不对称还原丙酮酸乙酯菌株的筛选及鉴定

以丙酮酸乙酯为底物,从成都某化工厂污水池及其附近土壤中分离到36株可将丙酮酸乙酯不对称还原成(S)-乳酸乙酯的菌株.经过多次复筛,最终获得了一株具有较高催化活性的酵母菌BTY18-6.在以该菌株静息细胞为催化剂,催化不对称还原丙酮酸乙酯合成(S)-乳酸乙酯的反应中,底物浓度为60 mmol/L时,底物转化率为86.9%,产物(S)-乳酸乙酯的ee值为88.7%.通过对其形态学、生理生化特征及其26S rDNA Dl/D2区域的分析表明,BTY18-6为胶红酵母.     

Modulation of steroidogenesis by coastal waters and sewage effluents of Hong Kong, China, using the H295R assay

BACKGROUND, AIM, AND SCOPE: The presence of a variety of pollutants in the aquatic environment that can potentially interfere with the production of sex steroid hormones in wildlife and humans has been of increasing concern. The aim of the present study was to investigate the effects of extracts from Hong Kong marine waters, and influents and effluents from wastewater treatment plants on steroidogenesis using the H295R cell bioassay. After exposing H295R cells to extracts of water, the expression of four steroidogenic genes and the production of three steroid hormones were measured. MATERIALS AND METHODS: Water samples were collected during the summer of 2005 from 24 coastal marine areas and from the influents and effluents of two major waste water treatment plants (WWTPs) in Hong Kong, China. Samples were extracted by solid phase extraction (SPE). H295R cells were exposed for 48 h to dilutions of these extracts. Modulations of the expression of the steroidogenic genes CYP19, CYP17, 3betaHSD2, and CYP11beta2 were determined by measuring mRNA concentrations by real-time polymerase chain reaction (Q-RT-PCR). Production of the hormones progesterone (P), estradiol (E2), and testosterone (T) was quantified using enzyme linked immunosorbent assays (ELISA). RESULTS: Extracts from samples collected in two fish culture areas inhibited growth and proliferation of H295R cells at concentrations greater or equal to 10(5) L equivalents. The cells were exposed to the equivalent concentration of active substances in 10,000 L of water. Thus, to observe the same level of effect as observed in vitro on aquatic organisms would require a bioaccumulation factor of this same magnitude. None of the other 22 marine samples affected growth of the cells at any dilution tested. Twelve of the marine water samples completely inhibited the expression of CYP19 without affecting E2 production; inhibition of CYP17 expression was observed only in one of the samples while expression of CYP11beta2 was induced as much as five- and ninefold after exposure of cells to extracts from two locations. The expression of the progesterone gene 3betaHSD2 was not affected by any of the samples; only one sample induced approximately fourfold the production of E2. Although more than twofold inductions were observed for P and T production, none of these values were statistically significant to conclude effects on the production of these two hormones. While influents from WWTPs did not affect gene expression, an approximately 30% inhibition in the production of E2 and a 40% increase in P occurred for the exposure with influents from the Sha Tin and Stonecutters WWTPs, respectively. Effluents from WWTPs did not affect the production of any of the studied hormones, but a decrement in the expression of the aldosterone gene CYP11beta2 was observed for the Sha Tin WWTP exposure. No direct correlation could be established between gene expression and hormone production. DISCUSSION: Observed cytotoxicity in the two samples from fish culture areas suggest the presence of toxic compounds; chemical analysis is required for their full identification. Although effluents from WWTPs did not affect hormone production, other types of endocrine activity such as receptor-mediated effects cannot be ruled out. Interactions due to the complexity of the samples and alternative steroidogenic pathways might explain the lack of correlation between gene expression and hormone production results. CONCLUSIONS: Changes observed in gene expression and hormone production suggest the presence in Hong Kong coastal waters of pollutants with endocrine disruption potential and others of significant toxic effects. The aromatase and aldosterone genes seem to be the most affected by the exposures, while E2 and P are the hormones with more significant changes observed. Results also suggest effectiveness in the removing of compounds with endocrine activity by the WWTPs studied, as effluent samples did not significantly affect hormone production. The H295R cell showed to be a valuable toll in the battery required for the analysis of endocrine disrupting activities of complex environmental samples. RECOMMENDATIONS AND PERSPECTIVES: Due to the intrinsic complexity of environmental samples, a combination of analytical tools is required to realistically assess environmental conditions, especially in aquatic systems. In the evaluation of endocrine disrupting activities, the H295R cell bioassay should be used in combination with other genomic, biological, chemical, and hydrological tests to establish viable modes for endocrine disruption and identify compounds responsible for the observed effects.     

