scholarly journals Indole-3-acetic acid regulates the central metabolic pathways in Escherichia coli

Microbiology ◽  
2006 ◽  
Vol 152 (8) ◽  
pp. 2421-2431 ◽  
Author(s):  
C. Bianco ◽  
E. Imperlini ◽  
R. Calogero ◽  
B. Senatore ◽  
P. Pucci ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Metabolic Pathways ◽  
Tricarboxylic Acid Cycle ◽  
Tricarboxylic Acid ◽  
Mrna Levels ◽  
Glyoxylate Shunt ◽  
Acetyl Coa ◽  
Acid Cycle ◽  
E Coli ◽  
K 12

The physiological changes induced by indoleacetic acid (IAA) treatment were investigated in the totally sequenced Escherichia coli K-12 MG1655. DNA macroarrays were used to measure the mRNA levels for all the 4290 E. coli protein-coding genes; 50 genes (1.1 %) exhibited significantly different expression profiles. In particular, genes involved in the tricarboxylic acid cycle, the glyoxylate shunt and amino acid biosynthesis (leucine, isoleucine, valine and proline) were up-regulated, whereas the fermentative adhE gene was down-regulated. To confirm the indications obtained from the macroarray analysis the activity of 34 enzymes involved in central metabolism was measured; this showed an activation of the tricarboxylic acid cycle and the glyoxylate shunt. The malic enzyme, involved in the production of pyruvate, and pyruvate dehydrogenase, required for the channelling of pyruvate into acetyl-CoA, were also induced in IAA-treated cells. Moreover, it was shown that the enhanced production of acetyl-CoA and the decrease of NADH/NAD+ ratio are connected with the molecular process of the IAA response. The results demonstrate that IAA treatment is a stimulus capable of inducing changes in gene expression, enzyme activity and metabolite level involved in central metabolic pathways in E. coli.

scholarly journals Role of Phosphoenolpyruvate in the NADP-Isocitrate Dehydrogenase and Isocitrate Lyase Reaction in Escherichia coli

2006 ◽  
Vol 189 (3) ◽  
pp. 1176-1178 ◽  
Author(s):  
Tadashi Ogawa ◽  
Keiko Murakami ◽  
Hirotada Mori ◽  
Nobuyoshi Ishii ◽  
Masaru Tomita ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Isocitrate Dehydrogenase ◽  
Isocitrate Lyase ◽  
Tricarboxylic Acid Cycle ◽  
Tricarboxylic Acid ◽  
Glyoxylate Shunt ◽  
Acid Cycle ◽  
Uncompetitive Inhibition

ABSTRACT Phosphoenolpyruvate inhibited Escherichia coli NADP-isocitrate dehydrogenase allosterically (Ki of 0.31 mM) and isocitrate lyase uncompetitively (Ki ′ of 0.893 mM). Phosphoenolpyruvate enhances the uncompetitive inhibition of isocitrate lyase by increasing isocitrate, which protects isocitrate dehydrogenase from the inhibition, and contributes to the control through the tricarboxylic acid cycle and glyoxylate shunt.

scholarly journals [13C]propionate oxidation in wild-type and citrate synthase mutant Escherichia coli: evidence for multiple pathways of propionate utilization

1993 ◽  
Vol 291 (3) ◽  
pp. 927-932 ◽  
Author(s):  
C T Evans ◽  
B Sumegi ◽  
P A Srere ◽  
A D Sherry ◽  
C R Malloy
Keyword(s):  
Escherichia Coli ◽  
Citrate Synthase ◽  
Tricarboxylic Acid Cycle ◽  
Tricarboxylic Acid ◽  
Spin Coupling ◽  
Wild Type ◽  
Acid Cycle ◽  
E Coli ◽  
Cell Extracts ◽  
In Cells

The metabolism of propionate was examined in wild-type Escherichia coli and cells lacking citrate synthase by high-resolution 13C n.m.r. Spectra of cell extracts from wild-type E. coli show that glutamate becomes highly enriched in 13C when 13C-enriched propionate is the sole carbon source. No glutamate labelling was detected when the tricarboxylic acid cycle was blocked either by deletion of citrate synthase or by inhibition of succinate dehydrogenase by malonate. The 13C fractional enrichment in glutamate C-2, C-3 and C-4 in wild-type cells was quantitatively and qualitatively different when [2-13C]propionate as opposed to [3-13C]propionate was supplied. Approximately equal labelling occurred in the C-2, C-3 and C-4 positions of glutamate when [3-13C]propionate was available, and multiplets due to carbon-carbon spin-spin coupling were observed. However, in cells supplied with [2-13C]propionate, very little 13C appeared in the glutamate C-4 position, and the remaining glutamate resonances all appeared as singlets. The unequal and non-identical labelling of glutamate in cells supplied with [2-13C]- as opposed to [3-13C]propionate is consistent with the utilization of propionate by E. coli via two pathways, oxidation of propionate to pyruvate and carboxylation of propionate to succinate. These intermediates are further metabolized to glutamate by the action of the tricarboxylic acid cycle. The existence of an organized tricarboxylic acid cycle is discussed as a consequence of the ability to block utilization of propionate in tricarboxylic acid-cycle-defective E. coli.

