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Biomedical subjects

G Gottschalk

Publications and source records attributed to G Gottschalk.

At least 91 records · Page 5Linked to original sources
PubMed1985

Sodium ions and an energized membrane required by Methanosarcina barkeri for the oxidation of methanol to the level of formaldehyde.

Methanogenesis from methanol by cell suspensions of Methanosarcina barkeri was inhibited by the uncoupler tetrachlorosalicylanilide. This inhibition was reversed by the addition of formaldehyde. 14C labeling experiments revealed that methanol served exclusively as the electron acceptor, whereas formaldehyde was mainly oxidized to CO2 under these conditions. These data support the hypothesis (M. Blaut and G. Gottschalk, Eur. J. Biochem. 141: 217-222, 1984) that the first step in methanol oxidation depends on the proton motive force or a product thereof. Cell extracts of M. barkeri converted methanol and formaldehyde to methane under an H2 atmosphere. Under an N2 atmosphere, however, formaldehyde was disproportionated to CH4 and CO2, whereas methanol was metabolized to a very small extent only, irrespective of the presence of ATP. It was concluded that cell extracts of M. barkeri are not able to oxidize methanol. In further experiments, the sodium dependence of methanogenesis and ATP formation by whole cells was investigated. Methane formation from methanol alone and the corresponding increase in the intracellular ATP content were strictly dependent on Na+. If, in contrast, methanol was utilized together with H2, methane and ATP were synthesized in the absence of Na+. The same is true for the disproportionation of formaldehyde to methane and carbon dioxide. From these experiments, it is concluded that in M. barkeri, Na+ is involved not in the process of ATP synthesis but in the first step of methanol oxidation.

Adenosine Triphosphate↗
PubMed1984

Coupling of ATP synthesis and methane formation from methanol and molecular hydrogen in Methanosarcina barkeri.

The addition of methanol to a cell suspension of Methanosarcina barkeri resulted in an increase of the intracellular ATP concentration from 1 nmol/mg to 10 nmol/mg protein and in the formation of a proton-motive force delta p of -130 mV. delta p consisted of more than 90% of the membrane potential delta psi. These values were similar under N2 and under H2. The addition of the uncoupler tetrachlorosalicylanilide to the above system under N2 led to a drastic decrease of both, the ATP concentration and the delta p and to a stop of methanogenesis. With methanol and H2, however, methane formation continued, although the effect of the uncoupler on the ATP pool and on delta p was a under N2. The proton-translocating ATPase inhibitor N,N'-dicyclohexylcarbodiimide caused a rapid exhaustion of the ATP pool and a discontinuation of methane synthesis, whereas delta p was unaffected. Inhibition of methane formation under these conditions could be relieved by the addition of the uncoupler tetrachlorosalicylanilide. These results demonstrate that methane formation according to the equation CH3OH + H2----H2----CH4 + H2O was coupled to ATP synthesis by a chemiosmotic mechanism and was under the control of delta psi: Methane formation only proceeded if the delta psi generated was used for ATP synthesis or if an uncoupler was present. Under N2, methane formation according to the equation 4CH3OH ----CO2 + 3CH4 + 2H2O was abolished by an uncoupler, because one step in the oxidation of methanol to 1 CO2 apparently depended on an energized state of the membrane.

Adenosine Triphosphate↗
PubMed1984

Formation of n-Butanol from d-Glucose by Strains of the "Clostridium tetanomorphum" Group.

A clostridial strain has been isolated that produced n-butanol, ethanol, butyrate, and acetate as major fermentation products from glucose but no acetone. At a pH of 6.6, n-butanol was formed by this microorganism only during growth. On the basis of its physiological characteristics and DNA-DNA homology data, the strain was assigned to the "Clostridium tetanomorphum" group (S. Nakamura, I. Okado, T. Abe, and S. Nishida, J. Gen. Microbiol. 113:29-35, 1979). All members of this group were shown to produce n-butanol from glucose as the major fermentation product, whereas C. cochlearium produced it in only minor amounts.

Journal Article↗
PubMed1983

Characterization of the cytochromes occurring in Methanosarcina species.

Several members of the genus Methanosarcina were investigated by room-temperature and low-temperature difference spectroscopy for the presence of cytochromes. In combination with potentiometric titrations two membrane-bound b-cytochromes and one membrane-bound c-cytochrome could be detected in cells grown on methanol or trimethylamine. Very probably acetate-grown cells contained an additional cytochrome b. The midpoint potentials of the two b-type cytochromes were Em1 = -325 mV and Em2 = -183 mV, respectively. The additional b cytochrome formed during growth on acetate exhibited a midpoint potential of Em3 = -250 mV.

Acetates↗
PubMed1983

How perchlorate improves excitation-contraction coupling in skeletal muscle fibers.

