R. S. Wolfe

Bio: R. S. Wolfe is an academic researcher from University of Illinois at Urbana–Champaign. The author has contributed to research in topics: Methanobacterium & Ferredoxin. The author has an hindex of 24, co-authored 38 publications receiving 2679 citations.

Methanobacillus omelianskii, a Symbiotic Association of Two Species of Bacteria*

Abstract: Two bacterial species were isolated from cultures of Methanobacillus omelianskii grown on media, containing ethanol as oxidizable substrate. One of these, the S organism, is a gram negative, motile, anaerobic rod which ferments ethanol with production of H2 and acetate but is inhibited by inclusion of 0.5 atm of H2 in the gas phase of the medium. The other organism is a gram variable, nonmotile, anaerobic rod which utilizes H2 but not ethanol for growth and methane formation. The results indicate that M. omelianskii maintained in ethanol media is actually a symbiotic association of the two species.

Methanococcus jannaschii sp. nov., an extremely thermophilic methanogen from a submarine hydrothermal vent

Abstract: A new extremely thermophilic methane-producing bacterium was isolated from a submarine hydrothermal vent sample collected by a research team from the Woods Hole Oceanographic Institution using the manned submersible ALVIN. The sample was obtained from the base of a “white smoker” chimney on the East Pacific Rise at 20° 50′ N latitude and 109° 06′ W longitude at a depth of 2600 m. The isolate was a motile irregular coccus with an osmotically fragile cell wall and a complex flagellar system. In defined medium with 80% H2 and 20% CO2, the isolate had a doubling time of 26 min at 85° C. The pH range for growth was 5.2 to 7.0 with an optimum near 6.0. NaCl was required for growth with an optimum of 2 to 3% (w/v). The mol % G+C was 31%. In cell-free extracts, methane formation from methylcoenzyme M was temperature-dependent, and H2 or formate served as electron donors. Methane formation from H2 and CO2 occurred at a much lower rate. Oligonucleotide cataloging of the 16S ribosomal RNA established the isolate as a new species of the genus Methanococcus and the name Methanococcus jannaschii is proposed. The isolation of M. jannaschii from a submarine hydrothermal vent provides additional evidence for biogenic production of CH4 from these deep-sea environments.

Genetic Engineering of Microorganisms for Chemicals

Abstract: Selection of petite strains of yeast (that is, strains unable to respire aerobically) on media containing allyl alcohol will result in enrichment for mutants at the ADCl locus. This locus codes for the constitutive alcohol dehydrogenase, ADH-I, which is primarily responsible for the production of ethanol in yeast. The mutant enzymes are functional, and confer resistance to allyl alcohol on the cell by shifting the NAD-NADH balance in the direction of NADH. These mutants exhibit altered Km's for cofactor, substrate, or both, and often have altered Vmax's. In this paper, the methodology for obtaining these mutants and for determining the amino acid substitutions responsible for these changes is presented. Several new mutants have been at least approximately localized, and one, DB-AA3Nl5, has been shown to be due to the substitution of an arginine for a tryptophan at position 54. This substitution would be expected, by analogy with the known tertiary structure of the horse liver alcohol dehydrogenase, to decrease the hydrophobic environment of the active site pocket. The substitution has a pronounced effect on the Km for ethanol, but far less on that for acetaldehyde. The current status of investigation of other classes of functional mutants of this enzyme, and the potential both for selection of useful variants of this molecule and for an increase understanding of its function are discussed.

Methanogenium, a new genus of marine methanogenic bacteria, and characterization ofMethanogenium cariaci sp. nov. andMethanogenium marisnigri sp. nov.

Abstract: A new genus of marine methanogenic bacteria and two species within this genus are described.Methanogenium is the proposed genus andMethanogenium cariaci the type species. Cells of the type species are Gram-negative, peritrichously flagellated, irregular cocci with a periodic wall surface pattern. Colonies formed by these bacteria are yellow, circular and umbonate with entire edges. The DNA base composition is 52 mol% guanine plus cytosine. Formate or hydrogen and carbon dioxide serve as substrates for growth. Cells ofMethanogenium marisnigri are of similar shape but smaller diameter thanM. cariaci. The colonies ofM. marisnigri are convex, and the DNA base composition is 61 mol % G+C. Formate or hydrogen and carbon dioxide are growth substrates. Sodium chloride is required for growth of both methanogens.

