一种5 重real-time PCR筛查转基因水稻方法的建立

以转基因水稻中最常用的CaMV35S启动子、NOS终止子、Cry1Ab/Ac基因、HPT基因及SPS水稻内标基因为研究对象，利用5 种不同的荧光信号（FAM、HEX、Taxas Red、Cy5、Cy5.5）进行多重实时聚合酶链式反应（real-time polymerase chain reaction，real-time PCR）检测方法的研究。通过引物组合筛选、反应体系优化、特异性测试、灵敏度测试、适用性测试等一系列实验，建立了5 重real-time PCR方法，灵敏度可达0.032%。此方法具有灵敏度高、结果准确、通量大等优点，可实现水稻中转基因成分的快速、高效检测。     

基于核磁共振代谢组学分析不同品种草莓的化学成分差异

目的采用基于核磁共振(nuclear magnetic resonance,NMR)的代谢组学方法对"丰香""久香""宁玉"和"甜查理"4个不同品种草莓进行指纹图谱比较和化学成分差异分析。方法采用主成分分析法综合评价4个不同品种草莓的整体分布状况;采用正交偏最小二乘判别分析法筛选并鉴别出引起不同草莓品种有所差异的潜在差异代谢物。结果研究发现,苹果酸在"丰香"草莓中含量最低,蔗糖含量则最高;蔗糖在"久香"草莓中的含量最低;丙氨酸、柠檬酸、天冬酰胺、D-葡萄糖和D-果糖在"宁玉"中含量最低;丙氨酸、苹果酸、柠檬酸、天冬氨酸和D-葡萄糖在"甜查理"中含量最高。结论基于NMR的代谢组学技术可以用于草莓品种的鉴别和化学成分的差异分析,并为草莓的品种溯源提供科学数据。     

High pressure CO2 reduces the wet heat resistance of Bacillus subtilis spores by perturbing the inner membrane

Spores of wild-type Bacillus subtilis PS533 were treated by wet heat at 75 °C for 30 min, and high pressure CO₂ (HPCD) at 6.5 MPa and 30 °C or 75 °C for 30 min. The spores were analyzed for wet heat resistance (85 °C, 90 °C, 95 °C) and typical germination events including DPA release and cortex hydrolysis, inner membrane permeability, and germination triggered by nutrient (L-valine and AGFK) or non-nutrient (dodecylamine and high pressure at 150 MPa or 550 MPa) germinants. The results showed that (i) HPCD-treated spores exhibited reduced wet heat resistance compared to the untreated or wet heat-treated spores; (ii) HPCD-treated spores did not undergo typical germination events such as DPA release or cortex hydrolysis compared to normally germinated spores; (iii) HPCD-treated spores released more metal ions and exhibited decreased ability to maintain DPA, indicating that the permeability of inner membrane of HPCD-treated spores was increased; (iv) HPCD-treated spores exhibited reduced germination rate when triggered by L-valine or 150 MPa, but increased germination rate when triggered by dodecylamine or 550 MPa, suggesting that the fluidity of the inner membrane of HPCD-treated spores might be increased. These results indicated that HPCD could reduce the wet heat resistance of spores, and this resistance decrease was probably due to the modification of the inner membrane caused by HPCD.Industrial relevanceThe extremely high wet heat resistance of spores makes them a significant problem in the thermal processing of foods. Thus, it of great interest to develop a process to reduce the wet heat resistance of spores. In this work, we found that HPCD can significantly reduce the wet heat resistance of B. subtilis spores, and this was achieved by perturbing the inner membrane of spores. These results can improve our understanding of the inactivation mechanism of spores by HPCD, and also provide an alternative approach for spore inactivation in foods.     

