Characterization of subterranean bacteria in the Hungarian Upper Permian Siltstone (Aleurolite) Formation

The main purpose of this work was to study the microbiology of the Hungarian Upper Permian Siltstone (Aleurolite) Formation, to assess the safety of future underground repositories for nuclear waste. Sixty-seven air, groundwater, technical water, rock, and surface samples were collected aseptically from different depths. The number of aerobic and anaerobic isolates was 277. The mesophilic minimum and maximum CFU counts of the air samples were 1.07-5.84 x 10(2).mL-1 (aerobic) and 0.22-1.04 x 10(2).mL-1 (anaerobic), respectively; those of the water samples were 0.39-1.25 x 10(5).mL-1 (aerobic) and 0.36-3.9 x 10(3).mL-1 (anaerobic); those of the technical water samples were 0.27-5.03 x 10(6).mL-1 (aerobic) and 4 x 10(5)-->10(6).mL-1 (anaerobic); and those of the aleurolite samples were 2.32 x 10(2)-2.47 x 10(5).g-1 (aerobic) and 0.45-9.5 x 10(2).g-1 (anaerobic). In the groundwater, the thermophilic aerobic bacteria count was 0-2.4 x 10(2).mL-1 and the thermophilic anaerobic bacteria count was 0.43-4.6 x 10(4).mL-1. The gases produced by the 16 gas-forming isolates were CO2 (aerobic isolates), and CO2 and H2 (anaerobic isolates). About 20% of the aerobic isolates produced siderophores. The proportions of organic acid producers were lowest in aerobic and anaerobic isolates from the aleurolite, 13% and 14%, respectively. The highest proportions of acid producers in the aerobic and anaerobic isolates from the air samples were 63% and 54%. Altogether 160 of the aerobic isolates and 52 of the anaerobic isolates were spore formers. The radiosensitivity of the aerobic isolates was also determined; the D10 values of the sporeformers ranged between 0.8-2.44 kGy. Our results indicate that the sulfate-reducing bacteria and the production of complexing agents (siderophores) may contribute to the mobilization of radionuclides from underground repositories. As well, microbial gas production can influence the environmental conditions. The variability in bacterial radiotolerance indicates the biodiversity at this potential disposal site. These facts must be considered during the planning of a nuclear waste repository.     

Factors controlling the resealing of the membrane of human erythrocyte ghosts after hypotonic hemolysis

Summary In accordance with former observations of Hoffman (1962a), ghost populations obtained by hypotonic hemolysis and subsequent restoration of isotonicity by the addition of alkali salts, were found to be composed of 3 types of ghosts. For our purposes it was useful to distinguish between: (1) ghosts which reseal immediately after hemolysis (type I); these ghosts are incapable of incorporating alkali ions which are added after hemolysis; (2) ghosts which reseal after the addition of alkali ions (type II); salt added to the hemolysate becomes trapped inside these ghosts in the course of the resealing process at temperatures above 0°C; and (3) ghosts which remain leaky regardless of the experimental condition (type III). The discrimination between the various types of ghosts was partly achieved by a kinetic method first devised by Hoffman (1962a), and partly by sucrose density gradient centrifugation.The relative sizes of the 3 fractions depend on the temperature at which hemolysis took place and on the time interval which elapsed between hemolysis and the addition of salt. At 37°C the resealing process is fast. Many of the ghosts reseal before salt can be added to the hemolysate. Hence, the fraction of type I ghosts is high after hemolysis at that temperature. At 0°C resealing is extremely slow. Hence, salt which has been added to the hemolysate at that temperature will enter the ghosts and become trapped during subsequent incubation at 37°C. There are no ghosts of type I and many ghosts of type II (about 60%). Regardless of the temperature at hemolysis, there are always ghosts which do not reseal even after prolonged incubation at 37°C. A method has been designed which permits the preparation of homogeneous populations of type II ghosts.Complexing agents (ATP, EDTA, 2,3-DPG) may prevent the resealing of the ghost membrane. However, they exert this effect only at elevated temperatures and when present in the medium at the instant of hemolysis. At 0°C, the presence of complexing agents in the medium at the instant of hemolysis has no effect on the subsequent resealing at 37°C. The recovery of the ghost membrane takes place in spite of the continued presence of the agents and eventually leads to trapping of these agents inside the resealed ghosts.The experiments support the contention that the complexing agents interact with a membrane constituent which is neither accessible from the inner nor from the outer surface of the cell membrane but becomes exposed during the hemolytic event when the complexing agents penetrate across the membrane. Apparently, at low tempertrures membrane ligands are more successful in competing with the added complexing agents for this constituent than at higher temperatures.Extending former observations of Hoffman, we found that not only Mg⁺⁺ but also Ca⁺⁺ facilitates the resealing process. Perhaps one or the other of the two alkaline earth ions is the membrane constituent which normally participates in the maintenance of the integrity of the red blood cell membrane.     

