氧化亚铁硫杆菌对金属铜的加工

材料加工的传统技术包括物理方法和化学方法。当今 ,生物技术已进入各个领域 ,也渗透到材料加工领域。因此 ,材料加工技术也包括生物方法。根据加工工件体积变化 ,生物方法加工分为生物去除加工 (Ｒｅｍｏｖａｌ)、生物沉积加工 (Ａｄｄｉｔｉｏｎ)和生物成形加工 (Ｄｅｆｏｒｍａｔｉｏｎ)。研究生物加工方法的最早报导是 1 993年日本冈山大学宇野义幸等人[1～ 3] ,证实了细菌对纯铁、纯铜去除加工的可能性以及附加电场的作用。国内的研究工作进一步证实了生物加工能力 ,并加工出微小齿轮[4～5] 。本文报导氧化亚铁硫杆菌 (Ｔｈｉｏｂａｃ…     

Innovative use of Thiobacillus ferrooxidans for the biological machining of metals

Preliminary results on the novel use of the bacterium Thiobacillus ferrooxidans (ATCCJ 3598 and ATCC33020) for the micro‐machining (or biomachinig) of metals are reported. Biomachning is a controlled microbiological process to selectively form microstrucutures on a metal work‐piece by metal removal (or dissolution) using microorganisms. Applying copper and mild steel as work‐pieces, it was shown that the mass removed increased proportionately with machining time. In another experiment, the work‐pieces were coated with organic photo‐resistive materials to mask (i.e. protect) certain regions of the metlas, thereby defining the microstructure to be formed. The unmasked regions were successfully biomachined; the final machined profile was shown to be similar to the coating image on the original metal. Although biomachining proceeded at a slower rate than chemical machining, the undesired leaching of the metal in the region under the masked area (termed undercutting) was not as severely encountered when compared with the latter. This work demonstrates the potential use of microorganisms for the biomachining of metals. As a “green process”, the innovative use of T. ferrooxidans for the micro‐machining of metals opens up the possibility of biomachining as an alternative to conventional metal processing.     

Studies on kinetics and thermodynamics of biomachining pure copper

The kinetics and thermodynamics of biomachining pure copper by Thiobacillus ferrooxidans were quantitatively analyzed. The kinetic effect of bacteria on the ion-cycle between Fe2 and Fe3 and the thermodynamic effect of hydrolysis on the changes of pH in biomachining processes were expounded through six series of experiment. Finally a model of ion-cycle in the biomachining processes was proposed, and the measures for maintaining the stable equilibrium of the ion-cycle were also discussed.     

Establishment of a simple and effective isolation method for cyanophycin from recombinant Saccharomyces cerevisiae

An efficient, time-saving, and cost-effective method for isolation of the polyamide cyanophycin from recombinant Saccharomyces cerevisiae was established. Due to its simple procedure, this isolation method may be also applicable at industrial scale and also to other intracellular compounds in this yeast. Production of cyanophycin gained preferential interest in the past, as degradation products thereof are of pharmaceutical and technical interest. Recently, it was shown that Saccharomyces cerevisiae represents a putative candidate for cyanophycin synthesis at industrial scale. For identification of optimal isolation procedures, several parameters such as heat stress, freeze drying, and freeze/thaw cycles of transgenic yeast cells were compared for their effectiveness of cyanophycin isolation. Additionally, optimal resuspension solutions for the applied cells and minimal required materials or chemicals were determined to make the process most environmentally and economically friendly. Maximal cyanophycin granule polypeptide yields of 21% (w/w) were obtained after incubation of dry cells at 70 °C or 80 °C and precipitation of the polymer with two volumes of ethanol.     

