Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “PENTOSES”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 1,297 records · Page 72Linked to original sources
PubMed1977

On arabinose as a component of brain hyaluronate. Confirmation by chromatographic, enzymatic and chemical ionization-mass spectrometric analyses.

The controversy about the presence of the pentose arabinose in brain hyaluronate was reinvestigated using modern analytical tehcnics. The purified bovine brain hyaluronate contained the neutral sugars: arabinose, 0.18%; glucose, 0.05%; and fucose, 0.22%. The confirmation of the presence of arabinose was obtained by paper and thin layer chromatography of the neutral sugars in deionized hyaluronate hydrolysate. Gas-liquid chromatography of the aldononitrile peracetate of the pentose isolated by preparative paper chromatography gave a single distinct peak, corresponding to standard arabinose on three columns packed with three different phases. Chemical ionization data and mass spectrum of the aldononitrile peracetate derivative agreed with those of the authentic arabinonitrile tetracetate. Analysis of the isolated pentose with the help of the enzymes L-arabinose isomerase and L-ribulose kinase, which are specific for their substrates, further established its identity as L-arabinose.

Animals↗
PubMed1981

The phosphogluconate pathway and synthesis of 5-phosphoribosyl-1-pyrophosphate in human fibroblasts.

The phosphogluconate pathway (pentose phosphate cycle) of normal fibroblasts was stimulated 20-fold by methylene blue and inhibited to 14% of the baseline rate by 6-aminonicotinamide. In fibroblasts deficient in glucose-6-phosphate dehydrogenase activity (an average of 1.5% of normal mean), the pentose phosphate cycle was unaffected by either methylene blue or 6-aminonicotinamide. In normal cells, neither the intracellular concentration nor the rate of generation of 5-phosphoribosyl-1-pyrophosphate was altered by the marked and opposite changes in the rate of the phosphogluconate pathway caused by methylene blue and 6-aminonicotinamide. Intracellular ribose 5-phosphate concentration was increase by methylene blue (an average increase of 83%) but not significantly altered by 6-aminonicotinamide. In fibroblasts deficient in glucose-6-phosphate dehydrogenase activity, the 5-phosphoribosyl-1-pyrophosphate concentration and rate of generation were higher rather than lower in comparison to normal cells under all conditions studied. The data suggest a predominant role for the nonoxidative branch of the phosphogluconate pathway in supplying ribose 5-phosphate for nucleotide biosynthesis. Pentose phosphate supply cannot be considered an essential function of the oxidative branch in fibroblasts.

Cell Line↗
PubMed1982

The formation of mixed disulphides in rat lung following paraquat administration. Correlation with changes in intermediary metabolism.

We have investigated the hypothesis that the formation of mixed disulphides between protein sulphydryl and glutathione may be responsible for controlling the activity of the pentose phosphate pathway and fatty acid synthesis in rat lung. Using lung slices, taken form rats 2 h after dosing with a range of concentrations (5-80 mg/kg) of the pulmonary toxin paraquat, the pentose phosphate pathway was found to be stimulated in direct proportion to a reduction in fatty acid synthesis. These effects were also linearly related to an increase in mixed (total) disulphide levels in the lung. This was quantitatively similar to an increase in mixed (glutathione) disulphides, although non-protein sulphydryl and oxidised glutathione levels remained normal. Thus, an early biochemical event in the mechanism of the paraquat toxicity in the lung involves an increased formation of mixed (glutathione) disulphides and simultaneous regulation of pentose phosphate pathway activity and fatty acid synthesis. These data support the concept that the formation of mixed disulphides of protein and glutathione is a mechanism for maintaining NADPH levels despite the 'redox' stress caused by the cyclical and NADPH dependent reduction and reoxidation of paraquat.

Animals↗
PubMed2006

Effect of vitamin C and folic acid on seed vigour response and phenolic-linked antioxidant activity.

