ESR技术应用于逆胶束酶体系性能的研究

采用鼠李糖脂构建逆胶束体系,并利用电子自旋共振（ESR）技术研究了鼠李糖脂逆胶束及鼠李糖脂逆胶束酶体系的性能.采用ESR光谱技术计算得出逆胶束体系中超精细分裂常数变化,研究表明鼠李糖脂在正己烷中的临界胶度为0.07mmol/L.通过分析ESR光谱中旋转相关时间的变化,探究电子自旋探针在逆胶束中运动受阻和翻转所需时间的情况同时,ESR光谱的峰值体现了样品中自由基含量的多少.通过对比逆胶束体系、逆胶束酶体系以及逆胶束酶-苯酚体系的ESR光谱,结果表明鼠李糖脂逆胶束-苯酚体系的自由基最多.通过研究对16-氮氧自由基硬脂酸对两个体系作用,推测出电子自旋探针自由基团定位于逆胶束水核中,并且推断电子自旋探针位于水核的边缘区域,即结合水水层.该研究为逆胶束酶体系的应用提供了坚实的理论基础.     

Validation of a conductometric bienzyme biosensor for the detection of proteins as marker of organic matter in river samples

This article describes a conductometric bi-layer based bienzyme biosensor for the detection of proteins as a marker of organic matter in rivers. Proteins were chosen to be used as indicators of urban pollution. The working mechanism of the bienzyme biosensor is based on the enzymatic hydrolysis of proteins into several fractions (peptides and amino acids), which results in a local conductivity change depending of the concentration of proteins. In this work, we began with the optimization of biosensor response using bovine serum albumin (BSA) as standard protein. For this objective seven enzymatic biosensors were prepared: four enzymatic sensors with only one layer of enzyme (proteinase K, trypsin, pronase or protease X) and three other enzymatic sensors with two layers (first layer: membrane containing proteinase K; second layer: one of the three other enzymes: trypsin, pronase or protease X). The biosensors were obtained through the deposition of enzymatic layers and the cross-linking process between enzymes and BSA in saturated glutaraldehyde vapour. The response of the various biosensors, described previously, were compared with the values of total organic carbon (TOC), and those of organic nitrogen (Norg), as determined by the laboratory accredits (CEMAGREF of Lyon) using the traditional method of analysis (NF EN 1484, infrared spectroscopy) and (NF EN 25663, mineralization/colorimetry assay) respectively for each water sample obtained from di erent sites in Lyon (France). The linear correlations obtained with the response of the seven biosensors showed the most important indices of correlations for the biosensor with two enzymatic layers: proteinase K + pronase (pkp). The optimum conditions for the preparation of the pkp biosensor increased the sensitivity and gave a limit of quantification of 0.583 g/L for TOC and 0.218 g/L for Norg in water samples. This sensor shows good reproducibility (2.28%), a capacity to be used at temperatures range 10– 30°C (depending on the season) and moreover a long lifetime (5 weeks).     

Biosensors: new approaches in drug discovery

The development of biosensors for analytical purposes has attracted a great deal of attention in recent years. A biosensor is defined as an analytical device consisting of a biological component (e.g., enzyme, antibody, entire cell, DNA) and a physical transducer (e.g., electrode, optical device). Biosensors are mostly designed for routine analysis, such as clinical diagnosis, quality control of food, in-process control of fermentations, and in environmental analysis. Many of these sensors are also suitable for screening purposes in order to find new drugs. Such systems should yield information either about compounds with known bioactivity or about the bioactivity of samples with known or unknown chemical composition. Biosensors intended for the latter purpose are essentially based on whole cells carrying receptors and ion channels at their surfaces. Miniaturization of structures, primarily based on silicon, allows integration of many sensors into arrays, which may be suitable for the screening of natural and chemical products as well as combinatorial libraries. Until now, no commercially available sensors of this kind exist but they are expected in the near future. Different biosensors, based on enzymes, antibodies, cells, artificial membranes and entire animal tissues, which can be used in drug discovery and may lead to efficient screening systems in the future, are described in this review.     

