A hemoprotein with a zinc-mirror heme site ties heme availability to carbon metabolism in cyanobacteria.

Heme has a critical role in the chemical framework of the cell as an essential protein cofactor and signaling molecule that controls diverse processes and molecular interactions. Using a phylogenomics-based approach and complementary structural techniques, we identify a family of dimeric hemoproteins comprising a domain of unknown function DUF2470. The heme iron is axially coordinated by two zinc-bound histidine residues, forming a distinct two-fold symmetric zinc-histidine-iron-histidine-zinc site. Together with structure-guided in vitro and in vivo experiments, we further demonstrate the existence of a functional link between heme binding by Dri1 (Domain related to iron 1, formerly ssr1698) and post-translational regulation of succinate dehydrogenase in the cyanobacterium Synechocystis, suggesting an iron-dependent regulatory link between photosynthesis and respiration. Given the ubiquity of proteins containing homologous domains and connections to heme metabolism across eukaryotes and prokaryotes, we propose that DRI (Domain Related to Iron; formerly DUF2470) functions at the molecular level as a heme-dependent regulatory domain.

Exploring metabolic responses and pathway changes in CHO-K1 cells under varied aeration conditions and copper supplementations using 1 H NMR-based metabolomics.

The optimization of bioprocess for CHO cell culture involves careful consideration of factors such as nutrient consumption, metabolic byproduct accumulation, cell growth, and monoclonal antibody (mAb) production. Valuable insights can be obtained by understanding cellular physiology to ensure robust and efficient bioprocess. This study aims to improve our understanding of the CHO-K1 cell metabolism using 1 H NMR-based metabolomics. Initially, the variations in culture performance and metabolic profiles under varied aeration conditions and copper supplementations were thoroughly examined. Furthermore, a comprehensive metabolic pathway analysis was performed to assess the impact of these conditions on the implicated pathways. The results revealed substantial alterations in the pyruvate metabolism, histidine metabolism, as well as phenylalanine, tyrosine and tryptophan biosynthesis, which were especially evident in cultures subjected to copper deficiency conditions. Conclusively, significant metabolites governing cell growth and mAb titer were identified through orthogonal partial least square-discriminant analysis (OPLS-DA). Metabolites, including glycerol, alanine, formate, glutamate, phenylalanine, and valine, exhibited strong associations with distinct cell growth phases. Additionally, glycerol, acetate, lactate, formate, glycine, histidine, and aspartate emerged as metabolites influencing cell productivity. This study demonstrates the potential of employing 1 H NMR-based metabolomics technology in bioprocess research. It provides valuable guidance for feed medium development, feeding strategy design, bioprocess parameter adjustments, and ultimately the enhancement of cell proliferation and mAb yield.

Substrate binding and catalytic mechanism of the Se-glycosyltransferase SenB in the biosynthesis of selenoneine.

Selenium is an essential multifunctional trace element in diverse organisms. The only Se-glycosyltransferase identified that catalyzes the incorporation of selenium in selenoneine biosynthesis is SenB from Variovorax paradoxus. Although the biochemical function of SenB has been investigated, its substrate specificity, structure, and catalytic mechanism have not been elucidated. Here, we reveal that SenB exhibits sugar donor promiscuity and can utilize six UDP-sugars to generate selenosugars. We report crystal structures of SenB complexed with different UDP-sugars. The key elements N20/T23/E231 contribute to the sugar donor selectivity of SenB. A proposed catalytic mechanism is tested by structure-guided mutagenesis, revealing that SenB yields selenosugars by forming C-Se glycosidic bonds via spontaneous deprotonation and disrupting Se-P bonds by nucleophilic water attack, which is initiated by the critical residue K158. Furthermore, we functionally and structurally characterize two other Se-glycosyltransferases, CbSenB from Comamonadaceae bacterium and RsSenB from Ramlibacter sp., which also exhibit sugar donor promiscuity.

CLPP-Null Eukaryotes with Excess Heme Biosynthesis Show Reduced L-arginine Levels, Probably via CLPX-Mediated OAT Activation.

