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.     

滨海滩涂围垦区不同围垦年限土壤酶活性变化及其与理化性质关系

土壤酶几乎参与所有的土壤生物化学过程,厘清土壤酶活性与土壤理化性质的关系能够揭示围垦后土壤质量变化状况,为土壤生态系统的科学调控提供理论依据.选择江苏中部如东不同围垦年限(7、32、40、63 a)的滩涂围垦区为研究对象,以未围垦光滩作为对照(0 a),利用经典统计学分析和冗余分析研究不同围垦年限土壤酶活性变化过程,及其与土壤理化性质之间的关系.经典统计学分析表明不同围垦年限间土壤酶活性具有显著性差异,淀粉酶、脱氢酶和酸性磷酸酶活性随着围垦年限的增长呈现出先上升后下降的过程,而脲酶和碱性磷酸酶活性则随着围垦年限的增长呈现出先上升-下降-再上升的过程.同时,不同围垦年限的表层土壤酶活性基本上高于深层.冗余分析表明5种理化性质在前两轴累积能够解释土壤酶活性的69.8%,pH、电导率、土壤有机碳、全氮和全磷与土壤酶活性呈极显著相关关系(P0.01).土壤理化性质对土壤酶活性影响重要性大小排序为全氮有机碳pH全磷电导率.     

真菌WZ-Ⅰ对有机磷杀虫剂毒死蜱的酶促降解

从高效降解真菌镰孢霉属WZ-Ⅰ(Fusarium LK.ex Fx)中提取了降解酶,研究了该降解酶的分离条件及对毒死蜱的降解特性,研究表明,其胞内酶对毒死蜱的降解率高达60.8%,细胞碎片对毒死蜱的降解率为48%,但由(NH₄)₂SO₄沉淀提取的胞外酶液对毒死蜱的降解率仅为11.3%,经8次非诱导条件下培养提取粗酶液,酶活力损失较少,判断WZ-Ⅰ菌株的毒死蜱降解酶为胞内酶且属于组成酶.以牛血清白蛋白为标准蛋白测得粗提酶中可溶性蛋白的含量为3.36mg·mL^-1;该酶对毒死蜱的酶促降解最适pH为6.8,在pH 6.0～9.0之间都有较高的活性;最适温度为40℃,在实验温度范围(20～50℃)内该降解酶均具有较好的降解活性,但在55℃时,酶活迅速降低,降低到最高酶活力的41%.测得粗提酶中其米氏常数K_m为1.049 26mmol·L^-1,v_max为0.253 5μmol·(mg·min)^-1.研究结果表明该酶具有较好的热稳定性和pH稳定性,对热和pH均具有较好的耐受力,高效降解真菌WZ-Ⅰ所产生的胞内酶对毒死蜱具有较好的降解效果.     

环境监测中的新工具--酶免疫检测技术

酶免疫检测技术，是根据抗原抗体反应具有高度的特异性，将酶标记物的抗体作为标准试剂来鉴定未知的抗原。ＥＩＡ技术是环境污染物筛选试验的良好工具，其与气相色谱。高效液相色谱结合，能极大地提高环境监测能力。将酶联免疫吸附检测试剂盒在环境监测中的应用结果，与ＧＣ，ＨＰＬＣ等传统分析方法相比，具有良好的相关性。     

Die molekulare Basis der Heterogenität der Phenylketonurie

The heterogeneity of phenylketonuria can be explained on the molecular basis. Because the phenylalanine-hydroxylating system consists of more than one enzyme, each enzyme can be mutated. Different mutations of the phenylalanine hydroxylase molecule resulting in a lack or reduction of enzyme activity can be anticipated. Enzyme activity lower than 1 % compared to normals is correlated to the severe form; enzyme activity higher than 5% is correlated to the mild form of the disease. Phenylketonuria can also be caused by a defective synthesis or defective reduction of the cofactor. Patients suffer severe neurological symptoms, because of an impaired synthesis of dopamine and serotonin.     

邻苯二甲酸二(2-乙基己基)酯酶促降解性的研究

从处理石化厂废水的活性污泥中分离出一株邻苯二甲酸酯降解菌FS1（Pseudomonas fluorescens FS1),测定了P.fluorescens FS1的邻苯二甲酸酯降解酸对邻苯二甲酸二（2－乙基已基）酯（DEHP）的降解特性。P.fluorescens FS1细胞中的颗粒部分、溶液部分对DEHP都具有降解作用，初步认为P.fluorescens FS1降解酶属胸内酶。P.fluorescens FS1降解酶对DEHP的最适酸度为pH6.5-8.0，温度为25－35℃，米氏常数Km为190.48nmol/mL。在好氧条件下，DEHP与P.fluorescens FS1降解酯酶作用，形成邻苯二甲酸单酯和邻苯二甲酸后，可进一步降解成苯甲酸、对羟基苯甲酸，最终转化为CO2与H2O。     

Long-term stabilization of enzyme activity by immobilizing enzyme colony in polymaleimidostyrene-modified micelles

Amperometric biosensors fabricated by immobilizing enzyme colony in polymaleimidostyrene-modified micelles exhibited good sensitivity and great long-term stability. To our knowledge, there has not been a general design for various enzyme biosensors. Enzyme micelle membrane which is different from any of other conventional immobilization methods, is an innovative way and will be a well-developed biosensor technology to provide rapid and reliable measurements. The potential of using enzyme micelle membrane to fabricate biosensors will be of great hope.     