Folic acid and vitamin C were used in the concentration range of 0-500muM as exogenous growth enhancers to stimulate pea (Pisum sativum) seedling vigour. The results suggest that a concentration of 50muM folic acid and 500muM vitamin C were optimum in maximally enhancing seed vigour and potentially seedling performance according to both agronomic and biochemical seed vigour parameters. Results indicated that germination percentage, shoot weight, shoot height, and root length were enhanced in folic acid and vitamin C treated plants compared to control plants. The levels of enhanced phenolic content in response to folic acid and vitamin C treatments were highest on days 8 and 10. Evaluation of critical biochemical parameters indicated that the average glucose-6-phosphate dehydrogenase (G6PDH) activity and proline content in response to treatments were higher than control and correlated to enhanced phenolic content and DPPH-based antioxidant activity. Key enzymes, guaiacol peroxidase (GPX), superoxide dismutase (SOD), and catalase (CAT) were also higher in response to treatments and correlated to enhanced phenolic content and DPPH-based antioxidant activity. Taken together, these studies support the hypothesis that the proline-linked pentose phosphate pathway stimulates phenolic synthesis and related free-radical scavenging antioxidant activity. Further, this proline-linked pentose phosphate pathway stimulation in response to folic acid and vitamin C was also correlated to antioxidant enzyme response indicated by the stimulation of GPX, SOD, and CAT activities. Therefore, this study indicates the enhancement of seed vigour response by folic acid and vitamin C as reflected in both agronomic and biochemical responses, and this occurred through the stimulation of phenolic-linked antioxidant response that is likely positively modulated through the proline-linked pentose phosphate pathway.

Analysis of Variance↗
PubMed2004

Metabolic flux analysis for a ppc mutant Escherichia coli based on 13C-labelling experiments together with enzyme activity assays and intracellular metabolite measurements.

The physiology and central metabolism of a ppc mutant Escherichia coli were investigated based on the metabolic flux distribution obtained by (13)C-labelling experiments using gas chromatography-mass spectrometry (GC-MS) and 2-dimensional nuclear magnetic resonance (2D NMR) strategies together with enzyme activity assays and intracellular metabolite concentration measurements. Compared to the wild type, its ppc mutant excreted little acetate and produced less carbon dioxide at the expense of a slower growth rate and a lower glucose uptake rate. Consequently, an improvement of the biomass yield on glucose was observed in the ppc mutant. Enzyme activity measurements revealed that isocitrate lyase activity increased by more than 3-fold in the ppc mutant. Some TCA cycle enzymes such as citrate synthase, aconitase and malate dehydrogenase were also upregulated, but enzymes of glycolysis and the pentose phosphate pathway were downregulated. The intracellular intermediates in the glycolysis and the pentose phosphate pathway, therefore, accumulated, while acetyl coenzyme A and oxaloacetate concentrations decreased in the ppc mutant. The intracellular metabolic flux analysis uncovered that deletion of ppc resulted in the appearance of the glyoxylate shunt, with 18.9% of the carbon flux being channeled via the glyoxylate shunt. However, the flux of the pentose phosphate pathway significantly decreased in the ppc mutant.

Carbon Isotopes↗
PubMed2005

The influence of different cultivation conditions on the metabolome of Fusarium oxysporum.

The two most widespread pentose sugars found in the biosphere are d-xylose and l-arabinose. They are both potential substrates for ethanol production. The purpose of this study was to better understand the redox constraints imposed to Fusarium oxysporum during utilization of pentoses. In order to increase ethanol yield and decrease by-product formation, nitrate was used as nitrogen source. The use of NADH, the cofactor in denitrification process when using nitrate as a nitrogen source, improved the ethanol yield on xylose to 0.89 mol mol(-1) compared to the ethanol yield achieved using ammonium as nitrogen source 0.44 mol mol(-1). The improved ethanol yield was followed by a 28% decrease in yield of the by-product xylitol. In order to investigate the metabolic pathway of arabinose and the metabolic limitations for the efficient ethanol production from this sugar, the extracellular and intracellular metabolite profiles were determined under aerobic and anaerobic cultivation conditions. The results of this study clearly show difficulties in channelling of glucose-1-P (G1P) to pentose phosphate pathway (PPP) and reduced NADPH regeneration, suggesting that NADPH becomes a limiting factor for arabinose conversion, resulting in excessive acetate production. Variations of the fungus intracellular amino and non-amino acid pool, under different culture conditions, were evaluated using principal component analysis (PCA). PCA projection of the metabolome data collected from F. oxysporum subjected to environmental perturbations succeeded to visualize different physiological states and the conclusions of this study were that the metabolite profile is unique according to: (1) the carbon source and (2) the oxygen supply, and to a lesser extent to the cultivation phase.

Adaptation, Physiological↗
PubMed2006

Estradiol stimulates the biosynthetic pathways of breast cancer cells: detection by metabolic flux analysis.

