Acidithiobacillus thiooxidans

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Acidithiobacillus thiooxidans
Acidithiobacillus thiooxidans CLST.webp
Acidithiobacillus thiooxidans strain CLST
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Domain: Bacteria
Phylum: Pseudomonadota
Class: Acidithiobacillia
Order: Acidithiobacillales
Family: Acidithiobacillaceae
Genus: Acidithiobacillus
Species:
A. thiooxidans
Binomial name
Acidithiobacillus thiooxidans
(Kelly and Wood 2000)
Type strain
DSM 17318
ATCC 19377T
DAMS
Synonyms

Thiobacillus concretivorus
Kelly & Harrison 1989
Thiobacillus thiooxidans
Kelly & Wood 2000

Contents

Acidithiobacillus thiooxidans, formerly known as Thiobacillus thiooxidans until its reclassification into the newly designated genus Acidithiobacillus of the Acidithiobacillia subclass of Pseudomonadota, [1] is a Gram-negative, rod-shaped bacterium that uses sulfur as its primary energy source. [2] It is mesophilic, with a temperature optimum of 28 °C. [2] This bacterium is commonly found in soil, sewer pipes, and cave biofilms called snottites. [2] A. thiooxidans is used in the mining technique known as bioleaching, where metals are extracted from their ores through the action of microbes.

Morphology

A. thiooxidans is a Gram-negative, rod-shaped bacterium with rounded ends that occurs in nature either as singlecells, as is the most common case, or sometimes in pairs, but rarely in triplets. [2] Its motility is due to a polar flagellum. [2] It is an obligate acidophile with an optimal pH less than 4.0, but it also qualifies as an obligate aerobe and chemolithotroph. [2] Described as a colorless, sulfur-oxidizing bacterium, A. thiooxidans does not accumulate sulfur either within or outside of its very small cells, which have an average size around 0.5  μm in diameter and 1 μm or less in length. [2]

Cultural characteristics

A. thiooxidans has so far not grown on agar or other solid media, instead it prefers liquid media with a strong, evenly dispersed clouding throughout, and it produces no sediment formation or surface growth. [2] Although it does not grow on traditional organic media, it will not be harmed by a medium containing peptone or glucose. [2] Media best suited for its growth are those that are inorganic and allow A. thiooxidans to use sulfur as a source of energy. [2] The following characteristic reactions accompany the growth of A. thiooxidans in the presence of tricalcium phosphate: the layer on the surface of the medium formed by sulfur tends to drop to the bottom, tricalcium phosphate is dissolved by the product of sulfur oxidation, sulfuric acid, giving soluble phosphate and CaSO4 + 2 H2O, and radiating monoclinic crystals that hang from the sulfur particles floating on the medium surface or protruding upward from the bottom are formed by the precipitation of calcium sulfate. [2] The medium becomes acidic with a pH around 2.8 and remains stationary until all the calcium phosphate has been dissolved. [2] Anything with the tendency to change the medium to an alkaline state would be considered harmful to the uniform growth of A. thiooxidans, but if it is left unharmed by an excess of acid or alkali, numerous consecutive generations may be kept alive on the liquid media. [2]

Temperature range

A.s thiooxidans thrives at an optimum temperature of 28-30 °C. [2] At lower temperatures (18 °C and below) and at 37 °C or higher, sulfur oxidation and growth are significantly slower, while temperatures between 55 and 60 °C are sufficient to kill the organism. [2]

Metabolism

A. thiobacillus, a strictly aerobic species, fixes CO2 from the atmosphere to meet its carbon requirements. [2] In addition, other essential nutrients are required in varying amounts. [2] A general lack of knowledge exists for acidophilic microorganisms in terms of the oxidation systems of reduced inorganic sulfur compounds (RISCs). [3] Fazzini et al.. (2013) presented the first experimentally validated stoichiometric model that was able to quantitatively assess the RISCs oxidation in A. thiooxidans (strain DSM 17318), the sulfur-oxidizing acidophilic chemolithotrophic archetype. By analyzing literature and by genomic analyses, a mix of formerly proposed models of RISCs oxidation were combined and evaluated experimentally, placing thiosulfate partial oxidation by the Sox system (SoxABXYZ), along with abiotic reactions, as the central steps of the sulfur oxidation model. [3] This model, paired with a detailed stoichiometry of the production of biomass, provides accurate predictions of bacterial growth. [3] This model, which has the potential to be used in biohydrometallurgical and environmental applications, constitutes an advanced instrument for optimizing the biomass production of A. thiooxidans. [3]

