Enzymes synthesized by hyperthermophiles (bacteria and archaea with optimal growth temperatures of >80°C), also called hyperthermophilic enzymes, are typically thermostable (i.e., resistant to irreversible inactivation at high temperatures) and are optimally active at high temperatures. These enzymes share the same catalytic mechanisms with their mesophilic counterparts. When cloned and expressed in mesophilic hosts, hyperthermophilic enzymes usually retain their thermal properties, indicating that these properties are genetically encoded. Sequence alignments, amino acid content comparisons, crystal structure comparisons, and mutagenesis experiments indicate that hyperthermophilic enzymes are, indeed, very similar to their mesophilic homologues. No single mechanism is responsible for the remarkable stability of hyperthermophilic enzymes. Increased thermostability must be found, instead, in a small number of highly specific alterations that often do not obey any obvious traffic rules. After briefly discussing the diversity of hyperthermophilic organisms, this review concentrates on the remarkable thermostability of their enzymes. The biochemical and molecular properties of hyperthermophilic enzymes are described. Mechanisms responsible for protein inactivation are reviewed. The molecular mechanisms involved in protein thermostabilization are discussed, including ion pairs, hydrogen bonds, hydrophobic interactions, disulfide bridges, packing, decrease of the entropy of unfolding, and intersubunit interactions. Finally, current uses and potential applications of thermophilic and hyperthermophilic enzymes as research reagents and as catalysts for industrial processes are described.

Hyperthermophilic Enzymes: Sources, Uses, and Molecular Mechanisms for Thermostability

For the first time a single experimental approach, 16S ribosomal RNA sequence characterization, has been used to develop an overview of phylogenetic relationships in the bacterial world. The techni...

https://science.sciencemag.org/content/sci/209/4455/457.full.pdf

The Phylogeny of Prokaryotes

The information available on the biological activity of hydrogen sulfide has been examined for present status of critical results pertaining to the toxicity of hydrogen sulfide. This review of the literature is intended as an evaluative report rather than an annotated bibliography of all the source material examined on hydrogen sulfide. The information was selected as it might relate to potential toxic effects of hydrogen sulfide to man and summarized, noting information gaps that may require further investigation. Several recommendations are listed for possible consideration for either toxicological research or additional short- and long-term tests. Two bibliographies have been provided to assist in locating references considered in this report: (1) literature examined but not cited and (2) reference citations. The majority of the references in the first bibliography were considered peripheral information and less appropriate for inclusion in this report.

A Critical Review of the Literature on Hydrogen Sulfide Toxicity

Barriers to dispersal between populations allow them to diverge through local adaptation or random genetic drift. High-resolution multilocus sequence analysis revealed that, on a global scale, populations of hyperthermophilic microorganisms are isolated from one another by geographic barriers and have diverged over the course of their recent evolutionary history. The identification of a biogeographic pattern in the archaeon Sulfolobus challenges the current model of microbial biodiversity in which unrestricted dispersal constrains the development of global species richness.

https://science.sciencemag.org/content/sci/301/5635/976.full.pdf

Geographic Barriers Isolate Endemic Populations of Hyperthermophilic Archaea

Thermophilic and hyperthermophilic Archaea and Bacteria have been isolated from marine hydrothermal systems, heated sediments, continental solfataras, hot springs, water heaters, and industrial waste They catalyze a tremendous array of widely varying metabolic processes As determined in the laboratory, electron donors in thermophilic and hyperthermophilic microbial redox reactions include H2, Fe(2+), H2S, S, S2O3(2-), S4O6(2-), sulfide minerals, CH4, various mono-, di-, and hydroxy-carboxylic acids, alcohols, amino acids, and complex organic substrates; electron acceptors include O2, Fe(3+), CO2, CO, NO3(-), NO2(-), NO, N2O, SO4(2-), SO3(2-), S2O3(2-), and S Although many assimilatory and dissimilatory metabolic reactions have been identified for these groups of microorganisms, little attention has been paid to the energetics of these reactions In this review, standard molal Gibbs free energies (DeltaGr(0)) as a function of temperature to 200 degrees C are tabulated for 370 organic and inorganic redox, disproportionation, dissociation, hydrolysis, and solubility reactions directly or indirectly involved in microbial metabolism To calculate values of DeltaGr(0) for these and countless other reactions, the apparent standard molal Gibbs free energies of formation (DeltaG(0)) at temperatures to 200 degrees C are given for 307 solids, liquids, gases, and aqueous solutes It is shown that values of DeltaGr(0) for many microbially mediated reactions are highly temperature dependent, and that adopting values determined at 25 degrees C for systems at elevated temperatures introduces significant and unnecessary errors The metabolic processes considered here involve compounds that belong to the following chemical systems: H-O, H-O-N, H-O-S, H-O-N-S, H-O-C(inorganic), H-O-C, H-O-N-C, H-O-S-C, H-O-N-S-C(amino acids), H-O-S-C-metals/minerals, and H-O-P For four metabolic reactions of particular interest in thermophily and hyperthermophily (knallgas reaction, anaerobic sulfur and nitrate reduction, and autotrophic methanogenesis), values of the overall Gibbs free energy (DeltaGr) as a function of temperature are calculated for a wide range of chemical compositions likely to be present in near-surface and deep hydrothermal and geothermal systems

http://rcn.montana.edu/Resources/LRES555/Manuscripts/energetics.pdf

Energetics of overall metabolic reactions of thermophilic and hyperthermophilic Archaea and Bacteria

Sulfolobus is a new genus of bacteria characterized as follows: 1. generally spherical cells producing frequent lobes; 2. facultative autotrophy with growth on sulfur or on a variety of simple organic compounds; 3. unusual cell wall structure devoid of peptidoglycan; 4. acidophilic, pH optimum of 2–3 and range from 0.9–5.8; 5. thermophilic with temperature optimum of 70–75°C and range from 55–80°C (one strain grew at 85°C). The DNA base composition of five strains was determined by cesium chloride density gradient centrifugation and found to be 60–68% guanine plus cytosine. Sulfolobus apparently has no close relationship with any previously described bacteria, either heterotrophic or autrotrophic. Techniques are presented for distinguishing Sulfolobus from Thermoplasma, another genus of acidophilic thermophilic spherically shaped organisms. Sulfolobus has been isolated from a variety of natural acidic thermal habitats, both terrestrial and aquatic. Most isolations have been from habitats in Yellowstone National Park, but strains were also isolated from Italy, Dominica and El Salvador. It is suggested that Sulfolobus may be an important geochemical agent in the production of sulfuric acid from sulfur in high temperature hydrothermal systems.

Robert T. Belly

Papers

Sulfolobus : A new genus of sulfur-oxidizing bacteria living at low pH and high temperature