两种启动子对聚γ-谷氨酸降解酶基因在地衣芽胞杆菌中的加强表达效果

聚γ-谷氨酸(γ-PGA)是一种应用前景良好的生物高分子材料.比较了蔗糖诱导的枯草芽胞杆菌果聚糖蔗糖酶基因(SacB)启动子和地衣芽胞杆菌α-淀粉酶基因启动子对γ-PGA降解酶基因ywtD在地衣芽胞杆菌中加强表达的影响.分别用SacB基因启动子和α-淀粉酶启动子构建了穿梭表达载体pHY300-SYT和pHY300-PYT,通过电转化地衣芽胞杆菌WX-02获得重组子SYT和PYT.酶活测定结果显示SYT和PYT中γ-PGA降解酶基因ywtD得到加强表达,摇瓶发酵结果显示两个重组菌株的γ-PGA相对分子质量都由1 000 000~1 200 000降低为800 000~900 000,PYT的γ-PGA产量较对照菌株PLK提高了33%,由13.50 g L-1提高到17.97 g L-1,而SYT的γ-PGA产量则降低为10.85 g L-1.因此,α-淀粉酶启动子更适合于在地衣芽胞杆菌WX-02菌株中表达γ-PGA降解酶基因,从而获得高产低分子量γ-PGA的工程菌.     

1株α-蒎烯降解菌的分离鉴定及降解特性研究

从处理废气的生物滤塔内筛选到1株能高效降解α-蒎烯的菌株PT.通过菌落形态、生理生化特征、16SrRNA基因序列相似性分析及Biolog鉴定等方法,确定该菌株属于荧光假单胞菌(Pseudomonas fluorescens).菌株PT最佳生长条件为: NaCl浓度0.00%、pH7.13、温度25.5℃,降解速率达到最大值5.22mg/(L×h).降解过程符合Haldane’s抑制生长动力学模型,最大比降解速率为0.0364h-1.菌株PT能不同程度地降解一些分子结构较为简单或与α-蒎烯结构相似的工业有机污染物.代谢产物分析表明,菌株PT在降解α-蒎烯的过程中产生柠檬油精、紫苏酸等结构较为简单的物质,它们最终被完全矿化为CO2或合成细胞自身组成物质.碳平衡分析表明,底物有机碳含量完全矿化和转化为细胞生物量的比例分别为64.83%和30.37%.     

Photosensitivity sources of dissolved organic matter from wastewater treatment plants and their mediation effect on 17α-ethinylestradiol photodegradation

● EE2 photodegradation behavior in the presence of four WWTPs’ DOM was explored. ● The ³DOM* played a major role in the EE2 photodegradation mediated by WWTPs’ DOM. ● The A²/O process DOM contained more aromatic and oxygen-containing substances. ● Possible photosensitivity sources of DOM in the A²/O process were proposed. Dissolved organic matter (DOM) from each treatment process of wastewater treatment plants (WWTPs) contains abundant photosensitive substances, which could significantly affect the photodegradation of 17α-ethinylestradiol (EE2). Nevertheless, information about EE2 photodegradation behavior mediated by DOM from diverse WWTPs and the photosensitivity sources of such DOM are inadequate. This study explored the photodegradation behavior of EE2 mediated by four typical WWTPs’ DOM solutions and investigated the photosensitivity sources of DOM in the anaerobic-anoxic-oxic (A²/O) process. The parallel factor analysis identified three varying fluorescing components of these DOM, tryptophan-like substances or protein-like substances, microbial humus-like substances, and humic-like components. The photodegradation rate constants of EE2 were positively associated with the humification degree of DOM (P < 0.05). The triplet state substances were responsible for the degradation of EE2. DOM extracted from the A²/O process, especially in the secondary treatment process had the fastest EE2 photodegradation rate compared to that of the other three processes. Four types of components (water-soluble organic matter (WSOM), extracellular polymeric substance, humic acid, and fulvic acid) were separated from the A²/O process DOM. WSOM had the highest promotion effect on EE2 photodegradation. Fulvic acid-like components and humic acid-like organic compounds in WSOM were speculated to be important photosensitivity substances that can generate triplet state substances. This research explored the physicochemical properties and photosensitive sources of DOM in WWTPs, and explained the fate of estrogens photodegradation in natural waters.     