Essential nutrients

Carbon

A. thiooxidans derives all of the energy needed to satisfy its carbon requirement from the fixation of CO2. [2] An important distinction can be made between sulfur-oxidizing and nitrifying bacteria by their response to the introduction of carbon to the culture in the form of carbonates and bicarbonates. [2] Carbonates keep the medium alkaline, thus preventing growth of A. thiooxidans which grows best under acidic conditions, while bicarbonates have been shown to allow a healthy growth if kept in small concentrations. [2] Bicarbonate, however, is unnecessary because the CO2 from the atmosphere appears to be sufficient to support growth of A. thiooxidans, and would actually have an injurious effect in that it would tend to make the medium less acidic. [2]

Nitrogen

A. thiooxidans requires only small amounts of nitrogen due to its small amount of growth, but the best sources are ammonium salts of inorganic acids, especially sulfate, followed by the ammonium salts of organic acids, nitrates, asparagine, and amino acids. [2] If no nitrogen source is introduced into the medium, some growth is observed, with A. thiooxidans deriving the necessary nitrogen from either traces of atmospheric ammonia, distilled water, or the contamination of other salts. [2]

Oxygen

A. thiooxidans is obligately aerobic because it uses atmospheric oxygen for the oxidation of sulfur to sulfuric acid. [2]

Influence of organic substances

In the presence of a good nitrogen source, organic substances like glucose, glycerol, mannitol, and alcohol seem to either act similarly to stimulants or take part in the organism's structural requirements, causing no harm to A. thiooxidans and appearing to have somewhat of a favorable effect on it. [2]

Energy source

A. thiooxidans uses elemental sulfur as its primary energy source and oxidizes it by the sulfide-quinone reductase and sox pathways. [2] Sulfur is oxidized to sulfuric acid by A. thiooxidans and the energy liberated is used for growth and maintenance. [2] In addition to sulfur, A. thiooxidans can use thiosulfate or tetrathionate as sources of energy, but growth in a liquid medium on thiosulfate is slow, generally taking about 10 to 12 days under favorable conditions as opposed to only 4 to 5 days for growth on elemental sulfur, as demonstrated by the change in pH and turbidity. [2] A. thiooxidans is incapable of oxidizing iron or pyrite, but it has been shown to grow on sulfur from pyrite when cocultured with the bacterium Leptospirillum ferrooxidans , a species that can oxidize iron but not sulfur. [2]

A. thiooxidans is completely autotrophic and, although glucose does not cause any harm and can be beneficial to some extent, the amount of acid produced and sulfur oxidized are not significantly different between cultures that either contained or did not contain glucose. [2]

Autotrophy

As an autotrophic bacterium, A. thiooxidans uses inorganic substances to fulfill its energy requirement, and atmospheric carbon to satisfy its carbon demands. [2] Because A. thiooxidans derives its energy from inorganic elemental sulfur, carbon directly from the atmosphere, and nitrogen from ammonium sulfate and other inorganic salts, and also because of its small mineral requirements, this autotrophic microorganism was likely among the first aerobes contributing to weathering through the formation of sulfuric acid, which interacted with insoluble phosphates, carbonates, and silicates.

