Co2NiO4@碳布复合电极的制备及其在葡萄糖传感器中的应用

为了获得传感性能优异的无酶葡萄糖传感器的电极材料，采用水热法在碳布(CC)表面原位生长纳米针状的前驱体，再通过热处理得到Co₂NiO₄纳米针状阵列，构建Co₂NiO₄@CC复合电极体系。采用XRD、SEM和BET对该复合电极材料进行物相、形貌与结构表征。在0.10 mol/L氢氧化钠溶液体系中，采用自主搭建的测试平台进行葡萄糖检测。结果表明：Co₂NiO₄@CC电极对葡萄糖表现出较高的灵敏度29.72μA/(mM·cm²)和良好线性响应0.01 mol/L～11.50 mmol/L;根据信噪比S/N=3,计算出检测限为1.65 mol/L,并且表现出良好的重复使用性和选择性。本文为复合型无酶葡萄糖传感材料的研究提供了新的设计思路。     

团聚状纳米AuPd修饰的H2O2生物传感器研究

采用微胶束法室温条件下制备团聚状的AuPd合金纳米粒子,使用紫外可见光谱(UV-vis),透射电镜(TEM),X-射线粉末衍射(XRD)和X-射线能量色散谱(EDS)表征团聚结构AuPd合金纳米粒子的形貌、尺寸、结构和组成。用制备的AuPd纳米粒子修饰辣根过氧化物酶玻碳电极,制备无电子媒介的过氧化氢生物传感器HRP/AuPd/GCE。使用循环伏安法和计时电流法表征了HRP/AuPd/GCE对H2O2的检测性能。实验结果表明:该传感器对H2O2具有良好的检测性能和稳定性,在H2O2浓度为1×10-7mol/L～5×10-3mol/L范围内检测电流与H2O2浓度有线性关系,线性相关系数R2=0.995 01,检出限为7.6×10-7mol/L。     

基于Au@PtNPs-Cu-MOF/GCE传感器检测果蔬中葡萄糖

制备了新型的金@铂纳米复合材料-铜金属有机骨架/玻碳电极传感器(Gold platinum nanocomposites-Copper metal organic skeleton/Glassy carbon electrode, Au@PtNPs-Cu-MOF/GCE)对水果中的葡萄糖进行快速检测分析。制备了二维的Cu-MOF纳米材料,并在其表面负载核壳结构的Au@PtNPs,合成了一种新型的Au@PtNPs-Cu-MOF纳米复合材料。利用Au@PtNPs-Cu-MOF构建了一种新型的Au@PtNPs-Cu-MOF/GCE,通过对电解质pH、计时安培电位的优化,确定Au@PtNPs-Cu-MOF/GCE的最佳工作条件,对梨中的葡萄糖进行定量分析。研究发现,葡萄糖浓度与其峰电流分别在0.1-10和10-2600 μmol/L范围内呈良好的线性关系,检出限(S/N=3)为0.0872 μmol/L,线性范围宽,检出限低,明显优于其它的葡萄糖无酶传感器。且Au@PtNPs-Cu-MOF/GCE的重复性和抗干扰能力较好,8次扫描的相对标准偏差为1.32%。对梨汁中的检测结果为1.8457 mmol/L与高效液相色谱法一致且偏差仅为5.46%,符合检测要求。     

基于金磁微粒模拟酶电化学增强体系检测抗坏血酸

通过自组装方法构建金磁微粒(Fe3O4@Au)复合纳米粒子,并表征其理化性能。基于复合纳米粒子的过氧化物模拟酶活性,构建出具有高灵敏度和选择性检测抗坏血酸的无酶增强型电化学传感器。采用循环伏安法对抗坏血酸进行测定,并优化电化学检测体系。结果表明,在最适检测体系下,所构建的无酶电化学传感器对抗坏血酸检测具有较高的灵敏度,抗坏血酸浓度在1~100 mmol/L范围内,电流响应与其具有良好线性关系,工作曲线方程为y=867.81x+3.860 8(R~2=0.991 9),检出限为0.011 7 mg/L。此外,所构建的无酶电化学传感器已成功地应用于实际样品检测。鉴于此,所提出的新型无酶电化学分析方法在食品质量控制和安全检测等领域有着巨大的应用前景。     

