黑曲霉内切聚半乳糖醛酸酶A基因在大肠杆菌中的表达及产物酶学性质研究

采用Trizol法提取黑曲霉(Aspergillus niger JL-15)的总RNA,RT-PCR扩增到黑曲霉内切聚半乳糖醛酸酶A基因(pgaA),pgaA全长1113bp,编码370个氨基酸残基.pgaA经EcoRI和NotI双酶切,定向插入到表达载体pET-32a(+)中,并转化Escherichia coli Rosetta-gami B(DE3),重组子经IPTG诱导,其表达产物(rePGA)果胶酶活性最高为637.0U/mg.酶学性质研究表明,rePGA的最适温度为60℃,热稳定性较差,最适pH为pH 5.0;Ca2+对其活性具有显著促进作用,而Fe3+和Mg2+对其活性具有显著抑制作用;SDS-PGAE结果表明,rePGA的相对分子质量约为40 500Da;rePGA的米氏常数(Km)和最大反应速度(Vmax)分别为4.05mg/mL和39.37μmol/(mL.min),rePGA能快速降低果胶溶液的粘度,符合其内切作用模式特点.     

1-MCP结合开孔包装对软枣猕猴桃果实软化的研究

目的 采后1-甲基环丙烯（1-MCP）处理缓解软枣猕猴桃开孔包装贮藏失水引起的果实软化问题。方法 以“龙城二号”软枣猕猴桃为材料,0.5μL/L的1-甲基环丙烯（1-MCP）熏蒸结合开孔包装进行贮藏,探究贮藏期间1-MCP结合开孔包装对软枣猕猴桃果实软化的影响。结果 1-MCP处理结合开孔包装与对照相比更有利于保持果实外观,明显抑制了果实中维生素C和总酚的降解;此外,1-MCP处理还延缓了果实硬度的下降,可能是由于抑制了细胞壁降解酶多聚半乳糖醛酸酶（PG）和纤维素酶（CX）的活性,从而推迟了原果胶和纤维素含量的降低,因此延缓了可溶性果胶含量的升高,与对照组相比,货架期延长了4d。结论 1-MCP结合开孔包装贮藏可以保持果实的感观品质,使细胞壁降解进程受到抑制,进而推迟了软枣猕猴桃开孔包装贮藏中轻度失水导致的软化进程,达到了改善果实软化的目的。     

气调包装对软枣猕猴桃品质的影响

目的 研究自发气调处理对采后软枣猕猴桃果实品质的影响。方法 将软枣猕猴桃置于塑料气调箱内,气调处理组果实放入气调箱封盖处理,对照组进行不封盖处理,于4 ℃下贮藏64 d,每16 d对软枣猕猴桃呼吸代谢及抗性相关指标进行测定,即气调箱内CO2和O2含量,可溶性固形物（TSS）、丙二醛（MDA）、总酚含量,及抗氧化相关酶活性等指标。结果 自发气调处理能够使CO2和O2含量维持在适宜的贮藏范围内,从而保持较高的TSS和总酚含量,抑制MDA含量及多酚氧化酶（PPO）活性,增强过氧化物酶（POD）、苯丙氨酸解氨酶（PAL）和过氧化氢酶（CAT）活性。结论 贮藏48~64 d,自发气调处理软枣猕猴桃的最适宜气体环境为体积分数3%左右的CO2和体积分数16%左右的O2,保持了软枣猕猴桃较好品质的机制,通过诱导果实的苯丙烷及抗氧化代谢实现。     

枸杞中甜菜碱的提取及其缓蚀性能研究

为了开发一种环保型天然缓蚀剂，采用纤维素酶酶解法提取枸杞中的甜菜碱，在单因素实验的基础上，采用响应面实验优化提取工艺，并研究了甜菜碱粗提液和粗提液与聚天冬氨酸复配物对Q235碳钢在硫酸溶液的缓蚀作用及其机理。结果表明，枸杞中甜菜碱的最佳提取工艺条件为：酶解时间为59 min,酶解温度为46℃,酶解pH值为3,纤维素酶用量为3 mg/g,液料比为25∶1 mL/g,甲醇体积分数为80%。在最优工艺参数下，枸杞中甜菜碱的提取率为20.34%,与模型预测值20.45%吻合良好。缓蚀实验表明，枸杞甜菜碱粗提液在硫酸介质中对碳钢具有良好的缓蚀效果，缓蚀效率随粗提液浓度的增大而升高，30℃下，当甜菜碱粗提液的浓度达到250 mg/L时，缓蚀效率可达到90%以上；与聚天冬氨酸复配后，缓蚀效率明显提高。甜菜碱粗提液及其复配物均为混合型缓蚀剂，可有效吸附在碳钢表面形成一层保护膜，吸附过程遵循Langmuir等温方程式，二者在碳钢表面的吸附均属于自发过程，属于以物理吸附为主的物理吸附和化学吸附共同作用的过程。     