黔西北土法炼锌废弃地植被重建的限制因子

以土法炼锌废弃地的废渣、污染土壤和背景土壤为基质材料,分别种植黑麦草(Lolium perenne)和三叶草(Trifolium pretense),分析各种基质的基本化学特性、重金属(Pb、Zn、Cd)含量及其赋存形态、两种植物生长特性.结果表明,土法炼锌废渣上植被重建的主要限制因子包括高盐碱胁迫、有机质含量低、养分缺乏(TN、碱解N、TK).废渣重金属含量高,有效态含量低,对植物毒性小,但存在潜在危害性.污染土壤重金属含量低于废渣,但生物有效态重金属含量高.污染土壤植被重建的限制因子包括重金属毒性、P和K的有效性.废渣与污染土壤混合是土法炼锌废弃地基质改良的有效途径.     

Bacterial Diversity in the Hyperalkaline Allas Springs (Cyprus), a Natural Analogue for Cementitious Radioactive Waste Repository

The biogeochemical gradients that will develop across the interface between a highly alkaline cementitious geological disposal facility for intermediate level radioactive waste and the geosphere are poorly understood. In addition, there is a paucity of information about the microorganisms that may populate these environments and their role in biomineralization, gas consumption and generation, metal cycling, and on radionuclide speciation and solubility. In this study, we investigated the phylogenetic diversity of indigenous microbial communities and their potential for alkaline metal reduction in samples collected from a natural analogue for cementitious radioactive waste repositories, the hyperalkaline Allas Springs (pH up to 11.9), Troodos Mountains, Cyprus. The site is situated within an ophiolitic complex of ultrabasic rocks that are undergoing active low-temperature serpentinization, which results in hyperalkaline conditions. 16S rRNA cloning and sequencing showed that phylogenetically diverse microbial communities exist in this natural high pH environment, including Hydrogenophaga species. This indicates that alkali-tolerant hydrogen-oxidizing microorganisms could potentially colonize an alkaline geological repository, which is predicted to be rich in molecular H₂, as a result of processes including steel corrosion and cellulose biodegradation within the wastes. Moreover, microbial metal reduction was confirmed at alkaline pH in this study by enrichment microcosms and by pure cultures of bacterial isolates affiliated to the Paenibacillus and Alkaliphilus genera. Overall, these data show that a diverse range of microbiological processes can occur in high pH environments, consistent with those expected during the geodisposal of intermediate level waste. Many of these, including gas metabolism and metal reduction, have clear implications for the long-term geological disposal of radioactive waste.     

