海藻糖对银杏叶过氧化物酶活性的影响

对银杏叶过氧化物酶热稳定性、最适pH值及海藻糖对酶热稳定性的保护作用和pH值的影响进行了初步研究.结果表明,用40℃和50℃分别处理30 min,过氧化物酶活性分别剩余78%和39%,在60,70,80℃下活性可分别维持27,18和3 min.银杏叶过氧化物酶最适pH值为6.4和9.2.加入海藻糖后酶活性略有降低(活性丧失约8%),但加热后海藻糖的保护优势得以显现,海藻糖浓度为0.14 mol/L时保护效果最好,而且不同温度处理下酶活的下降趋势均较对照组缓慢,尤其在50,60,70℃条件下,相比对照组高出12%~27%的活性,加热时间长短对海藻糖的保护效果无太大影响.经0.14 mol/L浓度的海藻糖处理后,酶活的最适pH值为6.6和9.4.     

基于伴刀豆球蛋白固定过氧化物酶无介体新型生物传感器的研制

以纳米金吸附辣根过氧化物酶,用活化的伴刀豆球蛋白(Con A)将其固定在裸金电极表面,研制成一种新型的无介体辣根过氧化物酶生物传感器。探讨了纳米金的尺寸、组装膜层数、工作电位和pH等实验条件对传感器性能的影响。在pH7.0,外加电压-150mV(vs.SCE)条件下,传感器对H2O2在5.0×10-6~1.2×10-2mol/L范围内呈线性关系;检出限为2.9×10-6mol/L。将传感器用于实际样品的测定,结果良好。     

Synthesis of an enzyme-like imprinted polymer with the substrate as the template, and its catalytic properties under aqueous conditions

Transition state analogues (TSAs) have long been regarded as ideal templates for the preparation of catalytically active synthetic imprinted polymers. In the current work, however, a new type of molecularly imprinted polymer (MIP) was synthesized with the substrate (homovanillic acid, HVA) as the template and hemin introduced as the catalytic center, with the use of plural functional monomers to prepare the active sites. The MIP successfully mimicked natural peroxidase, suggesting that it may not be imperative to employ a TSA as the template when preparing enzyme-like imprinted polymers and that the imprinted polymer matrix provided an advantageous microenvironment around the catalytic center (hemin), essentially similar to that supplied by apo-proteins in natural enzymes. Significantly, by taking advantage of the special structure of hemin and multiple-site interactions provided by several functional monomers, the intrinsic difficulties for MIPs in recognizing template molecules in polar solutions were overcome. The newly developed polymer showed considerable recognizing ability toward HVA, catalytic activity, substrate specificity and also stability, which are the merits lacked by the natural peroxidase. Meanwhile, the ease of recovery and reuse the MIP implies the potential for industrial application.     

A novel electrochemical aptasensor for highly sensitive detection of thrombin based on the autonomous assembly of hemin/G-quadruplex horseradish peroxidase-mimicking DNAzyme nanowires

In this work, a new signal amplified strategy was constructed based on isothermal exponential amplification reaction (EXPAR) and hybridization chain reaction (HCR) generating the hemin/G-quadruplex horseradish peroxidase-mimicking DNAzyme (HRP-mimicking DNAzyme) nanowires as signal output component for the sensitive detection of thrombin (TB). We employed EXPAR’s ultra-high amplification efficiency to produce a large amount of two hairpin helper DNAs within a minutes. And then the resultant two hairpin helper DNAs could autonomously assemble the hemin/G-quadruplex HRP-mimicking DNAzymes nanowires as the redox-active reporter units on the electrode surface via hybridization chain reaction (HCR). The hemin/G-quadruplex structures simultaneously served as electron transfer medium and electrocatalyst to amplify the signal in the presence of H₂O₂. Specifically, only when the EXPAR reaction process has occurred, the HCR could be achieved and the hemin/G-quadruplex complexes could be formed on the surface of an electrode to give a detectable signal. The proposed strategy combines the amplification power of the EXPAR, HCR, and the inherent high sensitivity of the electrochemical detection. With such design, the proposed assay showed a good linear relationship within the range of 0.1 pM–50 nM with a detection limit of 33 fM (defined as S/N = 3) for TB.     

Ferrocene and Ferricenium Ion as Versatile Photometric Titrants of H2O2 and D-Glucose in the Presence of Peroxidase and Glucose Oxidase. A Ferrocene-Peroxidase Stairway #

Abstract

Hydrogen peroxide can routinely be determined in the presence of ferrocene (FcH) and horseradish peroxidase by monitoring at 617 nm the enzymatically produced ferricenium dye. In contrast, D-glucose can be assayed by following the fading of the ferricenium dye FcH⁺PF₆ ^? in the presence of glucose oxidase. The change in absorbance in both cases corresponds to the amount of analyte. viz. H₂O₂ or D-glucose, in solution. The routine is very simple, invariant to the concentrations of both ferrocenes/ferricenium salt and enzyme and allows numerous “one pot” measuremeats with the detection limit of 10^?4 M for both the analytes. It takes 2–4 and 5–10 min to accomplish one analysis of H₂O₂ and D-glucose in the presence of peroxidase and glucose oxidase, respectively.

     

Rapid Selection of Phage Se-scFv with GPX Activity via Combination of Phage Display Antibody Library with Chemical Modification

IntroductionReactive oxygen species(ROS) are known to de-stroy biomacromolecules and cause cell injury[1]. Un-der normal circumstances, there is a balance betweenthe production of ROS and their destruction. Many dis-eases, such as brain ischemia, tumor, v…     

Enhanced Peroxidase‐Like Properties of Graphene–Hemin‐Composite Decorated with Au Nanoflowers as Electrochemical Aptamer Biosensor for the Detection of K562 Leukemia Cancer Cells

Graphene composites with hemin and gold nanoparticles show a better performance for hydrogen peroxide decomposition compared to that of the three components alone or duplex/hybrid complexes. Our previous studies showed that the morphology of the Au nanoparticles may greatly influence the catalytic activity of graphene‐family peroxidase mimics. Recently, we found that Au nanoflowers could grow in situ and form on the surface of hemin/RGO (reduced graphene oxide). The prickly morphology of this Au nanoflower brought a higher catalytic ability with enhanced kinetic parameters than traditional Au nanoparticles that showed a smooth surface. Therefore, based on this discovery, a smart electrochemical aptamer biosensor for K562 leukemia cancer cells was further presented with good performance in selectivity and sensitivity attributed to the excellent mimetic peroxidase catalytic activity of this newly synthesized Au nanoflower decorated graphene–hemin composite (H‐RGO‐Au NFs).     

不同耐肥性的小麦品种幼苗硝酸还原酶、过氧化物酶活力及同功酶的研究

在不同耐肥性的6个小麦品种幼苗中,硝酸还原酶(NR)活力差异显著,耐肥性弱的品种NR活力高,活力是负相关关系。过氧化物酶活力,耐肥性强的高于耐肥性弱的。根据聚丙烯酰胺凝胶电泳分离出的过氧化物酶的同工酶谱,耐肥性强的品种均有8条酶带,且着色深,耐肥性弱的品种则有7条,着色较浅。此外通过根系活力测定,耐肥性强的低于耐肥性弱的品种,而叶绿素含量则高于耐肥性弱的品种。