The serine peptidase CLPP is conserved among bacteria, chloroplasts, and mitochondria. In humans and mice, its loss causes Perrault syndrome, which presents with growth deficits, infertility, deafness, and ataxia. In the filamentous fungus Podospora anserina, CLPP loss leads to longevity. CLPP substrates are selected by CLPX, an AAA+ unfoldase. CLPX is known to target delta-aminolevulinic acid synthase (ALAS) to promote pyridoxal phosphate (PLP) binding. CLPX may also influence cofactor association with other enzymes. Here, the evaluation of P. anserina metabolomics highlighted a reduction in arginine/histidine levels. In Mus musculus cerebellum, reductions in arginine/histidine and citrulline occurred with a concomitant accumulation of the heme precursor protoporphyrin IX. This suggests that the increased biosynthesis of 5-carbon (C5) chain deltaALA consumes not only C4 succinyl-CoA and C1 glycine but also specific C5 delta amino acids. As enzymes responsible for these effects, the elevated abundance of CLPX and ALAS is paralleled by increased OAT (PLP-dependent, ornithine delta-aminotransferase) levels. Possibly as a consequence of altered C1 metabolism, the proteome profiles of P. anserina CLPP-null cells showed strong accumulation of a methyltransferase and two mitoribosomal large subunit factors. The reduced histidine levels may explain the previously observed metal interaction problems. As the main nitrogen-storing metabolite, a deficiency in arginine would affect the urea cycle and polyamine synthesis. Supplementation of arginine and histidine might rescue the growth deficits of CLPP-mutant patients.

Associations of diet quality and food consumption with serum biomarkers for lipid and amino acid metabolism in Finnish children: the PANIC study.

PURPOSE: To investigate the associations of overall diet quality and dietary factors with serum biomarkers for lipid and amino acid metabolism in a general population of children. METHODS: We studied 194 girls and 209 boys aged 6-8 years participating in the Physical Activity and Nutrition in Children study. Food consumption was assessed by 4-day food records and diet quality was quantified by the Finnish Children Healthy Eating Index (FCHEI). Fasting serum fatty acids, amino acids, apolipoproteins, as well as lipoprotein particle sizes were analyzed with high-throughput nuclear magnetic resonance spectroscopy. Data were analyzed using linear regression adjusted for age, sex, and body fat percentage. RESULTS: FCHEI was directly associated with the ratio of polyunsaturated (PUFA) to saturated fatty acids (SFA) (PUFA/SFA), the ratio of PUFA to monounsaturated fatty acids (MUFA) (PUFA/MUFA), the ratio of PUFA to total fatty acids (FA) (PUFA%), the ratio of omega-3-fatty acids to total FA (omega-3 FA%), and inversely associated with the ratio of MUFA to total FA (MUFA%), alanine, glycine, histidine and very-low density lipoprotein (VLDL) particle size. Consumption of vegetable oils and vegetable-oil-based margarine (≥ 60% fat) was directly associated with PUFA/SFA, PUFA/MUFA, PUFA%, the ratio of omega-6 FA to total FA (omega-6 FA%), and inversely associated with SFA, MUFA, SFA to total FA (SFA%), MUFA%, alanine and VLDL particle size. Consumption of high-fiber grain products directly associated with PUFA/SFA, PUFA/MUFA, omega-3 FA%, omega-6 FA%, PUFA% and inversely associated with SFA and SFA%. Fish consumption directly related to omega-3 FA and omega-3 FA%. Consumption of sugary products was directly associated with histidine and VLDL particle size. Vegetable, fruit, and berry consumption had direct associations with VLDL particle size and the ratio of apolipoprotein B to apolipoprotein A1. Consumption of low fat (< 1%) milk was directly associated with phenylalanine. A higher consumption of high-fat (≥ 1%) milk was associated with lower serum MUFA/SFA and higher SFA%. Sausage consumption was directly related to SFA% and histidine. Red meat consumption was inversely associated with glycine. CONCLUSIONS: Better diet quality, higher in intake of dietary sources of unsaturated fat and fiber, and lower in sugary product intake were associated with more favorable levels of serum biomarkers for lipid and amino acid metabolism independent of adiposity. TRIAL REGISTRATION: ClinicalTrials.gov: NCT01803776, registered March 3, 2013.

L-Lysine supplementation affects dietary protein quality and growth and serum amino acid concentrations in rats.

