。