Phylogeny

Most of the information about Acidithiobacillus comes from experimental and genome-based analyses of two other related species, Acidithiobacillus: A. ferrooxidans and A. caldus . The complete draft genome sequence of A. thiooxidans ATCC 19377 was determined using a whole-genome shotgun strategy and was revealed to contain a total of 3,019,868 base pairs in 164 contigs. [4] The GC ratio was found to be 53.1% to 46.9%; [4] 3,235 protein-coding genes were predicted in the genome of A. thiooxidans, which also contained 43 tRNAs, one complete and one partial 5S-16S-23S operon, and complete sets of genes for amino acid, nucleotide, inorganic sulfur compound, and central carbon metabolism. [4] The genome also contains the genes sulfur quinone oxidoreductase (sqr), tetrathionate hydrolase (tetH), and thiosulfate quinone oxidoreductase (doxD), along with the two gene clusters that encode the sulfur oxidation complex SOX (soxYZB-hyp-resB-saxAX-resC and soxYZA-hyp-soxB), which were previously found in A. caldus and Thiobacillus denitrificans, a neutrophilic sulfur oxidizer. [4] Acidithiobacillus thiooxidans strains have been differentiated from other related Acidithiobacilli, including A. ferrooxidans and A. caldus, by sequence analyses of the PCR-amplified 16S-23S rDNA intergenic spacer (ITS) and restriction fragment length polymorphism (RFLP). [5] The strains of A. thiooxidans that were investigated by these researchers (metal mine isolates) yielded RFLP patterns that were identical to the A. thiooxidans type strain (ATCC 19377T), except for strain DAMS, which had a distinct pattern for all enzymes tested. [5] All three Acidithiobacillus species were differentiated by phylogenetic analysis of the ITS sequences. [5] The size and sequence polymorphism of the ITS3 region contributed to the inter- and infraspecific genetic variations that were detected in this analysis. [5] No significant correlation was shown by Mantel tests between the similarity of ITS sequences and the geographical origin of strains. [5] Bergamo et al.. (2004) concluded that the 16S-23S rDNA spacer region is a useful target for developing molecular methods that focus on the detection, rapid differentiation and identification of Acidithiobacillus species.

Snottites

Snottites are highly acidic biofilms (pH 0-1) that form on the walls and ceilings of hydrogen sulfide-rich caves where sulfide-rich springs gas H2S into the cave air. [6] The snottite microbial communities have very low species diversity and are predominantly composed of sulfur-oxidizing microorganisms. [7] Sulfide oxidation produces sulfuric acid, which dissolves the limestone walls of the cave. [6] Microcrystalline gypsum precipitates as a corrosion residue that eventually limits pH buffering by the underlying limestone and enables the development of extremely acidic wall surfaces. [6] A. thiooxidans is known to inhabit these biofilms. Snottite morphology and distribution within caves depends on the availability of carbon, nitrogen, and energy substrates in the atmosphere. [6] Snottite formations are generally milky in color, suspended vertically from cave ceilings and walls, and have a phlegm-like consistency (hence the name). [8]

Frasassi cave system, Italy

Le Grotte di Frasassi (Frasassi Caves) are located in the Apennine Mountains in the Marches Region, central Italy. [7] This cave system was formed by the process of sulfuric acid speleogenesis due to sulfide-oxidizing microorganisms. [7] The snottites within the Frasassi Caves are very viscous with a pH range of 0-2.5. [6] The most abundant bacterial 16S rRNA sequences (>98% 16S rRNA similarity) in snottites collected throughout the Frasassi cave system are relatives of A. thiooxidans and the genera Acidimicrobium and Ferrimicrobium (family Acidimicrobiaceae, Actinobacteria). [6] FISH analyses of snottite samples have indicated that Acidithiobacillus and the Acidimicrobiaceae are the most abundant bacterial populations within the caves. [6] Populations of biofilms in the Frasassi cave system are dominated by A. thiooxidans (>70% of cell population) with smaller populations including an archaeon in the uncultivated G-plasma clade of Thermoplasmatales (>15%) and a bacterium in the family Acidimicrobiaceae (>5%). [6] Acidithiobacillus is believed to be the primary producer and the snottite architect. [6]

Bioleaching

Bioleaching is a mining technique in which metals are extracted from their insoluble ores through the use of living organisms by biological oxidation. This technique has progressed steadily in the past 20 years by taking advantage of bacteria such as A. thiooxidans. Biomining operations have enabled the solubilization of low-grade mineral ores. Compared to traditional smelting and extracting procedures, bioleaching is much less expensive and does not release as many environmental toxicants, but it does require a greater amount of time. Bioleaching involves at least three important subprocesses, viz., attack of the sulfide mineral, microbial oxidation of ferrous iron, and some sulfur moiety. [9] The overall process occurs via one of two pathways depending on the nature of the sulfide mineral, a pathway via thiosulfate resulting in sulfate being formed or a polythionate pathway resulting in the formation of elemental sulfur. [9]


Related Research Articles

Bioleaching is the extraction or liberation of metals from their ores through the use of living organisms. Bioleaching is one of several applications within biohydrometallurgy and several methods are used to treat ores or concentrates containing copper, zinc, lead, arsenic, antimony, nickel, molybdenum, gold, silver, and cobalt.

