Partitioning Of Amino Acids Research Articles

Description of the fasted serum metabolomic signature of lean and obese cats at maintenance and of obese cats under energy restriction.

This study aimed to investigate the serum metabolomic profile of obese and lean cats as well as obese cats before and after energy restriction for weight loss. Thirty cats, 16 obese (body condition score 8 to 9/9) and 14 lean (body condition score 4 to 5/9), were fed a veterinary weight loss food during a 4-week period of weight maintenance (L-MAINT and O-MAINT). The 16 obese cats were then energy restricted by a 60% energy intake reduction with the same food for a 10-week period (O-RESTRICT). Fasted serum metabolites were measured using nuclear magnetic resonance and direct infusion mass spectrometry after the maintenance period for L-MAINT and O-MAINT cats and after the energy restriction period for O-RESTRICT and compared between groups using a two-sided t-test. Obese cats lost 672 g ± 303 g over the 10-week restriction period, representing a weight loss rate of 0.94 ± 0.28% per week. Glycine, l-alanine, l-histidine, l-glutamine, 2-hydroxybutyrate, isobutryric acid, citric acid, creatine, and methanol were greater in O-RESTRICT compared to O-MAINT. There was a greater concentration of long-chain acylcarnitines in O-RESTRICT compared to both O-MAINT and L-MAINT, and greater total amino acids compared to O-MAINT. Glycerol and 3-hydroxybutyric acid were greater in O-MAINT compared to L-MAINT, as were several lysophosphatidylcholines. Thus, energy restriction resulted in increased dispensable amino acids in feline serum which could indicate alterations in amino acid partitioning. An increase in lipolysis was not evident, though greater circulating acylcarnitines were observed, suggesting that fatty acid oxidation rates may have been greater under calorie restriction. More research is needed to elucidate energy metabolism and substrate utilization, specifically fatty acid oxidation and methyl status, during energy restriction in strict carnivorous cats to optimize weight loss.

UMAMIT44 is a key player in glutamate export from Arabidopsis chloroplasts.

Selective partitioning of amino acids among organelles, cells, tissues, and organs is essential for cellular metabolism and plant growth. Nitrogen assimilation into glutamine and glutamate and de novo biosynthesis of most protein amino acids occur in chloroplasts; therefore, various transport mechanisms must exist to accommodate their directional efflux from the stroma to the cytosol and feed the amino acids into the extraplastidial metabolic and long-distance transport pathways. Yet, Arabidopsis (Arabidopsis thaliana) transporters functioning in plastidial export of amino acids remained undiscovered. Here, USUALLY MULTIPLE ACIDS MOVE IN AND OUT TRANSPORTER 44 (UMAMIT44) was identified and shown to function in glutamate export from Arabidopsis chloroplasts. UMAMIT44 controls glutamate homeostasis within and outside of chloroplasts and influences nitrogen partitioning from leaves to sinks. Glutamate imbalances in chloroplasts and leaves of umamit44 mutants impact cellular redox state, nitrogen and carbon metabolism, and amino acid (AA) and sucrose supply of growing sinks, leading to negative effects on plant growth. Nonetheless, the mutant lines adjust to some extent by upregulating alternative pathways for glutamate synthesis outside the plastids and by mitigating oxidative stress through the production of other amino acids and antioxidants. Overall, this study establishes that the role of UMAMIT44 in glutamate export from chloroplasts is vital for controlling nitrogen availability within source leaf cells and for sink nutrition, with an impact on growth and seed yield.

Daylength variation affects growth, photosynthesis, leaf metabolism, partitioning, and metabolic fluxes.

Daylength, a seasonal and latitudinal variable, exerts a substantial impact on plant growth. However, the relationship between daylength and growth is nonproportional, suggesting the existence of adaptive mechanisms. Thus, our study aimed to comprehensively investigate the adaptive strategies employed by plants in response to daylength variation. We grew false flax (Camelina sativa) plants, a model oilseed crop, under long-day (LD) and short-day (SD) conditions and used growth measurements, gas exchange measurements, and isotopic labeling techniques, including 13C, 14C, and 2H2O, to determine responses to different daylengths. Our findings revealed that daylength influences various growth parameters, photosynthetic physiology, carbon partitioning, metabolic fluxes, and metabolite levels. SD plants employed diverse mechanisms to compensate for reduced CO2 fixation in the shorter photoperiod. These mechanisms included enhanced photosynthetic rates and reduced respiration in the light (RL), leading to increased shoot investment. Additionally, SD plants exhibited reduced rates of the glucose 6-phosphate (G6P) shunt and greater partitioning of sugars into starch, thereby sustaining carbon availability during the longer night. Isotopic labeling results further demonstrated substantial alterations in the partitioning of amino acids and TCA cycle intermediates between rapidly and slowly turning over pools. Overall, the results point to multiple developmental, physiological, and metabolic ways in which plants adapt to different daylengths to maintain growth.