植物酶浓度对检测乐果灵敏度的影响研究

从小麦中提取植物酶，以农药乐果为抑制剂，采用分光光度法研究了用各种酶浓度小麦酯酶检测乐果的分辨率和灵敏度。研究显示，存在一最佳酶浓度，用此浓度的酶液检测乐果，分辨率和灵敏度最高。     

香溪河流域富磷区土壤酶活性与无机磷形态相关性研究

陆源氮磷不断向水体输入已经成为水体富营养化的主要诱因。香溪河流域内分布着鄂西著名的磷矿区,该区域丰富的磷对香溪河水质产生严重影响。选取流域内磷矿区为研究对象,旨在探讨磷矿区土壤酶活性与无机磷形态之间的内在关系,进而探求用土壤酶系统表征土壤磷污染程度的可行性。结果表明:土壤酶活性与无机磷形态之间有着密切关系。其中,过氧化氢酶、蔗糖酶、脲酶活性与无机磷形态呈正相关关系,而多酚氧化酶和碱性磷酸酶活性与无机磷形态呈负相关关系。土壤过氧化氢酶、多酚氧化酶、蔗糖酶、脲酶及碱性磷酸酶活性均与无机磷形态之间呈现出不同程度的相关性,可作为反映磷矿区土壤磷污染状况的指示酶。     

红松人工林土壤酶活性与化学性质对氮添加的响应

土壤酶参与土壤中进行的众多复杂的生物化学过程,对陆地生态系统物质循环具有重要作用.为了探索温带森林土壤酶活性和化学性质对氮沉降的响应,本研究依托于黑龙江凉水国家级自然保护区的红松（Pinus koraiensis）人工林氮添加实验样地,分析了4种土壤酶活性.结果表明,土壤中N-乙酰-氨基葡萄糖苷酶（NAG）和碱性磷酸酶（AKP）活性均随着施氮浓度增加显著升高,而β-葡糖苷酶（BG）以及酸性磷酸酶（ACP）活性在不同的施氮处理之间无显著差异;同一施氮水平下的上层（0~10 cm）土壤的全碳、全氮、全磷、有效氮含量以及4种酶活性显著高于下层（10~20 cm）土壤,pH值则无显著差异.全碳与NAG、BG、AKP、ACP之间均存在极显著正相关,全氮与BG、NAG、AKP、ACP存在显著或极显著正相关,有效氮与NAG、AKP间存在显著和极显著正相关,全磷与ACP、AKP存在显著和极显著正相关.施氮水平和土层对土壤酶活性及土壤化学性质存在不同程度的影响,氮的长期大量输入可直接或间接地改变土壤化学性质并影响土壤酶活性.     

长期施肥对黄土高原梯田土壤养分特征和微生物资源限制的影响

长期不同施肥处理会改变土壤的理化性质和微生物生物量,但长期不同施肥对黄土高原梯田土壤微生物胞外酶活性变化和养分特征的影响尚不清楚.本研究以中国科学院安塞水土保持综合试验站长期养分定位试验样地为基础,分析了不施肥(CK)、有机肥与氮肥混施(MN)、有机肥与磷肥混施(MP)、有机肥、氮肥与磷肥混施(MNP)、单施有机肥(M...     

Enzymatic pattern processing

 A table-top prototype has been constructed that uses the enzyme malate dehydrogenase to recognize input signal patterns. The device is controlled by the enzyme in response to injection of Mg²⁺ used as a signaling substance. Output is monitored spectroscopically. If Mg²⁺ is injected along either of two signal lines (i.e., if the input signal pattern is 10 or 01) the device emits an output of 1. Injection along neither or both lines results in an output of 0. The enzyme in effect is used as a transform that converts the linearly inseparable exclusive-or problem into a linearly separable problem. Received: 3 April 2000 / Accepted in revised form: 16 June 2000     

A screen-printed, amperometric biosensor for the determination of organophosphorus pesticides in water samples

An amperometric biosensor based on screen-printed electrodes (SPEs) was developed for the determination of organophosphorus pesticides in water samples. The extent of acetylcholinesterase (AChE) deactivation was determined and quantified for pesticide concentrations in water samples. An enzyme immobilization adsorption procedure and polyacrylamide gel matrix polymerization were used for fabrication of the biosensor, with minimal losses in enzyme activity. The optimal conditions for enzyme catalytic reaction on the SPEs surfaces were acetylthiocholine chloride (ATChCl) concentration of 5 mmol/L, pH 7 and reaction time of 4 min. The detection limits for three organophosphorus pesticides (dichlorvos, monocrotophs and parathion) were in the range of 4 to 7 g/L when an AChE amount of 0.1 U was used for immobilization.     