Applications of biosensors for bacteria and virus detection in food and water–A systematic review

Biosensors for sensitive and specific detection of foodborne and waterborne pathogens are particularly valued for their portability, usability, relatively low cost, and real-time or near real-time response. Their application is widespread in several domains, including environmental monitoring. The main limitation of currently developed biosensors is a lack of sensitivity and specificity in complex matrices. Due to increased interest in biosensor development, we conducted a systematic review, com...     

复配逆胶束体系中漆酶催化性能研究

在由鼠李糖脂和Tween-80构建而成的复配逆胶束媒介研究了漆酶的催化性能.采用紫外法考察了Tween-80摩尔含量、逆胶束含水量、水相酸碱度、水相盐度以及助表面活性剂种类等反应条件对漆酶催化性能的影响.研究结果表明,反应的最佳条件:Tween-80的摩尔含量为30%,体系含水量为20,缓冲溶液pH为4.5,KCI浓度为70mmol/L,助表面活性剂正己醇.并且在最佳条件下,漆酶在复配逆胶束体系中的酶活比在单相鼠李糖脂逆胶束中高1.44倍,比在水溶液中高4.35倍,研究结果表明复配表面活性剂在构建胶束酶学上具有很大的潜力.     

Enzymatic oxidation of aqueous pentachlorophenol

The influence of the nonionic surfactant Tween 80 on pentachlorophenol （PCP） oxidation catalyzed by horseradish peroxidase was studied. The surfactant was tested at concentrations below and above its critical micelle concentration （CMC）. Enhancement of PCP removal was observed at sub-CMCs. The presence of Tween 80 in the reaction mixture reduced enzyme inactivation which occurred through a combination of free radical attack and sorption by precipitated products. A simple first-order model was able to simulate time profiles for enzyme inactivation in the presence or absence of Tween 80. At supra-CMCs, the surfactant caused noticeable reductions in PCP removal, presumably through micelle partitioning of PCP which precluded the hydrophobic PCP molecule from interacting with the enzyme.     

四种酰胺类除草剂对土壤酶活性的影响

在实验室控制条件下,研究了酰胺类除草剂丁草胺、乙草胺、丙草胺和异丙甲草胺在田间推荐用量下,对土壤过氧化氢酶活性和脱氢酶活性的影响.结果表明,培养初期,4种除草剂对2种酶活性有不同程度的抑制作用,土壤过氧化氢酶活性于第7d就很快恢复,而脱氢酶活性在13d后得以逐渐恢复.添加稻秆可使土壤酶活性提高,土壤脱氢酶比过氧化氢酶活性变化大.4种除草剂对土壤生态环境影响不大,稻秆还田有利于提高土壤中生化过程的活性,缓冲外来污染物对土壤生态的影响.     

用细胞电泳率指标检测膜毒性物质的探讨

细胞电泳地是近期发展起来的一种检测细胞膜表面微小改变的生物学方法，当膜表面电荷密度分布受影响时，细胞在等强度电场作用下的移动速率会发生改变，根据电泳速率的改变，可以判断外源性物质对细胞膜是否具损伤作用。     

Microvillar enzyme assays in amniotic fluid and fetal tissues at different stages of development

A systematic study of microvillar enzyme activities in the amniotic fluid in correlation with their values in different fetal tissues during development has been undertaken. Microvillar enzymes appeared in the amniotic fluid at the time of disappearance of the anal membrane, 12–13 weeks, and declined from the 18th week until the 24th week. The study of fetal tissues and fluids has shown that gamma-glutamyltranspeptidase is mainly of liver origin. The significant decrease of the activities of these amniotic fluid enzymes has been the basis of prenatal diagnosis of cystic fibrosis. These assays may be useful for the diagnosis of certain digestive tract abnormalities at later stages of pregnancy.     

