Bioenergetic aspects of halophilism.

Examination of microbial diversity in environments of increasing salt concentrations indicates that certain types of dissimilatory metabolism do not occur at the highest salinities. Examples are methanogenesis for H2 + CO2 or from acetate, dissimilatory sulfate reduction with oxidation of acetate, and autotrophic nitrification. Occurrence of the different metabolic types is correlated with the free-energy change associated with the dissimilatory reactions. Life at high salt concentrations is energetically expensive. Most bacteria and also the methanogenic Archaea produce high intracellular concentrations of organic osmotic solutes at a high energetic cost. All halophilic microorganisms expend large amounts of energy to maintain steep gradients of NA+ and K+ concentrations across their cytoplasmic membrane. The energetic cost of salt adaptation probably dictates what types of metabolism can support life at the highest salt concentrations. Use of KCl as an intracellular solute, while requiring far-reaching adaptations of the intracellular machinery, is energetically more favorable than production of organic-compatible solutes. This may explain why the anaerobic halophilic fermentative bacteria (order Haloanaerobiales) use this strategy and also why halophilic homoacetogenic bacteria that produce acetate from H2 + CO2 exist whereas methanogens that use the same substrates in a reaction with a similar free-energy yield do not.     

Bioenergetic challenges of microbial iron metabolisms

Before cyanobacteria invented oxygenic photosynthesis and O(2) and H(2)O began to cycle between respiration and photosynthesis, redox cycles between other elements were used to sustain microbial metabolism on a global scale. Today these cycles continue to occur in more specialized niches. In this review we focus on the bioenergetic aspects of one of these cycles - the iron cycle - because iron presents unique and fascinating challenges for cells that use it for energy. Although iron is an important nutrient for nearly all life forms, we restrict our discussion to energy-yielding pathways that use ferrous iron [Fe(II)] as an electron donor or ferric iron [Fe(III)] as an electron acceptor. We briefly review general concepts in bioenergetics, focusing on what is known about the mechanisms of electron transfer in Fe(II)-oxidizing and Fe(III)-reducing bacteria, and highlight aspects of their bioenergetic pathways that are poorly understood.     

Thermophilic microbial leaching of heavy metals from municipal sludge using indigenous sulphur-oxidizing microbiota

 It was demonstrated in shake-flask experiments that the sulphur-oxidizing microbiota of municipal sludges can be used at 53°C for heavy-metal leaching. Five sludges were tested and the average final pH, oxidation/reduction potential and SO^2- ₄ concentration after 30 days were 2.8, 237 mV and 5668 mg/l respectively. Ferric chloride was added to enhance the redox potential and to lower pH, which resulted in average values of 409 mV and 1.86 respectively. The average solubilisation of Cd, Cu, Cr, Mn, Ni, Pb, Zn, K and P after ferric chloride addition was 35.6±20.6%, 65.9±15.1%, 28.5±13.5%, 74.0±10.0%, 60.3± 13.1%, 33.7±27.6%, 83.9±6.2%, 39.0±16.6 and 18.2±15.8% respectively. The present process enhanced the sludge dewaterability compared to the conventional thermophilic digestion. During the leaching batch process, the volatile and volatile suspended solids were degraded to the same level as observed when the conventional thermophilic digestion was used as control. Received: 7 June 1995/Received revision: 8 September 1995/Accepted: 29 September 1995     

Bioenergetic aspects of apoptosis, necrosis and mitoptosis

In this review I summarize interrelations between bioenergetic processes and such programmed death phenomena as cell suicide (apoptosis and necrosis) and mitochondrial suicide (mitoptosis). The following conclusions are made. (I) ATP and rather often mitochondrial hyperpolarization (i.e. an increase in membrane potential, ΔΨ) are required for certain steps of apoptosis and necrosis. (II) Apoptosis, even if it is accompanied by ΔΨ and [ATP] increases at its early stage, finally results in a ΔΨ collapse and ATP decrease. (III) Moderate (about three-fold) lowering of [ATP] for short and long periods of time induces apoptosis and necrosis, respectively. In some types of apoptosis and necrosis, the cell death is mediated by a ΔΨ-dependent overproduction of ROS by the initial (Complex I) and the middle (Complex III) spans of the respiratory chain. ROS initiate mitoptosis which is postulated to rid the intracellular population of mitochondria from those that are ROS overproducing. Massive mitoptosis can result in cell death due to release to cytosol of the cell death proteins normally hidden in the mitochondrial intermembrane space.     