The anaplerotic fixation of carbon dioxide by Escherichia coli

1966 ◽  
Vol 165 (999) ◽  
pp. 179-188 ◽  
Keyword(s):  
Escherichia Coli ◽  
Tricarboxylic Acid Cycle ◽  
Tricarboxylic Acid ◽  
Growth Media ◽  
Wild Type ◽  
Carboxylase Activity ◽  
Acid Cycle ◽  
K 12 ◽  
Tricarboxylic Acid Cycle Intermediates ◽  
Fixation Of Carbon Dioxide

The isolation of a mutant (AB 1622) of Escherichia coli K 12 is described, which differs from its parent organism (AB 1621) in lacking the ability to grow upon glucose, glycerol or pyruvate unless utilizable intermediates of the tricarboxylic acid cycle are also supplied in the growth media; both the mutant and its parent grow readily on acetate as sole carbon source. Washed suspensions of AB 1622 oxidized pyruvate only to the level of acetate, which accumulated; when catalytic quantities of L-malate were also supplied, pyruvate was oxidized further and the extent to which it was oxidized approached that observed with suspensions of the wild-type organisms. These observations suggest that the mutant is unable to effect the net formation of tricarboxylic acid cycle intermediates from pyruvate or phosphopyruvate. Analysis of extracts of the mutant, and of its parent organism, showed that the former lacked phosphopyruvate carboxylase activity (EC 4.1.1.31), although other enzymes capable in theory of catalysing the carboxylation of C 3 -acids were abundant in both extracts. It is thus concluded that the net formation of C 4 -acids through carboxylation of C 3 -precursors is necessarily achieved through the agency of phosphopyruvate carboxylase.

scholarly journals Growth Enhancement Facilitated by Gaseous CO2 through Heterologous Expression of Reductive Tricarboxylic acid Cycle Genes in Escherichia coli

Fermentation ◽  
2021 ◽  
Vol 7 (2) ◽  
pp. 98
Author(s):  
Shou-Chen Lo ◽  
En-Pei Isabel Chiang ◽  
Ya-Tang Yang ◽  
Si-Yu Li ◽  
Jian-Hau Peng ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Carbon Fixation ◽  
Tricarboxylic Acid Cycle ◽  
Anaerobic Respiration ◽  
Tricarboxylic Acid ◽  
Coli Strain ◽  
Growth Enhancement ◽  
Acid Cycle ◽  
E Coli ◽  
Reductive Tricarboxylic Acid Cycle

The enzymatic mechanisms of carbon fixation by autotrophs, such as the reductive tricarboxylic acid cycle (rTCA), have inspired biotechnological approaches to producing bio-based chemicals directly through CO2. To explore the possibility of constructing an rTCA cycle in Escherichia coli and to investigate their potential for CO2 assimilation, a total of ten genes encoding the key rTCA cycle enzymes, including α-ketoglutarate:ferredoxin oxidoreductase, ATP-dependent citrate lyase, and fumarate reductase/succinate dehydrogenase, were cloned into E. coli. The transgenic E. coli strain exhibited enhanced growth and the ability to assimilate external inorganic carbon with a gaseous CO2 supply. Further experiments conducted in sugar-free medium containing hydrogen as the electron donor and dimethyl sulfoxide (DMSO) as the electron acceptor proved that the strain is able to undergo anaerobic respiration, using CO2 as the major carbon source. The transgenic stain demonstrated CO2-enhanced growth, whereas the genes involved in chemotaxis, flagellar assembly, and acid-resistance were upregulated under the anaerobic respiration. Furthermore, metabolomic analysis demonstrated that the total concentrations of ATP, ADP, and AMP in the transgenic strain were higher than those in the vector control strain and these results coincided with the enhanced growth. Our approach offers a novel strategy to engineer E. coli for assimilating external gaseous CO2.

