scholarly journals 15N and ²H Intrinsic Labeling Demonstrate That Real Digestibility in Rats of Proteins and Amino Acids from Sunflower Protein Isolate Is Almost as High as That of Goat Whey

2019 ◽  
Vol 150 (3) ◽  
pp. 450-457 ◽  
Author(s):  
Romain Tessier ◽  
Nadezda Khodorova ◽  
Juliane Calvez ◽  
Romain Kapel ◽  
Alain Quinsac ◽  
...  
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Goat Milk ◽  
Protein Digestibility ◽  
Protein Isolate ◽  
Plant Protein Sources ◽  
Male Wistar Rats ◽  
Sunflower Protein ◽  
Meal Ingestion

ABSTRACT Background In the context of developing plant protein sources for humans, sunflower is a good candidate in its form as an oilseed coproduct. Objectives We aimed to compare the real digestibility in rats of a sunflower isolate to that of goat whey protein. We also studied the efficiency of 15N and 2H intrinsic labeling in this assessment. Methods Sunflower seeds and goat milk were labeled with 15N and 2H. Male Wistar rats (10 wk old) were fed a meal containing 12% of either sunflower isolate (n = 8) or whey (n = 8). Six hours after meal ingestion, protein and amino acid digestibility were assessed by measuring nitrogen, hydrogen, and amino acids in the digesta, as well as isotope enrichments in the bulk and individual amino acids. The differences between groups and isotopes were respectively tested with an unpaired and a paired t test. Results Protein isolate purity was 87% for whey and 94% for sunflower. 2H and 15N enrichments were, respectively, 0.12 atom % (AP) and 1.06 AP in sunflower isolate and 0.18 AP and 0.95 AP in whey. Fecal 15N protein digestibility was 97.2 ± 0.2% for whey and 95.1 ± 0.5% for sunflower isolate. The use of 2H resulted in a lower digestibility estimate than 15N for whey (96.9 ± 0.2%, P < 0.05) and sunflower (94.2 ± 0.5%, P < 0.01). For both isotopes, protein digestibility was about 2% higher for whey than for sunflower isolate. Mean 15N amino acid caecal digestibility was 97.5 ± 0.2% for whey and 96.3 ± 0.2% for sunflower isolate. The values obtained with 15N and 2H resulted in significant differences ranging from −0.1% to 3.5%. The DIAAS was >1.0 for whey and 0.84 for sunflower (lysine). Conclusions The protein and amino acid digestibility of sunflower isolate was high but its DIAAS reflected a moderate lysine imbalance. Despite slight differences with 15N, deuterium produced comparable results, making it suitable for in vivo digestion studies.

scholarly journals Advantages and limitations of the protein digestibility-corrected amino acid score (PDCAAS) as a method for evaluating protein quality in human diets

2012 ◽  
Vol 108 (S2) ◽  
pp. S333-S336 ◽  
Author(s):  
Gertjan Schaafsma
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Protein Quality ◽  
Protein Digestibility ◽  
Essential Amino Acids ◽  
Tissue Growth ◽  
Biological Efficiency ◽  
Amino Acid Supplementation ◽  
Reference Pattern ◽  
Plant Protein Sources

PDCAAS is a widely used assay for evaluating protein quality. It is a chemical score, which is derived from the ratio between the first limiting amino acid in a test protein and the corresponding amino acid in a reference amino acid pattern and corrected for true faecal N digestibility. Chemical scores exceeding 100 % are truncated to 100 %. The advantages of the PDCAAS are its simplicity and direct relationship to human protein requirements. The limitations are as follows: the reference pattern is based on the minimum amino acid requirements for tissue growth and maintenance and does not necessarily reflect the optimum intake. Truncated PDCAAS of high-quality proteins do not give any information about the power of these proteins to compensate, as a supplement, for low levels of dietary essential amino acids in low-quality proteins. It is likely that faecal N digestibility does not take into account the loss from the colon of indispensable amino acids that were not absorbed in the ileum. Anti-nutritional factors, such as lectins and trypsin inhibitors, in several plant protein sources can cause heightened endogenous losses of amino acids, an issue which is particularly relevant in animal feedstuffs. The assumption that amino acid supplementation can completely restore biological efficiency of the protein source is incorrect since the kinetics of digestion and absorption between supplemented free amino acids and amino acids present in dietary proteins, are different.