Single amino acid (AA) supplementations in foods are increasing, however their potential nutritional and physiological impacts are not fully understood. This study examined the effects of L-lysine (Lys) supplementation on protein quality of diets, serum AA concentrations and associations between the ratio of supplemental Lys to dietary protein (X) with body weight gain (BWG) in Sprague-Dawley male rats. Rats were fed one of 10 diets containing either 7% or 20% casein and supplemented with 0% (Control), 1.5%, 3%, 6% Lys or 6% Lys + 3% L-arginine (Arg) (8 rats/diet group) for 1 week. Lys supplementation reduced the protein quality of the casein-based diets (p < 0.01). BWG was reduced by supplemental Lys when X > 0.18. Free Lys supplementation dose-dependently increased serum Lys levels (p < 0.01), while increased protein-bound Lys (1.4% vs 0.52%) had little effect on serum Lys (p > 0.05). In the 7% casein diets, ≥ 1.5% supplemental Lys reduced serum alanine, asparagine, glycine, isoleucine, leucine, serine, tyrosine, valine, carnitine, ornithine, and increased urea. Supplementation of ≥ 3% Lys additionally reduced tryptophan and increased histidine, methionine and α-aminoadipic acid (α-AAA) compared to the Control (p < 0.05). In the 20% casein diets, addition of ≥ 1.5% Lys reduced serum asparagine and threonine, and ≥ 3% Lys reduced leucine, proline, tryptophan, valine, and ornithine, and 6% Lys reduced carnitine, and increased histidine, methionine, and α-AAA. Overall, this study showed that free Lys supplementation in a Lys-sufficient diet reduced the protein quality of the diets and modified the serum concentrations of many amino acids. Excess free Lys intake adversely affected growth and utilization of nutrients due to AA imbalance or antagonism. Overall lower protein intake increases susceptibility to the adverse effects of Lys supplementation.

Evaluation of crystalline amino acids as potent stimulatory chemoattractants for the slipper lobster Thenus orientalis.

Intensive research on the effectiveness of chemoattractants has been widely explored to improve the feed qualities in expanding crustacean farming. Taste preferences in slipper lobster remained unknown despite their significant contribution to the lobster fisheries. Chemoattractants allow better performance in aquaculture species by increasing food attractiveness and palatability. Amino acids (AA) have been leading in previous research on crustacean feeding behavior. Given that slipper lobster possesses chemoreceptors to detect and orient towards food, this study investigated an approach to identify the AA with the most potent chemoattractant in eliciting a response from slipper lobster. Behavioral assays were performed to evaluate the responses of slipper lobster Thenus orientalis (carapace length, 52.34 ± 1.52 mm) on 15 crystalline AA and three derivatives of AA (DAA) at three concentrations between 10-1 and 10-3 M as test substances (TS). Meretrix sp. extract was used as a positive control and clean filtered seawater as a negative control. The behavioral responses of 14 T. orientalis were evaluated based on their antennular flicking rate, third maxillipeds activity, and substrate probing by the pereiopods. T. orientalis responded to the solutions of single AA down to a concentration of 10-3 M, excluding histidine and serine. The behavioral activity displayed by T. orientalis increased with the TS concentrations. L-glutamic acid monosodium salt monohydrate, betaine, and glycine solutions elicited the most behavioral responses, whereas histidine exhibited the lowest behavioral responses. Conclusively, L-glutamic acid monosodium salt monohydrate, betaine, and glycine can be potential chemoattractants for T. orientalis.

Effect of carnosine synthesis precursors in the diet on jejunal metabolomic profiling and biochemical compounds in slow-growing Korat chicken.

The slow-growing Korat chicken (KR) has been developed to provide an alternative breed for smallholder farmers in Thailand. Carnosine enrichment in the meat can distinguish KR from other chicken breeds. Therefore, our aim was to investigate the effect of enriched carnosine synthesis, obtained by the ß-alanine and L-histidine precursor supplementation in the diet, on changes to metabolomic profiles and biochemical compounds in slow-growing KR jejunum tissue. Four hundred 21-day-old female KR chickens were divided into 4 experimental groups: a group with a basal diet, a group with a basal diet supplemented with 1.0% ß-alanine, 0.5% L-histidine, and a mix of 1.0% ß-alanine and 0.5% L-histidine. The feeding trial lasted 70 d. Ten randomly selected chickens from each group were slaughtered. Metabolic profiles were analyzed using proton nuclear magnetic resonance spectroscopy. In total, 28 metabolites were identified. Significant changes in the concentrations of these metabolites were detected between the groups. Partial least squares discriminant analysis was used to distinguish the metabolites between the experimental groups. Based on the discovered metabolites, 34 potential metabolic pathways showed differentiation between groups, and 8 pathways (with impact values higher than 0.05, P < 0.05, and FDR < 0.05) were affected by metabolite content. In addition, biochemical changes were monitored using synchrotron radiation-based Fourier transform infrared microspectroscopy. Supplementation of ß-alanine alone in the diet increased the ß-sheets and decreased the α-helix content in the amide I region, and supplementation of L-histidine alone in the diet also increased the ß-sheets. Furthermore, the relationship between metabolite contents and biochemical compounds were confirmed using principal component analysis (PCA). Results from the PCA indicated that ß-alanine and L-histidine precursor group was highly positively correlated with amide I, amide II, creatine, tyrosine, valine, isoleucine, and aspartate. These findings can help to understand the relationships and patterns between the spectral and metabolic processes related to carnosine synthesis.