The effect of the "chaotropic" anion, perchlorate, on the activation of contraction has been studied in voltage clamped frog skeletal muscle fibers. It was found that the voltage dependence of either the contractile force or the intramembrane charge movement was shifted towards more negative membrane potentials. The maximum values of force or charge movement attained with large depolarizing pulses did not change significantly. It is concluded that a specific perchlorate effect on the movement of charged particles can explain the potentiating effect of perchlorate anions on contractile force, strengthening the view that these charged particles serve as voltage sensors regulating Ca2+ release from the sarcoplasmic reticulum.

Animals↗
PubMed1983

Tritan pedigree without optic-nerve atrophy.

Results of several previous reports have questioned the occurrence of the tritan color deficiency independently of dominantly inherited optic atrophy. This report describes the results of testing 34 members of a pedigree (including four tritans) for whom optic atrophy can be ruled out according to criteria previously described by Krill et al.

Adolescent↗
PubMed1983

Perchlorate-induced alterations in electrical and mechanical parameters of frog skeletal muscle fibres.

The effect of the perchlorate anion (ClO4-) on the potential dependence of mechanical and electrical parameters was investigated in skeletal muscles fibres of the frog. Two main methods were employed: twitches and K contractures were induced in isolated fibres from the semitendinosus or iliofibularis muscle, and point voltage clamp was applied in sartorius and short toe muscle fibres. Twitch height was unaffected below 10(-4) M-ClO4-, it usually increased several-fold in the concentration range of 10(-3) to 10(-2) M-ClO4- and continued to rise slowly between 10(-2) and 10(-1) M-ClO4-. ClO4- caused a parallel shift of the activation curve, which relates peak force to membrane potential, towards more negative potentials by up to 40 mV (70 mM-ClO4-). The shift in force activation was not accompanied by a corresponding shift in the potential dependence of force inactivation. In the presence of ClO4-, maximum force development upon depolarization to -60 or -50 mV could be maintained for several minutes, suggesting that spontaneous relaxation after full depolarization is due to a potential-dependent inactivation process, and not to an exhaustion of Ca2+ release. ClO4- shifted the threshold for the initiation of the action potential only slightly towards more negative potentials (approximately 10 mV at 70 mM-ClO4-). Little or no shift was observed in the lower concentration range (less than 10 mM) where the threshold of force activation was shifted by about 20 mV. ClO4- slightly depressed the activation of the delayed rectifier without causing any distinct change in its threshold potential. Electrophoretic injection of ClO4- (internal ClO4- concentration ([ClO4-]i) approximately 1 mM) induced similar effects to those following external application of this anion, i.e. a shift of force activation towards more negative potentials. Of several other anions tested, only dichromate, which resembles ClO4-in its tetrahedal structure, similarly caused force activation after repolarization. We conclude that at low concentrations (less than 10 mM) ClO4- rather specifically improves excitation-contraction coupling by direct interference with the gating mechanism which activates Ca release from the sarcoplasmic reticulum. At higher concentrations, it may also influence potential-dependent membrane processes by adsorption to the outer surface of the membrane.

Action Potentials↗
PubMed1983

Methanogenesis from Choline by a Coculture of Desulfovibrio sp. and Methanosarcina barkeri.

A sulfate-reducing vibrio was isolated from a methanogenic enrichment with choline as the sole added organic substrate. This organism was identified as a member of the genus Desulfovibrio and was designated Desulfovibrio strain G1. In a defined medium devoid of sulfate, a pure culture of Desulfovibrio strain G1 fermented choline to trimethylamine, acetate, and ethanol. In the presence of sulfate, more acetate and less ethanol were formed from choline than in the absence of sulfate. When grown in a medium containing sulfate, a coculture of Desulfovibrio strain G1 and Methanosarcina barkeri strain Fusaro degraded choline almost completely to methane, ammonia, and hydrogen sulfide and presumably to carbon dioxide. Methanogenesis occurred in two distinct phases separated by a lag of about 6 days. During the first phase of methanogenesis choline was completely converted to trimethylamine, acetate, hydrogen sulfide, and traces of ethanol by the desulfovibrio. M. barkeri fermented trimethylamine to methane, ammonia, and presumably carbon dioxide via dimethyl- and methylamine as intermediates. Simultaneously, about 60% of the acetate expected was metabolized. In the second phase of methanogenesis, the residual acetate was almost completely catabolized.

Journal Article↗
PubMed1983

Betaine: New Oxidant in the Stickland Reaction and Methanogenesis from Betaine and l-Alanine by a Clostridium sporogenes-Methanosarcina barkeri Coculture.

