Kinetics of chromate reduction during naphthalene degradation in a mixed culture

A mixed culture of Bacillus sp. K1 and Sphingomonas paucimobilis EPA 505 was exposed to chromate and naphthalene. Batch experiments showed that chromate was reduced and naphthalene was degraded by the mixed culture. Chromate reduction occurred initially at a high rate followed by a decrease in rate until chromate reduction ceased. Chromate reduction decreased in the mixed culture when a lower ratio of S. paucimobilis EPA 505 to Bacillus sp. K1 was utilized. A kinetic model incoporating a term for the cell density ratio is proposed to describe chromate reduction in the mixed culture under both chromate limited and electron donor limited conditions. The validity of the model, and its parameter values, was verified by experimental data generated under a variety of initial population compositions and a broad range of chromate concentrations. The consistent result of experimental data with model predictions implies that the model is useful for evaluating the interactions and the use of mixed culture for chromate removal. (c) 1996 John Wiley & Sons, Inc.     

Long-term impact of dissolved O(2) on the activity of anaerobic granules

The impact of influent dissolved O(2) on the characteristics of anaerobic granular sludge was investigated at various dissolved O(2) concentrations (0.5-8.1 ppm) in 1- and 5-L laboratory-scale upflow anaerobic sludge bed (UASB)-like anaerobic/aerobic coupled reactors with a synthetic wastewater (carbon sources containing 75% sucrose and 25% acetate). The rate of dissolved O(2) supplied to the coupled reactor was as high as 0.40 g O(2)/L(rx).d, and the anaerobic/aerobic coupled reactors maintained excellent methanogenic performances at a COD loading rate of 3 g COD/L(rx).d even after the reactors had been operated with dissolved O(2) for 3 months. The activities of granular sludge on various substrates (glucose, propionate, and hydrogen) were not impaired, and acetate activity was even improved over a short term. However, after 3 months of operation, slight declines on the acetoclastic activities of granules were observed in the coupled reactor receiving the recirculated fluid containing 8.1 ppm dissolved O(2).Methane yield in the anaerobic control reactor and anaerobic/aerobic coupled reactors revealed that a significant aerobic elimination (up to 30%) of substrate occurred in the coupled reactors, as expected. The presence of dissolved O(2) in the recirculated fluid resulted in the development of fluffy biolayers on the granule surface, which imposed a negative impact on the settleability of granular sludge and caused a slightly higher sludge washout. This research shows that the anaerobic/aerobic coupled reactor can be successfully operated under O(2)-limited conditions and is an ideal engineered ecosystem integrating oxic and anaerobic niches. (c) 1996 John Wiley & Sons, Inc.     

Modeling simultaneous hexavalent chromium reduction and phenol degradation by a defined coculture of bacteria

Based on the kinetics of Cr(VI) reduction by Escherichia coli ATCC 33456 and phenol degradation by Pseudomonas putida DMP-1, a mathematical model is developed to describe simultaneous Cr(VI) reduction and phenol degradation in the coculture of the two species. The developed model incorporates the toxicity effects of Cr(VI) and phenol on phenol degradation and Cr(VI) reduction in the coculture. The model illustrates the inhibitory effects of phenol on Cr(VI) reduction and Cr(VI) toxicity toward phenol degradation. The model also reveals the recoveries of the activities of the repressed bacterial cells with continuous Cr(VI) reduction and phenol degradation in the coculture. The model is capable of predicting simultaneous Cr(VI) reduction and phenol degradation within a broad range of Cr(VI) and phenol concentrations and under an appropriate composition of populations. However, the model simulates lower concentrations of phenol than experimental observations once Cr(VI) is reduced to a low level (<7 mg/L). The model simulation for Cr(VI) also deviates from experimental data when P. putida is outnumbered by E. coli by a ratio of 1:5. (c) 1995 John Wiley & Sons, Inc.     

The induction of nitrate reductase and the preferential assimilation of ammonium in germinating rice seedlings

Nitrate reduotase is induced by nitrate in excised embryos and germinating intact seedlings of rice (Oryza sativa L.). The enzyme is induced 24 hr after imbibition. The rate of enzyme formation increases with the age of seedlings. There is a lag period of 30 to 40 min between the addition of substrate and the formation of nitrate reductase. Formation of the enzyme is promoted by the presence of ammonium. Chloramphenicol, actinomycin D and cycloheximide effectively inhibit the formation of nitrate reductase.     

固氮蓝藻的室内培养试验

固氮蓝藻是水稻田中的有效固氮微生物,在水稻田中人工大量培养繁殖,可提高水稻田的一定肥力,解决水稻的部分氮素肥源,使水稻获得增产(黎尚豪,1959;利卓燊,1961,1962)。室內培养是大田藻种来源的第一步,也是研究蓝藻的生理学和生态学的方法之一。近廿多年来已进行了許多的試驗研究(田宮Tamiya,1957;叶清泉、黎尚豪等,1959;藤原彰夫和奥津正彥,1958—1960),对固氮蓝藻的生理特性及培养方法,已有一定的基础。我們为了簡化培养的設置及加速固氮蓝藻的生长,采用不通气的靜置培养及对培养中的几个主要环节进行了以下試驗。     