红壤性水稻土铅环境容量研究

以湖南省桃林铅锌矿区为试验区,通过水稻盆栽、田间小区、大田、农田地表径流和淋溶等试验以及土壤酶和微生物试验,研究土壤铅对土壤-作物(水稻)、土壤-微生物和酶、土壤-水体三体系的各种效应,得到各单体系中铅的临界含量并确定了红壤性水稻土中铅的临界含量,通过铅的平衡试验,建立了土壤铅环境容量数学模型,得出红壤性水稻土的铅环境容量。     

Prenatal diagnosis of pyruvate carboxylase deficiency

Prenatal diagnosis of pyruvate carboxylase (PC) deficiency was performed in a family at risk for the acute neonatal form of this disease which manifests secondary citrullinemia. The diagnosis of an affected child was confirmed by enzyme assay and ³H-biotin labelling of proteins in cultured fetal skin fibroblasts. Sufficient amniocytes were cultured in 3–4 weeks for enzyme analysis in two centres. Citrulline concentration in amniotic fluid (AF) was normal in the affected fetus.     

含硫芳香族化合物降解酶的定域及胞内产物的鉴定

为研究含硫芳香族化合物的生物降解途径 ,采用静息细胞技术提取假单胞杆菌 (Pseudomonas sp.)不同部位的酶 .结果表明 ,对硝基苯砜基乙酸异丁酯降解酶主要定域在细胞膜周及膜内 .被试化合物在膜内酶的作用下 ,发生碳硫键断裂 ,降解产物是对硝基苯磺酸 .在膜周酶的作用下 ,发生碳硫键和酯的烷氧键 2种方式的断裂 ,生成对硝基苯磺酸和对硝基苯砜基乙酸 .     

苯甲酸厌氧降解机理及功能基因的研究进展

对芳香化合物降解重要中间体的苯甲酸在厌氧条件下的降解途径及机理。指出了降解过程中每一步骤所涉及的酶及其相关基因。并对其中关键的辅酶进行了的阐述     

高碘地下卤水中细菌产碘氧化酶的影响因素

高碘地下卤水的细菌碘氧化过程可以减轻工艺管线的生物污损危害。分离自高碘地下卤水中的细菌Roseovarius sp.IOB-7,其分泌的碘氧化酶可催化氧化碘离子生成能够抑制敏感菌生长的碘单质,从而减少生物污损。实验结果显示,加入16μmol/L的Cu~(2+)可促进IOB-7的生长,并将碘氧化酶活力提高了1.75倍。碘氧化酶的最适反应温度为50℃,最适pH为5.5。以碘离子为底物的催化氧化反应中,当pH值为9时,Trametes versicolor漆酶已经完全失活,而碘氧化酶则仍然保持了32%的相对酶活力,显示出更好的pH应用范围和稳定性。开展高碘地下卤水环境中细菌碘氧化酶的研究,对利用生物酶法进行工艺管线生物污损的防治有积极的指导意义。     

First trimester monitoring of a pregnancy at risk for glucose phosphate isomerase deficiency

First trimester prenatal diagnosis was offered to the mother of a child affected by severe haemolytic anaemia due to glucose phosphate isomerase (GPI) deficiency. The mutant enzyme was characterized by an increased thermal lability. Both parents had 50 per cent normal red cell GPI activity. We have shown that the homozygous and heterozygous genotypes can be clearly distinguished from each other and controls by combinations of the measurement of enzyme activity and enzyme thermal lability. Examination of trophoblast cells obtained at 9 weeks of gestation led to the diagnosis of a GPI heterozygous fetus. The result was confirmed by analysis on uncultured and cultured amniotic fluid cells sampled at 16 weeks and by red blood cell studies of the healthy newborn. Prenatal diagnosis of GPI deficiency is indicated in families with previous cases resulting in severe haemolysis and mainly with the conservative view of arranging appropriate therapeutic measures for affected fetuses.     

Cloning, expression in Escherichia coli, and enzymatic properties of laccase from Aeromonas hydrophila WL-11

A strain WL-11 with high laccase activity was isolated from activated sludge collected from the effluent treatment plant of a textile and dyeing industry.It was identified as Aeromonas hydrophila by physiological test and 16S rDNA sequence analysis.A gene encoding of laccase from a newly isolated Aeromonas hydrophila WL-11 was cloned and characterized.Nucleotide sequence analysis showed an open reading frame of 1605 bp encoding a polypeptide comprised of 534 amino acids.The primary structure of the enzyme predicted the structural features characteristic of other laccases,including the conserved regions of four histidine-rich copper-binding sites.The predicted amino acid sequence showed a high homology(more than 60%) with bacterial laccases in the genome and protein databases and the highest degree of similarity(61% identity) was observed with the multicopper oxidase of Klebsiella sp.601.When expressed in Escherichia coli,the recombinant enzyme was overproduced in the cytoplasm as soluble and active form.The purified enzyme had an optimum pH of 2.6 and 8.0 for ABTS(2,2'-azino-bis(3-ethylbenzthiazolinesulfonic acid) and DMP(2,6-dimethoxyphenol),respectively.The kinetic study on ABTS revealed a higher affinity of this enzyme to this substrate than DMP.