scholarly journals Pyruvate accumulation in growth-inhibited cultures of Aerobacter aerogenes

1968 ◽  
Vol 106 (2) ◽  
pp. 375-380 ◽  
Author(s):  
M. Webb

1. Accumulation of pyruvate occurs during the early stages of exponential growth of aerobic, anaerobic and static cultures of a strain of Aerobacter aerogenes. In normal cultures of this organism the content of pyruvate increases until most of the glucose of the medium has been consumed, and then declines rapidly. The presence of unconsumed sugar is not the sole reason for the accumulation of keto acid, since this is unaffected by the addition of extra glucose to either exponentialphase or stationary-phase cultures. 2. In aminopterin-inhibited cultures, the rate of glucose utilization is decreased greatly, and pyruvate continues to accumulate throughout the period of incubation. This prolonged phase of accumulation appears to be a consequence of the growth inhibition, and not to a specific action of aminopterin on the phosphoroclastic breakdown of pyruvate, since it occurs also when growth is restricted by the antibiotics streptomycin, chloramphenicol and neomycin. 3. A possible explanation is suggested for the accumulation of pyruvate in the inhibited cultures.

1974 ◽  
Vol 20 (12) ◽  
pp. 1709-1712
Author(s):  
K. Amemiya

The metabolic activity of Thiobacillus thiooxidans was found to decrease rapidly as stationary phase was approached. Keeping the culture at constant pH (4.0) and supplementation with CO2 did not effect the decrease in metabolic activity although growth was increased. The respiration rate of cells obtained from stationary phase was negligible. No growth was obtained when the pH was adjusted to pH 6.0. Measurement of pyruvic acid, an inhibitor of metabolic activity, showed that it reached only about 1.0 × 10−5 M during the early stages of growth and then decreased during exponential growth.


Genetics ◽  
1991 ◽  
Vol 128 (4) ◽  
pp. 695-701 ◽  
Author(s):  
J Cairns ◽  
P L Foster

Abstract Mutation rates are generally thought not to be influenced by selective forces. This doctrine rests on the results of certain classical studies of the mutations that make bacteria resistant to phages and antibiotics. We have studied a strain of Escherichia coli which constitutively expresses a lacI-lacZ fusion containing a frameshift mutation that renders it Lac-. Reversion to Lac+ is a rare event during exponential growth but occurs in stationary cultures when lactose is the only source of energy. No revertants accumulate in the absence of lactose, or in the presence of lactose if there is another, unfulfilled requirement for growth. The mechanism for such mutation in stationary phase is not known, but it requires some function of RecA which is apparently not required for mutation during exponential growth.


2010 ◽  
Vol 76 (14) ◽  
pp. 4905-4908 ◽  
Author(s):  
C. Phoebe Lostroh ◽  
Bruce A. Voyles

ABSTRACT Acinetobacter species encounter cycles of feast and famine in nature. We show that populations of A cinetobacter baylyi strain ADP1 remain dynamic for 6 weeks in batch culture. We created a library of lacZ reporters inserted into SalI sites in the genome and then isolated 30 genes with lacZ insertions whose expression was induced by starvation during long-term stationary phase compared with their expression during exponential growth. The genes encode metabolic, gene expression, DNA maintenance, envelope, and conserved hypothetical proteins.


2017 ◽  
Vol 83 (18) ◽  
Author(s):  
Nikolas Duszenko ◽  
Nicole R. Buan

ABSTRACT Many, but not all, organisms use quinones to conserve energy in their electron transport chains. Fermentative bacteria and methane-producing archaea (methanogens) do not produce quinones but have devised other ways to generate ATP. Methanophenazine (MPh) is a unique membrane electron carrier found in Methanosarcina species that plays the same role as quinones in the electron transport chain. To extend the analogy between quinones and MPh, we compared the MPh pool sizes between two well-studied Methanosarcina species, Methanosarcina acetivorans C2A and Methanosarcina barkeri Fusaro, to the quinone pool size in the bacterium Escherichia coli. We found the quantity of MPh per cell increases as cultures transition from exponential growth to stationary phase, and absolute quantities of MPh were 3-fold higher in M. acetivorans than in M. barkeri. The concentration of MPh suggests the cell membrane of M. acetivorans, but not of M. barkeri, is electrically quantized as if it were a single conductive metal sheet and near optimal for rate of electron transport. Similarly, stationary (but not exponentially growing) E. coli cells also have electrically quantized membranes on the basis of quinone content. Consistent with our hypothesis, we demonstrated that the exogenous addition of phenazine increases the growth rate of M. barkeri three times that of M. acetivorans. Our work suggests electron flux through MPh is naturally higher in M. acetivorans than in M. barkeri and that hydrogen cycling is less efficient at conserving energy than scalar proton translocation using MPh. IMPORTANCE Can we grow more from less? The ability to optimize and manipulate metabolic efficiency in cells is the difference between commercially viable and nonviable renewable technologies. Much can be learned from methane-producing archaea (methanogens) which evolved a successful metabolic lifestyle under extreme thermodynamic constraints. Methanogens use highly efficient electron transport systems and supramolecular complexes to optimize electron and carbon flow to control biomass synthesis and the production of methane. Worldwide, methanogens are used to generate renewable methane for heat, electricity, and transportation. Our observations suggest Methanosarcina acetivorans, but not Methanosarcina barkeri, has electrically quantized membranes. Escherichia coli, a model facultative anaerobe, has optimal electron transport at the stationary phase but not during exponential growth. This study also suggests the metabolic efficiency of bacteria and archaea can be improved using exogenously supplied lipophilic electron carriers. The enhancement of methanogen electron transport through methanophenazine has the potential to increase renewable methane production at an industrial scale.