Selective estrogen receptor (ER) modulators are highly successful breast cancer therapies, but they are not effective in patients with ER negative and selective estrogen receptor modulator (SERM)-resistant tumors. Understanding the mechanisms of estrogen-stimulated proliferation may provide a route to design estrogen-independent therapies that would be effective in these patients. In this study, metabolic flux analysis was used to determine the intracellular fluxes that are significantly affected by estradiol stimulation in MCF-7 breast cancer cells. Intracellular fluxes were calculated from nuclear magnetic resonance (NMR)-generated isotope enrichment data and extracellular metabolite fluxes, using a specific flux analysis algorithm. The metabolic pathway model used by the algorithm includes glycolysis, the tricarboxylic acid cycle (TCA cycle), the pentose phosphate pathway, glutamine catabolism, pyruvate carboxylase, and malic enzyme. The pathway model also incorporates mitochondrial compartmentalization and reversible trans-mitochondrial membrane reactions to more accurately describe the role of mitochondria in cancer cell proliferation. Flux results indicate that estradiol significantly increases carbon flow through the pentose phosphate pathway and increases glutamine consumption. In addition, intra-mitochondrial malic enzyme was found to be inactive and the malate-aspartate shuttle (MAS) was only minimally active. The inactivity of these enzymes indicates that glutamine is not oxidized within mitochondria, but is consumed primarily to provide biosynthetic precursors. The excretion of glutamine carbons from the mitochondria has the secondary effect of limiting nicotinamide adenine dinucleotide (NADH) recycle, resulting in NADH buildup in the cytosol and the excretion of lactate. The observed dependence of breast cancer cells on pentose phosphate pathway activity and glutamine consumption for estradiol-stimulated biosynthesis suggests that these pathways may be targets for estrogen-independent breast cancer therapies.

Algorithms↗
PubMed2000

In vivo measurements of control coefficients for hexokinase and glucose-6-phosphate dehydrogenase in Xenopus laevis oocytes.

Hexokinase and glucose-6-phosphate dehydrogenase activities were increased in Xenopus laevis oocytes by microinjection of commercial pure enzymes. The effect of increased fractional activities on glycogen synthesis or on the production of 14CO(2) (the oxidative portion of the pentose phosphate pathway) was investigated by microinjection of [1-(14)C]glucose and measurements of the radioactivity in glycogen and CO(2). Control coefficients calculated from the data show that hexokinase plays an important role in the control of glycogen synthesis (control coefficient=0.7) but its influence on the control of the pentose phosphate pathway is almost nil (control coefficient=-0.01). Glucose-6-phosphate dehydrogenase injections did not affect the production of 14CO(2) by the pentose phosphate pathway, indicating that other factors control the operation of this pathway. In addition, an almost null control of this enzyme on glycogen synthesis flux was observed.

Animals↗
PubMed1992

Evolution of metabolic pathways by chance assembly of enzyme proteins generated from sense and antisense strands of pre-existing genes.

In order to get an insight into the evolutionary aspect of metabolic pathways, especially of the ubiquitous glycolytic pathway, we have carried out an extensive search of sense-sense and sense-antisense similarities for enzyme proteins in the glycolytic pathway, the pentose phosphate cycle, alcohol and lactate fermentation pathways and the TCA cycle. This investigation of amino acid sequences reveals a curious pattern of similarity relations; no similarity can be found between the enzyme proteins in a section of the glycolytic pathway where the glyceraldehyde-3-phosphate or even glycerol-3-phosphate is converted into the pyruvate while many examples of sense-sense and sense-antisense similarities are found even between enzyme proteins in distant blocks, e.g. between the proteins in the TCA cycle and those in the pentose phosphate cycle, as well as between the functionally associated proteins in each of these blocks. Complementary to this characteristic pattern of amino acid sequence similarity, the search for similarities of nucleotide sequences also finds that the similarities of glycolytic enzyme genes, some sense-sense and others sense-antisense similarities, are concentrated on the nucleotide sequences of prokaryotic 16S or eukaryotic 18S ribosomal RNA gene with its flanks, although some of the copy sequences are also found in transfer RNA genes as well as in 23S or 26S ribosomal RNA gene. These results strongly suggest that the metabolic pathways have been developed by the chance assembly of enzyme proteins generated from the sense and antisense strands of pre-existing genes, e.g. the fermentation pathways and pentose phosphate cycle by the proteins from the genes of enzymes in the glycolytic pathway and the TCA cycle from all these successively increased genes, ascribing the origin of metabolic enzyme genes to the close relation between the glycolytic enzyme protein genes and the RNA gene cluster.