scholarly journals Challenging in Rats the Use of 13C Spirulina as Reference Protein for the Dual Isotope Method to Determine Amino Acid Bioavailability (P08-061-19)

2019 ◽  
Vol 3 (Supplement_1) ◽  
Author(s):  
Romain Tessier ◽  
Nadezda Khodorova ◽  
Juliane Calvez ◽  
Daniel Tomé ◽  
Claire Gaudichon
Keyword(s):  
Amino Acid ◽  
Goat Milk ◽  
Protein Digestibility ◽  
Male Rats ◽  
National Agency ◽  
Dual Isotope ◽  
Isotope Method ◽  
Reference Protein ◽  
Bonferroni Test

Abstract Objectives In order to establish DIAAS in humans, the FAO recommended to develop a new method to measure indispensable amino acid (IAA) digestibility. This method uses two isotopic labeling, one for the protein to test and one for a reference protein. Spirulina was chosen as the 13C reference protein due to its commercial availability and affordability. However, the real digestibility of spirulina protein has not been measured in vivo. This work aims to assess the digestibility of spirulina and its repeatability in different meal tests in rats. Methods 23 Wistar male rats were fed a test meal containing 0.5 g of 15 N protein from either spirulina (n = 7), sunflower n = 8) or goat milk isolate (n = 8) and 10 mg of 13C labeled spirulina. Rats were euthanized 6 h after the meal and their digestive luminal contents (stomach, small intestine, ileum, caecum, colon) were collected. Protein digestibility was determined for the test and the reference proteins by measuring 15 N and 13C enrichments in the digesta by EA-IRMS. Caecal IAA digestibility of 13C spirulina was determined by measuring the quantity of AA in the caecum by UPLC and the 13C enrichment in AA by GC-C-IRMS. Group effects were tested using one way ANOVA and differences between groups using Bonferroni test. Results Six hours after ingestion, most of the dietary 15 N and 13C were found in the caecum and colon. But there at least twice more 15 N nitrogen in the caecum and colon in the spirulina group than in the two other groups. Therefore, spirulina protein digestibility (86.0 ± 0.7%) was lower (P < 0.001) than sunflower (95.1 ± 0.5%) and goat milk digestibility (97.2 ± 0.2%). 13C spirulina digestibility tended to be different (P = 0.06) when mixed to spirulina (90.6 ± 0.6%), sunflower (88.8 ± 0.5%) or goat milk (89.0 ± 0.5%) isolates. The caecal IAA digestibility of 13C spirulina was lower in the spirulina group than in sunflower and goat milk groups for every IAA tested, and the mean was 91.6 ± 0.2% for sunflower, 91.4 ± 0.4% for goat milk and 85.4 ± 0.6% for spirulina. Conclusions Spirulina protein is of lower digestibility than other animal or plant proteins. Protein and amino digestibility of a tracer dose of 13C spirulina appears to vary depending on the protein component of the meal. These results question the use of spirulina as a reference protein for the dual isotope method. Funding Sources French Research National Agency (ANR), SOFIPROTEOL. Supporting Tables, Images and/or Graphs

scholarly journals Real ileal digestibility of sunflower protein and amino acids in humans

2020 ◽  
Vol 79 (OCE2) ◽  
Author(s):  
Romain Tessier ◽  
Juliane Calvez ◽  
Nadezda Khodorova ◽  
Alain Quinsac ◽  
Romain Kapel ◽  
...  
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Healthy Volunteers ◽  
Protein Isolate ◽  
Nitrogen Flow ◽  
Dual Isotope ◽  
Ileal Digestibility ◽  
Isotope Method ◽  
Indispensable Amino Acids ◽  
Sunflower Protein