<span class="mw-page-title-main">Green sulfur bacteria</span> Family of bacteria

The green sulfur bacteria are a phylum, Chlorobiota, of obligately anaerobic photoautotrophic bacteria that metabolize sulfur.

<span class="mw-page-title-main">Snottite</span> Microbial mat often found in caves

Snottite, also snoticle, is a microbial mat of single-celled extremophilic bacteria which hang from the walls and ceilings of caves and are similar to small stalactites, but have the consistency of nasal mucus. In the Frasassi Caves in Italy, over 70% of cells in Snottite have been identified as Acidithiobacillus thiooxidans, with smaller populations including an archaeon in the uncultivated 'G-plasma' clade of Thermoplasmatales (>15%) and a bacterium in the Acidimicrobiaceae family (>5%).

Bacteria biooxidation is an oxidation process caused by microbes where the valuable metal remains in the solid phase. In this process, the metal remains in the solid phase and the liquid can be discarded. Bacterial oxidation is a biohydrometallurgical process developed for pre-cyanidation treatment of refractory gold ores or concentrates. The bacterial culture is a mixed culture of Acidithiobacillus ferrooxidans, Acidithiobacillus thiooxidans and Leptospirillum ferrooxidans. The bacterial oxidation process comprises contacting refractory sulfide ROM ore or concentrate with a strain of the bacterial culture for a suitable treatment period under an optimum operating environment. The bacteria oxidise the sulfide minerals, thus liberating the occluded gold for subsequent recovery via cyanidation.

Thiobacillus is a genus of Gram-negative Betaproteobacteria. Thiobacillus thioparus is the type species of the genus, and the type strain thereof is the StarkeyT strain, isolated by Robert Starkey in the 1930s from a field at Rutgers University in the United States of America. While over 30 "species" have been named in this genus since it was defined by Martinus Beijerinck in 1904,, most names were never validly or effectively published. The remainder were either reclassified into Paracoccus, Starkeya ; Sulfuriferula, Annwoodia, Thiomonas ; Halothiobacillus, Guyparkeria, or Thermithiobacillus or Acidithiobacillus. The very loosely defined "species" Thiobacillus trautweinii was where sulfur oxidising heterotrophs and chemolithoheterotrophs were assigned in the 1910-1960s era, most of which were probably Pseudomonas species. Many species named in this genus were never deposited in service collections and have been lost.

<i>Acidithiobacillus</i> Genus of bacteria

Acidithiobacillus is a genus of the Acidithiobacillia in the phylum "Pseudomonadota". This genus includes ten species of acidophilic microorganisms capable of sulfur and/or iron oxidation: Acidithiobacillus albertensis, Acidithiobacillus caldus, Acidithiobacillus cuprithermicus, Acidithiobacillus ferrianus, Acidithiobacillus ferridurans, Acidithiobacillus ferriphilus, Acidithiobacillus ferrivorans, Acidithiobacillus ferrooxidans, Acidithiobacillus sulfuriphilus, and Acidithiobacillus thiooxidans.A. ferooxidans is the most widely studied of the genus, but A. caldus and A. thiooxidans are also significant in research. Like all "Pseudomonadota", Acidithiobacillus spp. are Gram-negative and non-spore forming. They also play a significant role in the generation of acid mine drainage; a major global environmental challenge within the mining industry. Some species of Acidithiobacillus are utilized in bioleaching and biomining. A portion of the genes that support the survival of these bacteria in acidic environments are presumed to have been obtained by horizontal gene transfer.