L-Phenylalanine Partitioning Mechanisms in Model Biological Membranes.

Time-resolved fluorescence spectroscopy in combination with differential scanning calorimetry (DSC) was used to study the chemical interactions that occur when l-phenylalanine is introduced to solutions containing phosphatidylcholine vesicles. Studies reported in this work address open questions about l-Phe's affinity for lipid vesicle bilayers, the effects of l-Phe partitioning on bilayer properties, l-Phe's solvation within a lipid bilayer, and the amount of l-Phe within that local solvation environment. DSC data show that l-Phe reduces the amount of heat necessary to melt saturated phosphatidylcholine bilayers from their gel to liquid-crystalline state but does not change the transition temperature (Tgel-lc). Time-resolved emission shows only a single l-Phe lifetime at low temperatures corresponding to l-Phe remaining solvated in aqueous solution. At temperatures close to Tgel-lc, a second, shorter lifetime appears that is assigned to l-Phe already embedded within the membrane that becomes hydrated as water starts to permeate the lipid bilayer. This new lifetime is attributed to a conformationally restricted rotamer in the bilayer's polar headgroup region and accounts for up to 30% of the emission amplitude. Results reported for dipalmitoylphosphatidylcholine (DPPC, 16:0) lipid vesicles prove to be general, with similar effects observed for dimyristoylphosphatidylcholine (DMPC, 14:0) and distearoylphosphatidylcholine (DSPC, 18:0) vesicles. Taken together, these results create a complete and compelling picture of how l-Phe associates with model biological membranes. Furthermore, this approach to examining amino acid partitioning into membranes and the resulting solvation forces points to new strategies for studying the structure and chemistry of membrane-soluble peptides and selected membrane proteins.

Partition of Amino Acids in Aqueous Two-Phase Systems Comprising Hydrophilic Organic Solvents and Salts

An aqueous two-phase system (ATPS) comprising a hydrophilic organic solvent and inorganic salt is advantageous because of its high polarity, low viscosity, and low cost. In this work, nine amino acids were partitioned in an ATPS that comprised hydrophilic organic solvents (2-methyl-2-propanol, tetrahydrofuran, and 2-propanol) and salts (Na2SO4, K2HPO4, and (NH4)2SO4). Amino acids were successfully extracted using different combinations of solvents and salts. The distribution ratio of amino acids in the ATPS containing ammonium sulfate increased with an increase in their hydrophobicity, except for that of alanine. The specific extraction of alanine was observed because of its salting-out effect in ammonium sulfate solution. The sulfate salts of amino acids exhibited higher extractability than amphoteric amino acids.

Review: Influence of postabsorptive metabolism on essential amino acid partitioning in lactating dairy cows

