Variation in the activity levels of selected enzymes of Erwinia amylovora 595 in response to changes in dissolved oxygen tension and growth rate of D-glucose-limited chemostat cultures

1975 ◽  
Vol 21 (3) ◽  
pp. 343-352 ◽  
Author(s):  
D. A. Farago ◽  
L. N. Gibbins
Keyword(s):  
Dissolved Oxygen ◽  
Dilution Rate ◽  
Malate Dehydrogenase ◽  
Oxidase Activity ◽  
Erwinia Amylovora ◽  
Specific Activity ◽  
Activity Levels ◽  
Chemostat Cultures ◽  
Succinate Oxidase

Chemostat cultures of Erwinia amylovora 595, grown in mineral salts – nicotinic acid medium at 30 °C, and limited by D-glucose concentrations in the presence of dissolved oxygen tensions (D.O.T.) greater than about 6 mm Hg, became limited by oxygen availability below about 4 mm Hg. This latter limitation was accompanied by a marked increase in acid production as the D.O.T. was depressed. The transition between D-glucose- and oxygen-limitation was also characterized by a maximum in succinate oxidase activity, and a minimum in the in situ respiration. D-Glyceraldehyde-3-phosphate dehydrogenase and D-fructose-1,6-diphosphate aldolase showed small reductions in specific activity in the region 4–6 mm Hg D.O.T., but further reduction to 2 mm Hg resulted in a marked increase in the specific activity of aldolase. Malate dehydrogenase followed the converse trend, and attained very low activity levels when the D.O.T. decreased beyond the lower limits of detection. The in situ respiration was maximal at 2 mm Hg D.O.T., while potential respiration values were minimal at 2 mm Hg, and maximal at about 8 mm Hg D.O.T. The in situ respiration rate was proportional to dilution rate (D), in presence of excess oxygen, up to 0.18 h−1, after which a marked diminution occurred and continued until the wash-out rate was attained. Succinate oxidase activity decreased with increase in dilution rate, but remained constant above D = 0.18 h−1. Malate dehydrogenase showed a persistent decline with increase in dilution rate, while D-glyceraldehyde-3-phosphate activity increased somewhat at higher dilution rates. The data are interpreted in terms of two transition points, at 6 and 2 mm Hg D.O.T., and of a change from respiratory to fermentative metabolism at low D.O.T., and at high dilution rates.

scholarly journals Quantifying Substrate Uptake by Individual Cells of Marine Bacterioplankton by Catalyzed Reporter Deposition Fluorescence In Situ Hybridization Combined with Microautoradiography

2006 ◽  
Vol 72 (11) ◽  
pp. 7022-7028 ◽  
Author(s):  
Eva Sintes ◽  
Gerhard J. Herndl
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Single Cell ◽  
Specific Activity ◽  
Substrate Uptake ◽  
Activity Levels ◽  
Uptake Rates ◽  
Catalyzed Reporter Deposition ◽  
Card Fish

ABSTRACT Catalyzed reporter deposition fluorescence in situ hybridization combined with microautoradiography (MICRO-CARD-FISH) is increasingly being used to obtain qualitative information on substrate uptake by individual members of specific prokaryotic communities. Here we evaluated the potential for using this approach quantitatively by relating the measured silver grain area around cells taking up 3H-labeled leucine to bulk leucine uptake measurements. The increase in the silver grain area over time around leucine-assimilating cells of coastal bacterial assemblages was linear during 4 to 6 h of incubation. By establishing standardized conditions for specific activity levels and concomitantly performing uptake measurements with the bulk community, MICRO-CARD-FISH can be used quantitatively to determine uptake rates on a single-cell level. Therefore, this approach allows comparisons of single-cell activities for bacterial communities obtained from different sites or growing under different ecological conditions.