<span class="mw-page-title-main">Sulfur-reducing bacteria</span> Microorganisms able to reduce elemental sulfur to hydrogen sulfide

Sulfur-reducing bacteria are microorganisms able to reduce elemental sulfur (S0) to hydrogen sulfide (H2S). These microbes use inorganic sulfur compounds as electron acceptors to sustain several activities such as respiration, conserving energy and growth, in absence of oxygen. The final product of these processes, sulfide, has a considerable influence on the chemistry of the environment and, in addition, is used as electron donor for a large variety of microbial metabolisms. Several types of bacteria and many non-methanogenic archaea can reduce sulfur. Microbial sulfur reduction was already shown in early studies, which highlighted the first proof of S0 reduction in a vibrioid bacterium from mud, with sulfur as electron acceptor and H
2
as electron donor. The first pure cultured species of sulfur-reducing bacteria, Desulfuromonas acetoxidans, was discovered in 1976 and described by Pfennig Norbert and Biebel Hanno as an anaerobic sulfur-reducing and acetate-oxidizing bacterium, not able to reduce sulfate. Only few taxa are true sulfur-reducing bacteria, using sulfur reduction as the only or main catabolic reaction. Normally, they couple this reaction with the oxidation of acetate, succinate or other organic compounds. In general, sulfate-reducing bacteria are able to use both sulfate and elemental sulfur as electron acceptors. Thanks to its abundancy and thermodynamic stability, sulfate is the most studied electron acceptor for anaerobic respiration that involves sulfur compounds. Elemental sulfur, however, is very abundant and important, especially in deep-sea hydrothermal vents, hot springs and other extreme environments, making its isolation more difficult. Some bacteria – such as Proteus, Campylobacter, Pseudomonas and Salmonella – have the ability to reduce sulfur, but can also use oxygen and other terminal electron acceptors.

Lithotrophs are a diverse group of organisms using an inorganic substrate to obtain reducing equivalents for use in biosynthesis or energy conservation via aerobic or anaerobic respiration. While lithotrophs in the broader sense include photolithotrophs like plants, chemolithotrophs are exclusively microorganisms; no known macrofauna possesses the ability to use inorganic compounds as electron sources. Macrofauna and lithotrophs can form symbiotic relationships, in which case the lithotrophs are called "prokaryotic symbionts". An example of this is chemolithotrophic bacteria in giant tube worms or plastids, which are organelles within plant cells that may have evolved from photolithotrophic cyanobacteria-like organisms. Chemolithotrophs belong to the domains Bacteria and Archaea. The term "lithotroph" was created from the Greek terms 'lithos' (rock) and 'troph' (consumer), meaning "eaters of rock". Many but not all lithoautotrophs are extremophiles.

Microbial metabolism is the means by which a microbe obtains the energy and nutrients it needs to live and reproduce. Microbes use many different types of metabolic strategies and species can often be differentiated from each other based on metabolic characteristics. The specific metabolic properties of a microbe are the major factors in determining that microbe's ecological niche, and often allow for that microbe to be useful in industrial processes or responsible for biogeochemical cycles.

Biomining ( phytomining) is the concept of extracting metals from ores and other solid materials typically using prokaryotes, fungi or plants (phytoextraction. These organisms secrete organic compounds that chelate metals from the environment. The proposed technology is often aimed at extraction of iron, copper, zinc, gold, uranium, and thorium. Large chemostats of microbes can be grown to leach metals from their media. If it were practical, biomining would be an environmentally friendly alternative to traditional mining.

<span class="mw-page-title-main">Thiosulfate dehydrogenase</span>

Thiosulfate dehydrogenase is an enzyme that catalyzes the chemical reaction:

Thermithiobacillus tepidarius is a member of the Acidithiobacillia isolated from the thermal groundwaters of the Roman Baths at Bath, Somerset, United Kingdom. It was previously placed in the genus Thiobacillus. The organism is a moderate thermophile, 43–45 °C (109–113 °F), and an obligate aerobic chemolithotrophic autotroph. Despite having an optimum pH of 6.0–7.5, growth can continue to an acid medium of pH 4.8. Growth can only occur on reduced inorganic sulfur compounds and elementary sulfur, but unlike some species in other genus of the same family, Acidithiobacillus, Thermithiobacillus spp. are unable to oxidise ferrous iron or iron-containing minerals.

