汞胁迫对齿肋赤藓生物结皮细胞超微结构的影响

细胞器结构是生物细胞功能的基础,而重金属会对细胞器结构产生毒害效应,进而破坏生物的组织器官完整性及生理代谢功能,汞对生物结皮细胞器超微结构的影响尚未见报道。为探究汞胁迫对齿肋赤藓生物结皮叶肉细胞超微结构的影响,将叶肉细胞暴露于不同浓度的汞溶液中(0、10、20、30、40、50和60 mmol·L~(-1)),连续培养7 d,采用透射电镜技术观察叶肉细胞的变化。与对照组相比,汞浓度在20～60 mmol·L~(-1)时,齿肋赤藓生物结皮的细胞器超微结构随汞浓度增加逐渐发生变化,细胞壁逐渐模糊,出现质壁分离现象,液泡出现一定的空泡化,叶绿体膜破损,类囊体、基粒及基质片层消失,细胞核解体及核仁消失。综上所述,齿肋赤藓生物结皮在低汞浓度时可维持有机体免受毒害,表明齿肋赤藓生物结皮有一定的重金属胁迫耐性,但超过阈值(20 mmol·L~(-1))时,出现汞胁迫伤害,且随汞浓度增加,胁迫毒害效应加剧。本研究结果可为干旱区非维管植物(齿肋赤藓)适应重金属胁迫环境理论及抗性物种培养提供参考。     

强氯精对尼罗罗非鱼精巢N-乙酰-β-D-氨基葡萄糖苷酶的失活动力学(英文)

强氯精是水产养殖过程中普遍使用的消毒剂,为了解强氯精能够杀菌却不会对养殖水产动物尤其是其繁殖功能产生危害的有效浓度,以分离自尼罗罗非鱼(Oreochromis niloticus)精巢的N-乙酰-β-D-氨基葡萄糖苷酶(EC3.2.1.52,NAGase)为材料,利用底物反应的动力学方法探讨强氯精对该酶的失活动力学.结果显示该酶在强氯精溶液中是一个可逆的反竞争型的失活过程,其对酶抑制的IC50为(0.25±0.02)mg/m L;而且强氯精能降低该酶的热稳定性和p H稳定性;通过建立失活模型,计算得知正向和逆向微观失活速度常数k′+0和k′-0值分别为3.84×10~(-3)/s和8.37×10~(-4)/s.本研究结果可为评价强氯精对罗非鱼繁殖功能的影响提供理论依据.     

渗透压调控启动子表达木糖醇脱氢酶基因XYL2对重组酿酒酵母木糖代谢的影响

渗透压调控启动子在高底物浓度或盐类物质诱导下可实现目标基因的高表达和可控表达,因此可用于构建新的代谢工程菌株.利用来源于可耐高渗的产甘油假丝酵母(Candida glycerinogenes)的渗透压调控启动子PCgSTL3、PCgZWF和PCgGPD,在酿酒酵母(Saccharomyces cerevisiae)中控制木糖醇脱氢酶基因XYL2的表达水平,并研究其在NaCl诱导下对木糖醇脱氢酶活性及重组酿酒酵母木糖代谢流的影响.结果显示,以组成型强启动子PTPI为对照,添加0.4 mol/L NaCl作为渗透压诱导剂后,渗透压调控启动子控制下的木糖醇脱氢酶活性均得到增强,其中在PCgGPD控制下其活性变化最为明显,酶活提高了2.8倍.同时,以PCgGPD控制XYL2基因表达的重组菌木糖消耗量、乙醇产量和产率均为最高,与无诱导剂相比分别提高了62.3%、30.7%和9.7%,副产物木糖醇产率最低,下降了53.3%.本研究表明,渗透压调控启动子PCgGPD可显著提高木糖醇脱氢酶活性,通过添加NaCl作为渗透压诱导剂可实现对XYL2基因的可控表达,显著降低木糖醇积累,提高乙醇产率.     