基于Pd/MWCNTs/玻碳电极的电化学传感器检测水样中的羟胺

目的 采用Pd/MWCNTs纳米复合材料修饰玻碳电极构建电化学传感器,检测养殖水样及水产饲料中羟胺（NH2OH）的含量。方法 采用乙二醇还原法制备Pd纳米颗粒负载于碳纳米管上制备Pd/MWCNTs纳米材料,运用XRD(X-ray diffraction)和TEM (transmission electron microscopy) 手段对材料进行表征,构建基于Pd/MWCNTs/GCE的电化学传感器检测羟胺。 结果 在优化的工作条件下,Pd/MWCNTs纳米复合材料修饰电极检测羟胺线性检测范围2.0 μmol/L-1.4 mmol/L,检测下限达0.60 μmmol/L (S/N = 3),灵敏度为61.48 μA/mmol/L,是MWCNTs修饰电极检测灵敏度的10倍左右。结论 复合材料制备的传感器检测羟胺具有灵敏度高、线性范围宽、检测下限低的特点,可用于抗氧化剂羟胺的检测。     

基于Fe_3O_4-PGA@Au构建无酶电化学生物传感器检测葡萄糖

基于聚谷氨酸(poly-(γ-glutamic acid),PGA)金磁微粒(Fe_3O_4-PGA@Au)电沉积修饰玻碳电极,构建具有高灵敏度和选择性的无酶型葡萄糖电化学生物传感器。采用绿色还原吸附法制备Fe_3O_4-PGA@Au,并利用X射线衍射、傅里叶变换红外光谱和透射电子显微镜表征Fe_3O_4-PGA@Au的理化性质。优化葡萄糖传感检测体系的反应温度、pH值、扫描速率和反应时间等检测参数。采用循环伏安法考察无酶传感器检测葡萄糖时的电化学行为。结果表明,Fe_3O_4-PGA@Au具有良好的电催化性能;在葡萄糖浓度分别为0.1～5.0μmol/L和10～250μmol/L范围内时,所构建的生物传感器峰电流密度与其葡萄糖浓度具有良好的线性关系,检出限为0.74μmol/L(RSN=3);此无酶型生物传感器具有好的重复性和抗干扰性能。     

基于镍基碱式碳酸盐修饰电极的无酶葡萄糖电化学传感器构建

在碳纸基底上采用一步水热法原位生长过渡金属镍基碱式碳酸盐（Ni-CHs）,构建高灵敏度和选择性的无酶葡萄糖传感器。利用X射线衍射、扫描电子显微镜对材料进行物相鉴定和形貌表征。采用循环伏安法和时间-电流曲线评估传感器的性能。结果表明,Ni-CHs/CP在0.5 V的低电势下对葡萄糖具有最佳检测能力,传感器的氧化峰电流在葡萄糖浓度为0.95 μmol/L～2.623 mmol/L范围内呈现良好的线性关系,相关系数为0.998,检出限为0.31 μmol/L（RSN=3）,响应时间为3.5 s。此传感器具有良好的稳定性并成功用于实际样品的检测,加标回收率在95.65%～105.56%之间。     