大水体生态工程技术及绿萍的养用效果

大水体生态工程技术是综合协调水面、水下、水中、空间的生态与生产各要素的相互关系,实行立体结构、优化组合的“萍-鸡-鱼”三业物质生产、循环利用的整体配套技术。生产应用表明,在水面上,采用生产绿萍的“浮盘”工艺,及筛选出适于饲用的良种,并提供新的栽培技术,绿萍产量高达495—547t·ha~(-1)·年~(-1);在水下2.5m深处,开展网箱养吃食性鱼,可提高饲料效价,增加鱼产量37.5—117.6％;在水中,放养滤食性鱼,利用网箱的鱼粪排出为饵料,水中鱼类增产4.29倍;在网箱上空间,设计出禽舍养鸡的工艺,用绿萍饲鸡,可增产2.5—7.5％和节省精料10—20％;再用鸡粪养绿萍和鱼等。形成节地、节粮、节肥、节能、节工的开发大水体生产的新型产业。     

Continuous Production of Thermostable beta-Amylase with Clostridium thermosulfurogenes: Effect of Culture Conditions and Metabolite Levels on Enzyme Synthesis and Activity

A beta-amylase-overproducing mutant of Clostridium thermosulfurogenes was grown in continuous culture on soluble starch to produce thermostable beta-amylase. Enzyme productivity was reasonably stable over periods of weeks to months. The pH and temperature optima for beta-amylase production were pH 6.0 and 60 degrees C, respectively. Enzyme concentration was maximized by increasing biomass concentration by using high substrate concentrations and by maintaining a low growth rate. beta-Amylase concentration reached 90 U ml at a dilution rate of 0.07 h in a 3% starch medium. A further increase in enzyme activity levels was limited by acetic acid inhibition of growth and low beta-amylase productivity at low growth rates.     

Proteomic studies of PP2A-B56gamma1 phosphatase complexes reveal phosphorylation-regulated partners in cardiac local signaling

Defects of kinase-phosphatase signaling in cardiac myocytes contribute to human heart disease. The activity of one phosphatase, PP2A, is governed by B targeting subunits, including B56gamma1, expressed in heart cells. As the role of PP2A/B56gamma1 on the heart function remains largely unknown, this study sought to identify protein partners through unbiased, affinity purification-based proteomics combined with the functional validation. The results reveal multiple interactors that are localized in strategic cardiac sites to participate in Ca2+ homeostasis and gene expression, exemplified by the Ca pump, SERCA2a, and the splicing factor ASF/SF2. These results are corroborated by confocal imaging where adenovirally overexpressed B56gamma1 is found in z-line/t-tubule region and nuclear speckles. Importantly, overexpression of B56gamma1 in cultured myocytes dramatically impairs cell contractility. These results provide a global view of B56gamma1-regulated local signaling and heart function.     

Identification and quantification of basic and acidic proteins using solution-based two-dimensional protein fractionation and label-free or 18O-labeling mass spectrometry

Reduction in sample complexity enables more thorough proteomic analysis using mass spectrometry (MS). A solution-based two-dimensional (2D) protein fractionation system, ProteomeLab PF 2D, has recently become available for sample fractionation and complexity reduction. PF 2D resolves proteins by isoelectric point (pI) and hydrophobicity in the first and second dimensions, respectively. It offers distinctive advantages over 2D gel electrophoresis with respects to automation of the fractionation processes and characterization of proteins having extreme pIs. Besides fractionation, PF 2D is equipped with built-in UV detectors intended for relative quantification of proteins in contrasting samples using its software tools. In this study, we utilized PF 2D for the identification of basic and acidic proteins in mammalian cells, which are generally under-characterized. In addition, mass spectrometric methods (label-free and 18O-labeling) were employed to complement protein quantification based on UV absorbance. Our studies indicate that the selection of chromatographic fractions could impact protein identification and that the UV-based quantification for contrasting complex proteomes is constrained by coelution or partial coelution of proteins. In contrast, the quantification post PF 2D chromatography based on label-free or 18O-labeling mass spectrometry provides an alternative platform for basic/acidic protein identification and quantification. With the use of HCT116 colon carcinoma cells, a total of 305 basic and 183 acidic proteins was identified. Quantitative proteomics revealed that 17 of these proteins were differentially expressed in HCT116 p53-/- cells.     

Expression, Purification and Functional Analysis of Hexahistidine-tagged Human Glutathione S-Transferase P1-1 and its Cysteinyl Mutants

The bacterial expression and purification of human glutathione S-transferase P1-1(hGST P1-1), as a hexahistidine-tagged polypeptide was performed. Site-directed mutagenesis was used to construct mutants in which alanine replaced two (C47A/C101A), three (C14A/C47A/C101A) or all four (C14A/C47A/ C101A/C169A) cysteine residues using the plasmid for the wild type enzyme. Analysis of their catalytic activities and kinetic parameters suggested that cysteins are not essential for the catalytic activity but may contribute to some extent to the catalytic efficiency. Moreover, on SDS-polyacrylamide gel electrophoresis (SDS-PAGE) under nonreducing conditions, hexahistidine-tagged hGST P1-1 (His₆-hGST P1-1) treated with 1 mM H₂O₂ showed at least three extra bands, in addition to the native His₆-hGST P1-1 subunit band. These extra bands were not detected in the cysteinyl mutants. Thus, it indicated that disulfide bonds were formed mainly within subunits between cysteine residues, causing an apparent reduction in molecular weight, only small amounts of binding between subunits being observed. These authors contributed equally to this work.