Multifactorial interaction of selenium, iron, xylose, and glycine on cordycepin metabolism in Cordyceps militaris.

Cordycepin, a nucleoside analog, is the main antioxidative and antimicrobial substance in Cordyceps militaris. To improve the metabolism of cordycepin, carbon sources, nitrogen sources, trace elements, and precursors were studied by single factor, Plackett-Burman, and central composite designs in C. militaris mycelial fermentation. Under the regulation of the multifactorial interactions of selenite, ferrous chloride, xylose, and glycine, cordycepin production was increased by 5.2-fold compared with the control. The gene expression of hexokinase, ATP phosphoribosyltransferase, adenylosuccinate synthetase, and cns1-3 in the glycolysis, pentose phosphate, and adenosine synthesis pathways were increased by 3.2-7.5 times due to multifactorial interactions, while the gene expression of histidine biosynthesis trifunctional protein and histidinol-phosphate aminotransferase in histidine synthesis pathway were decreased by 23.4%-56.2%. Increasing with cordycepin production, glucose uptake was accelerated, mycelia growth was inhibited, and the cell wall was damaged. Selenomethionine (SeMet), selenocysteine (SeCys), and selenium nanoparticles (SeNPs) were the major Se species in C. militaris mycelia. This study provides a new insight for promoting cordycepin production by regulating glycolysis, pentose phosphate, and histidine metabolism. KEY POINTS: • Cordycepin production in the CCDmax group was 5.2-fold than that of the control. • Glucose uptake of the CCDmax group was accelerated and cell wall was damaged. • The metabolic flux was concentrated to the cordycepin synthesis pathway.

Aqueous extract of Polygala japonica Houtt. ameliorated nonalcoholic steatohepatitis in mice through restoring the gut microbiota disorders and affecting the metabolites in feces and liver.

BACKGROUND: Polygala japonica Houtt. (PJ) has been demonstrated with several biological potentials such as lipid-lowering and anti-inflammatory effects. However, the effects and mechanisms of PJ on nonalcoholic steatohepatitis (NASH) remain unclear. PURPOSE: The aim of this study was to evaluate the effects of PJ on NASH and illustrate the mechanism based on modulating gut microbiota and host metabolism. MATERIALS AND METHODS: NASH mouse model was induced using methionine and choline deficient (MCD) diet and orally treated with PJ. The therapeutic, anti-inflammatory, and anti-oxidative effects of PJ on mice with NASH were firstly assessed. Then, the gut microbiota of mice was analyzed using 16S rRNA sequencing to assess the changes. Finally, the effects of PJ on the metabolites in liver and feces were explored by untargeted metabolomics. RESULTS: The results indicated that PJ could improve hepatic steatosis, liver injury, inflammatory response, and oxidative stress in NASH mice. PJ treatment also affected the diversity of gut microbiota and changed the relative abundances of Faecalibaculum. Lactobacillus, Muribaculaceae, Dubosiella, Akkermansia, Lachnospiraceae_NK4A136_group, and Turicibacter in NASH mice. In addition, PJ treatment modulated 59 metabolites both in liver and feces. Metabolites involved in histidine, and tryptophan metabolism pathways were identified as the key metabolites according to the correlation analysis between differential gut microbiota and metabolites. CONCLUSION: Our study demonstrated the therapeutic, anti-inflammatory and anti-oxidative potentials of PJ on NASH. The mechanisms of PJ treatment were related to the improvement of gut microbiota dysbiosis and the regulation of histidine and tryptophan metabolism.

Nutritional Supplementation and Enhanced Antioxidant Function by Dietary Intake of Selenoneine and Other Selenium Compounds in Red Seabream Pagrus major.