AbstractIntroduction:Sunflower protein is not used in human nutrition despite a relatively good amino acid composition. However, the bioavailability of sunflower isolate has never been measured in Humans. The goal of this work was to determine ileal digestibility of protein and amino acids from a sunflower isolate in healthy volunteers and to challenge newly developed dual isotope method.Materials and methods:Eight healthy volunteers were equipped with a naso-ileal tube. They received during four hours twelve doses of biscuits containing, in total, 25 g of 15N sunflower protein isolate together with 400 mg of a mixture of free 13C amino acids incorporated in chocolates. Polyethylene glycol was perfused as non-absorbable marker and ileal contents were collected during 8 hours after ingestion of the first meal. Real ileal digestibility was measured by assessing nitrogen and carbon content as well as 15N and 13C enrichments by EA-IRMS. Amino acid digestibility was determined by measuring 15N and 13C enrichments by GC-C-IRMS and quantity of amino acids by UPLC. Blood was collected for 8 h to determine 15N and 13C enrichments by GC-C-IRMS.Results:The ileal nitrogen flow was 2.7 ± 0.5 mL/min (mean ± SD). In average, 53.1 ± 12.0 mmol of exogenous nitrogen was recovered during the eight hours of experiment, resulting in an ileal digestibility of sunflower isolate was 85.6 ± 2.6 % of nitrogen ingested. 13C amino acids were also recovered at the ileal level. The mixture of free 13C revealed an ileal digestibility of 94.9 ± 0.9 %. Ileal indispensable amino acids digestibility and DIAAS are in progress.Discussion:Ileal digestibility of sunflower isolate incorporated in toasted biscuits was lower than the value found or a raw isolate in a rat model (94.5%). The study revealed that 5 % of free amino acids were not absorbed in the ileum. Amino acid digestibility will complete the study to evaluate the DIAAS of sunflower isolate and to compare values obtained with the standard method and with the dual isotope method.

Chemical and Nutritional Evaluation of Pumpkin (Cucurbita pepo) Seed Proteins

2019 ◽  
pp. 1-11 ◽  
Author(s):  
Adanma C. Innocent-Ukachi
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Chemical Properties ◽  
Seed Proteins ◽  
Protein Digestibility ◽  
Amino Acid Profile ◽  
Protein Isolate ◽  
Pumpkin Seed ◽  
Chemical Score

Chemical and nutritional properties of pumpkin (Curcubita pepo) seed proteins were studied. The seed was processed into defatted flour (CPF) which was further processed into Curcubita protein concentrate (CPC) and Curcubita protein isolate (CPI) by alkaline water/isoelectric precipitation. Chemical properties of the protein products were determined using standard methods of analysis. The amino acid profile was determined by an automated Technicon® liquid chromatography system. Protein digestibility was assessed in-vitro (IVPD) using trypsin-pepsin enzyme method while biological values were determined on the basis of their amino acid profile. Protein efficiency ratio (PER) was estimated according to a standard proposed regression equation. The seed proteins demonstrated high levels of crude protein (CPC=69.98% and CPI=74.15%), vitamin C (CPC=43.46 and CPI=52.36 mg/ml) and vitamin A (CPC=100.56 and CPI= 63.43 I.U/g) with low levels of thiamin and riboflavin. Both proteins showed low and similar (p>0.05) levels of sodium (0.14-0.18%), calcium (0.86-1.02%), magnesium (0.53-0.58%) and phosphorus (0.09-0.11%). Percentage ratios of essential to total amino acids obtained for CPC and CPI (44.24% and 45.50%, respectively) were greater than 36% which is considered adequate for an ideal protein. Protein biological values obtained for CPC and CPI respectively were: 95% and 53% (chemical score), 2.80 and 1.56 (PER} and 70.10% and 51.28% (essential amino acid index). CPC showed a better digestibility than CPI with IVPD value of 56.88%. Threonine and lysine were the most limiting amino acids in both protein products. All anti-nutrients evaluated were low and below allowable limits. In conclusion pumpkin seed proteins showed good biological values and could be used to improve the quality of other plant proteins or as a possible replacement for animal proteins in conventional foods.

scholarly journals Primary structure requirements for correct sorting of the yeast mitochondrial protein ADH III to the yeast mitochondrial matrix space.

