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Biomedical subjects

A J Stams

Publications and source records attributed to A J Stams.

At least 37 records · Page 2Linked to original sources
PubMed1999

Desulfobacca acetoxidans gen. nov., sp. nov., a novel acetate-degrading sulfate reducer isolated from sulfidogenic granular sludge.

A mesophilic sulfate reducer, strain ASRB2T, was isolated with acetate as sole carbon and energy source from granular sludge of a laboratory-scale upflow anaerobic sludge bed reactor fed with acetate and sulfate. The bacterium was oval-shaped, 1.3 x 1.9-2.2 microns, non-motile and Gram-negative. Optimum growth with acetate occurred around 37 degrees C in freshwater medium (doubling time: 1.7-2.2 d). Enzyme studies indicated that acetate was oxidized via the carbon monoxide dehydrogenase pathway. Growth was not supported by other organic acids, such as propionate, butyrate or lactate, alcohols such as ethanol or propanol, and hydrogen or formate. Sulfite and thiosulfate were also used as electron acceptors, but sulfur and nitrate were not reduced. Phylogenetically, strain ASRB2T clustered with the delta subclass of the Proteobacteria. Its closest relatives were Desulfosarcina variabilis, Desulfacinum infernum and Syntrophus buswellii. Strain ASRB2T is described as the type strain of Desulfobacca acetoxidans gen. nov., sp. nov.

Acetates↗
PubMed1999

High-rate anaerobic treatment of wastewater at low temperatures.

Anaerobic treatment of a volatile fatty acid (VFA) mixture was investigated under psychrophilic (3 to 8 degrees C) conditions in two laboratory-scale expanded granular sludge bed reactor stages in series. The reactor system was seeded with mesophilic methanogenic granular sludge and fed with a mixture of VFAs. Good removal of fatty acids was achieved in the two-stage system. Relative high levels of propionate were present in the effluent of the first stage, but propionate was efficiently removed in the second stage, where a low hydrogen partial pressure and a low acetate concentration were advantageous for propionate oxidation. The specific VFA-degrading activities of the sludge in each of the modules doubled during system operation for 150 days, indicating a good enrichment of methanogens and proton-reducing acetogenic bacteria at such low temperatures. The specific degradation rates of butyrate, propionate, and the VFA mixture amounted to 0.139, 0.110, and 0.214 g of chemical oxygen demand g of volatile suspended solids-1 day-1, respectively. The biomass which was obtained after 1.5 years still had a temperature optimum of between 30 and 40 degrees C.

Anaerobiosis↗
PubMed1999

Reductive dechlorination of tetrachloroethene to cis-1, 2-dichloroethene by a thermophilic anaerobic enrichment culture.

Thermophilic anaerobic biodegradation of tetrachloroethene (PCE) was investigated with various inocula from geothermal and nongeothermal areas. Only polluted harbor sediment resulted in a stable enrichment culture that converted PCE via trichloroethene to cis-1, 2-dichloroethene at the optimum temperature of 60 to 65 degrees C. After several transfers, methanogens were eliminated from the culture. Dechlorination was supported by lactate, pyruvate, fructose, fumarate, and malate as electron donor but not by H2, formate, or acetate. Fumarate and L-malate led to the highest dechlorination rate. In the absence of PCE, fumarate was fermented to acetate, H2, CO2, and succinate. With PCE, less H2 was formed, suggesting that PCE competed for the reducing equivalents leading to H2. PCE dechlorination, apparently, was not outcompeted by fumarate as electron acceptor. At the optimum dissolved PCE concentration of approximately 60 microM, a high dechlorination rate of 1.1 micromol h-1 mg-1 (dry weight) was found, which indicates that the dechlorination is not a cometabolic activity. Microscopic analysis of the fumarate-grown culture showed the dominance of a long thin rod. Molecular analysis, however, indicated the presence of two dominant species, both belonging to the low-G+C gram positives. The highest similarity was found with the genus Dehalobacter (90%), represented by the halorespiring organism Dehalobacter restrictus, and with the genus Desulfotomaculum (86%).