Selenoneine, 2-selenyl-Nα, Nα, Nα-trimethyl-L-histidine, is the major organic selenium compound in marine fish. To characterize biological antioxidant function of selenoneine in fish, the accumulation of selenoneine and other selenium compounds, i. e., sodium selenite and selenomethionine, in the muscle and other tissues of red seabream. We reared red seabream by feeding of 1% dry pellet containing of sodium selenite, selenomethionine, or selenoneine of body weight twice a day for 4 weeks. After that, we replaced to 1% of normal commercial dry pellet of body weight twice a day for 1 week from the selenium supplementation, and tissue distribution of total selenium was determined. Selenium supplementation with selenoneine, selenomethionine, and sodium selenite enhanced selenium accumulation in the white muscle, kidney, and hepatopancreas in comparison with the control group. By the dietary intake of selenoneine, total selenium concentrations were increased in the white muscle, heart, kidney, spleen, hepatopancreas, brain, and blood cells in a dose-dependent manner during the trials after 2 weeks. Dietary intake of selenoneine as well as sodium selenite and selenomethionine reduced oxidation-reduction potential (ORP). Selenoneine concentrations in the white muscle and blood cells were accumulated for 4 weeks by the selenoneine intake, whereas selenoneine concentration was not elevated by the intake of selenomethionine and sodium selenite, suggesting that tissue selenoneine levels might be derived from only selenoneine-containing diet. The uptake factor of selenoneine from the artificial feed containing selenoneine was calculated to be 0.0062 in the white muscle and 4.0 in the blood. The half-life of total selenium in the blood cells and white muscle were estimated to be 60 days in the white muscle and 30 days in the blood.

Lactational performance effects of supplemental histidine in dairy cows: A meta-analysis.

The objective of this meta-analysis was to examine the effects of supplemental His on lactational performance, plasma His concentration and efficiency of utilization of digestible His (EffHis) in dairy cows. The meta-analysis was performed on data from 17 studies published in peer-reviewed journals between 1999 and 2022. Five publications reported data from 2 separate experiments, which were included in the analyses as separate studies, therefore resulting in a total of 22 studies. In 10 studies, His was supplemented as rumen-protected (RP) His; in 1 study, 2 basal diets with different dHis levels were fed; and in the remaining experiments, free His was infused into the abomasum (4 studies), the jugular vein (3 studies) or deleted from a mixture of postruminally infused AA (4 studies). The main forages in the diets were corn silage in 14 and grass silage in 8 studies. If not reported in the publications, the supplies of dietary CP, metabolizable protein (MP), net energy of lactation, and digestible His (dHis) were estimated using NRC (2001). An initial meta-analysis was performed to test the standard mean difference (SMD; raw mean difference of treatment and control means divided by the pooled standard deviation of the means), that is, effect size, and the corresponding 95% confidence interval (CI) in production parameters between His-supplemented groups versus control. Further, regression analyses were also conducted to examine and compare the relationships between several response variables and dHis supply. Across studies, His supplementation increased plasma His concentration (SMD = 1.39; 95% CI: 1.17-1.61), as well as DMI (SMD = 0.240; 95% CI: 0.051-0.429) and milk yield (MY; SMD = 0.667; 95% CI: 0.468-0.866), respectively. Further, milk true protein concentration (MTP; SMD = 0.236; 95% CI: 0.046-0.425) and milk true protein yield (MTPY; SMD = 0.581; 95% CI: 0.387-0.776) were increased by His supplementation. Notably, the increase in MTP concentration and MTPY were 3.9 and 1.3 times greater for studies with MP-deficient (according to NRC 2001) diets compared with studies with MP-adequate diets. The regression analyses revealed that production parameters (DMI, MY, and MTPY) responded in a nonlinear manner to increasing His supply. Further, we detected a difference in the magnitude of change in MTPY and plasma His concentration with the level of His supply and between His supplementation methods, being greater for infused His compared with RPHis. Lastly, a linear and negative relationship between EffHis and the ratio of total digestible His to net energy for lactation supply was observed, indicating an important interaction between dHis and energy supply and EffHis (i.e., utilization of dHis to support protein export). Overall, these analyses confirm His as an important AA in dairy cattle nutrition.

Tolerable Upper Intake Level for Individual Amino Acids in Humans: A Narrative Review of Recent Clinical Studies.