Synthesis of symmetric dithiophosphinic acids for “minor actinide” extraction

New aromatic dithiophosphinic acid (R₂PS₂H; DPAH) derivatives were isolated using a synthetic pathway based on nucleophilic addition at phosphorus, which leads to regiospecific aromatic substituents on phosphorus. The synthesis improves DPAH designs that can provide insight into the differences in binding/chelation between DPAH and trivalent actinides/lanthanides. The synthesis gives good yields via isolation of the DPAH as the ammonium salt. Multinuclear NMR spectroscopy and X-ray structure determination are used for the identification of both the ammonium salt and free acid of the dithiophosphinic derivatives. The DPAH products are stable when exposed to the ambient atmospheric conditions for long periods. For instance, the bis(o-trifluoromethylphenyl) DPAH derivative showed very little degradation when contacted with water and nitric acid for prolonged periods. Furthermore, this derivative selectively extracts a trivalent actinide from a lanthanide with separation factors of ∼100 000 at low pH. Overall, these DPAH derivatives are exciting new compounds for potential application in actinide and lanthanide separations technologies.     

The greening process in cress seedlings. I. Pigment accumulation and ultrastructure after application of 5-aminolevulinate and complexing agents

A reduced rate of greening after continuous illumination was observed in dark-grown cress seedlings ( Lepidium sativum L.) incubated with 5-aminolevulinate (ALA) or the complexing agents 2,2'-bipyridyl, 8-hydroxyquinoline or 1,10-phenanthroline. This effect cannot be explained merely by photodynamic damage caused by chlorophyll precursors which are accumulated in the dark under these conditions. Flash light experiments revealed that photoconversion of protochlorophyll(ide) to chlorophyllide was not influenced by chelator treatment. The next step in the chlorophyll pathway, the esterification of chlorophyllide, however, was inhibited. Simultaneously applicated ALA and complexing agents did not result in a synergistic reponse; on the contrary, ALA seemed to render cress plants less susceptible to the treatment with complexing agents upon subsequent irradiation. Ultrastructural studies demonstrated that grana formation in light was inhibited after pretreatment with ALA or complexing agents.     

Influence of complexing agents on stability and activity.

Metal ion-complexing agents, like KCN, EDTA etc., inactivate alkaline phosphatase of pig kidney. This inactivation is reversible at low concentrations of the complexing agents and irreversible at high concentrations. The reversible inhibition is probably due to removal of Zn2+ ions from the active site, where they are necessary for catalytic action, whereas the irreversible inhibition results from the removal of Zn2+ ions necessary for preservation of the structure. The inactivation is pseudo-first order. It depends on the concentration, size and charge of the complexing agents. Beta-Glycerophosphate and Mg2+ ions protect the enzyme from inactivation by complexing agents. Quantitative examination of the effect of substrate leads to a model that is similar to the "sequential model" proposed by D.E. Koshland, G. Nemethy & D. Filmer (1966) (Biochemistry 5, 365-385) to explain allosteric behavior of enzymes. It describes the sequential addition of two substrate molecules at two active centres of the dimer enzyme. The binding of the substrate molecules is accompanied by changes in the conformation, which lead to stabilization of the enzyme against attack by complexing agents.     

Salt drying: a low‐cost,simple and efficient method for storing plants in the field and preserving biological repositories for DNA diversity research

Although a variety of methods have been optimized for the collection and storage of plant specimens, most of these are not suited for field expeditions for a variety of logistic reasons. Drying specimens with silica gel in polyethylene bags is currently the standard for field‐sampling methods that are suitable for subsequent DNA extraction. However, silica‐gel repositories are not readily available in remote areas, and its use is not very cost‐effective for the long‐term storage of collections or in developing countries with limited research budgets. Salting is an ancient and traditional drying process that preserves food samples by dehydrating tissues and inhibiting water‐dependent cellular metabolism. We compared salt and silica‐gel drying methods with respect to dehydration rates overtime, DNA quality and polymerase chain reaction(PCR) success to assess whether dry salting can be used as an effective plant preservation method for DNA analysis. Specimens from eleven plant species covering a variety of leaf structures, leaf thicknesses and water contents were analysed. Experimental work indicated that (i) levels of dehydration in sodium chloride were usually comparable to those obtained when silica gel was used, (ii) no spoilage, fungal or bacterial growth was observed for any of the species with all drying treatments and (iii) good yields of quality genomic DNA suitable for PCR applications were obtained in the salt‐drying treatments. The preservation of plant tissues in commercial table salt appears to be a satisfactory, and versatile method that may be suitable in remote areas where cryogenic resources and silica repositories are not available.     