1987 ◽  
Vol 7 (1) ◽  
pp. 294-304 ◽  
Author(s):  
D Pilgrim ◽  
E T Young
Keyword(s):  
Amino Acids ◽  
Enzyme Activity ◽  
Amino Acid ◽  
Proteolytic Processing ◽  
Mitochondrial Matrix ◽  
Wild Type ◽  
Mature Form ◽  
Amino Terminal ◽  
The Matrix

Alcohol dehydrogenase isoenzyme III (ADH III) in Saccharomyces cerevisiae, the product of the ADH3 gene, is located in the mitochondrial matrix. The ADH III protein was synthesized as a larger precursor in vitro when the gene was transcribed with the SP6 promoter and translated with a reticulocyte lysate. A precursor of the same size was detected when radioactively pulse-labeled proteins were immunoprecipitated with anti-ADH antibody. This precursor was rapidly processed to the mature form in vivo with a half-time of less than 3 min. The processing was blocked if the mitochondria were uncoupled with carbonyl cyanide m-chlorophenylhydrazone. Mutant enzymes in which only the amino-terminal 14 or 16 amino acids of the presequence were retained were correctly targeted and imported into the matrix. A mutant enzyme that was missing the amino-terminal 17 amino acids of the presequence produced an active enzyme, but the majority of the enzyme activity remained in the cytoplasmic compartment on cellular fractionation. Random amino acid changes were produced in the wild-type presequence by bisulfite mutagenesis of the ADH3 gene. The resulting ADH III protein was targeted to the mitochondria and imported into the matrix in all of the mutants tested, as judged by enzyme activity. Mutants containing amino acid changes in the carboxyl-proximal half of the ADH3 presequence were imported and processed to the mature form at a slower rate than the wild type, as judged by pulse-chase studies in vivo. The unprocessed precursor appeared to be unstable in vivo. It was concluded that only a small portion of the presequence contains the necessary information for correct targeting and import. Furthermore, the information for correct proteolytic processing of the presequence appears to be distinct from the targeting information and may involve secondary structure information in the presequence.

scholarly journals BIOSYNTHESIS IN ISOLATED ACETABULARIA CHLOROPLASTS

1972 ◽  
Vol 54 (2) ◽  
pp. 279-294 ◽  
Author(s):  
David C. Shephard ◽  
Wendy B. Levin
Keyword(s):  
Amino Acids ◽  
Protein Synthesis ◽  
Amino Acid ◽  
Technical Problems ◽  
Hydrolyzed Protein ◽  
Protein Amino Acids ◽  
Amino Acid Labeling ◽  
Labeling Pattern

The ability of chloroplasts isolated from Acetabulana mediterranea to synthesize the protein amino acids has been investigated. When this chloroplast isolate was presented with 14CO2 for periods of 6–8 hr, tracer was found in essentially all amino acid species of their hydrolyzed protein Phenylalanine labeling was not detected, probably due to technical problems, and hydroxyproline labeling was not tested for The incorporation of 14CO2 into the amino acids is driven by light and, as indicated by the amount of radioactivity lost during ninhydrin decarboxylation on the chromatograms, the amino acids appear to be uniformly labeled. The amino acid labeling pattern of the isolate is similar to that found in plastids labeled with 14CO2 in vivo. The chloroplast isolate did not utilize detectable amounts of externally supplied amino acids in light or, with added adenosine triphosphate (ATP), in darkness. It is concluded that these chloroplasts are a tight cytoplasmic compartment that is independent in supplying the amino acids used for its own protein synthesis. These results are discussed in terms of the role of contaminants in the observed synthesis, the "normalcy" of Acetabularia chloroplasts, the synthetic pathways for amino acids in plastids, and the implications of these observations for cell compartmentation and chloroplast autonomy.

scholarly journals Regulation of rat magnocellular neurosecretory system by D-aspartate: evidence for biological role(s) of a naturally occurring free D-amino acid in mammals

2000 ◽  
Vol 167 (2) ◽  
pp. 247-252 ◽  
Author(s):  
H Wang ◽  
H Wolosker ◽  
J Pevsner ◽  
SH Snyder ◽  
DJ Selkoe
Keyword(s):  
Gene Expression ◽  
Amino Acids ◽  
Amino Acid ◽  
Physiological Function ◽  
Neurosecretory System ◽  
Milk Ejection ◽  
Magnocellular Neurons ◽  
Rat Hypothalamus ◽  
Naturally Occurring

Little evidence is available for the physiological function of D-amino acids in species other than bacteria. Here we demonstrate that naturally occurring freed -aspartate (D-Asp) is present in all magnocellular neurons of rat hypothalamus. The levels of this naturally occurring D-amino acid were elevated during lactation and returned to normal thereafter in the magnocellular neurosecretory system, which produces oxytocin, a hormone responsible for milk ejection during lactation. Intraperitoneal injections of D-Asp reproducibly increased oxytocin gene expression and decreased the concentration of circulating oxytocin in vivo. Similar changes were observed in the vasopressin system. These results provide evidence for the role(s) of naturally occurring free D-Asp in mammalian physiology. The findings argue against the conventional concept that only L-stereoisomers of amino acids are functional in higher species.