Individual amino acids are widely popular as supplements because of various perceived and real health benefits. However, currently, there are no recommendations set by national health agencies for tolerable upper intake levels (UL) for amino acids because of a lack of well-conducted human dose-response trials. In the past decade, under the initiative of the International Council on Amino Acid Science, a nonprofit organization, a series of UL human clinical studies were conducted. The goal of this narrative review is to summarize the studies on 6 essential amino acids (leucine, tryptophan, methionine, lysine, histidine, and phenylalanine), 2 nonessential amino acids (arginine and serine), and 2 nonproteinogenic amino acids (ornithine and citrulline) and provide the first set of ULs. A brief background of the concept of the DRI framework of UL, the concept of UL for amino acids, and a perspective of the results are also provided. The data suggest that in relatively healthy adult individuals, the tested amino acids are well tolerated, and ULs, or the no-observed-adverse-effect-level (NOAEL), lowest-observed-adverse-effect-level (LOAEL), can be determined. The ULs were for leucine-young (35 g/d), tryptophan (4.5 g/d), and leucine-elderly (30 g/d); NOAEL and LOAEL for methionine at 3.2 and 6.4 g/d, respectively; NOAEL for arginine (30 g/d); NOAEL and LOAEL for lysine at 6 and 7.5 g/d, respectively; NOAEL and LOAEL for histidine at 8 and 12 g/d, respectively; and NOAEL for phenylalanine (12 g/d), serine (12 g/d), ornithine (12 g/d) and citrulline (24 g/d). This first set of human UL data are hoped to help national and international agencies set safety standards for supplemental amino acids.

Rapidly and accurately screening histidine decarboxylase inhibitors from Radix Paeoniae alba using ultrafiltration-high performance liquid chromatography/mass spectrometry combined with enzyme channel blocking and directional enrichment technique.

Histidine Decarboxylase (HDC), an unique enzyme responsible for the synthesis of histamine, which is an important mediator in allergy. Inhibition of HDC activity to decrease histamine production is one way to alleviate allergic symptoms. Traditional Chinese medicines (TCMs) with reported anti-allergy effect is one of important source to search for natural HDC inhibitor. Ultrafiltration combined with high-performance liquid chromatography/mass spectrometry (UF-HPLC/MS) is an effective method for screening HDC inhibitor from TCMs. Nevertheless, false-positive and false-negative results caused by the non-specific binding and the neglection of the trace active compounds are major problems in this method. In this study, an integrated strategy that combined UF-HPLC/MS with enzyme channel blocking (ECB) technique and directional enrichment (DE) technique was developed to seek natural HDC inhibitors from Radix Paeoniae alba (RPA), and at the same time, to reduce false-positive and false-negative results. HDC activity was detected to determine the validity of the screened compounds by RP-HPLC-FD in vitro. Molecular docking was applied to assay the binding affinity and binding sites. As a result, three compounds were screened from low content components of RPA after the DE. Among them, two non-specific compounds were eliminated by ECB, and the specific compound was identified as catechin, which has obvious HDC inhibition activity with IC50 0.52 mM. Furthermore, gallic acid (IC50 1.8 mM) and paeoniflorin (IC50>2 mM) from high content components of RPA were determined having HDC inhibitory activity. In conclusion, the integrated strategy of UF-HPLC/MS combined with ECB and DE technique is an effective mode for rapid and accurate screening and identification of natural HDC inhibitors from TCMs.

A tyrosine, histidine-selective bifunctional cross-linker for protein structure analysis.

Chemical cross-linking mass spectrometry (XL-MS) significantly contributes to the analysis of protein structures and the elucidation of protein-protein interactions. Currently available cross-linkers mainly target N-terminus, lysine, glutamate, aspartate, and cysteine residues in protein. Herein, a bifunctional cross-linker, named [4,4'-(disulfanediylbis(ethane-2,1-diyl)) bis(1-methyl-1,2,4-triazolidine-3,5-dione)] (DBMT) has been designed and characterized aiming to extremely expand the application of XL-MS approach. DBMT is capable of selectively targeting tyrosine residue in protein via an electrochemical click reaction, and/or targeting histidine residue in protein in the presence of 1O2 generated under photocatalytic reaction. A novel cross-linking strategy based on this cross-linker has been developed and demonstrated using model proteins, which provides a complementary XL-MS tool analyzing protein structure, protein complexes, protein-protein interactions, and even protein dynamics.

Effects of amino acids on the formation and distribution of glycerol core aldehydes during deep frying.

Glyceryl core aldehyde (GCAs) are hazard factors produced during the frying process using oils and fats, and GCAs control and mitigation research is very important. This study investigated the effects of adding amino acids (methionine, glycine, and histidine) at 2.5, 5, and 10 mM on the formation and distribution of four GCAs during frying. High oleic sunflower oil (HOSO) was selected as frying oil for French fries. After 12 h of frying, the content of GCAs in the tert-butylhydroquinone-treated group (0.02 wt%, 1.1 mM) decreased by 29 % compared with the control group. The addition of methionine, glycine, and histidine decreased the total GCAs by 51 %, 28 %, and 27 %, respectively. The total GCAs content was best inhibited by methionine, while glycine and histidine were not significantly different from TBHQ. Methionine addition significantly reduced GCAs (9-oxo), GCAs (10-oxo-8), and GCAs (11-oxo-9) by 39 %, 78 %, and 80 %, respectively, while histidine was the most potent inhibitor of GCAs (8-oxo), which decreased by 40 %. Methionine also proved effective in slowing degradation of frying oil quality. These results provide a new direction for decreasing GCAs in frying systems.