有机溶剂和变性剂对枯草芽胞杆菌溶栓酶活性的影响

以枯草芽胞杆菌LD-8547菌株发酵液为材料,提取分离溶栓酶,研究了几种有机溶剂及变性剂对枯草杆菌溶栓酶活性的影响.结果表明,甲醇对溶栓酶表现为抑制作用,在10%-15%浓度范围时,其抑制作用最强;乙醇在5%～15%浓度范围内对酶活有激活作用,但随浓度的增大则转变为较强的抑制作用;正丁醇和异丙醇对酶活力则表现出强烈的抑制作用;丙三醇对酶活力有轻微的抑制作用,随着浓度增加抑制作用逐渐增强;异戊醇在5%～15%浓度范围内有一定的激活作用,但当浓度大于15%时则表现为抑制作用.醛类对酶活也有较强的抑制作用;丙酮、DMSO、DMF、SDS和异氰硫酸胍对酶活均有不同程度的抑制作用;脲浓度＜0.01 mmol/L时略有激活作用,随浓度增大转变为抑制作用.实验同时对EDTA在不同浓度和不同时间内对酶活力的影响进行了研究,结果表明,浓度＜2mmol/L的EDTA基本不影响酶活力,但随着浓度的增大,其抑制作用明显增大.本实验还以牛血为材料,对酶液进行了体外抗血凝和溶血栓实验,结果表明,该酶具有抗血凝和溶血栓效果.     

苏云金芽胞杆菌(Bacillus thuringiensis)几丁质酶的分离纯化及酶学性质

以苏云金芽胞杆菌为材料 ,通过硫酸铵沉淀分级分离、DEAE - 32离子交换柱层析初步纯化 ,获得比活力为15 0 .5Umg-1的酶制剂 .研究酶催化胶体几丁质水解的反应时间和酶量对酶活力的关系 ,探讨酶催化胶体几丁质的最适条件 .温度和 pH对酶活力的影响和酶的热稳定性及酸碱稳定性的结果表明 :酶催化胶体几丁质水解的的最适pH为 5 .6、最适温度为 5 8℃ .该酶在pH 4 .5～ 6 .5区域稳定 ,而在pH >8能很快失活 ;在 5 5℃以下处理 30min ,酶活力保持不变 ;高于 5 5℃ ,酶快速失活 .图 5参 11     

土壤中金属阳离子对Cl-离子吸附的诱导效应

使用三种可变电荷土壤和一种恒电荷土壤 ,研究电解质溶液中金属阳离子对Cl-离子吸附的影响 .结果表明 ,在可变电荷土壤中 ,金属阳离子对Cl- 离子的吸附有明显的诱导效应 ,影响大小次序为 :AlCl3>CuCl2 >KCl>NaCl.而在恒电荷土壤中影响不大     

金属元素对厌氧氨氧化生化过程和工艺运行的影响

厌氧氨氧化为高效低耗生物脱氮技术,但金属元素对该工艺的作用存在两面性,一方面其促进作用可提高工艺处理效果,另一方面抑制作用使其工程应用受到一定限制.为此本文从生物化学和污水处理工艺的角度,系统地总结了国内外金属对厌氧氨氧化影响的研究现状,重点分析与讨论了不同金属元素对厌氧氨氧化细胞结构和相关酶蛋白活性等的影响和作用机理;综述了金属元素对厌氧氨氧化工艺运行的影响,讨论与分析了不同厌氧氨氧化工艺的半抑制浓度及活性修复方法.     

多点突变提高DhaA对芥子气的活性和热稳定性

为提高烷基卤脱卤酶Dha A对芥子气的活性和热稳定性,采用Autodock软件计算Dha A突变前后与芥子气的结合情况,利用重叠延伸PCR和DNA无缝拼接结合的方法,改变Dha A活性空腔进出口通道的大小,构建包含5个位点的Dha A突变体(Ile135Phe+Cys176Tyr+Val245Phe+Leu246Ile+Tyr273Phe);将Dha A及其突变体构建在p ET-28a载体上后,在Escherichia coli BL21(DE3)中进行表达,比较纯化后的野生型与突变体在酶学性质方面的变化情况.Autodock分子对接结果显示,突变后的Dha A与芥子气的结合能、结合效率和抑制常数均小于野生酶,突变体对芥子气的比活性提高了1.4倍,对10 mg/m L芥子气的降解率提高了近20%.热稳定性实验发现,Dha A突变体在50℃水浴1 h后残余酶活为76%,比野生型Dha A提高了19%.Dha A突变体的Tm值为56℃,比野生型Dha A提高了6℃.综上表明改变Dha A活性空腔内的进出口通道可以提高Dha A的热稳定性和对芥子气的催化活性.     