Animals↗
PubMed2001

Tracer studies with 13C-labeled carbohydrates in cultured plant cells. Retrobiosynthetic analysis of chelidonic acid biosynthesis.

The biosynthesis of chelidonic acid was studied in cell suspension cultures of Leucojum aestivum. Cell cultures were supplied with [U-13C]glucose, [l-13C]glucose or [U-13Cs]ribose/ribulose in standard medium containing unlabeled glucose. 13C labeling patterns of amino acids obtained by hydrolysis of biomass were determined by NMR spectroscopy and compared to the labeling pattern of chelidonic acid. The data document the incorporation of a contiguous 4-carbon fragment derived from the pentose phosphate pool into chelidonic acid. This suggests a biosynthetic pathway involving the condensation of phosphoenolpyruvate with a pentose phosphate followed by dehydration, dehydrogenation, ring closure and decarboxylation conducive to the loss of C-5 of the pentose precursor.

Carbohydrate Metabolism↗
PubMed1997

The influence of progesterone-induced proteins on glucose metabolism in early equine embryos.

The influence of different maternal plasma progesterone concentrations on embryonic glucose metabolism was studied. Uterine flushes were obtained after treating ovariectomized mares (n = 3) with 0 (control), 100 or 200 mg progesterone daily for 7 d. A group of progesterone-induced proteins (PIP) of Mr approximately 20,000 were identified in flushes from progesterone treatments by SDS-PAGE but were not observed in control flushes. Progesterone-induced proteins were removed from half the pooled flush in each treatment group by Sepharose blue CL-6B. In a 3 x 2 factorial (progesterone treatments, progesterone-induced proteins) experiment, 6 groups of Day 7 equine embryos (n = 6 per group) were incubated in culture media (MEM:DPBS; 1:3) containing radioactively-labeled glucose. Contributions of the Embden-Meyerhof pathway (EMP) and the pentose-phosphate pathway (PPP) to the total metabolism of glucose in early equine embryos were assessed separately. In the 200 mg progesterone treatment group, the presence of progesterone-induced proteins in the culture medium resulted in a 4-to 5-fold increase in the activities of the Embden-Meyerhof pathway and the pentose-phosphate pathway. These results lead to the following conclusions: 1)Addition of progesterone-induced uterine proteins from mares with high levels of circulating progesterone enhance the metabolic activities of the Embden-Meyerhof pathway and the pentose-phosphate pathway in Day 7.5 equine embryos in culture. 2)Uterine secretion of progesterone-induced proteins which is quantitatively and/or qualitatively adequate to modify embryonic glucose metabolism in vitro is dependent on a minimal concentration of maternal plasma progesterone.

Journal Article↗
PubMed1999

Blood sugar formation due to abnormally elevated gluconeogenesis: aberrant regulation in a parasitized insect, Manduca sexta Linnaeus.

Alterations of carbohydrate metabolism associated with parasitism were examined in an insect, Manduca sexta L. In insect larvae maintained on a low carbohydrate diet gluconeogenesis from [3-13C]alanine was established from the fractional 13C enrichment in trehalose, a disaccharide of glucose and the blood sugar of insects and other invertebrates. After transamination of the isotopically substituted substrate to [3-13C]pyruvate, the latter was carboxylated to oxaloacetate ultimately leading to de novo glucose synthesis and trehalose formation. Trehalose was selectively enriched with 13C at C1 and C6 followed by C2 and C5. 13C enrichment of blood sugar in insects parasitized by Cotesia congregata (Say) was significantly greater than was observed in normal animals. The relative contributions of pyruvate carboxylation and decarboxylation to trehalose labeling were determined from the 13C distribution in glutamine, synthesized as a byproduct of the tricarboxylic acid cycle. The relative contribution of carboxylation was significantly greater in parasitized larvae than in normal insects providing additional evidence of elevated gluconeogenesis due to parasitism. Despite the increased gluconeogenesis in parasitized insects the level of blood sugar was the same in all animals. Because de novo glucose synthesis does not normally maintain blood sugar level in insects maintained under these dietary conditions the findings suggest an aberrant regulation over gluconeogenesis. The 13C labeling in trehalose was nearly symmetric in all insects but the mean C1/C6 13C ratio was higher in parasitized animals suggesting a lower activity of the pentose phosphate pathway that brings about a redistribution of 13C in trehalose following de novo glucose synthesis. Additional studies with insects maintained on a high carbohydrate diet and administered [1,2-13C2]glucose confirmed a decreased level of pentose cycling during parasitism consistent with a lower level of lipogenesis. It is suggested, however, that the pentose pathway may facilitate the synthesis of trehalose from dietary carbohydrate by directing hexose phosphate cycled through the pathway to the production of energy.