Effect of Cycloheximide on In Vitro and In Vivo Protein Synthesis by Certain Tissues of Rats and Pigeons with Special Reference to Muscle

1973 ◽  
Vol 51 (12) ◽  
pp. 933-941 ◽  
Author(s):  
Njanoor Narayanan ◽  
Jacob Eapen
Keyword(s):  
Amino Acids ◽  
Body Weight ◽  
Protein Synthesis ◽  
Amino Acid ◽  
Amino Acid Incorporation ◽  
Contrast Administration ◽  
Acid Incorporation ◽  
Tissue Homogenates

The effect of cycloheximide in vitro and in vivo on the incorporation of labelled amino acids into protein by muscles, liver, kidneys, and brain of rats and pigeons was studied. In vitro incorporation of amino acids into protein by muscle microsomes, myofibrils, and myofibrillar ribosomes was not affected by cycloheximide. In contrast, administration of the antibiotic into intact animals at a concentration of 1 mg/kg body weight resulted in considerable inhibition of amino acid incorporation into protein by muscles, liver, kidneys, and brain. This inhibition was observed in all the subcellular fractions of these tissues during a period of 10–40 min after the administration of the precursor. Tissue homogenates derived from in vivo cycloheximide-treated animals did not show significant alteration in in vitro amino acid incorporation with the exception of brain, which showed a small but significant enhancement.

In vivo metabolizable branched poly(ester amide) based on inositol and amino acids as drug nanocarrier for cancer therapy

2021 ◽  
Author(s):  
Jun Wu ◽  
Qi-Juan Yuan ◽  
Li Wang ◽  
Jun Huang ◽  
Wei Zhao
Keyword(s):  
Mechanical Property ◽  
Amino Acids ◽  
Amino Acid ◽  
Cancer Therapy ◽  
Biomedical Applications ◽  
Bioactive Components ◽  
Good Biocompatibility

Amino acid-based poly(ester amide) (PEA) has been utilized for various biomedical applications for its tunable mechanical property, good biocompatibility, and biodegradability. However, bioactive components have rarely been incorporated into the...

Potential differences influence amino acid/Na+ symport rates in larval Manduca sexta midgut brush-border membrane vesicles.

1994 ◽  
Vol 189 (1) ◽  
pp. 55-67
Author(s):  
R Parthasarathy ◽  
W R Harvey
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Manduca Sexta ◽  
Brush Border ◽  
Brush Border Membrane ◽  
Membrane Vesicles ◽  
Brush Border Membrane Vesicles ◽  
Rate Determining Step ◽  
Half Saturation Constant

The time-dependent fluorescence intensity of an intravesicular potential-sensitive dye was used to probe the real-time kinetics of potential difference (PD)-dependent amino acid/Na+ symport at pH9 into brush-border membrane vesicles obtained from larval Manduca sexta midgut. Neutral amino acids (alanine, proline) are symported at higher rates as the vesicles are hyperpolarized. The symport rates of acidic (glutamate) and basic (arginine) amino acids are almost PD-independent. The half-saturation constant of alanine is PD-independent between -108 and -78 mV, although the maximal symport velocity increases by half as the voltage is increased. Amino acid throughput is evidently enhanced as the relatively high transmembrane PDs (&gt; 150 mV, lumen positive) measured in vivo are approached. The half-saturation concentrations of Na+ were in the range 15-40 mmol l-1 for most of the amino acids examined and increased with voltage for alanine. The Vmax observed as a function of cation or amino acid concentration increased as the vesicle was hyperpolarized in the case of leucine and alanine. The data support the hypothesis that carrier and substrates are at equilibrium inasmuch as substrate translocation seems to be the rate-determining step of symport.

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