Sulfur is metabolized by all organisms, from bacteria and archaea to plants and animals. Sulfur can have an oxidation state from -2 to +6 and is reduced or oxidized by a diverse range of organisms. The element is present in proteins, sulfate esters of polysaccharides, steroids, phenols, and sulfur-containing coenzymes.

<span class="mw-page-title-main">Acidophiles in acid mine drainage</span>

The outflow of acidic liquids and other pollutants from mines is often catalysed by acid-loving microorganisms; these are the acidophiles in acid mine drainage.

<span class="mw-page-title-main">Rusticyanin</span>

Rusticyanin (RCN) is a copper protein with a type I copper center that plays an integral role in electron transfer. It can be extracted from the periplasm of the gram-negative bacterium Thiobacillus ferrooxidans, also known as Acidithiobacillus ferrooxidans. Rusticyanin is also found in the membrane-bound form in the surface of T. ferrooxidans. It is a part of an electron transfer chain for Fe(II) oxidation.

Leptospirillum ferriphilum is an iron-oxidising bacterium able to exist in environments of high acidity, high iron concentrations, and moderate to moderately high temperatures. It is one of the species responsible for the generation of acid mine drainage and the principal microbe used in industrial biohydrometallurgy processes to extract metals.

Acidithiobacillus caldus formerly belonged to the genus Thiobacillus prior to 2000, when it was reclassified along with a number of other bacterial species into one of three new genera that better categorize sulfur-oxidizing acidophiles. As a member of the Gammaproteobacteria class of Pseudomonadota, A. caldus may be identified as a Gram-negative bacterium that is frequently found in pairs. Considered to be one of the most common microbes involved in biomining, it is capable of oxidizing reduced inorganic sulfur compounds (RISCs) that form during the breakdown of sulfide minerals. The meaning of the prefix acidi- in the name Acidithiobacillus comes from the Latin word acidus, signifying that members of this genus love a sour, acidic environment. Thio is derived from the Greek word thios and describes the use of sulfur as an energy source, and bacillus describes the shape of these microorganisms, which are small rods. The species name, caldus, is derived from the Latin word for warm or hot, denoting this species' love of a warm environment.

Acidithrix ferrooxidans is a heterotrophic, acidophilic and Gram-positive bacterium from the genus Acidithrix. The type strain of this species, A. ferrooxidans Py-F3, was isolated from an acidic stream draining from a copper mine in Wales. This species grows in a variety of acidic environments such as streams, mines or geothermal sites. Mine lakes with a redoxcline support growth with ferrous iron as the electron donor. "A. ferrooxidans" grows rapidly in macroscopic streamer, producing greater cell densities than other streamer-forming microbes. Use in a bioreactors to remediate mine waste has been proposed due to cell densities and rapid oxidation of ferrous iron oxidation in acidic mine drainage. Exopolysaccharide production during metal substrate metabolism, such as iron oxidation helps to prevent cell encrustation by minerals.

<span class="mw-page-title-main">Microbial oxidation of sulfur</span>

Microbial oxidation of sulfur is the oxidation of sulfur by microorganisms to build their structural components. The oxidation of inorganic compounds is the strategy primarily used by chemolithotrophic microorganisms to obtain energy to survive, grow and reproduce. Some inorganic forms of reduced sulfur, mainly sulfide (H2S/HS) and elemental sulfur (S0), can be oxidized by chemolithotrophic sulfur-oxidizing prokaryotes, usually coupled to the reduction of oxygen (O2) or nitrate (NO3). Anaerobic sulfur oxidizers include photolithoautotrophs that obtain their energy from sunlight, hydrogen from sulfide, and carbon from carbon dioxide (CO2).

Thiosocius is a genus of bacteria that lives in symbiosis with the giant shipworm Kuphus polythalamius. It contains a single species, Thiosocius teredinicola, which was isolated from the gills of the shipworm. The specific name derives from the Latin terms teredo (shipworm) and incola (dweller).

References

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