Availability of intestinal microbial lysine for whole body lysine homeostasis in human subjects

1999 ◽  
Vol 277 (4) ◽  
pp. E597-E607 ◽  
Author(s):  
Cornelia C. Metges ◽  
Antoine E. El-Khoury ◽  
Lidewij Henneman ◽  
Klaus J. Petzke ◽  
Ian Grant ◽  
...  
Keyword(s):  
De Novo ◽  
Human Subjects ◽  
Isotope Ratio Mass Spectrometry ◽  
Acid Synthesis ◽  
Whole Body ◽  
Microbial Protein ◽  
Amino Acid Synthesis ◽  
Adequate Diet ◽  
Appearance Rate ◽  
Intravenous And Oral Administration

We have investigated whether there is a net contribution of lysine synthesized de novo by the gastrointestinal microflora to lysine homeostasis in six adults. On two separate occasions an adequate diet was given for a total of 11 days, and a 24-h (12-h fast, 12-h fed) tracer protocol was performed on the last day, in which lysine turnover, oxidation, and splanchnic uptake were measured on the basis of intravenous and oral administration ofl-[1-13C]lysine andl-[6,6-2H2]lysine, respectively. [15N2]urea or15NH4Cl was ingested daily over the last 6 days to label microbial protein. In addition, seven ileostomates were studied with15NH4Cl. [15N]lysine enrichment in fecal and ileal microbial protein, as precursor for microbial lysine absorption, and in plasma free lysine was measured by gas chromatography-combustion-isotope ratio mass spectrometry. Differences in plasma [13C]- and [2H2]lysine enrichments during the 12-h fed period were observed between the two15N tracer studies, although the reason is unclear, and possibly unrelated to the tracer form per se. In the normal adults, after15NH4Cl and [15N2]urea intake, respectively, lysine derived from fecal microbial protein accounted for 5 and 9% of the appearance rate of plasma lysine. With ileal microbial lysine enrichment, the contribution of microbial lysine to plasma lysine appearance was 44%. This amounts to a gross microbial lysine contribution to whole body plasma lysine turnover of between 11 and 130 mg ⋅ kg−1 ⋅ day−1, depending on the [15N]lysine precursor used. However, insofar as microbial amino acid synthesis is accompanied by microbial breakdown of endogenous amino acids or their oxidation by intestinal tissues, this may not reflect a net increase in lysine absorption. Thus we cannot reliably estimate the quantitative contribution of microbial lysine to host lysine homeostasis with the present paradigm. However, the results confirm the significant presence of lysine of microbial origin in the plasma free lysine pool.

Whole Body De Novo Amino Acid Synthesis in Type I (Insulin-dependent) Diabetes Studied With Stable Isotope-labeled Leucine, Alanine, and Glycine

Diabetes ◽  
1985 ◽  
Vol 34 (1) ◽  
pp. 67-73 ◽  
Author(s):  
J. J. Robert ◽  
B. Beaufrere ◽  
J. Koziet ◽  
J. F. Desjeux ◽  
D. M. Bier ◽  
...  
Keyword(s):  
Amino Acid ◽  
Stable Isotope ◽  
De Novo ◽  
Acid Synthesis ◽  
Insulin Dependent Diabetes ◽  
Whole Body ◽  
Type I ◽  
Amino Acid Synthesis ◽  
Insulin Dependent

Whole body de novo amino acid synthesis in type I (insulin-dependent) diabetes studied with stable isotope-labeled leucine, alanine, and glycine

Diabetes ◽  
1985 ◽  
Vol 34 (1) ◽  
pp. 67-73 ◽  
Author(s):  
J. J. Robert ◽  
B. Beaufrere ◽  
J. Koziet ◽  
J. F. Desjeux ◽  
D. M. Bier ◽  
...  
Keyword(s):  
Amino Acid ◽  
Stable Isotope ◽  
De Novo ◽  
Acid Synthesis ◽  
Insulin Dependent Diabetes ◽  
Whole Body ◽  
Type I ◽  
Amino Acid Synthesis ◽  
Insulin Dependent

Effect of carbohydrate intake on de novo lipogenesis in human adipose tissue

1987 ◽  
Vol 253 (6) ◽  
pp. E664-E669 ◽  
Author(s):  
C. Chascione ◽  
D. H. Elwyn ◽  
M. Davila ◽  
K. M. Gil ◽  
J. Askanazi ◽  
...  
Keyword(s):  
Adipose Tissue ◽  
De Novo ◽  
Acid Synthesis ◽  
De Novo Lipogenesis ◽  
Whole Body ◽  
High Intake ◽  
Carbohydrate Diet ◽  
Triglyceride Synthesis ◽  
High Carbohydrate