13C isotopomer model for estimation of anaplerotic substrate oxidation via acetyl-CoA

1996 ◽  
Vol 271 (4) ◽  
pp. E788-E799 ◽  
Author(s):  
F. M. Jeffrey ◽  
C. J. Storey ◽  
A. D. Sherry ◽  
C. R. Malloy
Keyword(s):  
Biochemical Analysis ◽  
Tricarboxylic Acid Cycle ◽  
Tricarboxylic Acid ◽  
Acetyl Coa ◽  
Acid Cycle ◽  
Propionate Metabolism ◽  
13C Nuclear Magnetic Resonance ◽  
Isotopomer Analysis ◽  
Tricarboxylic Acid Cycle Intermediates ◽  
Anaplerotic Pathway

A previous model using 13C nuclear magnetic resonance isotopomer analysis provided for direct measurement of the oxidation of 13C-enriched substrates in the tricarboxylic acid cycle and/or their entry via anaplerotic pathways. This model did not allow for recycling of labeled metabolites from tricarboxylic acid cycle intermediates into the acetyl-CoA pool. An extension of this model is now presented that incorporates carbon flow from oxaloacetate or malate to acetyl-CoA. This model was examined using propionate metabolism in the heart, in which previous observations indicated that all of the propionate consumed was oxidized to CO2 and water. Application of the new isotopomer model shows that 2 mM [3-13C]propionate entered the tricarboxylic acid cycle as succinyl-CoA (an anaplerotic pathway) at a rate equal to 52% of tricarboxylic acid cycle turnover and that all of this carbon entered the acetyl-CoA pool and was oxidized. This was verified using standard biochemical analysis; from the rate (mumol.min-1.g dry wt-1) of propionate uptake (4.0 +/- 0.7), the estimated oxygen consumption (24.8 +/- 5) matched that experimentally determined (24.4 +/- 3).

Metabolic flux analysis of Escherichia coli in glucose-limited continuous culture. I. Growth-rate-dependent metabolic efficiency at steady state

Microbiology ◽  
2005 ◽  
Vol 151 (3) ◽  
pp. 693-706 ◽  
Author(s):  
Anke Kayser ◽  
Jan Weber ◽  
Volker Hecht ◽  
Ursula Rinas
Keyword(s):  
Escherichia Coli ◽  
Tricarboxylic Acid Cycle ◽  
Tricarboxylic Acid ◽  
Linear Increase ◽  
Carbon Substrate ◽  
Carbon Dioxide Evolution ◽  
Continuous Cultures ◽  
Acid Cycle ◽  
Biomass Formation ◽  
Carbon Supply

The Escherichia coli K-12 strain TG1 was grown at 28 °C in aerobic glucose-limited continuous cultures at dilution rates ranging from 0·044 to 0·415 h−1. The rates of biomass formation, the specific rates of glucose, ammonium and oxygen uptake and the specific carbon dioxide evolution rate increased linearly with the dilution rate up to 0·3 h−1. At dilution rates between 0·3 h−1 and 0·4 h−1, a strong deviation from the linear increase to lower specific oxygen uptake and carbon dioxide evolution rates occurred. The biomass formation rate and the specific glucose and ammonium uptake rates did not deviate that strongly from the linear increase up to dilution rates of 0·4 h−1. An increasing percentage of glucose carbon flow towards biomass determined by a reactor mass balance and a decreasing specific ATP production rate concomitant with a decreasing adenylate energy charge indicated higher energetic efficiency of carbon substrate utilization at higher dilution rates. Estimation of metabolic fluxes by a stoichiometric model revealed an increasing activity of the pentose phosphate pathway and a decreasing tricarboxylic acid cycle activity with increasing dilution rates, indicative of the increased NADPH and precursor demand for anabolic purposes at the expense of ATP formation through catabolic activities. Thus, increasing growth rates first result in a more energy-efficient use of the carbon substrate for biomass production, i.e. a lower portion of the carbon substrate is channelled into the respiratory, energy-generating pathway. At dilution rates above 0·4 h−1, close to the wash-out point, respiration rates dropped sharply and accumulation of glucose and acetic acid was observed. Energy generation through acetate formation yields less ATP compared with complete oxidation of the sugar carbon substrate, but is the result of maximized energy generation under conditions of restrictions in the tricarboxylic acid cycle or in respiratory NADH turnover. Thus, the data strongly support the conclusion that, in aerobic glucose-limited continuous cultures of E. coli TG1, two different carbon limitations occur: at low dilution rates, cell growth is limited by cell-carbon supply and, at high dilution rates, by energy-carbon supply.

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