基于纳米酶技术检测柑桔果实中葡萄糖

目的建立纳米酶检测体系快速检测葡萄糖含量的技术。方法采用甲烷氧化细菌的发酵代谢产物甲烷氧化菌素(methanobactin,MB)与Cu²⁺、纳米金(AuNPs)、葡萄糖氧化酶(glucose oxidase,Gox)进行配位结合形成具有过氧化物酶和葡萄糖氧化酶性质的纳米酶检测体系,并对柑桔果实中葡萄糖含量进行测定。结果Gox浓度0.81 U/mL、MB-Cu@AuNPs模拟酶浓度2×10^-5mol/L、对苯二酚浓度5×10^-4mol/L、反应温度60℃、pH 8为最佳检测条件;反应时间5 min,葡萄糖浓度线性范围为1×10^-3-5×10^-2mol/L。结论纳米酶检测体系对柑桔果实中葡萄糖含量的测定具有良好的稳定性,相比较常规快速血糖仪检测速度更快。     

辣根过氧化物酶/聚乙烯醇缩丁醛/碳纳米管修饰玻碳电极检测过氧化氢

制备了辣根过氧化物酶/聚乙烯醇缩丁醛/碳纳米管修饰玻碳电极的过氧化氢生物传感器。以对苯二酚为电子媒介,采用循环伏安法和电流时间法考察了该传感器对H2O2的催化性能。讨论了媒介体浓度、工作电位、温度、pH对电极响应的影响。结果表明传感器对H2O2表现出良好的电催化性能。在pH=7.0,对苯二酚浓度4.2mmol/L,工作电位-250mV的实验条件下,H2O2浓度在1.67×10-7～1.29×10-5mol/L及1.58×10-5～1.17×10-3mol/L范围内与传感器的电流响应呈线性关系,检出限(S/N=3)为5.554×10-8mol/L。该传感器制备方法简单,成本低,稳定性好,对HO有快速灵敏的响应。     

金磁微粒模拟酶检测食品中的葡萄糖

采用水热法制备Fe3O4纳米粒子,并通过对其表面氨基化与金纳米粒子自组装方法构建金磁微粒（Fe3O4@Au）,并表征其性能。在其具有模拟过氧化物酶活性的基础上结合葡萄糖在葡萄糖氧化酶作用下产生H2O2,并与过氧化物酶底物二胺盐产生显色反应的原理,建立可视化检测葡萄糖含量的简便方法,并优化葡萄糖检测体系,并对其选择性和回收率进行分析。结果表明,氨基化的Fe3O4纳米粒子可以有效固载金纳米粒子,Fe3O4@Au饱和磁化强度为43 emu/g。检测体系最优工艺组合为：Fe3O4@Au混悬液质量浓度0.15 g/mL、温度70 ℃、时间50 min。在优化条件下,葡萄糖在1～20 mmol/L范围内具有良好的线性关系,线性相关系数R2为0.992 5,检出限为2.45 μmol/L,加标回收率在96%～104%之间,并具有良好的选择性和稳定性。本研究将拓宽纳米材料模拟酶在食品检测的应用并为葡萄糖检测方法的改进提供一种新的思路。     

Thiomethylstilbenes as inhibitors of CYP1A1, CYP1A2 and CYP1B1 activities

Resveratrol (3,5,4'-trihydroxy-trans-stilbene) is a natural stilbene derivative occurring in grapes, peanuts and red wine. Its chemopreventive action has been established in studies on animal models. Recently, numerous classes of compounds with stilbene backbone have been investigated for their biological activity concerning cancer prevention; e. g. resveratrol methyl ethers appeared to be specific and potent inhibitors of cytochromes P450 (CYP) family 1 involved in the activation of procarcinogens. Since the replacement of the 4'-hydroxyl with a thiomethyl group is supposed to reduce toxicity of stilbene derivatives, the purpose of this study was the synthesis and evaluation of a series of 4-thiomethyl-trans-stilbene derivatives differing in a number and position of additional methoxy groups. Their inhibitory potency toward human recombinant CYPs: CYP1A1, CYP1A2 and CYP1B1 have been studied and compared with the effect of resveratrol and its analogues. Among compounds tested, 2-methoxy-4'-thiomethyl-trans-stilbene and 3-methoxy-4'-thiomethyl-trans-stilbene demonstrated the most potent and selective inhibitory effect on CYP1A1 and CYP1B1 activities. The results of our study indicate that modification of stilbene derivatives with thiomethyl group may influence the selectivity and inhibitory potency of these compounds toward P450 isozymes. Thus, it should be considered in developing new chemopreventive agents based on their mechanism of action.     