Bacteria, Anaerobic↗
PubMed1999

Purification and characterization of (per)chlorate reductase from the chlorate-respiring strain GR-1.

Strain GR-1 is one of several recently isolated bacterial species that are able to respire by using chlorate or perchlorate as the terminal electron acceptor. The organism performs a complete reduction of chlorate or perchlorate to chloride and oxygen, with the intermediate formation of chlorite. This study describes the purification and characterization of the key enzyme of the reductive pathway, the chlorate and perchlorate reductase. A single enzyme was found to catalyze both the chlorate- and perchlorate-reducing activity. The oxygen-sensitive enzyme was located in the periplasm and had an apparent molecular mass of 420 kDa, with subunits of 95 and 40 kDa in an alpha(3)beta(3) composition. Metal analysis showed the presence of 11 mol of iron, 1 mol of molybdenum, and 1 mol of selenium per mol of heterodimer. In accordance, quantitative electron paramagnetic resonance spectroscopy showed the presence of one [3Fe-4S] cluster and two [4Fe-4S] clusters. Furthermore, two different signals were ascribed to Mo(V). The K(m) values for perchlorate and chlorate were 27 and <5 microM, respectively. Besides perchlorate and chlorate, nitrate, iodate, and bromate were also reduced at considerable rates. The resemblance of the enzyme to nitrate reductases, formate dehydrogenases, and selenate reductase is discussed.

Amino Acid Sequence↗
PubMed1998

Investigation of the fumarate metabolism of the syntrophic propionate-oxidizing bacterium strain MPOB.

The growth of the syntrophic propionate-oxidizing bacterium strain MPOB in pure culture by fumarate disproportionation into carbon dioxide and succinate and by fumarate reduction with propionate, formate or hydrogen as electron donor was studied. The highest growth yield, 12.2 g dry cells/mol fumarate, was observed for growth by fumarate disproportionation. In the presence of hydrogen, formate or propionate, the growth yield was more than twice as low: 4.8, 4.6, and 5.2 g dry cells/mol fumarate, respectively. The location of enzymes that are involved in the electron transport chain during fumarate reduction in strain MPOB was analyzed. Fumarate reductase, succinate dehydrogenase, and ATPase were membrane-bound, while formate dehydrogenase and hydrogenase were loosely attached to the periplasmic side of the membrane. The cells contained cytochrome c, cytochrome b, menaquinone-6 and menaquinone-7 as possible electron carriers. Fumarate reduction with hydrogen in membranes of strain MPOB was inhibited by 2-(heptyl)-4-hydroxyquinoline-N-oxide (HOQNO). This inhibition, together with the activity of fumarate reductase with reduced 2,3-dimethyl-1,4-naphtoquinone (DMNH2) and the observation that cytochrome b of strain MPOB was oxidized by fumarate, suggested that menequinone and cytochrome b are involved in the electron transport during fumarate reduction in strain MPOB. The growth yields of fumarate reduction with hydrogen or formate as electron donor were similar to the growth yield of Wolinella succinogenes. Therefore, it can be assumed that strain MPOB gains the same amount of ATP from fumarate reduction as W. succinogenes, i. e. 0.7 mol ATP/mol fumarate. This value supports the hypothesis that syntrophic propionate-oxidizing bacteria have to invest two-thirds of an ATP via reversed electron transport in the succinate oxidation step during the oxidation of propionate. The same electron transport chain that is involved in fumarate reduction may operate in the reversed direction to drive the energetically unfavourable oxidation of succinate during syntrophic propionate oxidation since (1) cytochrome b was reduced by succinate and (2) succinate oxidation was similarly inhibited by HOQNO as fumarate reduction.

Adenosine Triphosphatases↗
PubMed1998

Detection and quantification of microorganisms in anaerobic bioreactors.

