酿酒酵母吸附Pb(Ⅱ)的表面特性研究

为深入探讨酿酒酵母与Pb(II)的相互作用机理,本文利用表面显微分析技术（SEM-EDS、TEM-EDS、AFM）研究了酿酒酵母细胞吸附重金属离子Pb(II)前后的细胞表面特性变化。研究结果表明,酿酒酵母细胞与Pb(II)作用后,细胞表面除吸附Pb(II)外,同时产生大量更高浓度的含Pb(II)沉淀,导致Pb(II)从溶液中被去除。酵母与Pb(II)反应前,酵母细胞表面可检测到的主要元素包括C、O、N、P、S、K、Mg;酵母与Pb(II)作用后,细胞表面始终保持C、O、P吸收峰,而N、K、Mg、S吸收峰随反应条件不同而减弱、消失或增强。P作为细胞表面组分可能与Pb(II)结合。酵母与Pb(II)作用过程中,重金属离子促进酵母细胞释放细胞内含物。原子力显微镜（AFM）证实,云母片表面对酵母吸附Pb(II)后细胞的铺展变形作用明显增大。     

Les Reciproqueteurs: post-regulatory corporatism

ABSTRACT

Reflexive governance can be understood as an emergent encapsulated trust-building corporatism where network participants are neither state functionaries nor market entrepreneurs but network reciproqueteurs. This paper argues that such reflexive network governance results in a post-regulatory corporatism (PRC)—a more adaptable, less formalized, and flexible mode of interest intermediation, policy-making, and policy-implementation than previous modes of corporatist intermediation. Functional differentiation processes engender ‘negotiated connected contracts' in rescaled space in between inter-regional assemblages, a mode of structurally coupling new social partners in the emergent transnational knowledge-based economy. This involves the building of new social capital of network trust-building manifested in the norms of reciprocity and reflexive law constituted as a new mode of protocolism: one associated with the social learning and policy designing necessary for ecological systems' autopoeisis, resilience, and sustainability. This paper conceptualizes reflexive network governance as protocolism in constellations of PRC and discusses examples from the area of environmental policy-making. PRC is understood as a new mode of negotiated rule-making: as a recursive protocolism of multi-stakeholder social pacts constituted by frame agreements and negotiated connected network contracts.     

Degradation kinetics of terephthalic acid wastewater with the hybrid strains constructed with Phanerochaete chrysosporium

The degradation kinetic parameters in terephthalic acid (TPA) wastewater for the hybrid strains of Fhh and Fhhh obtained through the protoplast fusion of the fungi Phanerochaete chrysosporium (PC) and Saccharomyces cerevisiae Y99, and the native bacteria YZ1 were measured in this research. The highest level of the specific degradation rate for Fhhh and Fhh during 20h reaction were 0.2238 and 0.2163 h^‐1, which were higher than that of their three parental strains and that of the anaerobic bacteria reported. It suggested that the abilities of growth and degradation for Fhh and Fhhh in TPA wastewater were better than their three parental strains. They could create potentials for the purification of TPA wastewater with higher efficiency.     

改进型重组基因酵母TR-GRIP1检测化合物甲状腺激素干扰活性

应用酵母双杂交技术构建重组TR(甲状腺激素受体)基因酵母,用以检测类/抗甲状腺激素化合物及环境样品的甲状腺激素干扰活性. 提取并纯化含有TR基因的酵母表达质粒pGBT9-TR以及含有TR共激活因子GRIP1基因的酵母表达质粒pGAD424-GRIP1,将pGBT9-TR和pGAD424-GRIP1同时转化至酵母细胞Y187,以营养缺陷型培养基(SD/-Trp/-Leu)筛选阳性菌落,构建TR-GRIP1双杂交酵母. 考察该酵母与天然甲状腺激素——T₃(三碘甲状腺原氨酸)的结合情况,确定最佳暴露时间,建立剂量－效应关系曲线. 结果表明:TR-GRIP1双杂交酵母能够与T₃结合诱导β-半乳糖苷酶活性,选择暴露时间为2 h,T₃诱导酶活性的EC₅₀值为1.1×10-7 mol/L;构建的重组基因酵母TR-GRIP1提供了检测化合物甲状腺激素干扰活性的新方法.      

Identification and characterization of integron mediated antibiotic resistance in pentachlorophenol degrading bacterium isolated from the chemostat

A bacterial consortium was developed by continuous enrichment of microbial population isolated from sediment core of pulp and paper mill e uent in mineral salts medium (MSM) supplemented with pentachlorophenol (PCP) as sole source of carbon and energy in the chemostat. The consortia contained three bacterial strains. They were identified as Escherichia coli, Pseudomonas aeruginosa and Acinetobacter sp. by 16S rRNA gene sequence analysis. Acinetobacter sp. readily degraded PCP through the formation of tetrachloro-phydroquinone (TecH), 2-chloro-1,4-benzenediol and products of ortho ring cleavage detected by gas chromatograph/mass spectrometer (GC-MS). Out of the three acclimated PCP degrading bacterial strains only one strain, Acinetobacter sp. showed the presence of integron gene cassette as a marker of its stability and antibiotic resistance. The strain possessed a 4.17 kb amplicon with 22 ORF’s. The plasmid isolated from the Acinetobacter sp. was subjected to shotgun cloning through restriction digestion by BamHI, HindIII and SalI, ligated to pUC19 vector and transformed into E. coli XLBlue1 , and finally selected on MSM containing PCP as sole source of carbon and energy with ampicillin as antibiotic marker. DNA sequence analysis of recombinant clones indicated homology with integron gene cassette and multiple antibiotic resistance genes.     