漆酶催化壳聚糖-阿魏酸接枝共聚提高产物抗氧化及抗菌性的研究

以阿魏酸(FA)为底物,利用漆酶作为催化剂催化壳聚糖与阿魏酸接枝反应。对FA-壳聚糖接枝产物分别进行线性电位滴定、红外光谱检测和热重分析,确定产物的接枝率和结构的变化,并进一步对其抗氧化性和抑菌性能进行研究。结果表明,FA-壳聚糖的接枝率约为11.4%;在漆酶催化过程中阿魏酸的酚羟基与壳聚糖上的氨基参与反应,FA与壳聚糖之间可能受到了席夫碱(Schiff)的修饰;与壳聚糖原样相比,FA-壳聚糖衍生物的热稳定性也得到了提高;FA-壳聚糖衍生物对DPPH和ABTS自由基的清除率的IC50值分别为1.88和0.61mg/mL,表明FA-壳聚糖的抗氧化性能得到改善;抑菌性能检测得出在浓度为1mg/mL时FA-壳聚糖较壳聚糖原样对大肠杆菌和沙门氏菌的抑菌圈分别提高了1.5和1.3mm,在浓度为5mg/mL时FA-壳聚糖较壳聚糖原样对枯草芽孢杆菌和金黄色葡萄球菌的抑菌圈分别提高了2.5和2.0mm。     

酶解木质素改性制备Cr(Ⅵ)吸附剂

酶处理甘蔗渣得到酶解木质素(EHL)原料,通过Mannich反应合成酶解木质素改性吸附剂,并对其吸附Cr(Ⅵ)的性能进行了初步研究。采用正交试验得到了酶解木质素改性制备重金属离子吸附剂的最佳工艺条件,并用红外光谱仪、X射线衍射仪和扫描电镜对吸附剂进行了结构表征。实验结果表明:在室温下,向200mL质量浓度为100mg/L的Cr(Ⅵ)溶液投入少量的改性产物,Cr(Ⅵ)去除率可达88.9%。     

黑蒜黑枸杞复合饮料制备过程中酶解工艺优化

目的以黑蒜、黑枸杞等为主要原料,以出汁率、透光率、饮料中花色苷含量和DPPH自由基清除率为依据,研究酶解条件对复合饮料产率和稳定性的影响。方法选择纤维素与果胶酶比例、复合酶添加量、酶解温度和酶解时间为自变量的4因素3水平的正交实验优化酶的最佳酶解条件。结果各因素对复合饮料的模糊综合评分值的影响主次顺序为复合酶添加量酶解温度酶解时间复合酶比例;最佳酶解工艺条件,复合酶质量分数0.06%,纤维素酶与果胶酶的质量比为2︰1,酶解温度为50℃,酶解时间为3 h。在此优化工艺条件下,出汁率为91.04%,透光率为77.27%,饮料中花色苷的质量浓度为167.11 mg/L,DPPH自由基清除率为58.81%。结论该工艺生产的复合饮料出汁率高,抗氧化活性强。     

甜玉米芯硒多糖的制备及其对淀粉酶抑制作用

目的 以甜玉米芯多糖为原料,以硒含量为指标,优化硒化甜玉米芯多糖(SeSCP-80-I)的制备工艺,探究甜玉米芯硒多糖对α-淀粉酶、α-葡萄糖苷酶这2种影响淀粉消化关键酶的抑制活性.方法 在单因素试验的基础上,通过响应面实验优化工艺条件,并利用氢化物发生-原子荧光光谱法测定硒含量,利用红外光谱法对其结构进行测定.结果 当反应温度为86℃、原料配比(SCP-80-I与Na2SeO3的质量比)为1:0.91、硝酸浓度为0.54％(体积分数)、反应时间为6.4 h时,硒含量值可达到3.717 mg/g.制备出的甜玉米芯硒多糖存在Se—O—C特征吸收峰和SeO32?特征吸收峰,说明结合形成了稳定的硒化衍生物.硒化修饰后的甜玉米芯多糖对α-淀粉酶、α-葡萄糖苷酶抑制的IC50值分别为5.495 mg/mL和2.687 mg/mL,均优于未经硒化修饰的甜玉米芯多糖对α-淀粉酶、α-葡萄糖苷酶的抑制作用.结论 甜玉米芯硒多糖具有良好的降血糖活性.     