Soil enzyme activities and their indication for fertility of urban forest soil

To reveal the biological characteristics of urban forest soil and the effects of soil enzyme on soil fertility as well as the correlation between physicochemical properties and enzyme activities, 44 urban forest soil profiles in Nanjing were investigated. Basic soil physicochemical properties and enzyme activities were analyzed in the laboratory. Hydrogen peroxidase, dehydrogenase, alkaline phosphatase, and cellulase were determined by potassium permanganate titration, TTC (C₁₉H₁₅N₄·Cl) colorimetry, phenyl phosphate dinatrium colorimetry, and anthrone colorimetry, respectively. The result showed that soil pH, organic carbon (C), and total nitrogen (N) had great effects on hydrogen peroxidase, dehydrogenase, and alkaline phosphatase activities in 0–20 cm thick soil. However, pH only had great effect on hydrogen peroxidase, dehydrogenase, and alkaline phosphatase activities in 20–40 cm thick soil. Hydrogen peroxidase, dehydrogenase, and alkaline phosphatase were important biological indicators for the fertility of urban forest soil. Both in 0–20 cmand 20–40 cmsoil, soil enzyme system (hydrogen peroxidase, dehydrogenase, alkaline phosphatase, and cellulase) had a close relationship with a combination of physicochemical indicators (pH, organic C, total N, available K, available P, cation exchange capacity (CEC), and microbial biomass carbon (C_mic)). The more soil enzyme activities there were, the higher the fertility of urban forest soil. __________ Translated from Urban Environment & Urban Ecology, 2007, 20(4): 4–6, 9 [译自: 城市环境与城市生态]     

双氰胺对污泥堆肥过程中酶活性和细菌群落演变的影响

鉴于氮肥增效剂在农田中的保氮经验,研究其对堆肥中氮素的保持具有重要意义.因此,本研究以氮肥增效剂-双氰胺（Dicyandiamide, DCD）为添加剂,脱水污泥为堆肥原料,进行了20 d的堆肥试验,探讨DCD对堆肥过程中氮素转化和酶活性变化及其细菌群落演变的影响.结果表明,DCD的添加降低了堆肥高温期的温度,延长了高温期的持续天数;DCD组显著降低了高温期NH₄-N的含量和整个时期NO₃-N的含量.堆肥结束时,总氮含量与对照相比提高了14.8%.此外,DCD的添加抑制了高温期纤维素分解酶和蛋白酶的活性,抑制了整个过程的硝酸还原酶的活性,而且高温期时,DCD组中厚壁菌门和绿弯菌门的丰度显著低于对照.冗余分析表明,NH₄-N、pH和温度主要影响高温期时酶的活性,NO₃-N和总氮主要影响稳定期时酶的活性.综上所述,DCD通过降低高温期的微生物丰度和相关酶活性,减缓了高温期的氨化作用和后期的硝化作用,减少了氮素损失,是一种可行的污泥堆肥添加剂.     

Direct electrochemistry of glucose oxidase and biosensing for glucose based on DNA/chitosan film

Glucose oxidase (GOD) is widely used in the glucose biosensor industry. The amperometric biosensors based on directly electron transfer (DET) between an electrode and immobilized GOD are especially promising. In this article, GOD was immobilized with a DNA/chitosan bio-material film on GC electrode, and the DET of GOD on DNA/chitosan was studied. The cyclic voltammetric results indicated that the GOD immobilized in the DNA/chitosan film underwent DET reaction, and the cyclic voltammogram displayed a pair of well-defined redox peaks with a formal potential of ?0.45 V (vs. Ag/AgCl) at pH 5.5. The response showed a surface-controlled electrode process with an electron transfer rate constant of 0.91 sec^?1 determined in the scan rate range from 10 to 100 mV/sec. The GOD immobilized in DNA/chitosan membrane retained its biocatalytic activity and stability. The immobilized GOD could electrocatalyze the reduction of dissolved oxygen and resulted in a great increase of the reduction peak current. Upon the addition of glucose, the reduction peak current decreased, which could be used for glucose detection with a sensitivity of 0.48 μA/(mmol/L), a linear range from 0.04 to 2.28 mmol/L and a detection limit of 0.04 mmol/L at a signal-to-noise ratio of 3. The sensor could exclude the interference of commonly coexisted uricacid and ascorbic acid.     