Characterization of the sec1-1 and sec1-11 mutations.

Sec1 proteins are implicated in positive and negative regulation of SNARE complex formation. To better understand the function of Sec1 proteins we have identified the nature of the temperature-sensitive mutations in sec1-1 and sec1-11. The sec1-1 mutation changes a conserved glycine(443) to glutamic acid. The sec1-11 mutation changes a highly conserved arginine(432) to proline. Based on homology and the crystal structure of the mammalian nSec1p, the corresponding amino acids localize to the 3b domain of nSec1p. Compared to the wild-type Sec1p the mutant proteins are less abundant even at the permissive temperature. Thus, the R432P and G443E mutations may cause structural alterations that affect folding and make the mutant proteins more susceptible to degradation. The remaining part is sufficient for growth and protein secretion at 24 degrees C and thus is likely to be properly folded. At 37 degrees C the mutant proteins become non-functional. In pulse-chase-type experiments the newly synthesized Sec1-1 and Sec1-11 proteins decayed similarly with the wild-type protein. Thus, the non-functionality of the mutant proteins cannot be explained by denaturation-induced degradation only. It is possible that the newly synthesized mutant proteins fold slowly and are susceptible to degradation before they have managed to fold and associate with other proteins. The mutant proteins were unable to interact with the Sec1p-interacting proteins Mso1p and Sso2p in the two-hybrid assay, even at the permissive temperature. These results localize sec1-1 and sec1-11 mutations to a domain of Sec1p and suggest a mechanism by which sec1-1 and sec1-11 cells become temperature-sensitive.     

1-L-谷氨酸-1-脱氧-D-果糖的热裂解分析研究

采用热重-微分热重技术研究了1-L-谷氨酸-1-脱氧-D-果糖的热失重和裂解温度,通过在线热裂解与气质联用技术分别分析研究了无氧和有氧条件下1-L-谷氨酸-1-脱氧-D-果糖在300℃、600℃、750℃和900℃四个温度的热裂解产物。研究结果表明1-L-谷氨酸-1-脱氧-D-果糖的裂解温度为161.3℃,在700℃时失重达到90.50%。无氧和有氧条件下裂解产物的种类和数量随着裂解温度升高而增多,有氧条件下裂解产物总数稍多于无氧条件,但品种有明显差异。 无氧裂解和有氧裂解产物主要为酮类、吡咯类、吡啶类、呋喃类、吡嗪类、吲哚类以及少量芳香族化合物。有氧热裂解产物的香韵分析结果表明1-L-谷氨酸-1-脱氧-D-果糖裂解产物具有烘烤香、坚果香、甜香、花香、奶香等香韵。      

Inhibition of human recombinant cytochromes P450 CYP1A1 and CYP1B1 by trans-resveratrol methyl ethers

CYP1A1 and CYP1B1 are the inducible forms of cytochrome P450 expressed in extrahepatic tissues, which are responsible for the biotransformation of polycyclic aromatic hydrocarbons, heterocyclic amines and estradiol to the carcinogenic intermediates. The aim of our research was to determine and compare the inhibitory effect of naturally occurring analogues of trans-resveratrol on the catalytic activities of human recombinant CYP1A1 and CYP1B1. Pinostilbene (3,4'-dihydroxy-5-methoxystilbene), desoxyrhapontigenin (3,5-dihydroxy-4'-methoxystilbene), and pterostilbene (3,5-dimethoxy-4'-hydroxystilbene) appeared to be very potent inhibitors of CYP1A1 catalytic activity with Ki values of 0.13, 0.16 and 0.57 microM, respectively. Results from this study indicate that trans-resveratrol analogues in which the hydroxy groups are substituted by methoxy groups exhibit a remarkably stronger inhibitory effect towards CYP1A1 in comparison to the parent compound. On the contrary, the potency of pinostilbene, desoxyrhapontigenin and pterostilbene towards CYP1B1 with Ki values of 0.90, 2.06 and 0.91 microM, respectively, was comparable to that of resveratrol. It appears that between these analogues, inhibition of CYP1A1 and CYP1B1 catalytic activities does not vary much regardless of the number and position of methylether substitution. The results suggest that the trans-resveratrol analogues: pinostilbene, desoxyrhapontigenin and pterostilbene, which occur in some food plants, might be considered as promising chemopreventive agents.     