煮制处理对绿豆中多酚含量及其抗氧化活性的影响

目的为明确煮制处理对绿豆中多酚类化合物含量及其抗氧化活性的影响,优化得出绿豆多酚的最佳提取条件和煮制工艺,使绿豆在煮制过程中维持较高多酚含量和抗氧化活性,为绿豆的开发利用提供有力的数据支撑。方法选用山西生产的明绿豆为原材料,采用超声-微波(U-M)协同萃取法提取绿豆多酚,采用传统煮制法进行煮制处理,采用福林酚法进行定量检测,以多酚含量为指标进行单因素及正交试验,得到最佳的提取条件,并在其基础上探讨最佳煮制工艺,对煮制后的绿豆进行感官评定分析。通过DPPH自由基清除测试和总抗氧化试剂盒检测煮制处理前后绿豆多酚提取物的抗氧化能力,并进行分析。结果当U-M处理时间为30min,处理温度为40℃,提取固液比(g/mL)为1︰15,乙醇提取剂体积分数为60%时为最佳提取参数,其多酚含量为3.14mg/g;当煮制功率为400W,煮制固液比(g/mL)为1︰25,煮制时间为30 min时得到煮制工艺的最佳参数,其多酚含量为2.45 mg/g,感官评定分数为83;对照组、煮制处理组的绿豆多酚清除DPPH能力分别为69.54%,56.30%,对照组、煮制处理组的总抗氧化能力分别为29.13 U/mL和23.57 U/mL,表明其均具有较强的抗氧化能力,且呈量效关系。结论初步明确了煮制处理对绿豆中多酚类物质含量和抗氧化活性的影响,即经煮制处理后,绿豆中的多酚含量较低,抗氧化能力较弱。在保障风味的基础上,优化了煮制工艺,为绿豆中多酚类物质的合理利用提供了科学的参考依据。     

不同浓度1-MCP熏蒸处理对“龙成2号”软枣猕猴桃品质的影响

目的 探究不同浓度1-MCP处理对软枣猕猴桃果实采后生理品质的影响。方法 分别使用0.5、1.0、1.5μL/L 1-MCP熏蒸处理“龙成二号”软枣猕猴桃24 h，以不做任何处理为对照组，测定果实贮藏期间的外观、质量损失率、呼吸强度、乙烯释放量、硬度、可溶性固形物（TSS）含量、口感、风味、相对电导率及丙二醛（MDA）含量等。结果 不同浓度1-MCP处理均可较好地保持果实的外观和风味，降低软枣猕猴桃的呼吸强度和乙烯释放量，抑制果实质量损失率和TSS含量的上升，延缓软枣猕猴桃硬度和可滴定酸含量的下降，及相对电导率和MDA含量的上升，从而延长果实的贮藏期，维持软枣猕猴桃采后的商品性。其中，以1.0μL/L 1-MCP处理的效果最佳。结论 不同浓度1-MCP熏蒸处理可以有效保持软枣猕猴桃的品质，减少膜损伤，延缓果实衰老，延长货架期。     

Green synthesis of gold nanoparticles using Parsley leaves extract and their applications as an alternative catalytic,antioxidant, anticancer,and antibacterial agents

For the first time in this study, gold nanoparticles (AuNPs) were biosynthesized by the eco-friendly and cost-effective procedure using Presley leaf (Petroselinum crispum) extract then used as antioxidant, anticancer, antibacterial and photocatalytic agents. Different four-volume of the extract 2.5 mL, 5 mL, 10 mL, and 20 ML named AuNPs(A), AuNPs(B), AuNPs(C) and AuNPs(D) were used to module the size and shape of AuNPs. The prepared NPs were characterized by various techniques including UV–Vis absorption spectroscopy, Transmission Electron Microscopy (TEM), dynamic light scattering (EDX), and X-ray diffractometric (XRD). TEM imaging confirmed the formation of spherical, semi-rod aggregates, and flower-shaped NPs. The reduction and stabilization effect of the plant extract to fabricate AuNPs were explained by FTIR analysis. The four AuNPs provided high antioxidant ability, while AuNPs(D) was the best one. The NPs showed an emerging antimicrobial activity against two gram-negative microbes and not effective against the gram-positive microbes. The photocatalytic capacity for degradation of methylene blue dye was achieved after only one minute (the four samples have an equal effect). The AuNPs(D) provide the best anticancer activity on the human cancerous colorectal cell line using MTT assay rather than the other three AuNPs. The results spotlight for using Presley as a common cheap plant for bio-fabricating AuNPs who possess huge multifunctional applications.     