A novel thermal biosensor based on enzyme reaction for pesticides measurement

A novel thermal biosensor based on enzyme reaction for pesticides detection has been developed. This biosensor is a flow injection analysis system and consists of two channels with enzyme reaction column and identical reference column, which is set for eliminating the unspecific heat, The enzyme reaction takes place in the enzyme reaction column at a constant temperature(40℃) realized by a thermoelectric thermostat. Thermosensor based on the thermoelectric module containing 127 serial BiTe-thermocouples is used to monitor the temperature difference between two effluents from enzyme reaction column and reference column. The ability of this biosensor to detect pesticides is demonstrated by the decreased degree of the hydrolytic heat in two types of thermosensor mode. The hydrolytic reaction is inhibited by 36% at 1mg/L DDVP and 50% at 10mg/L DDVP when cell-typed thermosensor is used. The percent inhibition is 30% at 1mg/L DDVP and 42% at 10mg/L DDVP in tube-typed thermosensor mode. The detection for real sample shows that this biosensor can be used for detection of organophosphate pesticides residue.     

生物传感器在渔业监测中的研究进展

论述了生物传感器在监测渔业环境中的BOD、N、P、病原体、农药和重金属等方面的国内外最新进展 ,分析了生物传感器在海洋渔业应用中存在的主要问题及原因 ,同时阐述了利用基因工程微生物技术、分子烙印技术、多成分实时分析技术等先进技术提高生物传感器的性能并扩大其在海洋渔业领域中的应用范围 ,在海洋渔业环境中为实现生物传感器与计算机在线自动监测与控制 ,开拓新的广阔前景。     

Relationship between metabolic enzyme activities and bioaccumulation kinetics of PAHs in zebrafish (Danio rerio)

Many studies have investigated bioaccumulation and metabolism of polycyclic aromatic hydrocarbons (PAHs) in aquatic organisms. However, lack of studies investigated both processes simultaneously, and the interaction between these two processes is less understood so far. This study investigated the bioaccumulation kinetics of PAHs and metabolic enzyme activities, including total cytochrome P450 (CYPs) and total superoxide dismutase (T-SOD), in zebrafish. Mature zebrafish was exposed to the mixture of phenanthrene and anthracene under constant concentration maintained by passive dosing systems for 16 days. The results showed that PAH concentrations in zebrafish experienced a peak value after exposure for 1.5 days, and then decreased gradually. The bioaccumulation equilibrium was achieved after exposure for 12 days. Both of the uptake rate constants (k_u) and the elimination rate constants (k_e) decreased after the peak value. The variation of PAH concentrations and metabolic enzyme activities in zebrafish had an interactive relationship. CYPs and T-SOD activities increased initially with the increase of PAH concentrations, but decreased to the lowest state when PAH concentrations reached the peak value. When the bioaccumulation equilibrium of PAHs was achieved, CYPs and T-SOD activities also reached the steady state. In general, CYPs and T-SOD activities were activated after exposure to PAHs. The decrease of PAH concentrations in zebrafish after the peak value may be attributed to the great drop of k_u and the variation of CYPs activities. This study suggests that an interactive relationship exists between bioaccumulation kinetics of PAHs and metabolic enzyme activities in aquatic organisms.     

Trends and Perspectives of Biosensors for Food and Environmental Virology

Food and environmental virology has become a very important and interesting area of research because of food safety and public health concerns. During the last few decades, increasing foodborne diseases and environmental generated illnesses are considered to be highly challenging issues. Biosensor technology holds great promise for the healthcare market, and the security sector. Similar to clinical diagnostic tools, biosensors are being developed for the rapid, reliable, yet inexpensive identification and enumeration of pathogenic viruses which are adulterating environment, food and feed commodities. In this modern era, bio-and nano-technologies play a pivotal role in virological diagnostics of food industry, environmental and veterinary samples. This review covers the recent advances and future prospects of nanotechnology-based bioanalytical microsystems for food and environmental virology.     