Animals↗
PubMed2003

Effect of carbohydrate overfeeding on whole body and adipose tissue metabolism in humans.

OBJECTIVE: To evaluate the effect of a 4-day carbohydrate overfeeding on whole body net de novo lipogenesis and on markers of de novo lipogenesis in subcutaneous adipose tissue of healthy lean humans. RESEARCH METHODS AND PROCEDURES: Nine healthy lean volunteers (five men and four women) were studied after 4 days of either isocaloric feeding or carbohydrate overfeeding. On each occasion, they underwent a metabolic study during which their energy expenditure and net substrate oxidation rates (indirect calorimetry), and the fractional activity of the pentose-phosphate pathway in subcutaneous adipose tissue (subcutaneous microdialysis with 1,6(13)C2,6,6(2)H2 glucose) were assessed before and after administration of glucose. Adipose tissue biopsies were obtained at the end of the experiments to monitor mRNAs of key lipogenic enzymes. RESULTS: Carbohydrate overfeeding increased basal and postglucose energy expenditure and net carbohydrate oxidation. Whole body net de novo lipogenesis after glucose loading was markedly increased at the expense of glycogen synthesis. Carbohydrate overfeeding also increased mRNA levels for the key lipogenic enzymes sterol regulatory element-binding protein-1c, acetyl-CoA carboxylase, and fatty acid synthase. The fractional activity of adipose tissue pentose-phosphate pathway was 17% to 22% and was not altered by carbohydrate overfeeding. DISCUSSION: Carbohydrate overfeeding markedly increased net de novo lipogenesis at the expense of glycogen synthesis. An increase in mRNAs coding for key lipogenic enzymes suggests that de novo lipogenesis occurred, at least in part, in adipose tissue. The pentose-phosphate pathway is active in adipose tissue of healthy humans, consistent with an active role of this tissue in de novo lipogenesis.

Adipose Tissue↗
PubMed1974

Regulation of hexose phosphate metabolism in Acetobacter xylinum.

The metabolism of glucose and fructose was studied in resting succinate-grown cells of Acetobacter xylinum. From fructose only cellulose and CO(2) were formed by the cells, whereas from glucose, gluconate was formed much more rapidly than these two products. The molar ratio of sugar converted into cellulose to sugar converted into CO(2) was significantly greater than unity for both hexoses. The pattern of label retention in the cellulose formed by the cells from specifically (14)C-labelled glucose, fructose or gluconate corresponded to that of hexose phosphate in a pentose cycle. On the other hand, the isotopic configuration of cellulose arising from variously singly (14)C-labelled pyruvate did not agree with the operation of a pentose cycle on gluconeogenic hexose phosphate. Readily oxidizable tricarboxylic acid-cycle intermediates such as acetate, pyruvate or succinate promoted cellulose synthesis from fructose and gluconate although retarding their oxidation to CO(2). The incorporation into cellulose of C-1 of fructose was greatly increased in the presence of these non-sugar substrates, although its oxidation to CO(2) was greatly diminished. It is suggested that the flow of hexose phosphate carbon towards cellulose or through the pentose cycle in A. xylinum is regulated by an energy-linked control mechanism.

Acetates↗
PubMed1974

The metabolism of macromolecules during the differentiation of Myxamoebae of the cellular slime mould Dictyostelium discoideum containing different amounts of glycogen.

1. Methods of obtaining myxamoebae of Dictyostelium discoideum strain Ax-2 (ATCC 24397) with glycogen contents in the range 0.05-5mg of glycogen/10(8) cells are described. The changes in cellular glycogen, protein and RNA content during the differentiation of such myxamoebae were determined. 2. Myxamoebal glycogen is not conserved during differentiation and gluconeogenesis may occur even in cells that contain a large amount of glycogen initially. 3. There is a marked net loss of cellular protein and RNA during differentiation and associated with this there are also marked decreases in the sizes of the intracellular pools of amino acids, acid-soluble proteins and pentose-containing materials. 4. During the early stages of development some protein and pentose(s) are excreted, but this cannot account for the decreased cellular content of protein and RNA. 5. There is a linear rate of production of NH(3) during development, and oxidation appears to be the fate of the major portion of the degraded protein and RNA. 6. However, provision of an alternative metabolizable energy source (glycogen) has little effect on the rate or extent of protein or RNA breakdown or on the changes in the sizes of the intracellular pools of amino acids, acid-soluble proteins and pentose-containing materials. 7. It is concluded that during development there is a requirement for the destruction of specific RNA and protein molecules for reasons other than the provision of oxidizable substrates. 8. The kinetic model of Wright et al. (1968) is discussed in relation to these changes in macromolecular content.