Microbial transformation of synthetic estrogen 17α-ethinylestradiol

Natural estrogens such as estrone, 17β-estradiol, estriol, and the particularly recalcitrant synthetic estrogen 17α-ethinylestradiol used as oral contraceptive, accumulate in the environment and may give rise to health problems. The processes participating in their removal from soil, wastewater, water-sediments, groundwater-aquifer material, and wastewater or sewage treatment plant effluents may involve the action of bacterial and microbial consortia, and in some cases fungi and algae. This review discusses the different efficiencies of bacterial degradation of 17α-ethinylestradiol under aerobic and anaerobic conditions, the role of sulfate-, nitrate-, and iron-reducing conditions in anaerobic degradation, and the role of sorption. The participation of autotrophic ammonia oxidizing bacteria and heterotrophic bacteria in cometabolic degradation of estrogens, the estrogen-degrading action of ligninolytic fungi and their extracellular enzymes (lignin peroxidase, manganese-dependent peroxidase, versatile peroxidase, laccase), and of algae are discussed in detail.     

Analysis of α, β, γ-hexachlorocyclohexanes in water by novel activated carbon fiber-solid phase microextraction coupled with gas chromatography--mass spectrometry

A fast and simple method for determination of α, β,γ-hexachlorocyclohexanes (HCHs) in water using activated carbon fibersolid phase microextraction(ACF-SPME) were studied. Results showed the performance of adsorption and desorption of three HCHs on ACF were excellent. A wide linear range from 10 to 100 μg/L and detection limits of the ng/L level were obtained using ACF-SPME with GC-MS in selected ion monitoring(SIM) acquisition mode. The proposed method was also successfully applied for determination of three HCHs in tap water. Compared to commercial fibers, ACF showed some advantages such as better resistance to solvents, higher thermal stability, longer lifetime and lower cost. The data demonstrated that GC-MS with ACF-SPME is well suitable for the analysis of HCHs in water.     

Alpha-thalassaemia prenatal diagnosis by two PCR-based methods

In Cyprus all couples carrying α⁰-thalassaemia mutations are detected in the course of the thalassaemia carrier screening program and prenatal diagnosis is offered to all of them. Prenatal diagnosis for α-thalassaemia is routinely done by two independent molecular methods. With the first method, the mutations of the parents are directly determined by gap-PCR and then the chorionic villus sample (CVS) is examined for the presence of these mutations. With the other method, a (CA)_n repeat polymorphic site located between the ψα₁- and α₂-globin genes is used for determining the presence or absence of the normal and mutant alleles. In the period from 1995 to 1999, molecular analysis of 46 couples in which haematological data were consistent with deletion of two α-globin genes in both partners indicated that only 13 of them were actually at risk for haemoglobin (Hb) Bart's hydrops fetalis and prenatal diagnosis was provided in 16 pregnancies. The molecular diagnosis was possible in all cases with the use of both gap-PCR and (CA)_n repeat polymorphisms analysis. No misdiagnosed cases for α-thalassaemia have been reported to date. Copyright © 2001 John Wiley & Sons, Ltd.     

Prenatal diagnosis of multiple acyl-CoA dehydrogenase deficiency: association with elevated α-fetoprotein and cystic renal changes

We report the occurrence of multiple acyl-CoA dehydrogenase deficiency (MADD) in two consecutive pregnancies in a young, Caucasian, non-consanguineous couple. In the first pregnancy, the maternal serum α-fetoprotein was elevated. A sonogram showed growth delay, cystic renal disease, and oligohydramnios; the parents decided to terminate the pregnancy. Postmortem examination confirmed the cystic renal disease and showed hepatic steatosis, raising the suspicion of a metabolic disorder. The diagnosis of MADD was made by immunoblot studies on cultured fibroblasts. In the subsequent pregnancy, a sonogram at 15 weeks' gestation showed an early growth delay but normal kidneys. The maternal serum and amniotic fluid concentrations of α-fetoprotein were elevated, and the amniotic fluid acylcarnitine profile was consistent with MADD. In vitro metabolic studies on cultured amniocytes confirmed the diagnosis. A follow-up sonogram showed cystic renal changes. These cases provide additional information regarding the evolution of renal changes in affected fetuses and show a relationship with elevated α-fetoprotein, which may be useful in counseling the couple at risk. MADD should be considered in the differential diagnosis of elevated α-fetoprotein and cystic renal disease. Early growth delay may be an additional feature. Copyright © 2001 John Wiley & Sons, Ltd.     