Bacterial leaching of metals from sewage sludge

Summary Thiobacillus thiooxidans is capable of oxidizing sulfur in digested sludge, while decreasing the pH value from about 5.5 to, say, 1.0 to 1.5. Insoluble metal sulfides can be solubilized through this acidification. Thiobacillus ferrooxidans oxidises pyritic ore in the presence of 6% centrifuged sludge if the pH value is adjusted to about 2.5. When mixing T. thiooxidans and T. ferrooxidans with sludge and 1% sulfur, the former acidifies the sludge and the latter oxidizes metal sulfides; together they solubilize more metal than T. thiooxidans alone. The following metals solubilized from their sulfides have been investigated so far: iron, copper, zinc, nickel, and cadmium. The possibility of recycling metals from sewage sludge with this method is discussed.     

Microbial leaching of metals from sulfide minerals

Microorganisms are important in metal recovery from ores, particularly sulfide ores. Copper, zinc, gold, etc. can be recovered from sulfide ores by microbial leaching. Mineral solubilization is achieved both by 'direct (contact) leaching' by bacteria and by 'indirect leaching' by ferric iron (Fe(3+)) that is regenerated from ferrous iron (Fe(2+)) by bacterial oxidation. Thiobacillus ferrooxidans is the most studied organism in microbial leaching, but other iron- or sulfide/sulfur-oxidizing bacteria as well as archaea are potential microbial agents for metal leaching at high temperature or low pH environment. Oxidation of iron or sulfur can be selectively controlled leading to solubilization of desired metals leaving undesired metals (e.g., Fe) behind. Microbial contribution is obvious even in electrochemistry of galvanic interactions between minerals.     

Microbial leaching of metals from solid industrial wastes

Biotechnological applications for metal recovery have played a greater role in recovery of valuable metals from low grade sulfide minerals from the beginning of the middle era till the end of the twentieth century. With depletion of ore/minerals and implementation of stricter environmental rules, microbiological applications for metal recovery have been shifted towards solid industrial wastes. Due to certain restrictions in conventional processes, use of microbes has garnered increased attention. The process is environmentally-friendly, economical and cost-effective. The major microorganisms in recovery of heavy metals are acidophiles that thrive at acidic pH ranging from 2.0–4.0. These microbes aid in dissolving metals by secreting inorganic and organic acids into aqueous media. Some of the well-known acidophilic bacteria such as Acidithiobacillus ferrooxidans, Acidithiobacillus thiooxidans, Leptospirillum ferrooxidans and Sulfolobus spp. are well-studied for bioleaching activity, whereas, fungal species like Penicillium spp. and Aspergillus niger have been thoroughly studied for the same process. This mini-review focuses on the acidophilic microbial diversity and application of those microorganisms toward solid industrial wastes.     

Selected aspects of microbial osmoregulation

Abstract Some salient characteristics of microbial osmoregulation are reviewed, with specific examples drawn from eukaryotes. As well as the need for an osmoregulatory solute to be 'compatible' with cellular processes under all conditions, the importance of the physiological method of regulating the content of the solute as a factor determining xerotolerance is emphasized. The significance of turgor/volume homeostasis is discussed and examples are cited in which, during exponential growth, there is apparently no homeostatic control of the cellular content of the major osmoregulatory solute. Some implications of this for the overall mechanism of osmoregulation are considered.
A recent experiment is described which raises questions about the timing of an osmoregulatory 'signal' in Saccharomyces cerevisiae . Other experiments are summarized which distinguish between osmoregulatory and compatible solutes in yeast. These experiments implicate trehalose as a non-osmoregulatory compatible solute in certain circumstances.     

Quantitative assessment of the effects of metals on microbial degradation of organic chemicals.