In the lactating cow, essential amino acids (EAAs) absorbed from the gut are partitioned to mammary and extra-mammary tissues via blood plasma circulation. There is also entry of EAA into plasma from the breakdown of proteins in the cow’s body. A balance model across plasma was solved to integrate entry rates of branched-chain (BCAA) and non-branched-chain EAA (NBAA) with their corresponding rate constants for clearance by mammary glands and the remainder of the body, for selected glucose and fat infusion experiments. Endogenous EAA entry from whole-body proteolysis was reduced by glucose and unchanged or increased by fat, the efficiency of net plasma BCAA clearance by mammary and extra-mammary tissues was elevated by glucose but slightly reduced by fat, and the efficiency of extra-mammary NBAA clearance may have decreased during glucose infusion but it was not affected or slightly increased by fat. These differences between glucose and fat responses can be accounted for by insulin and glucagon. Insulin suppresses endogenous EAA entry through mechanistic target of rapamycin complex 1, integrated stress response, and glycogen synthase kinase 3 signaling networks in skeletal muscle. While these networks can also regulate protein synthesis rates in muscle and the extra-mammary body, they exhibit low sensitivities to insulin in lactating ruminants. However, in the mammary glands, via these same networks, insulin stimulates clearance of EAA from plasma, although the drive to maintain a set point for milk protein yield takes precedence over nutritional signals. The glucose-induced increase in mammary BCAA clearance without an effect on NBAA clearance is due to a pronounced decrease in plasma BCAA concentrations. Because NBAAs do not experience a similar decline in concentration, the BCAA effect must be due to their metabolic transformation as opposed to sequestration in proteins. In adipose, the products of BCAA catabolism are lipogenic precursors. We propose that faster lipogenesis in adipose tissue, stimulated by glucose infusion, also promotes the uptake of precursor BCAA from plasma, causing a drop in their circulating concentrations. In addition, insulin stimulates BCAA oxidation in muscle as an alternative fuel to fatty acids. A lower efficiency of extra-mammary NBAA clearance during glucose infusion may be the consequence of decreased hepatic expression of AA-catabolizing enzymes in response to low glucagon concentration. The proportion of EAA entry partitioned to the mammary glands is a culmination of regulatory shifts at all of the points discussed above according to a regulated or unfair competition between mammary and extra-mammary processes.

Separation of glatiramer acetate and its constituent amino acids using aqueous two-phase systems composed of maltodextrin and acetonitrile

Glatiramer acetate (GA), FDA-approved therapeutic for multiple sclerosis (MS), is a random-sized synthetic polypeptide composed of four amino acids: tyrosine, glutamic acid, alanine, and lysine. This work aims to design an efficient protocol for the separation of GA from its broth. For this purpose, the feasibility of a carbohydrate-based aqueous two-phase system (ATPS) for separation of GA from the amino acids was investigated. Initially, the binodal curve and tie-lines for the ATPS composed of maltodextrin and acetonitrile were determined. Next, preliminary screening experiments for determining optimum conditions were conducted by introducing GA and amino acids into the ATPS individually and the effect of pH and carbohydrate concentration on partition coefficient were investigated. Results revealed that glutamic acid and alanine were migrated toward the acetonitrile rich phase while GA, tyrosine, and lysin showed the opposite trend. Besides, the partitioning of amino acids was correlated with their physicochemical and structural properties. The preliminary results suggested that the system composed of maltodextrin 15 wt%, acetonitrile 35 wt%, at pH = 6 can be considered as the optimum feed. In the next step, the partitioning of GA and amino acids broth was investigated in the optimum feed. Under these conditions, the partition coefficient and selectivity of GA were obtained to be 1.17 and 62%, respectively. Additionally, circular dichroism spectroscopy results proved that the structure of GA remains unchanged during the separation steps. Finally, a recovery method was proposed where the acetonitrile was evaporated and maltodextrin was precipitated by the addition of methanol.

Overexpression of GmAAP6a enhances tolerance to low nitrogen and improves seed nitrogen status by optimizing amino acid partitioning in soybean.

SummaryAmino acid transport via phloem is one of the major source‐to‐sink nitrogen translocation pathways in most plant species. Amino acid permeases (AAPs) play essential roles in amino acid transport between plant cells and subsequent phloem or seed loading. In this study, a soybean AAP gene, annotated as GmAAP6a, was cloned and demonstrated to be significantly induced by nitrogen starvation. Histochemical staining of GmAAP6a:GmAAP6a‐GUS transgenic soybean revealed that GmAAP6a is predominantly expressed in phloem and xylem parenchyma cells. Growth and transport studies using toxic amino acid analogs or single amino acids as a sole nitrogen source suggest that GmAAP6a can selectively absorb and transport neutral and acidic amino acids. Overexpression of GmAAP6a in Arabidopsis and soybean resulted in elevated tolerance to nitrogen limitation. Furthermore, the source‐to‐sink transfer of amino acids in the transgenic soybean was markedly improved under low nitrogen conditions. At the vegetative stage, GmAAP6a‐overexpressing soybean showed significantly increased nitrogen export from source cotyledons and simultaneously enhanced nitrogen import into sink primary leaves. At the reproductive stage, nitrogen import into seeds was greatly enhanced under both sufficient and limited nitrogen conditions. Collectively, our results imply that overexpression of GmAAP6a enhances nitrogen stress tolerance and source‐to‐sink transport and improves seed quality in soybean. Co‐expression of GmAAP6a with genes specialized in source nitrogen recycling and seed loading may represent an interesting application potential in breeding.