Rates of synthesis, from [14C]glucose, of fatty acids (de novo lipogenesis) and glycerol (triglyceride synthesis) were measured in biopsies of adipose tissue from nutritionally depleted patients given low- or high-carbohydrate intravenous nutrition. Simultaneously, energy expenditure and whole-body lipogenesis were measured by indirect calorimetry. Rates of whole-body lipogenesis were zero on the low-carbohydrate diet and averaged 1.6 g.kg-1.day-1 on the high-carbohydrate diet. In vitro rates of triglyceride synthesis increased 3-fold going from the low to the high intake; rates of fatty acid synthesis increased approximately 80-fold. In vitro, lipogenesis accounted for less than 0.1% of triglyceride synthesis on the low intake and 4% on the high intake. On the high-carbohydrate intake, in vitro rates of triglyceride synthesis accounted for 61% of the rates of unidirectional triglyceride synthesis measured by indirect calorimetry. In vitro rates of lipogenesis accounted for 7% of whole-body lipogenesis. Discrepancies between in vitro rates of fatty acid synthesis from glucose, compared with acetate and citrate, as reported by others, suggest that in depleted patients on hypercaloric high-carbohydrate diets, adipose tissue may account for up to 40% of whole-body lipogenesis.

scholarly journals Proteasome Inhibition in Brassica napus Roots Increases Amino Acid Synthesis to Offset Reduced Proteolysis

2020 ◽  
Vol 61 (6) ◽  
pp. 1028-1040
Author(s):  
Dan Pereksta ◽  
Dillon King ◽  
Fahmida Saki ◽  
Amith Maroli ◽  
Elizabeth Leonard ◽  
...  
Keyword(s):  
Amino Acid ◽  
Brassica Napus ◽  
De Novo ◽  
Metabolic Response ◽  
Sugar Metabolism ◽  
Proteasome Inhibition ◽  
Acid Synthesis ◽  
Amino Acid Synthesis ◽  
Amino Acid Depletion ◽  
Metabolic Activities

Abstract Cellular homeostasis is maintained by the proteasomal degradation of regulatory and misfolded proteins, which sustains the amino acid pool. Although proteasomes alleviate stress by removing damaged proteins, mounting evidence indicates that severe stress caused by salt, metal(oids), and some pathogens can impair the proteasome. However, the consequences of proteasome inhibition in plants are not well understood and even less is known about how its malfunctioning alters metabolic activities. Lethality causes by proteasome inhibition in non-photosynthetic organisms stem from amino acid depletion, and we hypothesized that plants respond to proteasome inhibition by increasing amino acid biosynthesis. To address these questions, the short-term effects of proteasome inhibition were monitored for 3, 8 and 48 h in the roots of Brassica napus treated with the proteasome inhibitor MG132. Proteasome inhibition did not affect the pool of free amino acids after 48 h, which was attributed to elevated de novo amino acid synthesis; these observations coincided with increased levels of sulfite reductase and nitrate reductase activities at earlier time points. However, elevated amino acid synthesis failed to fully restore protein synthesis. In addition, transcriptome analysis points to perturbed abscisic acid signaling and decreased sugar metabolism after 8 h of proteasome inhibition. Proteasome inhibition increased the levels of alternative oxidase but decreased aconitase activity, most sugars and tricarboxylic acid metabolites in root tissue after 48 h. These metabolic responses occurred before we observed an accumulation of reactive oxygen species. We discuss how the metabolic response to proteasome inhibition and abiotic stress partially overlap in plants.

scholarly journals Target of Rapamycin Inhibition in Chlamydomonas reinhardtii Triggers de Novo Amino Acid Synthesis by Enhancing Nitrogen Assimilation

2018 ◽  
Vol 30 (10) ◽  
pp. 2240.1-2254 ◽  
Author(s):  
Umarah Mubeen ◽  
Jessica Jüppner ◽  
Jessica Alpers ◽  
Dirk K. Hincha ◽  
Patrick Giavalisco
Keyword(s):  
Amino Acid ◽  
Chlamydomonas Reinhardtii ◽  
Nitrogen Assimilation ◽  
De Novo ◽  
Acid Synthesis ◽  
Target Of Rapamycin ◽  
Amino Acid Synthesis

Phenylbutyrate-induced glutamine depletion in humans: effect on leucine metabolism

1998 ◽  
Vol 274 (5) ◽  
pp. E801-E807 ◽  
Author(s):  
Dominique Darmaun ◽  
Susan Welch ◽  
Annie Rini ◽  
Brenda K. Sager ◽  
Astride Altomare ◽  
...  
Keyword(s):  
Protein Synthesis ◽  
De Novo ◽  
Oral Treatment ◽  
Healthy Adults ◽  
Whole Body ◽  
Dependent Manner ◽  
Glutamine Concentration ◽  
Body Protein ◽  
Healthy Humans ◽  
Appearance Rate