Mechanism of inhibition of succinate oxidase by products of oxidation of o-dihydroxycoumarins

1980 ◽  
Vol 45 (2) ◽  
pp. 641-652
Author(s):  
Petr Zbořil
Keyword(s):  
Inhibitory Activity ◽  
Oxidase Activity ◽  
Enzyme Preparation ◽  
Inhibitory Effects ◽  
Anaerobic Conditions ◽  
Long Period ◽  
Mechanism Of Inhibition ◽  
Intermediary Product ◽  
Succinate Oxidase ◽  
By Products

Semiquinone is an intermediary product of the oxidation of daphnetin (7,8-dihydroxycoumarin) and esculetin (6,7-dihydroxycoumarin) by diphenol oxidase; its concentration rapidly decreases. When the oxidation is effected by ferricytochrome c, the concentration of the semiquinone remains practically constant for a long period. Similarly, the ability of the products of daphnetin oxidation by diphenol oxidase to inhibit succinate oxidase activity in mitochondrial fragments rapidly decreases with time; the decrease is considerably slower in the case of cytochrome c. The inhibitory activity of the product decreases with time also during esculetin oxidation by ferricyanide. This indicates that the inhibitory effects must be ascribed predominantly to the semiquinone, the quinone is less efficient. The inhibition of succinate oxidase or succinate dehydrogenase was strongly decreased when the enzyme preparation of Keilin and Hartree was incubated with esculetin and ferricyanide in the presence of KCN or under anaerobic conditions. This demonstrates that the reaction of the inhibitor with the enzyme either involves subsequent oxidations or that the inhibitor preferentially reacts with the oxidized form of the sensitive component of the respiratory chain. The second alternative is very little probable since there is no correlation between the degree of inhibition and the binding of the inhibitor to mitochondrial fragments.

Interaction of aging and intermittent ethanol exposure on brain cytochrome c oxidase activity levels

Alcohol ◽  
2003 ◽  
Vol 29 (2) ◽  
pp. 91-100 ◽  
Author(s):  
Pia Jaatinen ◽  
Jarno Riikonen ◽  
Päivi Riihioja ◽  
Olli Kajander ◽  
Antti Hervonen
Keyword(s):  
Cytochrome C ◽  
Cytochrome C Oxidase ◽  
Oxidase Activity ◽  
Ethanol Exposure ◽  
Activity Levels

Expression of a novel FGF in the Xenopus embryo. A new candidate inducing factor for mesoderm formation and anteroposterior specification

Development ◽  
1992 ◽  
Vol 114 (3) ◽  
pp. 711-720 ◽  
Author(s):  
H.V. Isaacs ◽  
D. Tannahill ◽  
J.M. Slack
Keyword(s):  
Specific Activity ◽  
Biological Properties ◽  
The Body ◽  
Mesoderm Induction ◽  
Gastrula Stage ◽  
Blastula Stage ◽  
Mesoderm Formation ◽  
Low Levels

We have cloned and sequenced a new member of the fibroblast growth factor family from Xenopus laevis embryo cDNA. It is most closely related to both mammalian kFGF (FGF-4) and FGF-6 but as it is not clear whether it is a true homologue of either of these genes we provisionally refer to it as XeFGF (Xenopus embryonic FGF). Two sequences were obtained, differing by 11% in derived amino acid sequence, which probably represent pseudotetraploid variants. Both the sequence and the behaviour of in vitro translated protein indicates that, unlike bFGF (FGF-2), XeFGF is a secreted molecule. Recombinant XeFGF protein has mesoderm-inducing activity with a specific activity similar to bFGF. XeFGF mRNA is expressed maternally and zygotically with a peak during the gastrula stage. Both probe protection and in situ hybridization showed that the zygotic expression is concentrated in the posterior of the body axis and later in the tailbud. Later domains of expression were found near the midbrain/hindbrain boundary and at low levels in the myotomes. Because of its biological properties and expression pattern, XeFGF is a good candidate for an inducing factor with possible roles both in mesoderm induction at the blastula stage and in the formation of the anteroposterior axis at the gastrula stage.