Biodegradation inhibition of a benchmark chemical, 2,4-dichloro-phenoxyacetic acid methyl ester (2,4-DME), was used to quantify the inhibitory effects of heavy metals on aerobic microbial degradation rates of organic chemicals. This procedure used lake sediments and aufwuchs (floating mats) collected in the field or from laboratory microcosms. Effects of CuCl2, HgCl2, ZnCl2, Cd(NO3)2, and Cr(NO3)3 at initial concentrations ranging from 0.3 microM to 73 mM (approximately 0.1 to 10,000 mg liter-1) were investigated. In general, such metallic compounds appeared to be considerably more inhibitory to the biodegradation of an organic chemical than high concentrations of microbially toxic organics studied previously. Effects of various metal concentrations were evaluated based on the following: (i) estimated MICs, (ii) concentrations that caused a significant effect on biodegradation parameters (both a greater than 10% decrease in Vmax and a greater than 10% increase in t1/2 for 2,4-DME degradation), and (iii) concentrations that caused biodegradation half-life doublings (HLDs). The MICs of metals in sediment were lowest for Zn2+ (0.10 microM) and highest for Cd2+ and Cu2+ (0.9 and 1.2 microM, respectively). The MICs of metals in aufwuchs were lowest for Hg2+ (0.01 microM), intermediate for Cu2+ and Zn2+ (0.42 and 0.62 microM, respectively), and highest for Cr3+ and Cd2+ (3.4 and 5.6 microM, respectively). Compared with Cu2+ on aufwuchs, 70 times more Zn2+, 250 times more Cr3+, and 1,000 times more Cd2+ was required to significantly affect aufwuchs biodegradation rate parameters and coefficients (Vmax and t1/2). Aufwuchs was significantly affected by the lowest Hg2+ concentration tested (5 microM).(ABSTRACT TRUNCATED AT 250 WORDS)     

Rhizoremediation of metals: harnessing microbial communities

With the increasing successful stories of decontamination, different strategies for metal remediation are gaining importance and popularization in developing countries. Rhizoremediation, is one such promising option that harnesses the impressive capabilities of microorganisms associated with roots to degrade organic pollutants and transform toxic metals. Since it is a plant based in-situ phytorestoration technique it is proven to be economical, efficient and easy to implement under field conditions. Plants grown in metal contaminated sites harbor unique metal tolerant and resistant microbial communities in their rhizosphere. These rhizo-microflora secrete plant growth promoting substances, siderophores, phytochelators to alleviate metal toxicity, enhance the bioavailability of metals (phytoremediation) and complexation of metals (phytostabilisation). Selection of right bacteria/consortia and inoculation to seed/ roots of suitable plant species will widen the perspectives of rhizoremediation.     

Molecular aspects of microbial ice nucleation

Certain organisms nucleate the crystallization of ice. This requires a small volume of water to be induced, probably by lattice-matching with a solid template, to form an 'ice embryo'--a region sharing at least some of the characteristics of macroscopic ice. It is of particular interest to understand the structure and function of biological structures capable of lattice-matching (or otherwise inducing a quasi-crystalline state). Some strains of the Gram-negative eubacterial genera Erwinia, Pseudomonas, and Xanthomonas, and the mycobionts of certain lichens, display ice-nucleating activity. In bacteria, the activity is conferred by a protein that contains three nested periodicities of repetition, which probably reflects a hierarchy of three motifs of structural repetition. Thus the tertiary structure of the ice-nucleation protein is likely to be regular, consistent with the expectation of its forming a template for lattice-matching. Even within a clonal culture, the nucleating sites formed by bacteria and lichens vary considerably in the threshold temperatures at which they display activity; this indicates wide variations in either the size of the template, or its structural regularity, or both. However, ice-nucleating sites of lichen and bacterial origin are clearly differentiated by their sensitivities to experimental treatments.     

Plant uptake and the leaching of metals during the hot EDDS-enhanced phytoextraction process

Using pot experiments, the effect of the application of the biodegradable chelating agent S,S-ethylenediaminedisuccinic acid (EDDS) in hot solutions at 90 degrees C on the uptake of Cu, Pb, Zn, and Cd by corn (Zea mays L. cv. Nongda No. 108) and beans (P vulgaris L. white bean), and the potential leaching of metals from soil, were studied. When EDDS was applied as a hot solution at the rate of 1 mmol kg(-1), the concentrations and total phytoextraction of metals in plant shoots exceeded or approximated those in the shoots of plants treated with normal EDDS at the rate of 5 mmol kg(-1). On the other hand, the leaching of Cu, Pb, Zn, and Cd after the application of the hot EDDS solution at the rate of 1 mmol kg(-1) was reduced by 46%, 21%, 57%, and 35% in comparison with that from the application of normal EDDS at 5 mmol kg(-1), respectively. For treatment with 1 mmol kg(-1) of EDDS, the leached metals decreased to the levels of the control group (that without EDDS amendment) 14 d after the application of EDDS. The soil amendment with biodegradable EDDS in hot solutions may provide a good alternative to chelate-enhanced phytoextraction in enhancing metal uptake by plants and limiting metals from leaching out of the soil.     