Metabolomics reveals the defense mechanism of histidine supplementation on high-salt exposure-induced hepatic oxidative stress.

AIMS: This study mainly evaluated the protective mechanism of histidine against the hepatic oxidative stress after high-salt exposure (HSE) through combined analysis of non-targeted metabolomics and biological metabolic networks. MATERIALS AND METHODS: Dahl salt-sensitive (SS) rats were fed with normal-salt diet or HSE ± histidine in addition to drinking water for 14 days. Gas chromatography-mass spectrometry was used to analyze the hepatic metabolites. The metabolic profile was analyzed by SIMCA-14.1, the metabolic correlation network was performed using Gephi-0.9.2, and pathway enrichment was analyzed using MetaboAnalyst 5.0 online website. KEY FINDINGS: Results indicated that HSE disturbed the hepatic metabolic profile, generated abnormal liver metabolism and exacerbated oxidative stress. Histidine supplementation significantly reversed the hepatic metabolic profile. Of note, 14 differential metabolic pathways were enriched after histidine supplementation, most of which played an important role in ameliorating redox and nitric oxide (NO) metabolism. Histidine administration decreased the levels of hydroperoxide and malondialdehyde, and increased the activities of antioxidant enzymes (Catalase, Superoxide Dismutase, Glutathione S-transferase and Glutathione reductases). Histidine effectively enhanced the endogenous synthesis of glutathione by increasing the levels of glutamate and cysteine, thereby enhancing the antioxidant capacity of the glutathione system. After histidine administration, lysine, glutamate, and hypotaurine owned a higher metabolic centrality in the correlation network. In addition, histidine could also effectively increase the endogenous synthesis of NO by enhancing the L-arginine/NO pathway. SIGNIFICANCE: This study offers new insights into the metabolic mechanisms underlying the antioxidant protective effect of histidine on the liver.

Effect of supplementing rumen-protected methionine, lysine, and histidine to low-protein diets on the performance and nitrogen balance of dairy cows.

Lowering the dietary protein content can reduce N excretions and NH3 emissions from manure and increase milk N efficiency of dairy cows. However, milk yield (MY) and composition can be compromised due to AA deficiency. Methionine and Lys are known as first limiting EAA for dairy cows, and recently His is also mentioned as limiting, especially in grass-based or low-protein diets. To examine this, a trial was conducted with a 3-wk pre-experimental adaptation period (diet 16.5% crude protein), followed by a depletion period of 4 wk, in which 39 cows (average ± standard deviation: 116 ± 29.3 d in milk, 1.8 ± 1.2 lactations, 638 ± 73.2 kg of body weight, and 32.7 ± 5.75 kg MY/d) received a low-protein diet (CTRL) (14.5% crude protein). Then, taking into account parity, His plasma concentration, and MY, cows were randomly assigned to 1 of 3 treatment groups during the rumen-protected (RP) AA period of 7 wk; (1) CTRL; (2) CTRL + RP-Met + RP-Lys (MetLys); (3) CTRL + RP-Met + RP-Lys + RP-His (MetLysHis). Products were dosed, assuming requirements for digestible (d) Met, dLys, and dHis being, respectively, 2.4%, 7.0%, and 2.4% of intestinal digestible protein. In the cross-back period of 5 wk, all cows received the CTRL diet. During the last week of each period, a N balance was conducted by collecting total urine and spot samples of feces. Total feces production was calculated using the inert marker TiO2. Statistical analysis was performed with a linear mixed model with cow as random effect and data of the last week of the pre-experimental period used as covariate for the animal performance variables. No effect of supplementing RP-Met and RP-Lys nor RP-Met, RP-Lys, and RP-His on feed intake, milk performance, or milk N efficiency was observed. However, the plasma AA profile indicated additional supply of dMet, dLys, and dHis. Nevertheless, evaluation of the AA uptake relative to the cow's requirements showed that most EAA (exclusive Arg and Thr) were limiting over the whole experiment. Only dHis was sufficiently supplemented during the RP-AA period due to an overestimation of the diet's dMet and dLys supply in the beginning of the trial. The numerically increased milk urea N and urinary N excretion when RP-Met, RP-Lys, and RP-His were added to the low-protein diet suggest an increased catabolism of the excess His.