The presence of sulfate in anaerobic reactors can trigger competitive and syntrophic interactions between various groups of microorganisms, such as sulfate reducers, methanogens and acetogens. In order to steer the reactor process in the direction of sulfidogenesis or methanogenesis, it is essential to get insight into the population dynamics of these groups of microorganisms upon changes in the reactor operating conditions. Several methods exist to characterize and quantify the microbial sludge composition. Combining classical microbiological and modern molecular-based sludge characterization methods has proven to be a powerful approach to study the microbial composition of the anaerobic sludge.

Journal Article↗
PubMed1998

Anaerobic treatment of sulphate-rich wastewaters.

Until recently, biological treatment of sulphate-rich wastewater was rather unpopular because of the production of H2S under anaerobic conditions. Gaseous and dissolved sulphides cause physical-chemical (corrosion, odour, increased effluent chemical oxygen demand) or biological (toxicity) constraints, which may lead to process failure. Anaerobic treatment of sulphate-rich wastewater can nevertheless be applied successfully provided a proper treatment strategy is selected. The strategies currently available are discussed in relation to the aim of the treatment: i) removal of organic matter, ii) removal of sulphate or iii) removal of both. Also a whole spectrum of new biotechnological applications (removal of organic chemical oxygen demand, sulphur, nitrogen and heavy metals), recently developed based on a better insight in sulphur transformations, are discussed.

Bacteria, Anaerobic↗
PubMed1998

13C-NMR study of propionate metabolism by sludges from bioreactors treating sulfate and sulfide rich wastewater.

Applications of nuclear magnetic resonance (NMR) to study a variety of physiological and biochemical aspects of bacteria with a role in the sulfur cycle are reviewed. Then, a case-study of high resolution 13C-NMR spectroscopy on sludges from bioreactors used for treating sulfate and sulfide rich wastewaters is presented. 13C-NMR was used to study the effect of sulfate and butyrate on propionate conversion by mesophilic anaerobic (methanogenic and sulfate reducing) granular sludge and microaerobic (sulfide oxidizing) flocculant sludge. In the presence of sulfate, propionate was degraded via the randomising pathway in all sludge types investigated. This was evidenced by scrambling of [3-13C]propionate into [2-13C]propionate and the formation of acetate equally labeled in the C1 and C2 position. In the absence of sulfate, [3-13C]propionate scrambled to a lesser extend without being degraded further. Anaerobic sludges converted [2,3-13C]propionate partly into the higher fatty acid 2-methyl[2,3-13C]butyrate during the simultaneous degradation of [2,3-13C]propionate and butyrate. [4,5-13C]valerate was also formed in the methanogenic sludges. Up to 10% of the propionate present was converted via these alternative degradation routes. Labeled butyrate was not detected in the incubations, suggesting that reductive carboxylation of propionate does not occur in the sludges.

Biodegradation, Environmental↗
PubMed1998

Syntrophobacter fumaroxidans sp. nov., a syntrophic propionate-degrading sulfate-reducing bacterium.

A syntrophic propionate-oxidizing bacterium, strain MPOBT, was isolated from a culture enriched from anaerobic granular sludge. It oxidized propionate syntrophically in co-culture with the hydrogen- and formate-utilizing Methanospirillum hungateii, and was able to oxidize propionate and other organic compounds in pure culture with sulfate or fumarate as the electron acceptor. Additionally, it fermented fumarate. 16S rRNA sequence analysis revealed a relationship with Syntrophobacter wolinii and Syntrophobacter pfennigii. The G + C content of its DNA was 60.6 mol%, which is in the same range as that of other Syntrophobacter species. DNA-DNA hybridization studies showed less than 26% hybridization among the different genomes of Syntrophobacter species and strain MPOBT. This justifies the assignment of strain MPOBT to the genus Syntrophobacter as a new species. The name Syntrophobacter fumaroxidans is proposed; strain MPOBT (= DSM 10017T) is the type strain.

Anaerobiosis↗
PubMed1997

Understanding and advancing wastewater treatment.