Bioenergetic aspects of aerobic glucose metabolism of Escherichia coli K-12 under varying specific growth rates and glucose concentrations.

An attempt was made to find a bioenergetical explanation for the differential effect of specific growth rate and glucose concentration on glucose metabolism of Escherichia coli K-12 with the help of 2,4-dinitrophenol (DNP). The effect of DNP on biomass occurred only at high glucose concentrations. The presence of this uncoupler strongly stimulated glucose uptake rates and oxygen uptake rates, but repressed severly Yg values. Increase in glucose concentration, however, sharply decreased QO2. The amount of oxygen required for maintenance was not affected by DNP, but Yomax values were much lower in the presence of DNP. The results are discussed and it is suggested that aerobic fermentation is caused by a severe reduction of site 1 of the respiratory chain region, whereas biomass formation is affected by repression of the terminal cytochrome a2. In comparing the effect of glucose on biomass formation at similar Qglucose levels aerobic and anaerobic fermentation, repression occurred in both cases at glucose concentrations of 0.3% and above. Although the analyses of 15 enzymes established the metabolic differences, the repression of growth was common to both fermentation types.     

Continuous microbial leaching of a pyritic concentrate by Leptospirillum-like bacteria

Summary Continuous leaching of a pyritic flotation concentrate by mixed cultures of acidophilic bacteria was studied in a laboratory scale airlift reactor. Enrichment cultures adapted to the flotation concentrate contained Thiobacillus ferrooxidans and Thiobacillus thiooxidans. During the late stationary growth phase of these thiobacilli growth of Leptospirillum-like bacteria was observed, too. In discontinuous cultivation no significant influence of Leptospirillum-like bacteria on leaching rates could be detected. During continuous leaching at pH 1.5 Leptospirillum-like bacteria displaced Thiobacillus ferrooxidans. The iron leaching rate achieved by Leptospirillum-rich cultures was found to be up to 3.9 times higher than that by Leptospirillum-free cultures.     

Bioenergetic aspects of aerobic growth of Klebsiella aerogenes NCTC 418 in carbon-limited and carbon-sufficient chemostat culture

Carbon-limited chemostat cultures of Klebsiella aerogenes NCTC 418 consumed more oxygen per unit of cell synthesis when growing on mannitol or glycerol than when growing on glucose; and since the maintenance requirements were similar, this suggested that the extra reducing equivalents present in these compounds were oxidized wastefully. By comparison with carbon-limited cultures, carbon-sufficient cultures that were ammonia-, sulphate- or phosphate-limited generally consumed considerably more oxygen per unit of cell synthesis, particularly at low growth rates. Thus, according to the theory of Pirt, these carbon-sufficient cultures had a greatly increased maintenance energy requirement but nevertheless used the remaining energy with a much increased efficiency compared with carbon-limited cultures. This, we suggest, is a false conclusion which stems from the basic assumption that the maintenance requirement does not change with growth rate. Thus we propose an alternative theory which allows for this possibility, and present evidence to show that it may be applicable to both carbon-limited and carbon-sufficient chemostat cultures. Finally we offer an explanation of the high maintenance rate of oxygen consumption found with carbon-sufficient cultures, and consider this phenomenon in relation to the loose coupling between respiration and growth extant in most microbial cultures.     

Bioenergetic Aspects of Halophilism

Examinination of microbial diversity in environments of increasing salt concentrations indicates that certain types of dissimilatory metabolism do not occur at the highest salinities. Examples are methanogenesis for H₂ + CO₂ or from acetate, dissimilatory sulfate reduction with oxidation of acetate, and autotrophic nitrification. Occurrence of the different metabolic types is correlated with the free-energy change associated with the dissimilatory reactions. Life at high salt concentrations is energetically expensive. Most bacteria and also the methanogenic archaea produce high intracellular concentrations of organic osmotic solutes at a high energetic cost. All halophilic microorganisms expend large amounts of energy to maintain steep gradients of NA⁺ and K⁺ concentrations across their cytoplasmic membrane. The energetic cost of salt adaptation probably dictates what types of metabolism can support life at the highest salt concentrations. Use of KCl as an intracellular solute, while requiring far-reaching adaptations of the intracellular machinery, is energetically more favorable than production of organic-compatible solutes. This may explain why the anaerobic halophilic fermentative bacteria (order Haloanaerobiales) use this strategy and also why halophilic homoacetogenic bacteria that produce acetate from H₂ + CO₂ exist whereas methanogens that use the same substrates in a reaction with a similar free-energy yield do not.