Coordination chemical studies on metalloenzymes. Measurement of binding constant between apo-tyrosinase and copper ion

The behavior of complexing agents for the copper removal reaction was studied by the equilibrium dialysis method. In the copper removal reaction, complexing agents are divided into two types: those that are reducing agents and those that are not. Sodium cyanide and sodium thiosulfate are of the first type, and 8-hydroxyquinoline-5-sulfonic acid, 2,2′-bipyridyl, and picolinic acid are of the second type. From equilibrium dialysis with the first type of complexing agent, the apparent binding constant (pH 6.0) between cuprous ions and apotyrosinase was calculated to be 10¹⁵m^?1. Similarly, the apparent binding constant (pH 6.0) between cupric ions and apo-tyrosinase was about 10¹³m^?1, which was calculated from equilibrium dialysis with the second type of complexing agent. The apparent binding constant between cuprous ions and apo-tyrosinase was larger than that between cupric ions and apo-tyrosinase.     

Ligand-induced structural changes in amylose partially complexed with iodine

The influence of complexing agents such as methanol, ethanol, 1-propanol, 1-butanol, 1-pentanol, 1-hexanol, cyclohexanol and 2-octanol on the formation of a blue coloured amylose · iodine complex (pH 4.8), under suboptomum concentrations of iodine and in the absence of potassium iodide, is studied by recording the absorbance at 640 nm. A drop in absorbance at 640 nm accompanied by a blue shift in the spectrum (580–640 nm) was observed at higher concentration of the complexing agents. This behaviour of amylose partially complexed with iodine appears to be due to ligand-induced structural changes in the amylose chain. The fall in absorbance at 640 nm observed when the temeprature of amylose · oidine complex in the presence of complexing agents is raised, and the subsequent regeneration of the absorbance on cooling, indicates the possible helix to random coil transition of the amylose chain in an aqueous system.     

Effects of waterborne complexing agents on silver uptake and depuration in rainbow trout

Rainbow trout Oncorhynchus mykiss ( c . 60 g) were exposed for 1 week to 0·1 μM silver as AgNO₃ in ion poor water (Ca c . 150 μM, pH c . 8, water temperature 13° C) with or without waterborne organic matter (27 mg C l⁻¹ as Aldrich humic acid), thiosulphate (5 μM Na₂S₂O₃) or chloride (4 mM KCl). Organic matter decreased Ag accumulation by the gills initially, but did not decrease Ag accumulation by plasma or liver. Thiosulphate decreased the amount of Ag accumulated by the gills for the entire 1 week exposure but had no effect on Ag concentrations in the plasma, liver or bile. Chloride had no effect on Ag uptake in any of the tissues examined. All three complexing agents reduced the decreases in plasma Na and Cl concentrations caused by Ag. To study the effects of waterborne complexing agents on Ag depuration, rainbow trout were exposed to 0·1 μM AgNO₃ for 1 week then placed for 8 days in Ag‐free, ion poor water with or without waterborne organic matter (55 mg C l⁻¹) or thiosulphate (5 μM). These complexing agents did not alter depuration of Ag from the gills, plasma, liver or bile. Thus, once Ag has entered a fish, subsequent elimination of internal Ag is not affected by external complexing agents.     