基于中红外光谱技术的甘氨酸铁螯合物判别研究

目的研究基于中红外光谱(mid infrared spectroscopy,MIRS)技术定性判别有机微量元素添加剂掺假的可行性。方法以甘氨酸铁螯合物和硫酸亚铁为研究对象,分析样品的中红外光谱,解析不同样品光谱的特异性;建立偏最小二乘判别(partial least squares discriminant analysis,PLS-DA)校正模型对掺假样品进行判别,并比较不同预处理方法、全光谱与特征波段对校正模型判别效果的影响。结果甘氨酸铁螯合物与硫酸亚铁光谱差异显著,主要分布在(3500～3000) cm~(-1)、(1600～1300) cm~(-1)、(1300～1000) cm~(-1)、(660～550) cm~(-1),通过采用平滑(smoothing,SM)、归一化(normalization,Norm)与一阶导数(first derivative,FD)相结合的方法对光谱进行预处理,结合1300 cm~(-1)～1000 cm~(-1)波段建立的PLS-DA校正模型判别效果最优,校正集的判别正确率为97.5%,验证集的判别正确率为100.0%,对外部样品的判别正确率达到92.7%。结论中红外光谱技术结合化学计量学方法能够对甘氨酸铁螯合物的掺假进行准确判别。     

酶联免疫吸附法测定云南铁皮石斛中黄曲霉毒素B1

目的建立酶联免疫吸附法测定铁皮石斛中黄曲霉毒素B_1(afatoxin B_1, AFB_1)的分析方法,并研究酶联免疫吸附法(enzyme-linkedimmunosorbentassay,ELISA)和高效液相色谱法(highperformanceliquid chromatography, HPLC)测定云南铁皮石斛中AFB_1的结果差异性。方法用甲醇+水溶液(50+50, V/V)提取石斛中AFB_1,采用酶联免疫吸附法和高效液相色谱法同时检测云南铁皮石斛(铁皮枫斗)中AFB_1,进行准确性、精密度和回收率等方法学实验,比较2种方法的差异性。结果 ELISA法在测定范围内AFB_1与其对应的吸光度值之间呈良好的线性关系,回归方程Y=-34.798X+18.134,相关系数r~2为0.9967;添加1.0、4.0、10.0μg/kg3个浓度的平均加标回收率为85.3%～94.7%,相对标准偏差为2.57%～4.88%。2种方法的结果符合率为93.2%～117%,相对标准偏差均小于10%。结论 ELISA法具有操作简单、检测速度快、灵敏、特异性好等优点,可同时检测大批量样品中AFB_1的含量。     

蜂王浆蛋白肽的制备及其降血糖和抗氧化活性研究

目的:研究蜂王浆蛋白的最佳酶解工艺条件,并分析所制备酶解肽的氨基酸组成、分子量分布、降血糖及抗氧化活性。方法:以蜂王浆为原料,采用碱提酸沉法提取蜂王浆粗蛋白,以α-葡萄糖苷酶抑制率和ABTS自由基清除率为评价指标,筛选出酶解蜂王浆蛋白制备降血糖及抗氧化活性肽的最佳蛋白酶,并研究蜂王浆酶解肽的体外降血糖和抗氧化活性。结果:蜂王浆降血糖及抗氧化活性肽的最佳制备工艺为:以酸性蛋白酶为酶制剂,酶添加量为6000 U/g,酶解温度为43℃,酶解pH为4.0,酶解时间为4 h,料液比为1:10。在上述条件下,制备的蜂王浆蛋白肽的氨基酸总量为82.19%,相对分子质量集中在1000 Da以下。蜂王浆蛋白肽对α-葡萄糖苷酶的半抑制浓度(IC₅₀)为6.94 mg/mL,清除ABTS自由基、羟自由基、1,1-二苯基-2-三硝基苯肼(DPPH)自由基及超氧阴离子自由基的半抑制浓度(IC₅₀)分别为14.18、0.45、11.02和18.38 mg/mL。试验结果表明,蜂王浆蛋白肽具有一定的降血糖和抗氧化活性,可为蜂王浆高值化利用及活性肽产品开发提供理论依据。     

农业投入品中农药隐性成分的精准筛查方法

目的 建立了农业投入品中90种高频检出农药隐性成分的精准筛查方法。方法 样品采用乙腈超声提取、稀释并过膜净化后直接进样分析。采用气相色谱-质谱法（GC-MS）分析10种有机氯类和拟除虫菊酯类农药，采用液相色谱-串联质谱法（LC-MS/MS）分析其他80种非法添加农药成分，外标法定量。结果 90种农药的检出限在0.001 ~ 0.01 μg/mL；在各自的线性范围内，目标物的浓度与峰面积呈良好线性关系，相关系数大于0.99。利用本方法对天津市内抽检的45个农业投入品进行隐性成分筛查，共筛查出5个样品中含有未标注农药隐性成分。结论 该方法检测的农药隐性成分种类多、范围广，具有良好的可靠性和灵敏度，适用于农业投入品中农药隐性成分的快速筛查与分析。     