Partitioning of amino acids and proteins into decanol using phase transfer agents towards understanding life in non-polar liquids

Water has many roles in the context of life on Earth, however throughout the universe, other liquids may be able to support the emergence of life. We looked at the ability of amino acids, peptides, a depsipeptide, and proteins to partition into a non-polar decanol phase, with and without the addition of a phase transfer agent. Partitioning evaluated using UV detection, or with HPLC coupled to either charged aerosol detection or ESI-MS. For amino acids and short peptides, phase transfer agents were used to move the biomolecules to the decanol phase, and this transfer was pH dependent. For larger molecules, phase transfer agents did not seem to affect the transfer. Both the depsipetide, valinomycin, and the protein Taq DNA polymerase had solubility in the decanol phase. Additionally, valinomycin appeared to retain its biological ability to bind to potassium ions. These results show that most terrestrial biological molecules are not compatible with non-polar solvents, but it is possible to find and perhaps evolve polymers that are functional in such phases.

Manipulation of sucrose phloem and embryo loading affects pea leaf metabolism, carbon and nitrogen partitioning to sinks as well as seed storage pools

Seed development largely depends on the long-distance transport of sucrose from photosynthetically active source leaves to seed sinks. This source-to-sink carbon allocation occurs in the phloem and requires the loading of sucrose into the leaf phloem and, at the sink end, its import into the growing embryo. Both tasks are achieved through the function of SUT sucrose transporters. In this study, we used vegetable peas (Pisum sativum L.), harvested for human consumption as immature seeds, as our model crop and simultaneously overexpressed the endogenous SUT1 transporter in the leaf phloem and in cotyledon epidermal cells where import into the embryo occurs. Using this 'Push-and-Pull' approach, the transgenic SUT1 plants displayed increased sucrose phloem loading and carbon movement from source to sink causing higher sucrose levels in developing pea seeds. The enhanced sucrose partitioning further led to improved photosynthesis rates, increased leaf nitrogen assimilation, and enhanced source-to-sink transport of amino acids. Embryo loading with amino acids was also increased in SUT1-overexpressors resulting in higher protein levels in immature seeds. Further, transgenic plants grown until desiccation produced more seed protein and starch, as well as higher seed yields than the wild-type plants. Together, the results demonstrate that the SUT1-overexpressing plants with enhanced sucrose allocation to sinks adjust leaf carbon and nitrogen metabolism, and amino acid partitioning in order to accommodate the increased assimilate demand of growing seeds. We further provide evidence that the combined Push-and-Pull approach for enhancing carbon transport is a successful strategy for improving seed yields and nutritional quality in legumes.

Systematized biosynthesis and catabolism regulate citrulline accumulation in watermelon

Citrulline, a non-protein amino acid, is present in large amounts in watermelon (Citrullus lanatus (Thunb.) Matsum. & Nakai Cucurbitaceae) fruits. Amino acid profiling of various tissues of cv. Charleston Gray during plant development confirmed progressive accumulation of citrulline only in the fruit flesh and rind tissues. Citrulline content was positively correlated with precursor (ornithine) and by-product (arginine) amino acids during fruit ripening. Genetic variation in the partitioning of citrulline and related amino acids in the flesh and rind tissues was confirmed in a sub-set of watermelon cultivars. No correlation was established between morphological fruit traits (size and rind properties) and citrulline content. To understand the regulation of citrulline accumulation, we investigated the expression of genes associated with its biosynthesis and catabolism in flesh and rind tissues during fruit development. The expression of ornithine carbamoyltransferase (OTC) involved in the ultimate step of citrulline synthesis remained steady in both tissues. The expression of N-acetylornithine aminotransferase (N-AOA) involved in the production of N-acetylornithine and N-acetylornithine deacetylase (AOD-3) involved in ornithine synthesis coincided with increasing accumulation of citrulline in flesh and rind tissues during fruit development. Down-regulation N-acetylornithine-glutamate acetyltransferase (N-AOGA) suggests the subordinate role of the non-cyclic pathway in citrulline synthesis. Eccentricity between citrulline accumulation and expression of carbamoyl phosphate synthases (CPS-1, CPS-2) during fruit development suggest that the localized synthesis of carbamoyl phosphates may not be required for citrulline synthesis. Most genes involved in citrulline break-down (Argininosuccinate synthases - ASS-1, ASS-2, and ASS-3, Argininosuccinate lyases - ASL-1, Ornithine decarboxylase - ODC, Arginine decarboxylase - ADC) were consistently down-regulated during fruit development.