Microbiology ◽  
2014 ◽  
Vol 160 (2) ◽  
pp. 243-260 ◽  
Author(s):  
Öykü İrigül-Sönmez ◽  
Türkan E. Köroğlu ◽  
Büşra Öztürk ◽  
Ákos T. Kovács ◽  
Oscar P. Kuipers ◽  
...  

The lutR gene, encoding a product resembling a GntR-family transcriptional regulator, has previously been identified as a gene required for the production of the dipeptide antibiotic bacilysin in Bacillus subtilis. To understand the broader regulatory roles of LutR in B. subtilis, we studied the genome-wide effects of a lutR null mutation by combining transcriptional profiling studies using DNA microarrays, reverse transcription quantitative PCR, lacZ fusion analyses and gel mobility shift assays. We report that 65 transcriptional units corresponding to 23 mono-cistronic units and 42 operons show altered expression levels in lutR mutant cells, as compared with lutR + wild-type cells in early stationary phase. Among these, 11 single genes and 25 operons are likely to be under direct control of LutR. The products of these genes are involved in a variety of physiological processes associated with the onset of stationary phase in B. subtilis, including degradative enzyme production, antibiotic production and resistance, carbohydrate utilization and transport, nitrogen metabolism, phosphate uptake, fatty acid and phospholipid biosynthesis, protein synthesis and translocation, cell-wall metabolism, energy production, transfer of mobile genetic elements, induction of phage-related genes, sporulation, delay of sporulation and cannibalism, and biofilm formation. Furthermore, an electrophoretic mobility shift assay performed in the presence of both SinR and LutR revealed a close overlap between the LutR and SinR targets. Our data also revealed a significant overlap with the AbrB regulon. Together, these findings reveal that LutR is part of the global complex, interconnected regulatory systems governing adaptation of bacteria to the transition from exponential growth to stationary phase.


2021 ◽  
Author(s):  
Dimitar Plamenov Petrov ◽  
Steffen Kaiser ◽  
Stefanie Kaiser ◽  
Kirsten Jung

mRNA methylation is an important regulator of many physiological processes in eukaryotes but has not been studied in depth in prokaryotes. In contrast to the large number of eukaryotic mRNA modifications that have been described, N6-methyladenosine (m6A) is the only modification of bacterial mRNA identified to date. Here, we used a gel electrophoresis-based RNA separation method and quantitatively analyzed the mRNA-specific modification profile of Escherichia coli using mass spectrometry. In addition to m6A, we provide evidence for the presence of 7-methylguanosine (m7G), and we found first hints for 5-methylcytidine (m5C), N6,N6-dimethyladenosine (m6,6A), 1-methylguanosine (m1G), 5-methyluridine (m5U), and pseudouridine (Ψ) in the mRNA of E. coli, which implies that E. coli has a complex mRNA modification pattern. Furthermore, we observed changes in the abundance of some mRNA modifications during the transition of E. coli from the exponential growth to the stationary phase as well as upon exposure to stress. This study reveals a previously underestimated level of regulation between transcription and translation in bacteria.


mBio ◽  
2020 ◽  
Vol 11 (3) ◽  
Author(s):  
Yanyu Zhu ◽  
Mainak Mustafi ◽  
James C. Weisshaar