Nuclear waste repositories in salt mines: a new approach to safety assessment

The long-term safety of radioactive waste repositories in rock-salt mines in the deep underground benefits significantly from the barrier effect of overlying rocks. The concentrations of radioactive substances released from the repository and migrating in the aquifer up to the biosphere are greatly reduced during passage through these rocks. In former safety analyses of waste repositories this transport has generally been modelled as a combination of the involved phenomena, e.g. convection, dispersion, adsorption, etc. The data required for a numerical evaluation of the overall effect are obtained either as (conservative) estimates based on experience or are empirical, based mainly on laboratory experiments. The approach presented here is much simpler and entirely empirical, and therefore more transparent. It makes use of the fact that the groundwater in the overlying rocks always contains dissolved salt from the salt formation and carries it continuously into the receiving channels or the drainage system. The relation between the total amount of dissolved solids present in a certain subsurface catchment area and their steady-state concentration in the receiving channels is assumed to be equivalent to the relation between the given amount of radionuclides released from the repository and their concentration in the receiving channels, the latter leading to a certain radiation exposure of the population. Two versions of this approach are discussed: version (a) assumes a continuous stream of radionuclides released from the repository, and version (b) assumes a pulse release of radionuclides from the repository. A simple calculation using data from the Gorleben exploration leads to the inequality C _max(W)≤7.85 ⋅ 10^–15 W(Bq/l) (1) where C _max is the maximum radionuclide concentration (with respect to time) in the receiving channels and W (Bq) is the amount of radionuclides released from the repository in a very short time. C _max obtained from (1), is supposed to be an upper limit of the radionuclide concentration in the receiving channel and, therefore, a conservative estimate. In some catchment areas the salt concentration in a small region near the surface is higher by a factor of <2 to 3 than the concentration in the receiving channel. In order to be conservative, this empirical factor may be used to calculate the concentration of radioactive substances. Surprisingly, the values of radiation exposure resulting from both versions of the new approach are far below those calculated by applying elaborated models of the transport processes. The respective factors are 10–100 in case (a) and as much as 1000 in case (b). The reasons for these significant differences are discussed. Received: 21 September 1995 / Accepted in revised form: 13 June 1996     

Complexing of glycofuranosides with calcium ion,and its effect on their acid-catalysed solvolysis

Stability constants for the calcium-ion complexes of several methyl aldo-furanosides have been determined in aqueous solution by using a potentiostatic technique with an electrode that is selective for calcium ions. The results obtained have been verified by examining the chromatographic behaviour of the compounds on a strong cation-exchange resin in the Ca²⁺ form. The rate constants for the acid-catalyzed hydrolysis and methanolysis of a few 4-chlorophenyl aldofuranosides having different complexing abilities have been determined at various concentrations of calcium chloride. The dependences of the observed salt effects on the complexing ability of the substrate are discussed.     

Bile Salt Hydrolase (Bsh) Activity Screening of Lactobacilli: In Vitro Selection of Indigenous Lactobacillus Strains with Potential Bile Salt Hydrolysing and Cholesterol-Lowering Ability

The bile salt hydrolase (Bsh) activity of probiotic bacterium residing in gastrointestinal tract has often being associated with its cholesterol-lowering effects. Hence, Bsh activity was explored in this study as the criterion for the selection of most potential Bsh-active and cholesterol-lowering indigenous Lactobacillus strains. Forty lactobacilli were adjudged Bsh active after a preliminary screening of 102 lactobacilli and occurrence of Bsh activity correlated well with their natural habitats. Of the 40 shortlisted lactobacilli, fifteen putative Lactobacillus strains were selected and further tested for their comparative Bsh activity. In the end, indigenous Lactobacillus plantarum strains Lp91 and Lp21 were emerged as the promising Bsh-active lactobacilli with their substrate preference inclined more towards glycocholate than other bile acid amino conjugates. In addition, strains Lp91 and Lp21 also exhibited significantly high bile salt deconjugation, cholesterol assimilation and cholesterol co-precipitation ability in vitro. In conclusion, indigenous L. plantarum strains Lp91 and Lp21 may be the promising candidate probiotics to elucidate the ecological significance of probiotic Bsh activity in vivo.     