RyR1 exhibits lower gain of CICR activity than RyR3 in the SR: evidence for selective stabilization of RyR1 channel

2004 ◽  
Vol 287 (1) ◽  
pp. C36-C45 ◽  
Author(s):  
Takashi Murayama ◽  
Yasuo Ogawa
Keyword(s):  
Binding Sites ◽  
Skeletal Muscles ◽  
Specific Activity ◽  
Adenine Nucleotides ◽  
Minor Effect ◽  
Mammalian Skeletal Muscle ◽  
Selective Stabilization ◽  
Underlying Mechanisms ◽  
A Minor

We showed that frog α-ryanodine receptor (α-RyR) had a lower gain of Ca2+-induced Ca2+ release (CICR) activity than β-RyR in sarcoplasmic reticulum (SR) vesicles, indicating selective “stabilization” of the former isoform (Murayama T and Ogawa Y. J Biol Chem 276: 2953–2960, 2001). To know whether this is also the case with mammalian RyR1, we determined [3H]ryanodine binding of RyR1 and RyR3 in bovine diaphragm SR vesicles. The value of [3H]ryanodine binding (B) was normalized by the number of maximal binding sites (Bmax), whereby the specific activity of each isoform was expressed. This B/Bmax expression demonstrated that ryanodine binding of individual channels for RyR1 was <15% that for RyR3. Responses to Ca2+, Mg2+, adenine nucleotides, and caffeine were not substantially different between in situ and purified isoforms. These results suggest that the gain of CICR activity of RyR1 is markedly lower than that of RyR3 in mammalian skeletal muscle, indicating selective stabilization of RyR1 as is true of frog α-RyR. The stabilization was partly eliminated by FK506 and partly by solubilization of the vesicles with CHAPS, each of which was additive to the other. In contrast, high salt, which greatly enhances [3H]ryanodine binding, caused only a minor effect on the stabilization of RyR1. None of the T-tubule components, coexisting RyR3, or calmodulin was the cause. The CHAPS-sensitive intra- and intermolecular interactions that are common between mammalian and frog skeletal muscles and the isoform-specific inhibition by FKBP12, which is characteristic of mammals, are likely to be the underlying mechanisms.

scholarly journals Simple quality control technique to identify dissolved oxygen diffusion issues with biochemical oxygen demand bottle incubations

2017 ◽  
Author(s):  
Johnathan Daniel Maxey ◽  
Neil David Hartstein ◽  
Dorathy Penjinus ◽  
Alan Kerroux
Keyword(s):  
Dissolved Oxygen ◽  
Water Column ◽  
Biochemical Oxygen Demand ◽  
Oxygen Demand ◽  
Freshwater Ecosystems ◽  
Control Technique ◽  
Community Respiration ◽  
Parallel Diffusion ◽  
Quality Control Technique

Stratified estuaries are home to expanding aquaculture activities whose ecological footprints can be observed through trends in microbial community respiration in the water column. Bottle incubations are widely used to measure water column community respiration in marine and freshwater ecosystems by measuring the flux of dissolved oxygen occurring in the bottle over a period of time. When in situ dissolved oxygen (DO) concentrations are markedly different than DO concentration of the incubation medium the potential for diffusion of oxygen across the bottle opening is great and may be especially pronounced in strongly stratified systems with relatively low rates of pelagic oxygen consumption. We incubated 60 Biochemical Oxygen Demand (BOD) bottles filled with sterilized water with DO concentrations ranging from 2.51 mg O2 L-1 to 10.03 mg O2 L-1 for 24 hours in a temperature controlled water bath. There was a significant relationship when DO flux was set as a function of initial DO (DO Flux = -0.0017x + 0.0085, r2 = 0.72, p < 2.2 e-16). DO fluxes ranged from -0.012 mg O2 L-1 hour-1 to 0.005 mg O2 L-1 hour-1 for bottles incubated with initial DO ranging from 10.03 mg O2 L-1 to 3.31 mg O2 L-1, respectively. These results suggest that diffusion across the ground glass seal of BOD bottles is possible and that extra precaution through parallel diffusion controls should be considered when measuring pelagic respiration using BOD bottle incubations in systems with relatively low or relatively high in situ DO concentrations.