膜生物反应器（MBR）技术研究及其在国内应用现状

摘要：膜生物反应器（MBR）是一种膜分离单元与生物处理单元相结合的新型污水处理技术,在污水处理与曰用、垃圾渗滤液处理方面有良好的应用前景。本文阐述了MBR技术特点、分类、在生活污水处理、工业废水处理、垃圾渗滤液处理等方面的应用、存在的问题、以及研究发展方向。预测MBR将有巨大的发展潜力。     

Responses of antioxidant enzyme and photosynthesis in rape seedling to the combined stresses from simulated acid rain and elevated ultraviolet-B radiation

Effects of the simulated acid rain（AR） and ultraviolet-B（UV-B, 280-320 nm） radiation with a single or two ways simultaneously （AR ＋ UV-B） on the antioxidant enzyme and photosynthesis of the rape seedlings were investigated by the hydroponic culture. The results of static experiment indicated that the tolerance of rape seedling to single stress（AR or UV-B） is stronger than that to dual stresses（AR ＋ UV-B）. Furthermore, the dual stresses had additive effect on catalase activity, and a synergistic effect on MDA content, net photosynthesis rate, water use efficiency as well as intercellular CO2 concentration. Meanwhile, it has an independent effect on chlorophyll content, stomatal conductance, and transpiration rate as well as membrane permeability. During 64 h restoration course, the dynamic change in the curves of physiological and biochemical indices were not identical, and none of them show a simple linear variation. According to the static and dynamic experiments, it was found that a responsive sequence of catalase activity, membrane permeability, MDA content and photosynthetic characteristics to the above-mentioned stresses was as follows： AR ＋ UV-B 〉 UV-B 〉 AR.     

Bacterial diversity and distribution in the southeast edge of the Tengger Desert and their correlation with soil enzyme activities

The nature of microbial communities and their relation to enzyme activities in desert soils is a neglected area of investigation. To address this, the bacterial diversity and distribution and soil physico-chemical factors were investigated in the soil crust, the soil beneath the crust and rhizosphere soil at the southeast edge of the Tengger Desert, using the denaturing gradient gel electrophoresis of 16S rRNA genes amplified by the polymerase chain reaction. Phylogenetic analysis of the sequenced DGGE bands revealed a great diversity of bacteria. The Proteobacteria, consisting of the α, β, and γ subdivisions, were clearly the dominant group at all depths and in rhizosphere soil. Analysis of the enzyme activities indicated that the rhizosphere soil of Caragana korshinskii exhibited the highest protease and polyphenol oxidase activities, and in the soil crust there were increased activities of catalase, urease, dehydrogenase and sucrase. The bacterial community abundance closely correlated with soil enzyme activities in different soils. The presence of Cyanobacteria correlated with significant increases in protease, catalase and sucrase in the soil crust, and increased urease in the rhizosphere soil of Artemisia ordosica. The occurrence of Acidobacteria was associated with significant increases in urease, dehydrogenase, and sucrase in the rhizosphere soil of C. korshinski. The presence of γ-Proteobacteria correlated with a significant increase in polyphenol oxidase in the rhizosphere soil of A. ordosica. The study indicated a close relationship between the soil bacterial community and soil enzymes, suggesting the necessity of further investigations into bacterial function in this desert ecosystem.     

Immobilization of an enzyme from a Fusarium fungus WZ-I for chlorpyrifos degradation

The free enzyme extracted from WZ-I,which was identified as Fusarium LK.ex Fx,could effectively degrade chlorpyrifos,an organophosphate insecticide.The methods of immobilizing this free enzyme and determined its degradation-related characteristics were investigated.The properties of the immobilized enzyme were compared with those of the free enzyme.The optimal immobilization of the enzyme was achieved in a solution of 30 g/L sodium alginate at 4°C for 4-12 hr.The immobilized enzyme showed the maximal activity at pH 8.0,45°C.The maximum initial rate and the substrate concentration of the immobilized enzyme were less than that of the free enzyme.The immobilized enzyme,therefore,had a higher capacity to withstand a broader range of temperatures and pH conditions than the free enzyme.With varying pH and temperatures,the immobilized enzyme was more active than the free enzyme in the degradation reaction.In addition,the immobilized enzyme exhibited only a slight loss in its initial activity,even after three repeated uses.The results showed that the immobilized enzyme was more resistant to different environmental conditions,suggesting that it was viable for future practical use.