1-L-亮氨酸-1-脱氧-D-果糖和1-L-异亮氨酸-1-脱氧-D-果糖的热裂解分析

采用热重-差热法对1-L-亮氨酸-1-脱氧-D-果糖（Ⅰ）和1-L-异亮氨酸-1-脱氧-D-果糖（Ⅱ）的热失重和热裂解温度进行了研究,通过在线裂解GC-MS联用技术分别对（Ⅰ）和（Ⅱ）在350℃、450℃、550℃、650℃、750℃和850℃条件下的裂解产物进行了鉴定。研究结果表明2种Amadori化合物的的热失重曲线相似,（Ⅰ）的裂解温度为144.67℃,（Ⅱ）的裂解温度为164.26℃,600℃时（Ⅰ）和（Ⅱ）失重约80%;二者裂解产物主要为吡嗪类、吡啶类、吡咯类、喹啉类和呋喃类等杂环化合物,芳香族化合物以及醛类和酮类,其中以吡嗪类为主;裂解产物的数量随着裂解温度的升高而增多,（Ⅰ）的裂解产物数量与（Ⅱ）的裂解产物数量相当。对（Ⅰ）和（Ⅱ）的主要裂解产物形成途径进行了初步探讨,可为研究其在卷烟燃烧过程中的转化行为提供参考。      

Molecular cloning of CaYRB1, the Candida albicans RanBP1/YRB1 homologue.

The yeast Ran binding protein 1 (Yrb1p) is a small protein of 23 kDa that is highly conserved among eukaryotes. It stimulates the GTPase activity of Gsp1p in the presence of the GTPase activating protein Rna1p. In addition to its role in nucleocytoplasmic transport of macromolecules, YRB1/RanBP1 could be involved in the regulation of microtubules structure and dynamics. Since microtubules are tightly associated with morphological changes, we have been interested to study the role and function of YRB1 in the pathogenic fungus Candida albicans, where there is regulated change in cellular morphology. The gene product of CaYRB1 encodes a 212 amino acid protein displaying 73% homology to the S. cerevisiae homologue. The bacterially expressed gene product has an apparent molecular weight of 35.7 kDa. We show that it can complement a S. cerevisiae yrb1 null mutant and that its mRNA does not appear to be regulated in response to conditions inducing morphological changes in C. albicans.     

Effects of Cry1F and Cry34Ab1/35Ab1 on storage pests

Two crystalline protoxins from Bacillus thuringiensis (Bt), Cry1Fa1 and Cry34Ab1/Cry35Ab1 (Cry1F, Cry34/35Ab1), were evaluated for efficacy against lepidopteran and coleopteran storage pests. Cry1F was tested against the lepidopterans Sitotroga cerealella (Angoumois grain moth) and colonies of Plodia interpunctella (Indian mealmoth) that are susceptible or resistant to Bt Cry1Ab and Cry1Ac toxins, Bt subspecies entomocidus, and the commercial formulation Dipel^®. Cry1F was also tested against the coleopterans Cryptolestes pusillus (flat grain beetle) and Tribolium castaneum (red flour beetle). Cry34/35Ab1 was tested against S. cerealella, C. pusillus, and T. castaneum, and against additional coleopteran storage pests, including Tenebrio molitor (yellow mealworm), Trogoderma variabile (warehouse beetle), Oryzaephilus surinamensis (sawtoothed grain beetle), Rhyzopertha dominica (lesser grain borer), and Sitophilus oryzae (rice weevil). Strains of Bt-susceptible or -resistant P. interpunctella generally were more sensitive to Cry1A protoxin or toxin than either Cry1F protoxin or Dipel. Despite difficulties with the bioassay of S. cerealella larvae, the data suggest that Cry1F and Cry34/35Ab1 caused increased larval mortality, and a developmental delay was observed and no pupae emerged with 0.9% Cry1F. Neither Cry1F nor the corn rootworm-active toxin Cry34/35Ab1 significantly affected the biological parameters of the coleopteran species evaluated.     