ABSTRACT In nature, bacteria must survive long periods of nutrient deprivation while maintaining the ability to recover and grow when conditions improve. This quiescent state is called stationary phase. The biochemistry of Escherichia coli in stationary phase is reasonably well understood. Much less is known about the biophysical state of the cytoplasm. Earlier studies of harvested nucleoids concluded that the stationary-phase nucleoid is “compacted” or “supercompacted,” and there are suggestions that the cytoplasm is “glass-like.” Nevertheless, stationary-phase bacteria support active transcription and translation. Here, we present results of a quantitative superresolution fluorescence study comparing the spatial distributions and diffusive properties of key components of the transcription-translation machinery in intact E. coli cells that were either maintained in 2-day stationary phase or undergoing moderately fast exponential growth. Stationary-phase cells are shorter and exhibit strong heterogeneity in cell length, nucleoid volume, and biopolymer diffusive properties. As in exponential growth, the nucleoid and ribosomes are strongly segregated. The chromosomal DNA is locally more rigid in stationary phase. The population-weighted average of diffusion coefficients estimated from mean-square displacement plots is 2-fold higher in stationary phase for both RNA polymerase (RNAP) and ribosomal species. The average DNA density is roughly twice as high as that in cells undergoing slow exponential growth. The data indicate that the stationary-phase nucleoid is permeable to RNAP and suggest that it is permeable to ribosomal subunits. There appears to be no need to postulate migration of actively transcribed genes to the nucleoid periphery. IMPORTANCE Bacteria in nature usually lack sufficient nutrients to enable growth and replication. Such starved bacteria adapt into a quiescent state known as the stationary phase. The chromosomal DNA is protected against oxidative damage, and ribosomes are stored in a dimeric structure impervious to digestion. Stationary-phase bacteria can recover and grow quickly when better nutrient conditions arise. The biochemistry of stationary-phase E. coli is reasonably well understood. Here, we present results from a study of the biophysical state of starved E. coli. Superresolution fluorescence microscopy enables high-resolution location and tracking of a DNA locus and of single copies of RNA polymerase (the transcription machine) and ribosomes (the translation machine) in intact E. coli cells maintained in stationary phase. Evidently, the chromosomal DNA remains sufficiently permeable to enable transcription and translation to occur. This description contrasts with the usual picture of a rigid stationary-phase cytoplasm with highly condensed DNA.


1975 ◽  
Vol 148 (2) ◽  
pp. 253-258 ◽  
Author(s):  
J A Hackett ◽  
P J Brennan

Besides the monomannophosphoinositide previously reported in Corynebacterium aquaticum small amounts of other, apparently more glycosylated, mannophosphoinositides have been identified in stationary phase cells. Moreover, by labelling cells with [32P]Pi, phosphatidylinositol was found, comprising about 1.5% of the stationary-phase phospholipids. 2. Pulse-chase experiments performed on cells in the late exponential phase of growth further suggested the sequence phosphatidylinositol leads to monomannophosphoinositide as the first step in the biosynthesis of the mannophosphoinositides. 3. Di-and tri-mannophosphoinositides are apparently the main mannophosphoinositides present during exponential growth. Monomannophosphoinositide predominates only in late stationary phase; in the earlier stationary phase, phosphatidylinositol comprises 50% of the phosphoinositide lipid, and tetramannophosphoinositide constitutes much of the remainder. 4. The metabolism and functions of the mannophosphoinositides are discussed, particularly in relation to changes in their composition throughout the growth cycle.


1982 ◽  
Vol 100 (3) ◽  
pp. 327-332 ◽  
Author(s):  
John S. G. van den Bosch ◽  
Anthony G. H. Smals ◽  
Gerlach F. F. M. Pieters ◽  
Ignace M. Valk ◽  
Peter W. C. Kloppenborg

Abstract. A major problem in the androgen treatment of escessive height in boys is acceleration of growth velocity especially in the early stages of therapy. Oestrogen treatment in tall girls, in contrast, instantly decelerates growth velocity, probably by its plasma somatomedin lowering effect. As oestrogen administration in male subjects causes a similar somatomedin depression and immediate growth inhibition is also wanted in the treatment of excessive height in boys, the effect of short-term low dose oestrogen therapy (ethinyloestradiol, Ee, Lynoral®, 0.050 mg daily) on growth was studied in 10 constitutionally tall boys. During oestrogen therapy three week ulnar growth rate (TUG-rate) dropped instantly from 0.84 ± 0.42 to 0.33 ± 0.27 mm (P < 0.02) within 6 weeks. Three week body growth rate also changed significantly from 0.48 ± 0.23 to 0.12 ± 0.37 cm during oestrogen loading (P < 0.05). The magnitude of the latter changes, however, allows only evaluation of the whole group, whereas changes in TUG-rates far exceeded the limits of confidence in most individual boys. Growth deceleration during Ee was accompanied by a significant decrease in serum alkaline phosphatase activities (from 299 ± 72 U/l before to 240 ± 79 U/l during Ee, P < 0.01), plasma calcium (from 2.45 ± 0.06 to 2.35 ± 0.05 mmol/l during Ee, P < 0.05) and plasma testosterone levels (from 392 ± 128 ng/100 ml before to 27 ± 7 ng/100 ml during Ee, P < 0.005). Within 2 months after stopping Ee administration plasma testosterone levels were normal again (432 ± 282 ng/100 ml). Testicular size was not affected. Mild reversible gynaecomastia, however, was present in all boys. The results demonstrate an instant growth decelerating effect of low dose oestrogen administration in tall boys reminiscent to the findings in tall girls under the same low dose regimen. Furthermore these data provide a theoretical base for combining androgens and oestrogens in the early stages of treatment of excessive height in boys in order to antagonize the initial growth accelerating effect of androgens alone.


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