Electrochemical determination of anaerobic microbial decay coefficients

The biokinetics of attached and suspended bacteria are an essential component of activated sludge models, anaerobic digestion models and biofilm models. These parameters are often assumed or “confirmed” based on the goodness-of-fit of the bioprocess models. Using a microbial fuel cell with a baffled reactor chamber, the attached- and mixed-growth microbial decay coefficients were evaluated under anaerobic conditions. The capability for real-time voltage recording allows easy and accurate measurement of the anaerobic microbial decay coefficients (b_L, lysis-regrowth approach), which were determined to be 0.11 ± 0.01 and 0.15 ± 0.01 d⁻¹ for attached (to anode) and mixed (present in the anode chamber) growth microorganisms, respectively. The corresponding half-saturation constants using glucose as a substrate were 204 ± 10 and 123 ± 1 mg COD l⁻¹. Hence, like an oxygen uptake rate-based approach to measure the microbial kinetics under aerobic conditions, the electrochemical recording provides an attractive method to measure anaerobic microbial decay coefficients.     

Efficient photoelectrochemical oxidation of rhodamine B on metal electrodes without photocatalyst or supporting electrolyte

We designed photoelectrochemical cells to achieve efficient oxidation of rhodamine B (RhB) without the need for photocatalyst or supporting electrolyte. RhB, the metal anode/cathode, and O₂ formed an energy-relay structure, enabling the efficient formation of O ₂ ^– species under ultraviolet illumination. In a single-compartment cell (S cell) containing a titanium (Ti) anode, Ti cathode, and 10 mg·mL^–1 RhB in water, the zero-order rate constant of the photoelectrochemical oxidation (k_PEC) of RhB was 0.049 mg·L^–1·min^–1, while those of the photochemical and electrochemical oxidations of RhB were nearly zero. k_PEC remained almost the same when 0.5 mol·L^–1 Na₂SO₄ was included in the reactive solution, regardless of the increase in the photocurrent of the S cell. The k_PEC of the illuminated anode compartment in the two-compartment cell, including a Ti anode, Ti cathode, and 10 mg·mL^–1 RhB in water, was higher than that of the S cell. These results support a simple, eco-friendly, and energysaving method to realize the efficient degradation of RhB.

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Recent advances in three typical configurations and applications of bioelectrochemical systems北大核心CSCD

As the world's freshwater resources and available energy are alarmingly decreasing, the bioelectrochemical system (BES) is a cutting-edge technology for the resolution of the resource and energy issue. Researchers have paid much attention to t he application of t he BES configuration. Based on t he brief i ntroduction of m icrobial f uel cell a nd m icrobial electrolytic cell structure, principles, and domestic and foreign research, the BES and its influencing factors are introduced, specifically including: microbial activity, electrode materials, and configuration. Three important aspects (i.e., the electrode chamber, the reaction chamber, and micro-sensor) are summarized, and the advantages and disadvantages of single-electrode and multi-electrode chambers are compared, based on the microbial desalination cell. Microbial electrolysis desalination cell: Microbial electrolysis desalination and chemical-production cell have been discussed to introduce increasing reaction chamber configuration; this review focuses on the research of BES monitoring with regards to biochemical oxygen demand. The potential applications of the research progress are explored. The results show that the configuration of multi-chamber microbial fuel cell is complex and its efficiency is low, while the single chamber configuration is advantageous. The reaction chamber added is mainly aimed at desalination, and the study of the desalination pool still needs to be focused on optimizing the cation exchange membrane to maintain the anode pH balance and reduce the air cathode dissolved oxygen. Microbial electrode sensor can be applied in more areas, and its sensitivity and long-term stability need to be further improved. However, there is relatively less research on the abundance and activity of electricigen communities; the configurations and scopes of application of BES are still the research priority. © 2018 Science Press. All rights reserved.     

Oxidative and genotoxic effects of bisphenol A on primary gill cell culture of Lake Van Fish (Alburnus tarichi Güldenstädt, 1814)

Lake Van is the largest lake in Turkey. The lake limits lifespan due to its high pH and brackish water. For this reason, only a single species of fish (Van Fish) is living in the lake that has been adapted to these conditions. In the present study, we investigated the total oxidant status (TOS), total antioxidant status (TAS), malondialdehyde (MDA) level and DNA damage effect of bisphenol A (BPA) (10^?7, 10^?6 and 10^?5?M) on primary gill cell culture of Van Fish for 24 and 48?h of incubation periods. TAS levels were not changed when compared to those of the control group, but TOS levels were decreased in both 24 and 48?h. The MDA level increased only at the highest concentration (10^?5) at the end of 12 and 24?h (p?.05). DNA damage increased only at the 10^?5?M concentration after 48?h. At the end of the experiment, BPA exposure caused lipid peroxidation and genotoxic effect. These results indicate that high levels of BPA exposure induced oxidative stress and DNA damage by time- and concentration-dependent fashion in the gill cell culture of Van Fish. Gill cell culture is a useful model for the rapid identification of the harmful effects of chemicals in the aquatic environment.     

