缓冲液盐度对热提取活性污泥胞外聚合物的影响

采用热提取法提取污泥中微生物细胞外的胞外聚合物(Extracellular Polymeric Substance,EPS),分析了不同提取时间、提取温度、缓冲液盐度对污泥中EPS提取效果的影响,并采用平行因子模型(PARAFAC模型)分析了不同缓冲液盐度下EPS的荧光组分及特性,探究了缓冲液盐度对热提取污泥EPS的影响.结果表明,缓冲液盐度对热提取EPS的组分和荧光物质的强度均产生影响,0.50%的氯化钠缓冲液盐度条件下松散型EPS(loosely bound EPS,LB-EPS)组分含量及荧光物质最佳,0.05%氯化钠缓冲液盐度条件下紧密型EPS(tightly bound EPS,TB-EPS)的提取效果最佳;当提取时间为40 min,提取温度为70℃时,总EPS含量达(93.26±2.07)mg·g~(-1)(以VSS计);三维荧光结果显示,不同缓冲液盐度条件下污泥中EPS均包含类蛋白质(C1:λ_(Ex)/λ_(Em)=290 nm,302/350 nm)、类色氨酸(C2:λ_(Ex)/λ_(Em)=278 nm,340 nm)、类胡敏酸(C3:λ_(Ex)/λ_(Em)=310 nm,410 nm)和类腐殖酸(C4:λ_(Ex)/λ_(Em)=380/420 nm,474/514 nm)4类荧光组分峰,在溶解性EPS(SMP)和LB-EPS中类色氨酸的荧光强度最高,TB-EPS中类蛋白质荧光强度最高.因此,合适的热提取缓冲液盐度,有利于污泥EPS中各组分的高效提取.     

鸡蛋与豆浆降低蛋白质吸收

生豆浆中含有胰蛋白酶抑制物，它能抑制人体蛋白酶的活性，影响蛋白质在人体内的消化和吸收，鸡蛋的蛋清里含有黏性蛋白，可以同豆浆中的胰蛋白酶结     

母体铅暴露对仔鼠海马组织中P-tau表达的影响

为了探讨母体铅染毒对子一代(F1)仔鼠海马组织中Tau蛋白磷酸化(P-tau)表达的影响,对雌性小鼠自妊娠第1d开始经饮水染铅(0.3、1.0、3.0g·L-1,对照组饮蒸馏水)至仔鼠出生后21d断乳为止,随机抽取各组仔鼠,在出生后第21d分别测其血液和海马组织中铅的含量,并通过免疫组化方法和Western Blot法测定各组仔鼠海马组织中Tau蛋白磷酸化的表达.结果发现,孕哺期不同剂量铅暴露后,仔鼠血铅、海马铅水平均明显高于对照组(p<0.05).免疫组化染色结果显示,P-tau免疫组化阳性反应主要定位于胞浆,与对照组相比,各个铅暴露组仔鼠海马组织的CA1区域P-tau免疫组化阳性反应物的面密度显著增加,但平均灰度值明显降低(p<0.05).Western Blot结果显示,中、高剂量铅暴露组仔鼠海马组织中P-tau的表达明显高于对照组(p<0.05),但低剂量铅暴露组仔鼠海马组织中P-tau的表达与对照组相比差异不显著(p>0.05).母体铅暴露使铅在仔鼠体内蓄积,增强了仔鼠海马组织中Tau蛋白磷酸化水平,高磷酸化的Tau蛋白可能通过诱导微管功能紊乱、神经细胞死亡等,损伤仔鼠的学习记忆能力.     

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).     

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.     