Advanced model‐based control strategies for the intensification of upstream and downstream processing in mAb production

Current industrial trends encourage the development of sustainable, environmentally friendly processes with minimal energy and material consumption. In particular, the increasing market demand in biopharmaceutical industry and the tight regulations in product quality necessitate efficient operating procedures that guarantee products of high purity. In this direction, process intensification via continuous operation paves the way for the development of novel, eco‐friendly processes, characterized by higher productivity and lower production costs. This work focuses on the development of advanced control strategies for (i) a cell culture system in a bioreactor and (ii) a semicontinuous purification process. More specifically, we consider a fed‐batch culture of GS‐NS0 cells and the semicontinuous Multicolumn Countercurrent Solvent Gradient Purification (MCSGP) for the purification process. The controllers are designed following the PAROC framework/software platform and their capabilities are assessed in silico, against the process models. It is demonstrated that the proposed controllers efficiently manage to increase the system productivity, returning strategies that can lead to continuous, stable process operation. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:966–988, 2017     

A novel iron‐ and copper‐binding protein in the Lyme disease spirochaete

Iron and copper are transition metals that can be toxic to cells due to their abilities to react with peroxide to generate hydroxyl radical. Ferritins and metallothioneins are known to sequester intracellular iron and copper respectively. The Lyme disease pathogen Borrelia burgdorferi does not require iron, but its genome encodes a ferritin‐like Dps (D NA‐binding p rotein from s tarved bacteria) molecule, which has been shown to be important for the spirochaete's persistence in the tick and subsequent transmission to a new host. Here, we show that the c arboxyl‐terminal c ysteine‐r ich (CCR) domain of this protein functions as a copper‐binding metallothionein. This novel fusion between Dps and metallothionein is unique to and conserved in all Borrelia species. We term this molecule BicA for B orrelia i ron‐ and c opper‐binding protein A . An isogenic mutant lacking BicA had significantly reduced levels of iron and copper and was more sensitive to iron and copper toxicity than its parental strain. Supplementation of the medium with iron or copper rendered the spirochaete more susceptible to peroxide killing. These data suggest that an important function of BicA is to detoxify excess iron and copper the spirochaete may encounter during its natural life cycle through a tick vector and a vertebrate host.     

Genomic insights into the iron uptake mechanisms of the biomining microorganism Acidithiobacillus ferrooxidans

Commercial bioleaching of copper and the biooxidation of gold is a cost-effective and environmentally friendly process for metal recovery. A partial genome sequence of the acidophilic, bioleaching bacterium Acidithiobacillus ferrooxidans is available from two public sources. This information has been used to build preliminary models that describe how this microorganism confronts unusually high iron loads in the extremely acidic conditions (pH 2) found in natural environments and in bioleaching operations. A. ferrooxidans contains candidate genes for iron uptake, sensing, storage, and regulation of iron homeostasis. Predicted proteins exhibit significant amino acid similarity with known proteins from neutrophilic organisms, including conservation of functional motifs, permitting their identification by bioinformatics tools and allowing the recognition of common themes in iron transport across distantly related species. However, significant differences in amino acid sequence were detected in pertinent domains that suggest ways in which the periplasmic and outer membrane proteins of A. ferrooxidans maintain structural integrity and relevant protein-protein contacts at low pH. Unexpectedly, the microorganism also contains candidate genes, organized in operon-like structures that potentially encode at least 11 siderophore systems for the uptake of Fe(III), although it does not exhibit genes that could encode the biosynthesis of the siderophores themselves. The presence of multiple Fe(III) uptake systems suggests that A. ferrooxidans can inhabit aerobic environments where iron is scarce and where siderophore producers are present. It may also help to explain why it cannot tolerate high Fe(III) concentrations in bioleaching operations where it is out-competed by Leptospirillum species.