Ammonia↗
PubMed1980

The utilization of glucose for the synthesis of milk components in the fed and starved lactating goat in vivo.

1. [U-14C]Glucose and [3-3H]glucose were infused into fed and starved lactating goats in order to study glucose metabolism in the mammary gland. 2. Glucose carbon was oxidized and metabolizet to milk lactose, citrate and triacylglycerol in the lactating goat udder. 3. Recycling of glucose carbon in the lactating animal accounted for 10-20% of the total glucose turnover in the whole animal. Recycling of glucose 6-phosphate in the udder accounted for about 25% of the glucose 6-phosphate metabolized. 4. Flux of glucose 6-phosphate through the pentose phosphate pathway was sufficient to account for 34% of the NADPH required for fatty acid synthesis in the gland in the fed animal. 5. Net metabolism of glucose 6-phosphate via the pentose phosphate pathway accounted for 17.8 and 1.2% of the glucose phosphorylated by the mammary gland in the fed and starved animal respectively. Metabolism of glucose 6-phosphate via the pentose phosphate pathway was sufficient to account for all the CO2 produced from glucose in the fed animal, but only 17% of the CO2 produced from glucose in the starved animal.

Animals↗
PubMed1982

Transfer of 1-pyrroline-5-carboxylate as oxidizing potential from hepatocytes to erythrocytes.

The interconversions of proline and 1-pyrroline-5-carboxylate form an intercellular cycle that is the basis of a metabolic interaction between hepatocytes and erythrocytes. The cycle transfers oxidizing potential from hepatocytes to erythrocytes, which stimulates pentose phosphate pathway in erythrocytes. This interaction depends on the differential metabolism of proline and 1-pyrroline-5-carboxylate in erythrocytes and hepatocytes and consists of the following: in hepatocytes proline oxidase converts proline into 1-pyrroline-5-carboxylate, which is released into the medium and taken up by erythrocytes; erythrocyte 1-pyrroline-5-carboxylate reductase converts 1-pyrroline-5-carboxylate into proline and concomitantly generates NADP+; the generated oxidizing potential drives glucose metabolism through the pentose phosphate pathway in erythrocytes; finally, erythrocytes release proline into the medium, enabling it to re-enter hepatocytes and repeat the cycle. The increased activity of the pentose phosphate pathway in erythrocytes may enhance the production of 5-phosphoribosyl pyrophosphate, a necessary moiety for the processing of purines.

Animals↗
PubMed2003

The soluble and membrane-bound transhydrogenases UdhA and PntAB have divergent functions in NADPH metabolism of Escherichia coli.

Pentose phosphate pathway and isocitrate dehydrogenase are generally considered to be the major sources of the anabolic reductant NADPH. As one of very few microbes, Escherichia coli contains two transhydrogenase isoforms with unknown physiological function that could potentially transfer electrons directly from NADH to NADP+ and vice versa. Using defined mutants and metabolic flux analysis, we identified the proton-translocating transhydrogenase PntAB as a major source of NADPH in E. coli. During standard aerobic batch growth on glucose, 35-45% of the NADPH that is required for biosynthesis was produced via PntAB, whereas pentose phosphate pathway and isocitrate dehydrogenase contributed 35-45% and 20-25%, respectively. The energy-independent transhydrogenase UdhA, in contrast, was essential for growth under metabolic conditions with excess NADPH formation, i.e. growth on acetate or in a phosphoglucose isomerase mutant that catabolized glucose through the pentose phosphate pathway. Thus, both isoforms have divergent physiological functions: energy-dependent reduction of NADP+ with NADH by PntAB and reoxidation of NADPH by UdhA. Expression appeared to be modulated by the redox state of cellular metabolism, because genetic and environmental manipulations that increased or decreased NADPH formation down-regulated pntA or udhA transcription, respectively. The two transhydrogenase isoforms provide E. coli primary metabolism with an extraordinary flexibility to cope with varying catabolic and anabolic demands, which raises two general questions: why do only a few bacteria contain both isoforms, and how do other organisms manage NADPH metabolism?

Down-Regulation↗