Nickel accumulation and its effect on biomass, protein content and antioxidative enzymes in roots and leaves of watercress （Nasturtium officinale R. Br.）

In order to understand its response towards nickel stress, watercress (Nasturtium o cinale R. Br.) was exposed to nickel (1–25 mg/L) for 1, 3, 5 and 7 days. The accumulation and translocation of nickel were determined and the influence of nickel on biomass, protein content and enzymatic antioxidants was examined for both roots and leaves. It was determined that N. o cinale could accumulate appreciable amounts of Ni in both roots and leaves. Nickel accumulated particularly in the roots of plants. Biomass increased at low nickel concentrations but certain measurable change was not found at high concentrations. Under stress conditions the antioxidant enzymes were up-regulated compared to control. An increase in protein content and enzyme activities was observed at moderate exposure conditions followed by a decline at both roots and leaves. The maximum enzyme activities were observed at di erent exposure conditions. Our results showed that N. o cinale had the capacity to overcome nickel-induced stress especially at moderate nickel exposure. Therefore, N. o cinale may be used as a phytoremediator in moderately polluted aquatic ecosystems.     

好氧颗粒污泥胞外聚合物（EPS）的生化性研究

将人工培养的好氧颗粒污泥进行饥饿试验,研究EPS的可生化性.结果表明,好氧颗粒污泥EPS是由40%可生物降解EPS和60%不可生物降解EPS组成的,其中可生物降解EPS可作为颗粒污泥自身的能源,而不可生物降解EPS则不行,但其对颗粒污泥的立体结构有巨大作用.将从新鲜好氧颗粒污泥中提取的EPS投加到好氧颗粒污泥和活性污泥中,提取的EPS均可被两者作为碳源利用,且活性污泥对EPS的利用速率是好氧颗粒污泥的1.5倍;而从饥饿好氧颗粒中提取的EPS作为两者碳源时,均没有明显的利用效果,说明不可生物降解的EPS不能作为两者的能源.     

不同组成活性污泥胞外聚合物吸附Cd2+、Zn2+特征

以蛋白质和糖含量比分别为2.5∶1、7∶1和9∶1的3种活性污泥胞外聚合物(extracellular polymeric substances, EPS)EPS1、EPS2、EPS3作为吸附剂,研究其对水中Cd²⁺、Zn²⁺的吸附效能.结果表明,EPS对Cd²⁺和Zn²⁺的吸附与其组成有关,EPS1、EPS2和EPS3对Cd²⁺和Zn²⁺吸附量分别约为19.5、　27、　17　mg/g和40.5、 47.5、 37　mg/g. 3种胞外聚合物对Cd²⁺、Zn²⁺吸附过程可在1 h内快速平衡.动力学拟合结果表明,EPS1对2种重金属的吸附速率最快,而EPS2对Cd²⁺、Zn²⁺的平衡吸附容量最高.Freundlich和Langmuir方程均可描述3种EPS对Cd²⁺、Zn²⁺的吸附过程,方程参数拟合结果表明2种金属同EPS之间存在多种作用方式.3种EPS的吸附热力学方程拟合系数均表明EPS对Zn²⁺的吸附稳定性、吸附能力和亲和力均比对Cd²⁺的吸附强;当EPS中糖所占比例增加时,其对重金属的吸附能力也提高,表明糖在吸附过程中起着重要作用.     

生物炭对污泥特征及污水脱氮除磷的影响探究

为探究生物炭对活性污泥特征及脱氮除磷的影响,该文在中温条件下,采用序批式反应器,考察了不同类型基质生物炭对活性污泥处理城镇低C/N废水的影响。结果表明,生物炭降低了污泥体积指数,提高了混合液挥发性悬浮固体,利于微生物的增殖。此外,生物炭提高了活性污泥胞外聚合物(EPS)含量,在鸡粪生物炭、玉米秸秆生物炭和污泥生物炭存在的组别内,EPS的含量分别升高至41.3、46.5和39.8 mg/g,显著高于对照组。生物炭主要提高了EPS内蛋白质(PN)的含量,而对多糖(PS)含量影响不显著,进而提高了PN/PS。生物炭强化了活性污泥对氨氮及总磷的去除,而对化学需氧量的影响不显著。生物炭作为载体微生物提供良好附着进而提高微生物活性,此外,生物炭能吸附氨氮与总磷。