Solid-phase extraction and HPLC analysis of methylparaben and propylparaben in a concentrated antibiotic suspension

An accurate and precise solid-phase extraction coupled with high performance liquid chromatography (SPE/HPLC) method developed for the quantification of antimicrobial preservatives (methylparaben and propylparaben) in oxytetracycline injectable suspension is described in this article. The SPE technique was necessary to quantify the preservatives since the high concentration of the drug and excipients was masking low levels of preservatives, making quantification difficult. This developed HPLC method was stability-indicating and found to be linear between 1.3 to 2.4 mg/mL for methylparaben and 0.15 to 0.27 mg/mL for propylparaben in this concentrated antibiotic suspension formulation. The extraction recoveries were 98.8-101.6%. System precision and sample extraction precision (RSD) were less than 1%.     

Solid-Phase Extraction and HPLC Analysis of Methylparaben and Propylparaben in a Concentrated Antibiotic Suspension

ABSTRACT

An accurate and precise solid-phase extraction coupled with high performance liquid chromatography (SPE/HPLC) method developed for the quantification of antimicrobial preservatives (methylparaben and propylparaben) in oxytetracycline injectable suspension is described in this article. The SPE technique was necessary to quantify the preservatives since the high concentration of the drug and excipients was masking low levels of preservatives, making quantification difficult. This developed HPLC method was stability-indicating and found to be linear between 1.3 to 2.4 mg/mL for methylparaben and 0.15 to 0.27 mg/mL for propylparaben in this concentrated antibiotic suspension formulation. The extraction recoveries were 98.8–101.6%. System precision and sample extraction precision (RSD) were less than 1%.     

轻度失水对软枣猕猴桃机械伤和品质的影响

目的 探究轻度失水对软枣猕猴桃果实机械伤和品质的影响。方法 以软枣猕猴桃为材料,通过比较失水对运输模拟振动后果实机械伤的影响,以不失水模拟运输振动为对照,测定了处理后软枣猕猴桃果实贮藏过程中感官品质、营养品质、膜脂过氧化和活性氧代谢的变化情况。结果 与对照组相比,4%轻度失水处理有利于保持果实感官品质,使其腐烂率降低了10%、总酚含量提高了18.2%,抑制了其组织内丙二醛(MDA)含量和相对电导率的升高,保持贮藏过程中过氧化氢酶(CAT)、过氧化物酶(POD)、抗坏血酸过氧化物酶(APX)活性。结论 4%轻度失水可能是通过降低了果实硬度从而降低腐烂率,保持较高活性氧清除能力,抑制成熟进程,从而利于缓解软枣猕猴桃果实采后运输机械伤发生,保持果实品质。     

软枣猕猴桃果醋发酵工艺

以软枣猕猴桃为实验材料,通过单因素试验确定正交试验各因素的作用的最佳水平,进行醋酸发酵工艺条件的正交试验,结果表明：温度为33℃接种量为0．5％,酒精度为7°,pH值为4．4。通过软枣猕猴桃醋杀菌方式的选择试验,最佳杀菌方式为瞬时巴氏杀菌及95℃保持杀菌1min。     

4种植物精油对草莓致病菌的抑制作用研究

通过研究小茴香、百里香、罗勒和迷迭香4种植物精油对灰霉、根霉及黑曲霉等草莓主要致病菌的抑制效果,筛选出对草莓有良好贮藏保鲜效果的植物精油。采用培养基稀释法进行筛选,并确定了其最低抑菌浓度。最后对该精油进行气相色谱-质谱联用技术(GC-MS)分析和主要抑菌成分的气相色谱(GC)定量分析。结果表明,所选用的4种精油中,百里香精油具有广谱高效的抑菌性能,其对灰霉、根霉和黑曲霉的最低抑菌浓度均为2μL/L;百里香精油中的主要抑菌成分为百里香酚和香芹酚,且其相对质量分数分别为23.52%和3.76%,绝对质量浓度分别为181.594 9 mg/m L和27.665 6 mg/m L。     