Proteomics analysis reveals that CirA in Aeromonas hydrophila is involved in nutrient uptake.

The colicin I receptor (CirA) is a well-studied outer membrane protein that has been reported to play important roles in antibiotic resistance, virulence, and iron homeostasis, although its exact physiological roles require further investigation. In this study, differentially expressed proteins between the ΔahcirA and wild-type (WT) strains of Aeromonas hydrophila were compared using quantitative proteomics. Bioinformatics analysis revealed that the expression of peptide, histidine, and arginine ATP-binding cassette (ABC) transporter system-related proteins was significantly higher in the ΔahcirA strain. Subsequent growth assays revealed that ΔahcirA grew slower than the WT strain in nutrient-limited medium when supplemented with dipeptide, histidine, and arginine as the carbon source. Far-western blot analysis further confirmed that AhCirA can directly bind to histidine/arginine and dipeptide small-molecule substrates in addition to their periplasmic-binding proteins, AhDppA and AhHisJ, respectively. These results indicate that AhCirA may play an important role in the uptake of amino acids and peptides as a channel-forming porin while also directly interacting with ABC transporters to transport nutrient substances into the plasma membrane. Overall, this study demonstrates that AhCirA is a multifunctional protein in A. hydrophila and extends our understanding of known nutrient transport mechanisms among bacteria.

The effects of standardized ileal digestible His to Lys ratio on growth performance, intestinal health, and mobilization of histidine-containing proteins in pigs at 7 to 11 kg body weight.

The objectives were to evaluate the effects of standardized ileal digestible (SID) His:Lys ratio above the current NRC requirement on growth performance, intestinal health, and mobilization of His-containing proteins, including hemoglobin, carnosine, and trypsinogen, in nursery pigs from 7 to 11 kg body weight (BW). Forty pigs (26 d of age; initial BW of 7.1 ±â€…0.5 kg) were allotted to 5 dietary treatments based on a randomized complete block design with sex and initial BW as blocks. Dietary treatments were supplemented with varying SID His to Lys ratios of 26%, 32%, 38%, 43%, and 49% and fed to pigs for 14 d (SID Lys = 1.22%). Feed intake and BW were recorded at d 0, 7, and 14 to measure growth performance. Blood samples were collected on d 12. Pigs were euthanized on d 14 to collect pancreas, longissimus dorsi muscles, mid-jejunum, and jejunal mucosa. Data were analyzed using the Proc Mixed of SAS. Growth performance was not affected, whereas varying SID His to Lys ratio affected hemoglobin (P < 0.05, max: 12 g/dL at 36%), immunoglobulin A (IgA, P < 0.05, min: 1.25 µg/mg at 35%) in jejunal mucosa, villus height (P = 0.065, max: 536 µm at 40%) in jejunum, trypsinogen (P = 0.083, max: 242 pg/mg at 41%) in pancreas, and carnosine (P = 0.051, max: 4.7 ng/mg at 38%) in muscles. Varying SID His to Lys ratios linearly increased (P < 0.05, from 1.95 to 2.80 nmol/mg) protein carbonyl in muscles and decreased (P < 0.05, from 29.1% to 26.9%) enterocyte proliferation. In conclusion, SID His to Lys ratio between 35% and 41% in diets fed to nursery pigs at 7 to 11 kg enhanced intestinal health and maximized concentrations of His-containing proteins, indicating that His-containing proteins are effective response criteria when determining His requirement.

Histidine is an essential amino acid for protein synthesis, but it also plays a vital role in the metabolic system of pigs. An accurate assessment of His requirement provides pivotal information for efficient growth and health of pigs. Growth performance and plasma His concentration have been used to assess His requirement, but they may not be the effective parameters due to the contribution of His from mobilization of His-containing proteins, such as hemoglobin, carnosine, and pancreatic enzymes. Hemoglobin is a transport protein and the main component in red blood cells, enabling oxygen transport throughout the body. Most carnosine is stored in muscles at 3 to 4 g/kg wet weight and has antioxidative effects to prevent cells from oxidative damages. In addition, His has a critical role in serine peptidases as a part of the catalytic triad. In this study, growth performance did not respond to His deficiency due to the compensation of His from His-containing proteins and potentially due to a short experimental period. Standardized ileal digestible His to Lys ratio between 35% and 41% maximized concentrations of His-containing proteins and enhanced intestinal health in pigs at 7 to 11 kg body weight. This study indicated that hemoglobin, carnosine, and trypsinogen are effective response criteria when determining His requirement.