     

Effect of zinc supplementation on the antioxidant,copper, and iron status of physically active adolescents

Puberty associated with intense physical activity results in oxidation stress. Zinc supplementation may benefit antioxidant capacity although it may also affect iron and copper status. This study evaluated the effect of zinc supplementation on antioxidant, zinc and copper status of physically active male football players (13 years ± 0.4 years), divided in two groups and studied during 12 weeks: Zn‐supplemented (Zn‐SUP, 22 mg Zn d^?1 as zinc gluconate, n = 21) and placebo (PLA, n = 26). At baseline, there was no significant difference in biochemical indices between the two groups. After treatment, plasma zinc and erythrocyte iron increased in both groups (p < 0.001); urinary zinc increased (p < 0.001) only in Zn‐SUP, and erythrocyte zinc decreased (p = 0.002) only in PLA. Plasma iron and copper decreased (p = 0.01 and p = 0.015, respectively) only in Zn‐SUP. Plasma ferric‐reducing ability and plasma conjugated dienes increased, and erythrocyte osmotic fragility decrease in both groups, although the latter two were significantly lower in Zn‐SUP compared to PLA (p < 0.01). In conclusion, our study indicates that the use of 22 mg d^?1 of supplemental zinc during 12 week in adolescent athletes did not affect growth, improved markers of antioxidant status but reduced plasma iron and copper. Therefore, it appears that the use of zinc supplementation by healthy adolescent athletes benefits their antioxidant capacity but impairs copper and iron nutritional status. Copyright © 2009 John Wiley & Sons, Ltd.     

Life‐cycle and cost of goods assessment of fed‐batch and perfusion‐based manufacturing processes for mAbs

Life‐cycle assessment (LCA) is an environmental assessment tool that quantifies the environmental impact associated with a product or a process (e.g., water consumption, energy requirements, and solid waste generation). While LCA is a standard approach in many commercial industries, its application has not been exploited widely in the bioprocessing sector. To contribute toward the design of more cost‐efficient, robust and environmentally‐friendly manufacturing process for monoclonal antibodies (mAbs), a framework consisting of an LCA and economic analysis combined with a sensitivity analysis of manufacturing process parameters and a production scale‐up study is presented. The efficiency of the framework is demonstrated using a comparative study of the two most commonly used upstream configurations for mAb manufacture, namely fed‐batch (FB) and perfusion‐based processes. Results obtained by the framework are presented using a range of visualization tools, and indicate that a standard perfusion process (with a pooling duration of 4 days) has similar cost of goods than a FB process but a larger environmental footprint because it consumed 35% more water, demanded 17% more energy, and emitted 17% more CO₂ than the FB process. Water consumption was the most important impact category, especially when scaling‐up the processes, as energy was required to produce process water and water‐for‐injection, while CO₂ was emitted from energy generation. The sensitivity analysis revealed that the perfusion process can be made more environmentally‐friendly than the FB process if the pooling duration is extended to 8 days. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1324–1335, 2016     

Effects of metal quantity and quality to the removal of zinc and copper by two common green microalgae (Chlorophyceae) species

Appearance of metals as pollutants in the environment is an increasing global problem. Microalgae as subjects of biological remediation methods may provide a cost‐effective and environmentally friendly alternative to the removal of metals during wastewater treatment. Despite the high number of data in the topic, there is still little information on how the type and the concentration of the metal affect the process of removal. In this study, correlations among the algal species, quality and quantity of metals and characteristics of metal removal mechanism were investigated at lower metal concentrations (0.2–5.0 mg L^?1) during zinc and copper removal of the green algae Desmodesmus communis and Monoraphidium pusillum. Analyses of the results proved that there is a statistically significant interaction (P < 0.05) between algal species and quality and concentration of the metals, that is, they have a significant effect on the mode and extent of removal. Both metals were mainly extracellularly bound, but at concentrations of 0.2–1.4 mg L^?1, intracellular proportion could exceed the extracellular adsorption. Although there were differences between the two algae, generally copper appeared in a higher intracellular proportion than zinc in the whole studied concentration range. Overall, the quality and initial concentration of the metal is decisive for the way of removal, the knowledge of which is useful for planning post treatment retention times or post treatment processes of the used biomass during wastewater treatment.     