Consistent proteinaceous organic matter partitioning into mineral and organic soil fractions during pedogenesis in diverse ecosystems

Proteinaceous compounds are critical in soil organic matter (SOM) formation and persistence, but the partitioning into mineral-associated and organic forms during hundreds and thousands of years of pedogenesis are poorly understood across multiple climates and vegetation types. We investigated the partitioning of amino acids (AA) into mineral-bound (MB) and non-mineral associated organic (NMO) soil fractions to discern whether consistent patterns during ecosystem development were observed across two different climates (cool temperate continental, USA; and moist oceanic forests, New Zealand). Although each ecosystem retained unique soil AA signatures, consistent patterns in both systems were observed with three main findings. (1) Regardless of differences in climate and vegetation between the two ecosystems, AA consistently partitioned in similar ways into MB and NMO soil fractions. For example, Thr, Ser, and Asx were relatively more dominant in the NMO fraction while Arg, Lys, Cys, and Met were relatively more dominant in the MB soil fraction. (2) AA change, showed similar trends related to chemical groupings of positively-charged, polar aromatic, sulfur containing, and non-polar AAs across both ecosystems consistent with changing patterns of soil Fe and Al bearing minerals, such as an increasing weathering index (WI; Fe dithionite/total Fe) and losses of Fe and Al from surface soils during ecosystem development. (3) The pedogenic patterns of AA change, in each system paralleled biological transitions in bacterial communities, suggesting a linkage between the AA sources and the soil sink that contributes to soil organic N. This latter point contrasts with the potential for complete change in soil AA composition that could occur upon processing and binding in soil. Overall, the consistency in the types of AAs that partition into either MB or NMO soil fractions across locations provide evidence of similar processes that contribute to soil organic N accrual across soils. Some AA also appeared to be retained in soil by electrostatic forces, and AA–organic matter interactions, that need further study. Mineral–metal complexation of AAs with sulfur side chain groups, and non-polar interactions, respectively, provide examples of these potential mechanisms for study. The results not only support mechanisms of soil proteinacous organic matter chemistry being driven by the interactions with the soil matrix, acting as a “sink”, but also draw attention to the sources of organic matter (microbes, plants) in determining the composition of organic N in soil during pedogenesis.

Function of pea amino acid permease AAP6 in nodule nitrogen metabolism and export, and plant nutrition.

Legumes fix atmospheric nitrogen through a symbiotic relationship with bacteroids in root nodules. Following fixation in pea (Pisum sativum L.) nodules, nitrogen is reduced to amino acids that are exported via the nodule xylem to the shoot, and in the phloem to roots in support of growth. However, the mechanisms involved in amino acid movement towards the nodule vasculature, and their importance for nodule function and plant nutrition, were unknown. We found that in pea nodules the apoplasmic pathway is an essential route for amino acid partitioning from infected cells to the vascular bundles, and that amino acid permease PsAAP6 is a key player in nitrogen retrieval from the apoplasm into inner cortex cells for nodule export. Using an miRNA interference (miR) approach, it was demonstrated that PsAAP6 function in nodules, and probably in roots, and affects both shoot and root nitrogen supply, which were strongly decreased in PsAAP6-miR plants. Further, reduced transporter function resulted in increased nodule levels of ammonium, asparagine, and other amino acids. Surprisingly, nitrogen fixation and nodule metabolism were up-regulated in PsAAP6-miR plants, indicating that under shoot nitrogen deficiency, or when plant nitrogen demand is high, systemic signaling leads to an increase in nodule activity, independent of the nodule nitrogen status.

Force Field Benchmark of Amino Acids. 2. Partition Coefficients between Water and Organic Solvents.

2018 , 58 , 1037 - 1052 ), may aid in the development and systematic improvements of classical force fields to model proteins in aqueous and nonaqueous phases accurately.