Influence of the glass packing on the contamination of pharmaceutical products by aluminium. Part II: Amino acids for parenteral nutrition

The presence of aluminium in amino acids parenteral nutrition solutions can be related to the affinity of the amino acids for aluminium present in glass containers used for storage. For this study solutions of 19 amino acids used in parenteral nutrition were stored individually in glass flasks and the aluminium measured at determined time intervals. Solutions of complexing agents for aluminium, as ethylene-diaminetetraacetic acid, nitrilotriacetic acid, citrate, oxalate and fluoride ions were also stored in the same flasks and the aluminium measured during the same time interval. The measurements were made by electrothermal atomic absorption spectrometry. The aluminium content of the glass containers was also measured. The results showed that the glasses have from 0.6% to 0.8% Al. Only solutions of cysteine, cystine, aspartic acid and glutamic acid became contaminated by aluminium. As the same occurred with the complexing agents, aluminum can be released from glass due to an affinity of the substances for aluminium. Comparing the action of complexing agents and amino acids for which the stability constants of aluminium complex are known, it is possible to relate the magnitude of the stability constant with the aluminium leached from glass, the higher the stability constant, the higher the aluminium released. The analysis of commercial formulations with and without cysteine, cystine, glutamic acid or aspartic acid stored in glass containers confirms that the presence of these amino acids combined with the age of the soLution are, at least partially, responsible for the aluminium contamination. The resuLts demonstrated that the contamination is an ongoing process due to the presence of aluminium in glass combined with the affinity of some amino acids for this element.     

Temporal dynamics and degradation activity of an bacterial inoculum for treating waste metal-working fluid

In order for established bioreactors to be effective for treating chemically mixed wastes such as metal working fluids (MWF) it is essential that they harbour microbial populations that can maintain sufficient active biomass and degrade each of the chemical constituents present. In this study we investigated the effectiveness of a bacterial consortium composed of four species (Clavibacter michiganensis, Methylobacterium mesophilicum, Rhodococcus erythropolis and Pseudomonas putida), assembled on the basis of their apparent ubiquity in waste MWF, degradation ability and tolerance to fluctuating chemistry of the waste. The temporal dynamics of the inoculum and its effects on the fate of individual chemical components of the waste were studied, by regular sampling, over 400 h. Using a complementary approach of culture with chemotaxonomic (FAME) analysis and applying group specific probes (FISH), the inoculum was found to represent a significant component of the community in bioreactors with and without presence of indigenous MWF populations. In addition, the reduction in the COD by the consortium was approximately 85% of the total pollution load, and 30-40% more effectively than any other treatment (indigenous MWF community alone or activated sludge). Furthermore, all the chemical constituents, including the biocide (a formaldehyde release agent) demonstrated > 60% reduction. Many chemical components of the MWF proved to be recalcitrant in the other treatments. The results of this study confirm that assemblage of an inoculum, based on a comprehensive knowledge of the indigenous microbial community, in the target habitat, is a highly effective way of selecting microbial populations for bioaugmentation of bioreactors.     

Assessment of the performance of exotic and indigenous tree and shrub species for rehabilitating saline soils of Northern Kenya

In the semi‐desert zone of North Horr, northern Kenya, soil salinity reaches toxic levels, with extensive salt crusts at the surface. Environmental groups are involved in tree and shrub planting trials with the objective of stabilizing sand dunes and rehabilitating land. However, the performance of the various indigenous and exotic tree and shrub species in rehabilitating these saline soils has not been examined. Therefore this study aimed at evaluating the performance of various exotic and indigenous tree and shrub species in rehabilitating saline soils in North Horr. Mortality and relative growth rates for both exotic and indigenous tree and shrub species were assessed. Compared to indigenous species, exotic tree species (not naturally growing in the Marsabit ecosystem) failed to sustain an improved growth performance in saline soils of North Horr. The results of this study demonstrate that survival rates for indigenous plants in North Horr are higher than those of exotic ones. Thus, to rehabilitate saline soils and stabilize sand dunes, the use of indigenous tree and shrub species adapted to such saline conditions should be encouraged.     