果糖对莆田黑猪精液中NAGase的抑制动力学

为了解精液中不同类型糖类物质对精子细胞中N-乙酰-β-D-氨基葡萄糖苷酶(NAGase)的影响,以莆田黑猪精液NAGase为材料,研究精液和精液稀释液主要成分之一果糖对酶的抑制作用.结果显示,果糖对NAGase具有浓度效应(IC50=575 mmol L-1).果糖对NAGase的抑制类型是可逆的非竞争性抑制,抑制常数是1.27×10-2 mmol L-1.应用底物反应的动力学方法研究果糖对该酶的抑制动力学,建立了抑制动力学模型,证明了果糖对该酶的影响是快速结合再缓慢失活的过程.本研究表明,在高浓度的抑制剂溶液中NAGase将完全失活,底物对其无保护作用.     

假单胞茵胞内酶粗提液对藻毒素MCLR的降解

对比研究了假单胞菌M-6及其细胞内外提取液对微囊藻毒素-LR(MCLR)的降解效率.结果表明,细胞外提取液对藻毒素没有降解作用,胞内酶粗提液能在24h内降解MCLR,日均MCLR降解率是纯菌株M-6的4.7倍.进而研究了酶蛋白浓度、底物MCLR浓度、pH值及环境温度对胞内酶粗提液降解藻毒素效率的影响.当MCLR浓度为15.0mg·l~(-1)时,MCLR适宜的酶促降解反应条件为:酶蛋白浓度280mg·l~(-1),温度为30℃,反应pH值为7.0.MCLR液相色谱结构变化图表明,至少有3种酶参与了胞内酶粗提液降解MCLR的分子过程.     

Simulation of germination of pioneer species along an experimental drought gradient

The germination of ten plant species from the Iberian Peninsula was assessed along a water deficit gradient between -0. 1652 (moist) and -0.4988 MPa (dry) of osmotic potential, created by addition of increasing concentrations of polyethylene glycol (PEG 6000) to distilled water in which plants were grown hydroponically. The level and rate of germination of Daucus carota and Thapsia villosa significantly decreased with decreasing psi. Seeds of Dactylis glomerata and Dittrichia viscosa had positive germination responses to low osmotic potentials; germination of Epilobium hirsutum was not affected by osmotic potential. Germination of Medicago arabica, Cynosurus cristatus, Cistus ladanifer and Cistus albidus, was no favored by the addition of polyethylene glycol (PEG). Germination of Foeniculum vulgare and Thapsia villosa was inhibited by PEG.     

双负载双金属催化剂催化有机卤化物水相脱卤研究

通过把起相转移作用的聚乙二醇链固载在硅胶上（SiO2-PEG600）,再将聚乙稀吡咯烷酮（PVP）络合双金属Pd-Cu(PVP-PdCl2-CuCl2)后负载其中,制成一种新型双负载双金属水相脱卤催化剂PVP－PdCl2-CuCl2/SiO2-PEG600。以甲酸钠为氢转移试制,在水相中催化有机卤化物脱卤,研究结果表明：反应温度为800℃,Pd∶Cu=2∶1（摩尔比）,反应介质pH≈11.7左右时,该催化剂对芳香氯化物及α－氯代酮、酯具有良好的催化脱氯和重复使用性能。     

Preparation and application of a phosphorous free and non-nitrogen scale inhibitor in industrial cooling water systems