The present study was designed to determine whether sodium phenylbutyrate (ΦB) acutely induces a decrease in plasma glutamine in healthy humans, and, if so, will decrease estimates of whole body protein synthesis. In a first group of three healthy subjects, graded doses (0, 0.18, and 0.36 g ⋅ kg−1 ⋅ day−1) of ΦB were administered for 24 h before study: postabsorptive plasma glutamine concentration declined in a dose-dependent manner, achieving an ≈25% decline for a dose of 0.36 g ΦB ⋅ kg−1 ⋅ day−1. A second group of six healthy adults received 5-h infusions ofl-[1-14C]leucine andl-[1-13C]glutamine in the postabsorptive state on two separate days: 1) under baseline conditions and 2) after 24 h of oral treatment with ΦB (0.36 g ⋅ kg−1 ⋅ day−1) in a randomized order. The 24-h phenylbutyrate treatment was associated with 1) an ≈26% decline in plasma glutamine concentration from 514 ± 24 to 380 ± 15 μM (means ± SE; P < 0.01 with paired t-test) with no change in glutamine appearance rate or de novo synthesis; 2) no change in leucine appearance rate (Ra), an index of protein breakdown (123 ± 7 vs. 117 ± 5 μmol ⋅ kg−1 ⋅ h−1; not significant); 3) an ≈22% rise in leucine oxidation (Ox) from 23 ± 2 to 28 ± 2 μmol ⋅ kg−1 ⋅ h−1( P < 0.01), resulting in an ≈11% decline in nonoxidative leucine disposal (NOLD = Ra − Ox), an index of protein synthesis, from 100 ± 6 to 89 ± 5 μmol ⋅ kg−1 ⋅ h−1( P < 0.05). The data suggest that, in healthy adults, 1) large doses of oral phenylbutyrate can be used as a “glutamine trap” to create a model of glutamine depletion; 2) a moderate decline in plasma glutamine does not enhance rates of endogenous glutamine production; and 3) a short-term depletion of plasma glutamine decreases estimates of whole body protein synthesis.

scholarly journals Mechanism of De Novo Branched-Chain Amino Acid Synthesis as an Alternative Electron Sink in Hypoxic Aspergillus nidulans Cells

2010 ◽  
Vol 76 (5) ◽  
pp. 1507-1515 ◽  
Author(s):  
Motoyuki Shimizu ◽  
Tatsuya Fujii ◽  
Shunsuke Masuo ◽  
Naoki Takaya
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Aspergillus Nidulans ◽  
De Novo ◽  
Acid Synthesis ◽  
Branched Chain Amino Acids ◽  
Amino Acid Synthesis ◽  
Lactate Dehydrogenase Activity ◽  
Branched Chain Amino Acid ◽  
Branched Chain

ABSTRACT Although branched-chain amino acids are synthesized as building blocks of proteins, we found that the fungus Aspergillus nidulans excretes them into the culture medium under hypoxia. The transcription of predicted genes for synthesizing branched-chain amino acids was upregulated by hypoxia. A knockout strain of the gene encoding the large subunit of acetohydroxy acid synthase (AHAS), which catalyzes the initial reaction of the synthesis, required branched-chain amino acids for growth and excreted very little of them. Pyruvate, a substrate for AHAS, increased the amount of hypoxic excretion in the wild-type strain. These results indicated that the fungus responds to hypoxia by synthesizing branched-chain amino acids via a de novo mechanism. We also found that the small subunit of AHAS regulated hypoxic branched-chain amino acid production as well as cellular AHAS activity. The AHAS knockout resulted in higher ratios of NADH/NAD+ and NADPH/NADP+ under hypoxia, indicating that the branched-chain amino acid synthesis contributed to NAD+ and NADP+ regeneration. The production of branched-chain amino acids and the hypoxic induction of involved genes were partly repressed in the presence of glucose, where cells produced ethanol and lactate and increased levels of lactate dehydrogenase activity. These indicated that hypoxic branched-chain amino acid synthesis is a unique alternative mechanism that functions in the absence of glucose-to-ethanol/lactate fermentation and oxygen respiration.

scholarly journals Microbial amino acid synthesis and utilization in rats: the role of coprophagy

1996 ◽  
Vol 76 (5) ◽  
pp. 701-709 ◽  
Author(s):  
David Torrallardona ◽  
C. Ian Harris ◽  
Malcolm F. Fuller
Keyword(s):  
Body Weight ◽  
Isotope Ratio Mass Spectrometry ◽  
Acid Synthesis ◽  
Exchange Chromatography ◽  
The Body ◽  
Amino Acid Synthesis ◽  
Low Protein ◽  
N Enrichment ◽  
Fermentable Carbohydrates