Experimental investigation and thermodynamic modeling of amino acids partitioning in a water/ionic liquid system

Amino acids partitioning including phenylalanine, glutamic acid, and tryptophan in aqueous and ionic liquid phases at temperature of 298.15K and atmospheric pressure were measured. 1-Butyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide used in this work can produce two phases with water immediately. The effect of aqueous solution pH on amino acids partitioning was studied and revealed that amino acid partitioning coefficient was decreased with increasing pH. This phenomenon pertains to the electrostatic interaction between cations of amino acid and the anions of ionic liquid which is decreased when pH increases. Considering the effect of pH, liquid-liquid equilibrium data of amino acids were obtained in a pH owning maximum partitioning coefficient. It was observed that chemical structure of amino acid has an important effect on partitioning of amino acids between two phases. Experimental data indicated that glutamic acid is mostly transferred to aqueous phase while tryptophan moves to the ionic liquid phase. In the modeling section, equilibrium behavior of water, amino acid and ionic liquid system were predicted by SAFT equation of state. Necessary parameters of water and pure ionic liquid for equation of state were measured and amino acid parameters were obtained from literature. Binary interactions of components were optimized by equilibrium data. NRTL model was used in order to analyze cabapility of SAFT EoS to predict the behavior of system. The results indicate that SAFT model anticipates liquid-liquid equilibrium of the system better than NRTL model. The percentage of average absolute deviation for SAFT equiation is <1.2% while for NRTL model is >1.9%.

Partition Coefficient of Arginine between Translocon Interior and Lipid Phase

Depending on its hydrophobicity, the protein translocation complex sorts polypeptide segments into the membrane or into the aqueous solution. Its short residence time in the channel limits polypeptide sampling between the aqueous lumen and the membrane interior, giving rise to an out-of-equilibrium condition. Moreover, reduced mobility and orientational restrictions of confined intraluminal water molecules are bound to further bias peptide partition into the membrane, rendering good agreement between the physico-chemical and biological hydrophobicity scales all the more surprising. Here we report the equilibrium partition coefficient of arginine derived on the single molecule level by monitoring -repetitive lateral movement of a stalled polypeptide segment between the SecYEG lumen and the lipid interior. Polypeptide movements in the direction normal to the membrane were inhibited by sandwiching the transmembrane sequence between a SecA binding sequence and the sequence of the calmodulin binding protein. With calmodulin and SecA bound to opposite sides of the membrane, polypeptide residence within the SecYEG pore was indicated by channel opening, whereas episodes of residence in the hydrophobic membrane interior corresponded to channel closings. Translocon purification and reconstitution into planar lipid bilayers were performed as described previously.1-2 Comparison to amino acid partitioning in a biphasic system (physico-chemical hydrophobicity scale) allowed us to address the effects of water confinement by the SecYEG pore, while comparison to the biological hydrophobicity scale reveals out-of-equilibrium effects. 1. Knyazev, D. G.; ⋯; Pohl, P., The bacterial translocon SecYEG opens upon ribosome binding. J. Biol. Chem. 2013, 288 (25), 17941-6. 2. Knyazev, D. G.;⋯ ; Pohl, P., Ion conductivity of the bacterial translocation channel SecYEG engaged in translocation. J. Biol. Chem. 2014, 289 (35), 24611-6.

Partitioning of amino acids in the novel biphasic systems based on environmentally friendly ethyl lactate

For the first time, we report on the performance of biphasic system composed of ethyl lactate, water and inorganic salt (K3PO4, K2HPO4 and K2CO3) for the separation of amino acids (L-phenylalanine, L-tryptophan and L-tyrosine) from their aqueous solutions. Cloud points (solubility curve) and tie-lines for three ternary (ethyl la ctate + water + inorganic salt) systems at 298.2 K and 313.2 K at atmospheric pressure were determined. For certain composition range, these mixture exhibit biphasic systems – top and bottom phases rich in ethyl lactate and salt, respectively. Partition coefficients of amino acids and their extraction efficiencies, as essential parameters for design of any separation process, were measured at two temperatures – 298.2 K and 313.2 K. The maximum values of partition coefficients were observed for the system containing K3PO4: 3.5, 3.7 and 11.9 for L-phenylalanine at 313.2 K, L-tyrosine at 298.2 K and L-tryptophan at 313.2 K, respectively. The obtained results clearly showed that the biphasic systems based on ethyl lactate are suitable for the efficient and sustainable recovery of amino acids from solutions with water.