化学物质的Ah受体效应重组基因酵母检测法的优化

接有芳香烃受体(AhR)基因和芳香烃受体核转位子蛋白(ARNT)基因的酵母Saccharomyces cerevisiae菌株是测定化学物质的Ah受体效应的方法之一.本实验通过优化各个实验条件,发现用0.2%葡萄糖作前培养液培养菌24 h,用2%半乳糖作化学物质暴露时的培养液,暴露在玻璃管时,暴露时间从原来的18 h缩短到8 h.并用已经优化的方法测定了六氯苯和五氯苯的Ah受体效应,得出六氯苯和五氯苯相对于TCDD的毒性等当量值(TEF)分别为0.018629和0.000294.     

重金属离子的生物吸附容量与离子性质之间的关系

利用金属离子毒性评价和预测领域中的QSAR方法,探讨了重金属离子性质对生物吸附容量的影响．选用啤酒工业废弃的酿酒酵母为生物吸附剂,进行了10种金属离子Ag^＋,Cs^＋,Zn^＋,Pb^2＋,Ni^2＋,Cu^2＋,CO^2＋,Sr^2＋,Cd^2＋,Cr^3＋的生物吸附实验．选用22种参数来表征金属离子的物理化学性质,建立了金属的离子特性与生物吸附容量之间的关系．利用Langmuir方程计算得到酵母吸附金属离子的理论最大吸附量qmax,由大到小排序为Pb^2＋〉Ag^＋〉Cr^3＋〉Cu^2＋〉Zn^2＋〉Cd^2＋〉Co^2＋〉Sr^2＋〉Ni^2＋〉Cs^＋．离子性质与吸附量之间的线性拟合分析结果表明,共价指数Xm^2r与qmax具有良好的线性关系,共价指数越高,离子吸附量越大,金属离子与吸附剂表面官能团共价结合所占比重越大,键结合越牢固．对金属离子进行分类（按价态或离子的软硬性质）可以改善拟合效果．极化力Z^2／r、水解常数｜lgKOH｜、电离势,尸等多种物化性质与不含软离子的离子之间的理论最大吸附量也表现出良好的线性关系．     

4-壬基酚臭氧氧化中雌激素活性变化的影响因素

采用有效的重组基因酵母检测方法,研究了不同影响因素下水中典型内分泌干扰物4-壬基酚(4-NP)在臭氧氧化过程中雌激素活性的变化规律.结果表明,臭氧能迅速氧化4-NP,并有效地去除其雌激素活性;臭氧氧化过程中,水中高臭氧浓度、低pH值、低悬浮物含量、低·OH抑制剂浓度以及适量的腐殖酸浓度均有利于降低4-NP的雌激素活性.     

木聚糖酶基因xynB在不同大肠杆菌表达系统中的表达比较

从黑曲霉(Aspergillus niger)nl-1中克隆获得木聚糖酶基因xynB.该基因全长745 bp,含有67 bp内含子,与公布的黑曲霉xynB基因有较高的同源性.将PCR扩增的木聚糖酶成熟肽基因和含有信号肽的基因分别连接到表达载体肠杆菌Top10 F’、DH10B和两种BL21宿主中获得重组菌株.通过IPTG诱导,xynB基因在重组菌株中获得特异性表达.表达产物以胞内可溶性蛋白和不溶性包涵体形式存在.诱导4 h,重组菌株pTrc-99a-xynB[BL21 condon plus(DE3)]表达量最高,胞内酶活达到299 IU/L.重组质粒pTrc-99a-xyn B(S)在不同宿主胞内和胞外均能分泌目的蛋白,诱导10 h,胞外酶活pTrc-99a-xynB(S)[BL21 condon plus(DE3)]达到347 IU/L.而pET-20b-xynB(S)在两种BL21宿主中均不表达.经SDS-PAGE分析,以pTrc-99a和pET-20b作为表达载体时,重组蛋白相对分子质量分别约为45×103和30×103.与pET-20b相比,pTrc-99a以及自身信号肽的引导更有利于重组木聚糖酶的表达.     

代谢活化作用对多溴代联苯类污染物类/抗甲状腺激素活性的影响

选用大鼠肝匀浆(S9)和鼠肝癌细胞(H4ⅡE)两种体系代谢活化多溴代联苯醚混标(BDEs)和十溴联苯醚(BDE209),采用重组甲状腺激素受体基因酵母检测BDEs和BDE209母体及其代谢产物的类/抗甲状腺激素效应.结果表明,BDEs和BDE209母体均不表现甲状腺激素效应(p>0.05);但是经S9和H4ⅡE细胞代谢活化后,其代谢产物表现出明显的类甲状腺激素活性和抗甲状腺激素活性(p<0.05),BDEs和BDE209的干扰甲状腺激素效应需要经过代谢活化步骤.比较不同代谢活化体系,重组酵母细胞本身的代谢活化作用并不显著,而H4ⅡE细胞和S9代谢活化体系均能够导致活性中间体.