A novel environmentally friendly type of calcium carbonate, zinc (II) and iron (III) scale inhibitor Acrylic acid- allylpolyethoxy carboxylate copolymer (AA-APEL) was synthesized. The anti-scale property of the AA-APEL toward CaCO₃, zinc (II) and iron (III) in the artificial cooling water was studied through static scale inhibition tests. The observation shows that both calcium carbonate, zinc (II) and iron (III) inhibition increase with increasing the dosage of AA-APEL. The effect on formation of CaCO₃ was investigated with combination of scanning electronic microscopy (SEM), transmission electron microscopy (TEM), X-ray powder diffraction (XRD) analysis and fourier transform infrared spectrometer, respectively. The results showed that the AA-APEL copolymer not only influenced calcium carbonate crystal morphology and crystal size but also the crystallinity. The crystallization of CaCO₃ in the absence of inhibitor was rhombohedral calcite crystal, whereas a mixture of calcite with vaterite crystals was found in the presence of the AA-APEL copolymer. Inhibition mechanism is proposed that the interactions between calcium or iron ions and polyethylene glycol (PEG) are the fundamental impetus to restrain the formation of the scale in cooling water systems.     

Stabilization of the As-contaminated soil from the metal mining areas in Korea

The stabilization efficiencies of arsenic (As) in contaminated soil were evaluated using various additives such as limestone, steel mill slag, granular ferric hydroxide (GFH), and mine sludge collected from an acid mine drainage treatment system. The soil samples were collected from the Chungyang area, where abandoned Au-Ag mines are located. Toxicity characteristic leaching procedure, synthetic precipitation leaching procedure, sequential extraction analysis, aqua regia digestion, cation exchange capacity, loss on ignition, and particle size distribution were conducted to assess the physical and chemical characteristics of highly arsenic-contaminated soils. The total concentrations of arsenic in the Chungyang area soil ranged up to 145 mg/kg. After the stabilization tests, the removal percentages of dissolved As(III) and As(V) were found to differ from the additives employed. Approximately 80 and 40% of the As(V) and As(III), respectively, were removed with the use of steel mill slag. The addition of limestone had a lesser effect on the removal of arsenic from solution. However, more than 99% of arsenic was removed from solution within 24 h when using GFH and mine sludge, with similar results observed when the contaminated soils were stabilized using GFH and mine sludge. These results suggested that GFH and mine sludge may play a significant role on the arsenic stabilization. Moreover, this result showed that mine sludge can be used as a suitable additive for the stabilization of arsenic.     

Stabilization of the As-contaminated soil from the metal mining areas in Korea

The stabilization efficiencies of arsenic (As) in contaminated soil were evaluated using various additives such as limestone, steel mill slag, granular ferric hydroxide (GFH), and mine sludge collected from an acid mine drainage treatment system. The soil samples were collected from the Chungyang area, where abandoned Au–Ag mines are located. Toxicity characteristic leaching procedure, synthetic precipitation leaching procedure, sequential extraction analysis, aqua regia digestion, cation exchange capacity, loss on ignition, and particle size distribution were conducted to assess the physical and chemical characteristics of highly arsenic-contaminated soils. The total concentrations of arsenic in the Chungyang area soil ranged up to 145 mg/kg. After the stabilization tests, the removal percentages of dissolved As(III) and As(V) were found to differ from the additives employed. Approximately 80 and 40% of the As(V) and As(III), respectively, were removed with the use of steel mill slag. The addition of limestone had a lesser effect on the removal of arsenic from solution. However, more than 99% of arsenic was removed from solution within 24 h when using GFH and mine sludge, with similar results observed when the contaminated soils were stabilized using GFH and mine sludge. These results suggested that GFH and mine sludge may play a significant role on the arsenic stabilization. Moreover, this result showed that mine sludge can be used as a suitable additive for the stabilization of arsenic.     