Microbial Methylotrophic Metabolism: Recent Metabolic Modeling Efforts and Their Applications In Industrial Biotechnology

Developing methylotrophic bacteria into cell factories that meet the chemical demand of the future could be both economical and environmentally friendly. Methane is not only an abundant, low‐cost resource but also a potent greenhouse gas, the capture of which could help to reduce greenhouse gas emissions. Rational strain design workflows rely on the availability of carefully combined knowledge often in the form of genome‐scale metabolic models to construct high‐producer organisms. In this review, the authors present the most recent genome‐scale metabolic models in aerobic methylotrophy and their applications. Further, the authors present models for the study of anaerobic methanotrophy through reverse methanogenesis and suggest organisms that may be of interest for expanding one‐carbon industrial biotechnology. Metabolic models of methylotrophs are scarce, yet they are important first steps toward rational strain‐design in these organisms.     

Maintenance of biodegradation capacities of aerobic bacteria during long-term preservation

Six strains of aerobic Gram negative bacteria degrading toluene, 2,4-dichlorophenoxyacetate, 2,2-dichloropropionate or 3-chlorobenzoate were freeze-dried and liquid-dried in the presence or absence of a protective agent. Survival and maintenance of the biodegradation capability was checked before and after drying, and after storage of the ampoules for one year at 4° or 25°C. In many cases, stability of the degradation potential was low although viability was high. Survival and stability of all strains was always highest after preservation by liquid drying in the presence of myo-inositol and activated charcoal as protective agents. Losses of biodegradation abilities were highest after freeze-drying using no protective agents. Cells grown on complex medium were less sensitive to drying than cells grown under selective pressure (on mineral medium with a special compound as the sole carbon source). A choice of the most appropriate preservation method and the use of an effective protectant is recommended to avoid genetic alterations, and to maintain biodegradation capacities during long-term preservation.     

Isolation and Characterization of Iron and Sulfur-Oxidizing Bacteria from Maiduk Copper Mine at Shahrbabk Province in Iran

Microbial oxidation of iron and sulfur are important steps in biogeochemical cycles in mining environments. The aim of this study was the enrichment and identification of two important groups of bacteria that are involved in bioleaching of copper ores. Some soil samples were collected from the Maiduk copper mine. Iron-oxidizing bacteria were enriched in 9K medium containing ferrous sulfate, and sulfur oxidizers were enriched in 9K medium containing powdered sulfur instead of ferrous sulfate as energy source. After three subcultures, autotrophic bacteria were isolated on 9K agarose medium with appropriate energy sources. The autotrophic bacteria from the enrichments were identified by amplification of 16S rRNA gene and sequencing. Bioleaching experiments were performed in 100 ml of 9K medium containing 5 g of low-grade copper ore instead of ferrous sulfate. Twelve different iron and sulfur-oxidizing bacteria were isolated from the collected soil samples of Maiduk copper mine. Molecular identification indicated that two prevalent strains in the ore enrichments could be assigned to the Acidithiobacillus ferooxidans strain HGM and the Thiobacillus thioparus strain HGE. These two strains reached their logarithmic phase of growth after 8 days of incubation in their respective media at 30°C. Of these two cultures, strain HGM leached more copper ore (300 ppm) from the Maiduk copper ore than did strain HGE (200 ppm). Application of these two strains to the Maiduk copper ore in situ and to ore heaps should improve the leaching process.     

Tissue preservation biases in stable isotopes of fishes and molluscs from Patagonian lakes

Field work commonly involves preserving samples for later use; however, most preservation methods distort stable‐isotope (SI) signatures that are of interest to ecologists. Although preservation of muscle samples with table salt and rubbing alcohol affected the SI (δ¹³C and δ¹⁵N) of important consumers in Patagonian lakes (molluscs and fishes), variation among individuals and lakes generally exceeded that among preservation treatments. Mathematical corrections for these preservation biases are provided, and a potentially bias‐free preservation by air‐drying is suggested.     