鮸鱼肌肉和副产物营养组成分析及评价

对鮸鱼肌肉和副产物营养成分进行测定,鮸鱼肌肉和副产物水分含量分别为79.65%与69.15%,蛋白质含量分别为18.65%与19.36%,粗脂肪含量分别为0.24%与3.90%,灰分含量分别为1.15%与3.75%。鮸鱼肌肉和副产物的氨基酸总量(TAA)分别为18.58%与15.47%,必需氨基酸(EAA)质量分数分别为7.65%与5.52%,但只有鮸鱼肌肉的EAA/TAA(41.17%)与EAA/非必需氨基酸(69.99%)符合FAO/WHO优质蛋白质标准。氨基酸评分和化学评分的结果表明鮸鱼肌肉和副产物的第一限制性氨基酸均为蛋氨酸。鮸鱼肌肉必需氨基酸指数(EAAI)为89.07,副产物的EAAI为61.78。鮸鱼肌肉和副产物都含有丰富的丁酸、棕榈油酸、油酸、神经酸、二十二碳六烯酸、Mg、Ca、K和Fe。鮸鱼肌肉和副产物是高蛋白质、低脂肪、富含矿物质的食品源。     

Quality assurance in pepper and orange juice blend treated by high pressure processing and high temperature short time

The impact of mixing ratio of pepper juice/orange juice (v/v) at 1:5, 1:2, 1:1, 1:0.5 and 1:0.2 on sensory attributes of pepper and orange juice blend (POJB) was studied, and the ratio of 1:0.5 achieved the highest ratings in mouthfeel and overall acceptability and was chosen for the following study. Effects of high pressure processing (HPP, 550 MPa, 5 min, ambient temperature) and high temperature short time (HTST, 110 °C, 8.6 s) on quality attributes of the POJBs were compared. Reduction of total aerobic bacteria (TAB) and molds and yeasts (M&Y) in the POJBs was > 4 log cycles after HPP and HTST, and the two POJBs were microbiologically safe during the whole refrigerated storage. The color, flavor, appearance, mouthfeel and overall acceptability of HPP-treated POJB were closer to untreated POJB than the HTST-treated one. After 25 days, 77.3% and 75.3% of total phenols content, 90.8% and 90.7% of ascorbic acid, and more than 80% of antioxidant capacity in two POJBs were retained, respectively. Particle size distribution (PSD) of HPP-treated POJB was consistent with untreated POJB (1 ~ 76 μm, 3 peaks at 4.2, 17, 52 μm), while HTST changed the PSD (1 to 33 μm, 2 peaks at 2.4 and 17 μm). Higher level of sedimentation in HPP-treated POJB during storage was interpreted by higher residual PME activity (67.0%) and larger and more unstable pulp particles. The POJBs behaved as Newtonian fluids, their viscosity right after processing were ranked as HPP > untreated > HTST, and the values were slightly reduced during storage.Industrial RelevanceThis study was intended to develop yellow sweet pepper and orange juice blend (POJB), which are not available on the market. Further this study was also intended to explore the application of high pressure processing (HPP) and high temperature short time (HTST) on quality assurance of the POJB. This study would provide technical support for commercialization of juice blend products treated by high pressure.     

微生物源单宁酶的研究进展

单宁酶可将单宁水解成没食子酸和葡萄糖，是一种重要的工业用酶。植物、动物和微生物中都含单宁酶，其中微生物是单宁酶的主要来源。基于单宁酶的国内外研究成果，该文归纳总结了不同微生物来源的单宁酶、酶学性质、作用机制，阐述了单宁酶在食品、饲料、制革、精细化工等实际生产中的应用，并对单宁酶今后的研究方向和应用前景进行了展望。