Leaching characteristics of heavy metals during cement stabilization of fly ash from municipal solid waste incinerators

The leaching characteristics of heavy metals in products of cement stabilization of fly ash from a municipal solid waste incinerator were investigated in this paper. The stabilization of heavy metals such as Cd, Pb, Cu, and Zn in fly ash from such incinerators was examined through the national standard method in China based on the following factors: additive quantity of cement and Na₂S, curing time, and pH of leaching liquor. The results showed that as more additives were used, less heavy metals were leached except for Pb, which is sensitive to pH of the leachate, and the worse effect was observed for Cd. The mass ratio of cement to fly ash = 10% is the most appropriate parameter according to the national standard method. When the hydration of cement was basically finished, stabilization of heavy metals did not vary after curing for 1 d. The mixtures of cement and fly ash had excellent adaptability to environmental pH. The pH of leachate was maintained at 7 when pH of leaching liquor varied from 3 to 11.     

一株产耐热纤维素酶菌株的筛选及酶学性质

采用纤维素刚果红平板法从大田蘑菇种植场建堆的稻草样中分离得到了1株能产具有较好温度耐受性和pH稳定性的纤维素酶的放线菌DY3.综合形态、生理生化特征以及16S rDNA序列分析,将其初步鉴定为嗜热裂孢菌(Thermobifida fusca).对该菌所产纤维素酶的性质研究表明:最适催化温度为65℃,在70℃保温60 min后仍有75%以上的活力;最适催化pH为7.5,在pH 5.5～10.0之间该酶的稳定性较好,pH 10.0的条件下仍有80%的活力;最适作用底物是羧甲基纤维素钠.研究可为木质纤维素的生物预处理提供一定参考价值.     

Highly efficient microwave-assisted one-pot synthesis of 4-aryl-2-aminothiazoles in aqueous medium

The thiazole ring system is one of the most important heterocycles in nature, as it represents an important structural motif of many biological compounds, including vitamin B₁ (Thiamin), carboxylase and penicillin. There is, therefore, an urgent need to design rapid, efficient and environmentally benign protocols for the synthesis of thiazoles. Herein, we have developed a one-pot protocol for the synthesis of 4-aryl-2-aminothiazoles from the reaction of aromatic ketones, NBS (N-Bromosuccinimide) and thioureas under microwave irradiation at 80–85 °C in PEG (polyethylene glycol)-400 and water as a green reaction medium. The products were obtained in 84–89% yields in 28–32 min. The method has several advantages such as use of green solvent, easy work-up, excellent yield and avoiding use of lachrymatric α-haloketones.     

Antioxidant effect of lignite humic acids and its salts on the thermo-oxidative stability/degradation of polyvinyl alcohol blends

We demonstrate the antioxidant potential of humic acids in (green) polymer chemistry. Lignite humic acids and its sodium and ammonium salts were mixed at different concentrations with polyvinyl alcohol and thermogravimetry was used to assess the influence on the thermo-oxidative stability of resulted blends. Both protonized form and ammonium salt of humic acids caused increased stability of investigated polymers in the studied concentration range (0.5–10% of humic acids in polymer). In contrast to protonized form, the ammonium salt also showed moderating effect on polymer degradation kinetics. Sodium salts caused substantially lower stabilizing effect and presence of 10% of humic matter caused even destabilization. In all cases, however, when the degradation has already started, the presence of humic acids and its salts caused more intensive polymer decomposition, which resulted in lower weight of char.     

Environmental stress-induced extracellular isoperoxidase RC3 from rice

Effects of various environmental stresses such as heavy metals, salts and low (high) temperature on the secretion of peroxidase isozyme into the medium were examined in rice (Oryza sativa cv. Nak-Dong) suspension culture. The major extracellular peroxidases secreted into the medium by various stresses were cationic isoperoxidases. A far migrating cationic isoperoxidase RC3 was isolated from the medium after application of CaCl2, the effective stimulator for peroxidase secretion. Isolation of extracellular isoperoxidase RC3 was accomplished by ammonium sulfate fractionation, CM-cellulose cation-exchange chromatography, and Sephacryl S-100 gel filtration. The enzyme was a glycoprotein having molecular weight of approximately 34 KDa as determined by SDS-PAGE and 38 KDa by Sephacryl S-100 gel filtration. The pI value of the enzyme was 8.9. Kinetic studies revealed that the optimum pH of the enzyme was 6.0 for guaiacol and H2O2, and the Km values for guaiacol and H2O2 were 10.5 mM and 3.2 mM, respectively.