In bioreactors used for the purification of wastewater, microorganisms are active in biofilms or aggregates. Insight into the factors that determine the structure and function of aggregated biomass is increasing steadily. Besides conventional techniques, modem molecular techniques are used increasingly to get a better understanding of the complex microbial communities in wastewater treatment systems. In recent years, the combined use of these techniques has led to a good insight into the population dynamics of different types of microbes in bioreactors.

Bacteria, Anaerobic↗
PubMed1997

Taxonomic description of Methanococcoides euhalobius and its transfer to the Methanohalophilus genus.

A sequence analysis of the 16S-rRNA of Methanococcoides euhalobius revealed that this organism was highly related to members of the genus Methanohalophilus. On the basis of sequence data, an oligonucleotide probe specific to Methanohalophilus species was designed. Hybridization studies with this probe confirmed close relationship of Methanococcoides euhalobius to Methanohalophilus species. Therefore, we propose that Methanococcoides euhalobius should be transferred to the genus Methanohalophilus as Methanohalophilus euhalobius.

Classification↗
PubMed1997

Caloramator proteoclasticus sp. nov., a new moderately thermophilic anaerobic proteolytic bacterium.

A new moderately thermophilic proteolytic anaerobe, strain UT, was isolated from mesophilic granular methanogenic sludge. The cells were spore-forming, motile rods that were 0.4 micron wide and 2.4 to 4 microns long and stained gram negative. Electron micrographs of thin sections revealed the presence of an atypical gram-positive cell wall. Optimum growth occurred at 55 degrees C and at pH values between 7.0 and 7.5, with a doubling time of 30 min. The DNA base ratio of guanine plus cytosine was 31 mol%. The bacterium fermented proteins mainly to acetate, hydrogen, formate, and branched-chain fatty acids. Several amino acids, including glutamate, aspartate, arginine, histidine, threonine, methionine, and branched-chain amino acids, were also utilized. Glutamate was degraded to acetate, formate, hydrogen, and alanine. In addition, the strain degraded carbohydrates, including glucose, fructose, mannose, cellobiose, and starch, to acetate, ethanol, formate, lactate, and hydrogen. The results of a 16S rRNA sequence analysis phylogenetically placed strain UT in the low-guanine-plus-cytosine-content subgroup of the gram-positive phylum. We propose to classify the described strain in the genus Caloramator as a new species, Caloramator proteoclasticus. The type strain of C. proteoclasticus, strain U, has been deposited in the Deutsche Sammlung von Mikroorganismen as strain DSM 10124.

Bacteria, Anaerobic↗
PubMed1996

Purification and characterization of malate dehydrogenase from the syntrophic propionate-oxidizing bacterium strain MPOB.

Malate dehydrogenase from the syntrophic propionate-oxidizing bacterium strain MPOB was purified 42-fold. The native enzyme had an apparent molecular mass of 68 kDa and consisted of two subunits of 35 kDa. The enzyme exhibited maximum activity with oxaloacetate at pH 8.5 and 60 degrees C. The Ka for oxaloacetate was 50 microM and for NADH 30 microM. The Km values for L-malate and NAD were 4 and 1.1 mM, respectively. Substrate inhibition was found at oxaloacetate concentrations higher than 250 microM. The N-terminal amino acid sequence of the enzyme was similar to the sequences of a variety of other malate dehydrogenases from plants, animals and micro-organisms.

Amino Acid Sequence↗
PubMed1996

Purification and characterization of fumarase from the syntrophic propionate-oxidizing bacterium strain MPOB.