Four rats were housed in cages with mesh floors; another four rats were housed in tubular anti-coprophagy cages, in which they could not turn round to reach their own faeces. Both groups were fed for 6 d on a low-protein diet containing fermentable carbohydrates and 15NH4Cl. At the end of the experiment the rats were killed and their carcasses were homogenized, lysine was isolated by ion-exchange chromatography and its 15N enrichment measured by isotope-ratio mass spectrometry. The 15N enrichment in the lysine of the microbial fraction of faeces and the total amount of lysine in the body were also determined in order to estimate the amount of microbial lysine absorbed. The 15N enrichment in body lysine of non-coprophagic rats was not different from that previously measured in rats given unlabelled NH4Cl, but in coprophagic rats it was significantly higher. The daily absorption of microbial lysine by the coprophagic rats accounted for 20·7 (SE 2·55) mg/kg body weight0·75 but was only 0·5 (SE 1·04) mg/kg body weight0·75 for the non-coprophagic rats. This value was not significantly different from zero. The utilization of microbial amino acids via coprophagy resulted in a higher weight gain (adjusted for intake) in the coprophagic group (15·5 g/6 d) than in the non-coprophagic rats (3·1 g/6 d). It was concluded that, in rats, the utilization of microbial lysine occurred exclusively via coprophagy.

scholarly journals Microbial amino acid synthesis and utilization in rats: incorporation of15N from15NH4Cl into lysine in the tissues of germ-free and conventional rats

1996 ◽  
Vol 76 (5) ◽  
pp. 689-700 ◽  
Author(s):  
David Torrallardona ◽  
C. Ian Harris ◽  
Marie E. Coates ◽  
Malcolm F. Fuller
Keyword(s):  
Isotope Ratio Mass Spectrometry ◽  
Acid Synthesis ◽  
Exchange Chromatography ◽  
The Body ◽  
Amino Acid Synthesis ◽  
Germ Free ◽  
N Enrichment ◽  
Fermentable Carbohydrates ◽  
Measurement Artifact ◽  
Protein Free Diet

The absorption of lysine synthesised by the gastrointestinal microflora was estimated by comparing the15N incorporated into body lysine in four germ-free (15N-GF) and four conventional (15N-CV) rats. They were fed for 10d on a protein-free diet containing fermentable carbohydrates and15NHM4Cl; another four conventional rats (control), fed on the same diet but with unlabelled NH4Cl, were used to estimate the natural abundance of15N. The eviscerated carcass of each rat was homogenized and a sample hydrolysed. Lysine was isolated by ion-exchange chromatography and its15N enrichment was measured by isotope-ratio mass spectrometry. The15N-CV rats significantly incorporated15N into their body lysine. The15N-GF rats had a statistically significant, although small, incorporation of15body lysine, probably arising from a measurement artifact. It was concluded, therefore, that all [15N]lysine was of microbial origin. The total lysine content in the body and the15N enrichment of lysine in the microbial fraction of the faeces of the15N-CV rats were also determined. The amount of microbial lysine absorbed by the15N-CV rats was estimated by dividing the total amount of [15N]lysine in the body by the enrichment of microbial lysine. It was estimated that the daily absorption of microbial lysine by the conventional rats was 21·3 (SE 2·04) mg/kg body weight0·75

Glutamine nitrogen kinetics in insulin-dependent diabetic humans

1991 ◽  
Vol 261 (6) ◽  
pp. E713-E718 ◽  
Author(s):  
D. Darmaun ◽  
M. Rongier ◽  
J. Koziet ◽  
J. J. Robert
Keyword(s):  
Insulin Infusion ◽  
De Novo ◽  
Significant Rise ◽  
Whole Body ◽  
Type I ◽  
Insulin Deficiency ◽  
Appearance Rate ◽  
Insulin Dependent ◽  
Insulin Dependent Diabetic ◽  
Infusion System

To assess the effect of insulin deficiency on whole body glutamine kinetics, five young adults with type I (insulin-dependent) diabetes received 4-h primed continuous infusions of L-[1-13C]leucine and L-[2-15N]glutamine in the postabsorptive state after blood glucose had been clamped overnight at either a normoglycemic level (approximately 85 mg/dl) or a moderate hyperglycemic level (approximately 260 mg/dl) by means of an automated glucose control insulin infusion system. The hyperglycemic state was associated with a significant rise in leucine level [from 165 +/- 23 to 242 +/- 62 (SD) microM], appearance rate (from 125 +/- 11 to 142 +/- 17 mumol.kg-1.h-1), and oxidation (from 27 +/- 10 to 31 +/- 10 mumol.kg-1.h-1). In contrast, neither the plasma level nor the appearance rate of glutamine (333 +/- 51 vs. 318 +/- 58 mumol.kg-1.h-1) was affected. We conclude that insulin deficiency resulting in moderate hyperglycemia induces a 13% rise in whole body proteolysis and yet does not stimulate glutamine de novo synthesis, despite increased precursor availability.

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