桑皮纤维的提取方法及其应用和发展

目的综述桑皮纤维组成成分、结构特征、提取工艺及其研究进展。方法化学脱胶法和生物脱胶法是研究桑皮纤维提取最常用的方法。结果桑皮与其他造纸原料相比具有高果胶含量、低木素含量的特点,果胶的质量分数最高为11.16%,木素的质量分数最低为8.74%;不同地域桑皮的果胶含量相差30%左右,木素含量相差约为50%。桑皮纤维的横截面呈现椭圆或不规则状,纤维表面粗糙、不平整。桑皮纤维提取的重点在于果胶的脱除,桑皮脱除果胶的方法主要有化学法和生物法2种。结论纺织方向上的研究近乎完善,造纸方向上的研究还刚刚起步,而开发桑皮纤维相关产品以提高其应用价值是下一阶段研究的主要方向。     

Liquid-liquid extraction in the 96-well plate format with SRM LC/MS quantitative determination of methotrexate and its major metabolite in human plasma.

A method involving the semirobotic liquid-liquid extraction (LLE) in deep-well 96-well plates was developed for the quantitation of the anti-cancer/antiinflammatory drug methotrexate (MTX) and its major metabolite, 7-hydroxymethotrexate (7OH-MTX) in human plasma. The extraction time for the sample preparation was relatively short with four 96-well plates (384 samples) prepared in approximately 90 min by one person. The sample extracts were each analyzed within 1.2 min using a positive ion turbo-ionspray selected reaction monitoring liquid chromatography/mass spectrometric (SRM LC/MS) method in which 768 samples were easily analyzed within 22 h (maximum of 820 samples in 24 h). Deuterated internal standards, MTX-d3 and 7OH-MTX-d3, were used. The calibration curves for MTX and 7OH-MTX were linear (R2 > 0.997) and ranged from 0.5 to 250 and 0.75 to 100 ng/mL, respectively. The limit of quantitation (LOQ) for MTX and 7OH-MTX was 0.5 and 0.75 ng/mL, respectively; persistent carryover from the autosampler limited the LOQ achievable. The limit of detection (LOD) was 0.05 ng/mL for MTX and 0.1 ng/mL for 7OH-MTX. The intra- and inter-assay precision and accuracy did not exceed 15% for both MTX and 7OH-MTX. The recoveries were 61% for MTX and 47% for 7OH-MTX. The method was validated and demonstrated to be robust with high precision and accuracy.     

Determination of anionic and nonionic surfactants, their degradation products, and endocrine-disrupting compounds in sewage sludge by liquid chromatography/mass spectrometry

A comprehensive analytical method based on reversed-phase liquid chromatography and mass spectrometry using both atmospheric pressure chemical ionization and electrospray ionization has been developed for the simultaneous determination of anionic and nonionic surfactants, their polar degradation products, and endocrine-disrupting compounds (EDCs) in sewage sludge. Extraction of target compounds, with recovery rates from 86% to nearly 100% for polyethoxylates and from 84 to 94% for polar degradation products, was achieved applying ultrasonic solvent extraction with a mixture of methanol/ dichloromethane (7:3, v/v). Cleanup of sample extracts was performed on octadecyl solid-phase extraction cartridges. Determination of less polar compounds: alcohol ethoxylates (AEOs), nonylphenol ethoxylates (NPEOs), coconut diethanol amides, poly(ethylene glycol)s, and phthalate esters was accomplished by reversed-phase LC-APCI-MS in positive ionization mode, while more polar compounds: nonylphenolcarboxylates, nonylphenol (NP), octylphenol, and bisphenol Awere analyzed by ion-pair LC-ESI-MS under negative ionization conditions. This protocol was successfully applied to the trace determination of anionic and nonionic surfactants, polar degradation products, and EDCs in sewage sludge collected from different sewage treatment plants. The analysis revealed the presence of NP at high concentration levels ranging from 25 to 600 mg/kg. Polyethoxylates (AEOs and NPEOs) were also found in all samples at parts-per-million levels (10-190 mg/kg AEOs and 2-135 mg/kg NPEOs, respectively).