Fumarase from the syntrophic propionate-oxidizing bacterium strain MPOB was purified 130-fold under anoxic conditions. The native enzyme had an apparent molecular mass of 114 kDa and was composed of two subunits of 60 kDa. The enzyme exhibited maximum activity at pH 8.5 and approximately 54 degrees C. The Km values for fumarate and L-malate were 0.25 mM and 2.38 mM, respectively. Fumarase was inactivated by oxygen, but the activity could be restored by addition of Fe2+ and &beta;-mercaptoethanol under anoxic conditions. EPR spectroscopy of the purified enzyme revealed the presence of a [3Fe-4S] cluster. Under reducing conditions, only a trace amount of a [4Fe-4S] cluster was detected. Addition of fumarate resulted in a significant increase of this [4Fe-4S] signal. The N-terminal amino acid sequence showed similarity to the sequences of fumarase A and B of Escherichia coli (56%) and fumarase A of Salmonella typhimurium (63%).

Amino Acid Sequence↗
PubMed1996

Detection and localization of syntrophic propionate-oxidizing bacteria in granular sludge by in situ hybridization using 16S rRNA-based oligonucleotide probes.

In situ hybridization with fluorescent oligonucleotides was used to detect and localize microorganisms in the granules of two lab-scale upflow anaerobic sludge blanket reactors that had been fed for several months with either sucrose or a mixture of volatile fatty acids. Sections of the granules were hybridized with 16S rRNA-targeted oligonucleotide probes for Bacteria, Archaea, specific phylogenetic groups of methanogens, and two syntrophic propionate-oxidizing strains, MPOB and KOPROP1. Cells of the syntrophic strain KOPROP1 were not detected in either type of sludge granules. Hybridizations of the sucrose-fed granules showed an outer layer of mainly bacterial microcolonies with different morphologies. More inwards of these granules, a layer of different methanogenic microcolonies mixed with large colonies of the syntrophic strain MPOB could be detected. The MPOB colonies were intertwined with hydrogen- or formate-consuming methanogens, indicating their syntrophic growth. The granules fed with volatile fatty acids showed an outer layer of mainly bacteria and then a thick layer of Methanosaeta-like methanogens mixed with a few bacteria and a layer of methanogens mixed with syntrophic MPOB microcolonies. The centers of both sludge types consisted of large cavities and methanogenic microcolonies. These results indicate a juxtapositioning of syntrophic bacteria and methanogens and provide additional evidence for a layered microbial architecture of anaerobic granular sludge.

Bacteria↗
PubMed1996

Population dynamics of propionate-oxidizing bacteria under methanogenic and sulfidogenic conditions in anaerobic granular sludge.

Laboratory-scale upflow anaerobic sludge-bed reactors were inoculated with industrial granular sludge and fed with either propionate or propionate and sulfate. The population dynamics of the propionate-oxidizing bacteria Desulfobulbus sp. and the syntrophically growing strain SYN7 were studied in reactors by dot blot and in situ hybridization with 16S rRNA-based oligonucleotide probes.

Bacteria, Anaerobic↗
PubMed1995

Purification and characterization of a novel ADP-dependent glucokinase from the hyperthermophilic archaeon Pyrococcus furiosus.

Pyrococcus furiosus uses a modified Embden-Meyerhof pathway during growth on poly- or disaccharides. Instead of the usual ATP-dependent glucokinase, this pathway involves a novel ADP-dependent (AMP-forming) glucokinase. The level of this enzyme and some other glycolytic enzymes appeared to be closely regulated by the substrate. Growth on cellobiose resulted in a high specific activity of 0.96 units mg-1, whereas on pyruvate a 10-fold lower activity was found. The ADP-dependent glucokinase was purified 1350-fold to homogeneity. The oxygen-stable enzyme had a molecular mass of 93 kDa and was composed of two identical subunits (47 kDa). The glucokinase was highly specific for ADP, which could not be replaced by ATP, phosphoenolpyruvate, GDP, PPi, or polyphosphate. D-Glucose could be replaced only by 2-deoxy-D-glucose, albeit with a low efficiency. The Km values for D-glucose and ADP were 0.73 and 0.033 mM, respectively. An optimum temperature of 105 degrees C and a half-life of 220 min at 100 degrees C are in agreement with the requirements of this hyperthermophilic organism. The properties of the glucokinase are compared to those of less thermoactive gluco/hexokinases.

Adenosine Diphosphate↗