Epigenetic effect of long-term bisphenol A exposure on human breast adenocarcinoma cells

In this research, epigenetic effects of bisphenol A (BPA) on human breast cancer MCF-7 cells were analyzed. Genome-wide DNA methylation and gene expression were analyzed in MCF-7 cells exposed to BPA (10^?5 and 10^?6 mol/L for 5 weeks). No significant changes in the global level of 5-methyl-2′-deoxycytidine and 5-hydroxymethyl-2′-deoxycytidine were observed. DNA methylation profiling analysis indicated that BPA exposure resulted in the hypermethylation of FOXK2, LKB1, LMX1A and CUGBP2 and the hypomethylation of PTPRN2, TRIM27, BCAS3 and ZNF423. Decreased expression of apoptosis genes (P38 and BCL2L1) and increased expression of chemokine (Cxcl2 and ccl20) were detected. Changes of these genes were speculated to affect the ERα-related cell growth as well as cell apoptosis.     

3种兽药及饲料添加剂对鱼类的毒理效应

采用斑马鱼、鲤鱼作为生物材料,通过斑马鱼急性毒性试验、胚胎发育试验和鲤鱼肾细胞单细胞凝胶电泳试验,分别从生物个体水平、细胞水平和分子水平,对兽药及饲料添加剂喹乙醇、阿散酸和土霉素进行了生态毒理学效应研究。急性毒性试验结果表明,3种兽药及饲料添加剂在有生态学意义的浓度范围内对斑马鱼的急性毒性均不大;胚胎发育试验结果表明,喹乙醇和阿散酸对斑马鱼胚胎发育72 h的EC50值分别为221.20和239.54 mg.L-1,具有明显的致畸效应;单细胞凝胶电泳试验结果表明,喹乙醇和土霉素均具有一定的遗传毒性,能引起鲤鱼肾细胞DNA的明显损伤,并呈现出一定的剂量-效应关系。     

Cultured human-cell-based bioassay for environmental risk management

Among bioassays for evaluating various impacts of chemicalson humans and ecosystems, those based on culturedmammalian-cells can best predict acute lethal toxicity to humans. Weexpect them to be employed in the future in environmentalrisk management alongside mutagenicity tests and endocrine-disrupting activity tests. We recently developed adisposable bioassay device that immobilizes humanhepatocarcinoma cells in a small micropipette tip. Thisenables very quick (within 2 h) evaluation of acute lethaltoxicity to humans. For bioassay-based environmentalmanagement, 2 promising approaches have been demonstrated bythe US-EPA: toxicity identification evaluation (TIE) andtoxicity reduction evaluation (TRE). The Japanese Ministryof Environment has been supporting a multi-center validationproject, aimed at assembling a bioassay database. To makefull use of these resources, we present a numerical modelthat describes contribution of individual chemical toobserved toxicity. This will allow the selection of the mosteffective countermeasure to reduce the toxicity. Bioassay-based environmental risk management works retrospectively,whereas impact assessment using substance flow models andtoxicity databases works prospective. We expect that these 2approaches will exchange information, act complementarily,and work effectively in keeping our environment healthy inthe 21st century.     

厌氧折流板式微生物燃料电池堆影响因素研究

微生物燃料电池(MFC)中输出电压/电流的提升,以及反应器体积的扩展放大是其工程化应用的关键。本文构建了一个总体积为6.4 L的新型厌氧折流板式微生物燃料电池堆(ABSMFC)。以葡萄糖作为底物,探讨了阳极材料、液面高程差和水力停留时间(HRT)等因素对ABSMFC性能的影响。结果表明,碳纤维毡作为阳极时,电池单体外电路平均分压(R_(ex)=1 000Ω)为210 mV,填充石墨颗粒后增加到319.8 mV。格室间存在液面高程差时,电池单体、串联和并联的功率密度分别为207.1、181.1和215.7 mW/m~2,当无液面高程差(即水力相连)时为205.8、69.5和151.5 mW/m~2。4个电池单体串联和并联连接时,HRT对ABSMFC的产电稳定性无影响,溶解性COD的去除率和库仑效率均随HRT的增加而升高,且并联效果优于串联。