TiO2-NB/pg-C3N4可见光催化降解17α-乙炔雌二醇的机理

通过煅烧自制的TiO₂纳米带（TiO₂-NB）和多孔氮化碳（pg-C₃N₄）制备出新型的TiO₂纳米带/多孔氮化碳（TiO₂-NB/pg-C₃N₄）光催化剂,优化了其制备条件,并用于可见光条件下催化降解雌激素活性最强的17α-乙炔雌二醇（EE2）.通过SEM、TEM、XRD、UV-vis DRS、PL、XPS和BET等手段对催化剂结构和表面物理化学性质进行表征和分析.结果表明,B型TiO₂纳米带均匀负载至多孔氮化碳薄片上,形成TiO₂-NB/pg-C₃N₄复合光催化剂.最优条件下制备的TiO₂-NB/pg-C₃N₄降解EE2的速率常数（0.02537min^-1）分别是TiO₂-NB （0.00197min^-1）和pg-C₃N₄（0.0077min^-1）的12.88倍和3.29倍.自由基捕获实验结果显示超氧自由基（·O₂^-）是TiO₂-NB/pg-C₃N₄可见光催化降解EE2的主要活性物种.HPLC-MS中间体检测结果显示,可能的降解中间体主要有4个,提出可能的降解途径,而且通过MCF-7雌激素活性测试法可知降解过程产物的雌激素活性基本消除.     

二甲苯、蒽、苯并[α]芘对虾夷扇贝幼贝血清SOD、CAT活性的影响

以虾夷扇贝(Mizuhopecten yessoensis)为受试生物,设置3种不同浓度的二甲苯、蒽、苯并[α]芘处理健康的虾夷扇贝,检测血清中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性变化情况,结果发现:0.01~1.0 mg·L~(-1)的二甲苯、0.002~0.2 mg·L~(-1)的蒽、0.001~0.1 mg·L~(-1)的苯并[α]芘处理6、12 h后,虾夷扇贝血清SOD活性升高,处理12 h后对血清SOD活性诱导率均高于处理6 h的;处理6 h后,血清CAT活性降低,而处理12 h后,血清CAT活性升高;具有显著的剂量-效应关系和时效关系,表明二甲苯、蒽、苯并[α]芘对虾夷扇贝具有氧化胁迫作用,可能导致其氧化损伤。相关性分析发现,3种污染物处理6 h后,血清SOD活性与CAT活性变化呈显著负相关;而处理12 h后,血清SOD活性与CAT活性变化呈显著正相关。上述结果为开展二甲苯、蒽、苯并[α]芘对海洋贝类毒性评价提供基础数据。     

不同硝化负荷MBR去除类固醇雌激素的研究

以雌酮(E1)和17α-乙炔基雌二醇(EE2)2种典型类固醇雌激素为研究对象,考察了不同硝化负荷好氧MBR对其的去除特性.结果表明,硝化负荷为6.19,0.35,10.58mg/(gMLSS·d)的MBR对E1平均去除率分别为72.33%、64.06%和71.77%,去除负荷分别为29.98,23.06,27.66mg/(gMLSS·d);对EE2平均去除率分别为55.40%、42.87%和58.90%,去除负荷分别为22.39,16.54,23.58mg/(gMLSS·d).E1去除效果均优于EE2.MBR硝化负荷与氨氧化细菌之间有较好的线性关系;而SEs去除负荷与氨氧化细菌及硝化负荷之间不满足线性关系.硝化负荷为0.35mg/(gMLSS·d)时,SEs去除负荷较低,硝化负荷达到6.19mg/(gMLSS·d)时,SEs的去除负荷显著提高,而当硝化负荷增加到10.58mg/(gMLSS·d)时,SEs去除负荷不再有效提高.