Growing and nongrowing cells of Clostridium sporogenes fermented betaine with l-alanine, l-valine, l-leucine, and l-isoleucine as electron donors in a coupled oxidation-reduction reaction (Stickland reaction). For the substrate combinations betaine and l-alanine and betaine and l-valine balance studies were performed; the results were in agreement with the following fermentation equation: 1 R- CH(NH(2))-COOH + 2 betaine + 2 H(2)O --> 1 R-COOH + 1 CO(2) + 1 NH(3) + 2 trimethylamine + 2 acetate. Growth and production of trimethylamine were strictly dependent on the presence of selenite in the medium. With cell suspensions it was shown that C. sporogenes was unable to catabolize betaine as a single substrate. Betaine, however, was reduced to trimethylamine and acetate under an atmosphere of molecular hydrogen. For the reduction of betaine by cell extracts of C. sporogenes, dimercaptans such as 1,4-dithiothreitol could serve as electron donors. No betaine reductase activity was detected in cells grown in a complex medium without betaine. The pH optimum of betaine reductase was at pH 7.3. When C. sporogenes was cocultured with Methanosarcina barkeri strain Fusaro on betaine together with l-alanine, an almost complete conversion of the two substrates to CH(4), NH(3), and presumably CO(2) was observed.

Journal Article↗
PubMed1982

Cell and ATP yields of Citrobacter freundii growing with fumarate and H2 or formate in continuous culture.

Out of 19 strains belonging to the family Enterobacteriaceae only Escherichia coli and Citrobacter strains fermented fumarate exclusively to succinate. This fermentation was dependent on the presence of molecular hydrogen or formate. The inability of these micro-organisms to convert fumarate to succinate, acetate and CO2 correlated with their lack, or low activity, of oxaloacetate decarboxylase. Continuous culture experiments were performed with Citrobacter freundii in minimal or complex medium with fumarate + H2 of formate, and the growth parameters were determined. From the data obtained, a Ymax fumarate dissimilated value of 10.5 +/- 0.8 g dry wt per mol fumarate dissimilated was calculated. This value demonstrates that, per mol fumarate reduced, at least 0.6 +/- 0.05 mol ATP is produced and subsequently used for biosynthetic purposes.

Adenosine Triphosphate↗
PubMed1982

Purification of L-glutamate-dependent citrate lyase from Clostridium sphenoides and electron microscopic analysis of citrate lyase isolated from Rhodopseudomonas gelatinosa, Streptococcus diacetilactis and C. sphenoides.

Citrate lyase from Clostridium sphenoides was purified 72-fold with a yield of 11%. In contrast to citrate lyase from other sources the activity of this enzyme was strictly dependent on the presence of L-glutamate. The purified enzyme was only stable in the presence of 150 mM L-glutamate or 7 mM L-glutamate plus glycerol, sucrose or bovine serum albumin. Changes of the L-glutamate pool and of enzyme activity in growing cells of C. sphenoides indicated that citrate lyase activity in this organism was regulated by the intracellular L-glutamate concentration. Citrate lyase isolated from C. sphenoides, Rhodopseudomonas gelatinosa and Streptococcus diacetilactis was investigated by electron microscopy using the negative staining technique. Three different projections of enzyme molecules were observed: 'star' form, 'ring' form and 'triangle' form. In samples from R. gelatinosa and S. diacetilactis, star and ring forms occurred in a ratio of about 1:9. Using the enzyme from S. diacetilactis it was demonstrated that this ratio could be altered in favour of the star form by the addition of citrate or tricarballylate. The triangle form was observed in less than 1% of all evaluated molecules and may represent a transition form. In lyase samples from C. sphenoides there existed a correlation between enzyme activity and the proportion of stars and rings at varying concentrations of L-glutamate.

Bacteria↗
PubMed1982

Copurification of citrate lyase and citrate lyase ligase from Rhodopseudomonas gelatinosa and subsequent separation of the two enzymes.

A procedure has been worked out which allowed the purification and crystallization of a citrate lyase/citrate lyase ligase complex from Rhodopseudomonas gelatinosa. The complex was subsequently separated to yield two homogeneous enzymes. Citrate lyase ligase was purified 365-fold with a yield of 3.23%. The molecular weight of the enzyme was estimated to be 39500, the enzyme consisted of one polypeptide chain. The reaction rates for ATP, acetate and citrate lyase (sulfhydryl form) followed Michaelis-Menten kinetics (Km values: 0.14 mM, 5 mM and 37 nM respectively). Citrate lyase ligase exhibited a high substrate specificity and could not react with citrate lyases from nonphototrophic microorganisms. In contrast to the ligase from Streptococcus diacetilactis, the enzyme from R. gelatinosa was extremely labile; however, it could be stabilized by nucleotides, the most potent stabilizing one being ADP.

Carbon-Sulfur Ligases↗
PubMed1982

Characterization of ATP citrate lyase from Chlorobium limicola.

ATP citrate lyase (EC 4.1.3.8) from Chlorobium limicola was partially purified. It was established that the consumption of substrates and the formation of products proceeded stoichiometrically and that citrate cleavage was of the si-type. ADP and oxaloacetate inhibited enzyme activity. Oxaloacetate also inhibited the growth of C. limicola.

ATP Citrate (pro-S)-Lyase↗