Acetogenium kivui, a new thermophilic hydrogen-oxidizing acetogenic bacterium

Abstract: Hydrogen-oxidizing acetogenic bacteria in pure culture are presently represented by the two mesophilic species, Acetobacterium woodii and Clostridium aceticum. From Lake Kivu we have isolated a Gram negative, chemolithotrophic, thermophilic anaerobe (LKT-1) that oxidizes hydrogen and reduces carbon dioxide to acetic acid. It is a non-motile, non-sporeforming rod, about 0.7μm in width and 2–7.5μm in length, often occuring in pairs or chains. The cell wall has a banded appearance; the surface layer contains a regular array of particles with six-fold rotational symmetry. No outer membrane is present. The temperature optimum for growth is 66°C, and the pH optimum is 6.4. Organic growth substrates include glucose, mannose, fructose, pyruvate, and formate; acetate is the principal product. The doubling time for growth on hydrogen and carbon dioxide is about 2h. Vitamins are neither required nor stimulatory. Yeast extract and Trypticase enhance the final yield but do not affect the growth rate. Cysteine or sulfide are required and cannot be replaced by thioglycolate or dithiothreitol. LKT-1 was mass cultured on hydrogen and carbon dioxide in a 24.1 fermentor with a yield of 34g (wet weight) of cells. The DNA base composition as determined by buoyant density is 38 mol % guanine plus cytosine. LKT-1 appears only distantly related to physiologically similar bacteria. A new genus Acetogenium is proposed, and the species is Acetogenium kivui.

Microbial cellulose utilization: fundamentals and biotechnology.

Abstract: Fundamental features of microbial cellulose utilization are examined at successively higher levels of aggregation encompassing the structure and composition of cellulosic biomass, taxonomic diversity, cellulase enzyme systems, molecular biology of cellulase enzymes, physiology of cellulolytic microorganisms, ecological aspects of cellulase-degrading communities, and rate-limiting factors in nature. The methodological basis for studying microbial cellulose utilization is considered relative to quantification of cells and enzymes in the presence of solid substrates as well as apparatus and analysis for cellulose-grown continuous cultures. Quantitative description of cellulose hydrolysis is addressed with respect to adsorption of cellulase enzymes, rates of enzymatic hydrolysis, bioenergetics of microbial cellulose utilization, kinetics of microbial cellulose utilization, and contrasting features compared to soluble substrate kinetics. A biological perspective on processing cellulosic biomass is presented, including features of pretreated substrates and alternative process configurations. Organism development is considered for "consolidated bioprocessing" (CBP), in which the production of cellulolytic enzymes, hydrolysis of biomass, and fermentation of resulting sugars to desired products occur in one step. Two organism development strategies for CBP are examined: (i) improve product yield and tolerance in microorganisms able to utilize cellulose, or (ii) express a heterologous system for cellulose hydrolysis and utilization in microorganisms that exhibit high product yield and tolerance. A concluding discussion identifies unresolved issues pertaining to microbial cellulose utilization, suggests approaches by which such issues might be resolved, and contrasts a microbially oriented cellulose hydrolysis paradigm to the more conventional enzymatically oriented paradigm in both fundamental and applied contexts.

Energy conservation in chemotrophic anaerobic bacteria.

Abstract: [This corrects the article on p. 100 in vol. 41.].

Microbial Biofilms: from Ecology to Molecular Genetics

Abstract: Biofilms are complex communities of microorganisms attached to surfaces or associated with interfaces. Despite the focus of modern microbiology research on pure culture, planktonic (free-swimming) bacteria, it is now widely recognized that most bacteria found in natural, clinical, and industrial settings persist in association with surfaces. Furthermore, these microbial communities are often composed of multiple species that interact with each other and their environment. The determination of biofilm architecture, particularly the spatial arrangement of microcolonies (clusters of cells) relative to one another, has profound implications for the function of these complex communities. Numerous new experimental approaches and methodologies have been developed in order to explore metabolic interactions, phylogenetic groupings, and competition among members of the biofilm. To complement this broad view of biofilm ecology, individual organisms have been studied using molecular genetics in order to identify the genes required for biofilm development and to dissect the regulatory pathways that control the plankton-to-biofilm transition. These molecular genetic studies have led to the emergence of the concept of biofilm formation as a novel system for the study of bacterial development. The recent explosion in the field of biofilm research has led to exciting progress in the development of new technologies for studying these communities, advanced our understanding of the ecological significance of surface-attached bacteria, and provided new insights into the molecular genetic basis of biofilm development.