Chemical leaching of copper-zinc concentrate with ferric iron biosolution

The process of leaching of copper-zinc concentrate with a solution containing biogenic iron, which is a product of the metabolism of iron-oxidizing microorganisms, was studied. The dependence of leaching rate of metals on temperature and pH was determined. It was shown that up to 98% of zinc and 70% of iron could be removed from the concentrate, while up to 7 and 4 g/L of zinc and copper, respectively, were accumulated in the liquid phase, which was sufficient for metal recovery. It was established that a copper concentrate with copper content up to 16% and only 0.5% of zinc could be obtained after chemical leaching for 340 min at 80°C.     

Characterization of biosurfactants as insecticidal metabolites produced by Bacillus subtilis Y9

Biosurfactants have been used for crop protection in integrated pest management programs, as they are generally biodegradable and environmentally friendly. While screening for insecticidal bacteria against green peach aphid, Myzus persicae, we found that a Bacillus isolate, Y9, exhibited strong surface tension activity and produced insecticidal metabolites. The insecticidal metabolites with surface tension activity were extracted from the cell‐free supernatant of Y9 and subjected to repetitive column chromatography and preparative high performance liquid chromatography (HPLC). Based on data from instrumental analyses, the metabolites were identified as surfactin isomers consisting of C₁₄[Leu₇], C₁₄[Val₇] and C₁₅[Leu₇]. The isomers with leucine showed higher insecticidal activity than the isomers with valine. Y9 was capable of producing biosurfactants as insecticidal metabolites against M. persicae, and showed potential for use as a biocontrol agent in an integrated pest management program.     

3‐D analysis of bacterial cell‐(iron)mineral aggregates formed during Fe(II) oxidation by the nitrate‐reducing Acidovorax sp. strain BoFeN1 using complementary microscopy tomography approaches

The formation of cell‐(iron)mineral aggregates as a consequence of bacterial iron oxidation is an environmentally widespread process with a number of implications for processes such as sorption and coprecipitation of contaminants and nutrients. Whereas the overall appearance of such aggregates is easily accessible using 2‐D microscopy techniques, the 3‐D and internal structure remain obscure. In this study, we examined the 3‐D structure of cell‐(iron)mineral aggregates formed during Fe(II) oxidation by the nitrate‐reducing Acidovorax sp. strain BoFeN1 using a combination of advanced 3‐D microscopy techniques. We obtained 3‐D structural and chemical information on different cellular encrustation patterns at high spatial resolution (4–200 nm, depending on the method): more specifically, (1) cells free of iron minerals, (2) periplasm filled with iron minerals, (3) spike‐ or platelet‐shaped iron mineral structures, (4) bulky structures on the cell surface, (5) extracellular iron mineral shell structures, (6) cells with iron mineral filled cytoplasm, and (7) agglomerations of extracellular globular structures. In addition to structural information, chemical nanotomography suggests a dominant role of extracellular polymeric substances (EPS) in controlling the formation of cell‐(iron)mineral aggregates. Furthermore, samples in their hydrated state showed cell‐(iron)mineral aggregates in pristine conditions free of preparation (i.e., drying/dehydration) artifacts. All these results were obtained using 3‐D microscopy techniques such as focused ion beam (FIB)/scanning electron microscopy (SEM) tomography, transmission electron microscopy (TEM) tomography, scanning transmission (soft) X‐ray microscopy (STXM) tomography, and confocal laser scanning microscopy (CLSM). It turned out that, due to the various different contrast mechanisms of the individual approaches, and due to the required sample preparation steps, only the combination of these techniques was able to provide a comprehensive understanding of structure and composition of the various Fe‐precipitates and their association with bacterial cells and EPS.     