Lipid production by Cryptococcus albidus using biowastes hydrolysed by indigenous microbes

The efficiency of Cryptococcusalbidus was evaluated for its abilities to assimilate onion and apple hydrolysates as a medium for lipid production. Onion waste (OW) and apple waste (AW) were hydrolysed at an organic load of 2% total solids by indigenous microbes under mesophilic conditions. The indigenous microbes effectively hydrolysed both wastes giving the highest reducing sugar content of 4.8 g/L and 10.8 g/L with OW and AW hydrolysates, respectively. The microbiome analysis revealed that most of the indigenous microbes belonged to genus Bacillus and a significant population of α-proteobacteria and γ-proteobacteria were also present. Cell retention culture of C. albidus at a dilution rate of 0.01 h⁻¹ resulted in a total dry cell weight (DCW) of 13.5 g/L with an intracellular lipid content of 20.0% at 168 h, corresponding to an enhancement of 3.48-folds and 2.37-folds in DCW and lipid concentration, respectively, as compared to batch fermentation.

     

Diversity of azospirillum strains isolated from rice plants grown in saline and nonsaline sites of coastal agricultural ecosystem

The diversity of indigenous Azospirillum spp. associated with rice cultivated along the coastline of Tamil Nadu was analyzed. Twelve sites with varying soil characteristics such as salinity, texture, and the host variety were chosen. Of the 402 strains isolated using NFB media, 302 were confirmed to be Azospirillum spp. and subjected to DNA polymorphism analysis using PCR-RFLP of 16S rDNA. They were also screened for their salt tolerance and microaerobic N2-fixing-dependent growth. On species identification, all the strains were found to be A. brasilense, A. lipoferum, or unidentified. On comparing the influence of the previously noted variability on the indigenous population, soil salinity was found to play a dominant role. This was revealed by PCR-RFLP studies and salt tolerance studies. A high association between soil salinity and the distribution of Azospirillum genotypes reveals that soil salinity should be taken into consideration while developing biofertilizers specifically for the coastal agricultural ecosystem.     

Bioremediation 3.0: Engineering pollutant-removing bacteria in the times of systemic biology

Elimination or mitigation of the toxic effects of chemical waste released to the environment by industrial and urban activities relies largely on the catalytic activities of microorganisms—specifically bacteria. Given their capacity to evolve rapidly, they have the biochemical power to tackle a large number of molecules mobilized from their geological repositories through human action (e.g., hydrocarbons, heavy metals) or generated through chemical synthesis (e.g., xenobiotic compounds). Whereas naturally occurring microbes already have considerable ability to remove many environmental pollutants with no external intervention, the onset of genetic engineering in the 1980s allowed the possibility of rational design of bacteria to catabolize specific compounds, which could eventually be released into the environment as bioremediation agents. The complexity of this endeavour and the lack of fundamental knowledge nonetheless led to the virtual abandonment of such a recombinant DNA-based bioremediation only a decade later. In a twist of events, the last few years have witnessed the emergence of new systemic fields (including systems and synthetic biology, and metabolic engineering) that allow revisiting the same environmental pollution challenges through fresh and far more powerful approaches. The focus on contaminated sites and chemicals has been broadened by the phenomenal problems of anthropogenic emissions of greenhouse gases and the accumulation of plastic waste on a global scale. In this article, we analyze how contemporary systemic biology is helping to take the design of bioremediation agents back to the core of environmental biotechnology. We inspect a number of recent strategies for catabolic pathway construction and optimization and we bring them together by proposing an engineering workflow.