Effect of thermal injury on the TCA cycle enzymes of Staphylococcus aureus MF 31 and Salmonella typhimurium 7136

1971 ◽  
Vol 17 (6) ◽  
pp. 759-765 ◽  
Author(s):  
Richard I. Tomlins ◽  
Merle D. Pierson ◽  
Z. John Ordal
Keyword(s):  
Lactate Dehydrogenase ◽  
Malate Dehydrogenase ◽  
Thermal Injury ◽  
Specific Activity ◽  
Fumarate Hydratase ◽  
Tca Cycle ◽  
Tca Cycle Enzymes ◽  
The Tca Cycle ◽  
Oxoglutarate Dehydrogenase ◽  
Metabolic Activities

The heating of S. aureus MF-31 and S. typhimurium 7136 at 52C and 48C respectively, produced a sublethal heat injury. When injured cells were placed in fresh growth medium they recovered. The recovery of S. aureus was not inhibited by chloramphenicol. The metabolic activities of tricarboxylic acid (TCA) cycle enzymes, as well as other selected enzymes in crude extracts of normal and heat-injured cells of both microorganisms were assayed. In extracts from S. typhimurium there was some loss of specific activity with fumarate hydratase, glutamate dehydrogenase, fructose diphosphate aldolase, lactate dehydrogenase, and the NAD(P) oxidases as a result of heating. In extracts from S. aureus oxoglutarate dehydrogenase, malate dehydrogenase and lactate dehydrogenase were severely inactivated after heating. Other enzymes in comparison were only moderately sensitive to heat. No significant increase in enzyme activity was observed in extracts from injured cells of either microorganism. Re-naturation of lactate dehydrogenase and malate dehydrogenase occurred during the recovery of S. aureus both in the presence and absence of chloramphenicol. No renaturation of oxoglutarate dehydrogenase was found under the same conditions.

scholarly journals Stimulation of the alternative oxidase of Neurospora crassa by Nucleoside phosphates

1980 ◽  
Vol 188 (1) ◽  
pp. 141-144 ◽  
Author(s):  
J Vanderleyden ◽  
C Peeters ◽  
H Verachtert ◽  
H Bertrand
Keyword(s):  
Neurospora Crassa ◽  
Oxidase Activity ◽  
Alternative Oxidase ◽  
Purine Nucleoside ◽  
Submitochondrial Particles ◽  
Fold Stimulation ◽  
Succinate Oxidase ◽  
Stimulation Of

The alternative-oxidase-mediated succinate oxidase activity of Neurospora crassa decreases drastically when mitochondria are fractionated into submitochondrial particles or treated with deoxycholate. The activity, however, can be completely restored in the presence of nucleoside 5′-monophosphates. The purine nucleoside 5′-monophosphates are more effective than the pyrimidine homologues. 5′-GMP gives a 10-fold stimulation of the alternative-oxidase-mediated succinate oxidase activity in submitochondrial particles. A comparison is made with the results obtained earlier with Moniliella tomentosa [Hanssens & Verachtert (1976) J. Bacteriol. 125, 825–835; Vanderleyden, Van Den Eynde & Verachtert (1980) Biochem. J. 186, 309–316].

Activity patterns in relation to body mass and ambient temperature among overwintering cotton rats (Sigmodon hispidus)

1998 ◽  
Vol 76 (4) ◽  
pp. 668-672 ◽  
Author(s):  
Maria A Eifler ◽  
Norman A Slade
Keyword(s):  
Specific Activity ◽  
Activity Patterns ◽  
General Linear Model ◽  
Time Of Day ◽  
Sigmodon Hispidus ◽  
Activity Levels ◽  
Class Time ◽  
Time Class ◽  
Cotton Rats ◽  
Probability Of Capture

We examined mass-specific activity patterns among overwintering cotton rats (Sigmodon hispidus) in northeastern Kansas. We livetrapped animals for a 24-h period, checking traps every 2 h. Trapping occurred every 2 weeks for 5 months. We estimated probability of capture for each mass class, time class, and date, then tested for differences in probability of capture (i.e., activity levels) using a General Linear Model with temperature as a covariate. Large cotton rats were significantly less active than small and intermediate-sized cotton rats. Activity of small cotton rats increased with decreasing temperature, whereas larger cotton rats were less responsive to temperature. Finally, activity levels of large and small cotton rats did not vary significantly with time of day, but intermediate-sized cotton rats were significantly less likely to be captured during the night than at dusk.

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