Environmental Assessment of Waste-Solvent Treatment Options

A comparison of various waste‐solvent treatment technologies, such as distillation (rectification) and incineration in hazardous‐waste‐solvent incinerators and cement kilns, is presented for 45 solvents with respect to the environmental life‐cycle impact. The environmental impact was calculated with the ecosolvent tool that was previously described in Part I of this work. A comprehensive sensitivity analysis was performed, and uncertainties were quantified by stochastic modeling in which various scenarios were considered. The results show that no single treatment technology is generally environmentally superior to any other but that, depending on the solvent mixture and the process conditions, each option may be optimal in certain cases. Nevertheless, various rules of thumb could be derived, and a results table is presented for the 45 solvents showing under which process conditions and amount of solvent recovery distillation is environmentally superior to incineration. On the basis of these results and the ecosolvent tool, an easily usable framework was developed that helps decision makers in chemical industries reduce environmental burdens throughout the solvent life cycle. With clear recommendations on the environmentally optimized waste‐solvent treatment technology, the use of this framework contributes to more environmentally sustainable solvent management and thus represents a practical application of industrial ecology.     

A New Chiral Residue Analysis Method for Triazole Fungicides in Water Using Dispersive Liquid‐Liquid Microextraction (DLLME)

A rapid, simple, reliable, and environment‐friendly method for the residue analysis of the enantiomers of four chiral fungicides including hexaconazole, triadimefon, tebuconazole, and penconazole in water samples was developed by dispersive liquid–liquid microextraction (DLLME) pretreatment followed by chiral high‐performance liquid chromatography (HPLC)‐DAD detection. The enantiomers were separated on a Chiralpak IC column by HPLC applying n‐hexane or petroleum ether as mobile phase and ethanol or isopropanol as modifier. The influences of mobile phase composition and temperature on the resolution were investigated and most of the enantiomers could be completely separated in 20 min under optimized conditions. The thermodynamic parameters indicated that the separation was enthalpy‐driven. The elution orders were detected by both circular dichroism detector (CD) and optical rotatory dispersion detector (ORD). Parameters affecting the DLLME performance for pretreatment of the chiral fungicides residue in water samples, such as the extraction and dispersive solvents and their volume, were studied and optimized. Under the optimum microextraction condition the enrichment factors were over 121 and the linearities were 30–1500 µg L^?1 with the correlation coefficients (R²) over 0.9988 and the recoveries were between 88.7% and 103.7% at the spiking levels of 0.5, 0.25, and 0.05 mg L^?1(for each enantiomer) with relative standard deviations varying from 1.38% to 6.70% (n = 6) The limits of detection (LODs) ranged from 8.5 to 29.0 µg L^?1(S/N = 3). Chirality 25:567‐574, 2013. © 2013 Wiley Periodicals, Inc.     

A role for copper in protozoan grazing – two billion years selecting for bacterial copper resistance

The Great Oxidation Event resulted in integration of soft metals in a wide range of biochemical processes including, in our opinion, killing of bacteria by protozoa. Compared to pressure from anthropologic copper contamination, little is known on impacts of protozoan predation on maintenance of copper resistance determinants in bacteria. To evaluate the role of copper and other soft metals in predatory mechanisms of protozoa, we examined survival of bacteria mutated in different transition metal efflux or uptake systems in the social amoeba Dictyostelium discoideum. Our data demonstrated a strong correlation between the presence of copper/zinc efflux as well as iron/manganese uptake, and bacterial survival in amoebae. The growth of protozoa, in turn, was dependent on bacterial copper sensitivity. The phagocytosis of bacteria induced upregulation of Dictyostelium genes encoding the copper uptake transporter p80 and a triad of Cu(I)‐translocating P_IB‐type ATPases. Accumulated Cu(I) in Dictyostelium was monitored using a copper biosensor bacterial strain. Altogether, our data demonstrate that Cu(I) is ultimately involved in protozoan predation of bacteria, supporting our hypothesis that protozoan grazing selected for the presence of copper resistance determinants for about two billion years.