Energetics of side-chain partitioning of β-signal residues in unassisted folding of a transmembrane β-barrel protein

The free energy of water-to-interface amino acid partitioning is a major contributing factor in membrane protein folding and stability. The interface residues at the C terminus of transmembrane β-barrels form the β-signal motif required for assisted β-barrel assembly in vivo but are believed to be less important for β-barrel assembly in vitro. Here, we experimentally measured the thermodynamic contribution of all 20 amino acids at the β-signal motif to the unassisted folding of the model β-barrel protein PagP. We obtained the partitioning free energy for all 20 amino acids at the lipid-facing interface (ΔΔG0w,i(φ)) and the protein-facing interface (ΔΔG0w,i(π)) residues and found that hydrophobic amino acids are most favorably transferred to the lipid-facing interface, whereas charged and polar groups display the highest partitioning energy. Furthermore, the change in non-polar surface area correlated directly with the partitioning free energy for the lipid-facing residue and inversely with the protein-facing residue. We also demonstrate that the interface residues of the β-signal motif are vital for in vitro barrel assembly, because they exhibit a side chain–specific energetic contribution determined by the change in nonpolar accessible surface. We further establish that folding cooperativity and hydrophobic collapse are balanced at the membrane interface for optimal stability of the PagP β-barrel scaffold. We conclude that the PagP C-terminal β-signal motif influences the folding cooperativity and stability of the folded β-barrel and that the thermodynamic contributions of the lipid- and protein-facing residues in the transmembrane protein β-signal motif depend on the nature of the amino acid side chain.

An overview of energy and protein utilisation during growth in simple-stomached animals

The biological processes underlying the partitioning of amino acids and energy during animal growth are well understood qualitatively. However, if a deeper mechanistic understanding is to be achieved, such as to allow generalised predictions of growth outcomes, these biological processes need to be described quantitatively, along with critical control points. Concepts and rules can be formulated at mechanistic and semi-mechanistic levels, and often reflecting causation, to allow nutrient intake and partitioning to be described in a quantitative manner for different animal and environmental conditions. An overview is given of amino acid and energy partitioning during growth in monogastric animals, in terms of causation and quantitatively based descriptors. Current knowledge is far from complete, and areas requiring new insights and a more in-depth understanding of causative mechanisms include voluntary food-intake control, dynamics of nutrient uptake, temporary post-prandial nutrient storage, relationships among nutrient intakes, protein turnover and maintenance-energy requirement, colonic amino acid uptake in poultry, bioavailability of amino acids other than lysine, diet effects on gut endogenous amino acid loss, inevitable amino acid catabolism, preferential amino acid catabolism, and diet, age and genotype effects on body protein synthesis and degradation.

Connecting Source with Sink: The Role of Arabidopsis AAP8 in Phloem Loading of Amino Acids.

Allocation of large amounts of nitrogen to developing organs occurs in the phloem and is essential for plant growth and seed development. In Arabidopsis (Arabidopsis thaliana) and many other plant species, amino acids represent the dominant nitrogen transport forms in the phloem, and they are mainly synthesized in photosynthetically active source leaves. Following their synthesis, a broad spectrum of the amino nitrogen is actively loaded into the phloem of leaf minor veins and transported within the phloem sap to sinks such as developing leaves, fruits, or seeds. Controlled regulation of the source-to-sink transport of amino acids has long been postulated; however, the molecular mechanism of amino acid phloem loading was still unknown. In this study, Arabidopsis AMINO ACID PERMEASE8 (AAP8) was shown to be expressed in the source leaf phloem and localized to the plasma membrane, suggesting its function in phloem loading. This was further supported by transport studies with aap8 mutants fed with radiolabeled amino acids and by leaf exudate analyses. In addition, biochemical and molecular analyses revealed alterations in leaf nitrogen pools and metabolism dependent on the developmental stage of the mutants. Decreased amino acid phloem loading and partitioning to sinks led to decreased silique and seed numbers, but seed protein levels were unchanged, demonstrating the importance of AAP8 function for sink development rather than seed quality. Overall, these results show that AAP8 plays an important role in source-to-sink partitioning of nitrogen and that its function affects source leaf physiology and seed yield.