Thermodynamic characterization of the PR-10 allergens Bet v 1, Api g 1 and Dau c 1 and pH-dependence of nApi g 1 and nDau c 1

Natural and recombinant Bet v 1, the major birch pollen allergen, and homologous allergens, Api g 1 and Dau c 1, from celery and carrot, respectively, were studied by CD spectroscopy under conditions of varying denaturant concentration, pH and temperature to determine fundamental thermodynamic parameters for conformational stability. Thermodynamic studies increase basic knowledge regarding differences between birch pollen-related allergens and are of importance in choosing processing conditions. The conformational stability determined from guanidine hydrochloride denaturation curves was similar for rBet v 1.0101 and rApi g 1.0101. Conformational responses to chaotropic salt were different for recombinant allergens from different species, but were similar for the natural isoform mixtures. The conformational stabilities of nApi g 1 and nDau c 1, were shown to be similar to rBet v 1.2801 at pH > 4.4 [Mogensen, J. E., Ipsen, H., Holm, J., & Otzen, D. E. (2004). Elimination of a misfolded folding intermediate by a single point mutation. Biochemistry, 43(12), 3357-3367], but nApi g and nDau c 1 were stable to heating at lower pH-values.     

Characterization of the adr1-1 nonsense mutation identifies the translational start of the yeast transcriptional activator ADR1

We have characterized a nonsense mutation in the ADR1 gene that identifies the translational start of the ADR1 protein. The ADR1 gene of Saccharomyces cerevisiae is required for synthesis of the glucose-repressible alcohol dehydrogenase (ADH2). The adr1-1 mutation, which inhibits ADH2 expression, was identified as a C to G transversion at base pair +32. This alteration would result in a UGA nonsense codon in place of a serine codon that would lead to termination of the ADR1 polypeptide after the 10th amino acid. The effect of the adr1-1 mutation was partially reversed by UGA-tRNA suppressors, indicating that the adr1-1 mutation affects ADR1 expression at the translational level. These observations establish that the first available AUG in the ADR1 sequence is used as the translational start site of ADR1. Tyrosine or leucine UGA-tRNA-suppressors resulted in levels of adr1-1 activity similar to that found for a serine UGA-tRNA-suppressor, suggesting that serine residue-11 is not essential to ADR1 function. Northern analyses showed that the 5.1 kb ADR1 mRNA was two- to three-fold more abundant when isolated from a strain carrying the ADR1 allele than from an isogenic strain containing the adr1-1 allele. These data confirm that the 5.1 kb mRNA is the ADR1 mRNA and suggest that inhibition of adr1-1 mRNA translation results in more rapid degradation of the adr1-1 mRNA.     

Physical mapping of a centromere-proximal region of chromosome IV-l defines the placement of genes USO1, MBP1, PSA1 and SLC1

A physical map of a 14·5 kb region close to the centromere on the left arm of chromosome IV of Saccharomyces cerevisiae is presented. This map has been constructed by restriction analysis of a clone from a YCp50 genomic library and by use of pre-existing and new sequence data from this region. The map reveals the following gene order (reading from the most centromere-distal to the most centromere-proximal locus): USO1/INT1–MBP1–PSA1–SLC1–YLA1 and defines the size of the open reading frames and intergenic regions.