Exploring antithrombotic mechanisms and effective constituents of Lagopsis supina using an integrated strategy based on network pharmacology, molecular docking, metabolomics, and experimental verification in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Thrombosis is a common cause of morbidity and mortality worldwide. Lagopsis supina (Stephan ex Willd.) Ikonn.-Gal. ex Knorring is an ancient Chinese herbal medicine used for treating thrombotic diseases. Nevertheless, the antithrombotic mechanisms and effective constituents of this plant have not been clarified. AIM OF THE STUDY: This work aimed to elucidate the pharmacodynamics and mechanism of L. supina against thrombosis. MATERIALS AND METHODS: Systematic network pharmacology was used to explore candidate effective constituents and hub targets of L. supina against thrombosis. Subsequently, the binding affinities of major constituents with core targets were verified by molecular docking analysis. Afterward, the therapeutic effect and mechanism were evaluated in an arteriovenous bypass thrombosis rat model. In addition, the serum metabolomics analysis was conducted using ultra-high performance liquid chromatography coupled with Q-Exactive mass spectrometry. RESULTS: A total of 124 intersected targets of L. supina against thrombosis were predicted. Among them, 24 hub targets were obtained and their mainly associated with inflammation, angiogenesis, and thrombosis approaches. Furthermore, 9 candidate effective constituents, including (22E,24R)-5α,8α-epidioxyergosta-6,22-dien-3ß-ol, aurantiamide, (22E,24R)-5α,8α-epidioxyergosta-6,9 (11),22-trien-3ß-ol, lagopsinA, lagopsin C, 15-epi-lagopsin C, lagopsin D, 15-epi-lagopsin D, and lagopsin G in L. supina and 6 potential core targets (TLR-4, TNF-α, HIF-1α, VEGF-A, VEGFR-2, and CLEC1B) were acquired. Then, these 9 constituents demonstrated strong binding affinities with the 6 targets, with their lowest binding energies were all less than -5.0 kcal/mol. The antithrombotic effect and potential mechanisms of L. supina were verified, showing a positively associated with the inhibition of inflammation (TNF-α, IL-1ß, IL-6, IL-8, and IL-10) and coagulation cascade (TT, APTT, PT, FIB, AT-III), promotion of angiogenesis (VEGF), suppression of platelet activation (TXB2, 6-keto-PGF1α, and TXB2/6-keto-PGF1α), and prevention of fibrinolysis (t-PA, u-PA, PAI-1, PAI-1/t-PA, PAI-1/u-PA, and PLG). Finally, 14 endogenous differential metabolites from serum samples of rats were intervened by L. supina based on untargeted metabolomics analysis, which were closely related to amino acid metabolism, inflammatory and angiogenic pathways. CONCLUSION: Our integrated strategy based on network pharmacology, molecular docking, metabolomics, and in vivo experiments revealed for the first time that L. supina exerts a significant antithrombotic effect through the inhibition of inflammation and coagulation cascade, promotion of angiogenesis, and suppression of platelet activation. This paper provides novel insight into the potential of L. supina as a candidate agent to treat thrombosis.

Effect of temperature regulation on microbial community, volatile flavours, amino acid profiles, and iridoid glycosides during noni (Morinda citrifolia L.) fruit fermentation.

Noni fruit has an unpleasant flavour but is highly bioactive. Therefore, it is necessary to clarify the effect of temperature regulation on quality of fermented noni fruit. In the present study, the formation of flavours, amino acid profiles, and iridoid glycosides during noni fruit fermentation at different temperatures were investigated. We initially found that different temperatures affected core microbial communities. The general evolutionary trends of Acetobacter and Gluconobacter were influenced by different temperatures. Furthermore, high temperature helped maintain low octanoic and hexanoic acids. Subsequently, we found that high temperature improved total amino acids and iridoid glycosides. The correlation network analysis revealed that bacterial communities impacted the quality (volatile flavours, amino acid profiles, and iridoid glycosides) of fermented noni fruit. Overall, altering the temperature induced variations in microbial communities and quality during the noni fruit fermentation process. These results are instrumental in the pursuit of quality control in natural fermentation processes.

Facile synthesis of magnetic ionic covalent organic framework and dispersive magnetic solid phase extraction of aromatic amino acid oxidation products in thermally processed foods.

Aromatic amino acid oxidation products (AAAOPs) are newly discovered risk substances of thermal processes. Due to its significant polarity and trace level in food matrices, there are no efficient pre-treatment methods available to enrich AAAOPs. Herein, we proposed a magnetic cationic covalent organic framework (Fe3O4@EB-iCOF) as an adsorbent for dispersive magnetic solid-phase extraction (DMSPE). Benefiting from the unique charged characteristics of Fe3O4@EB-iCOF, AAAOPs can be enriched through electrostatic interaction and π-π interactions. Under the optimal DMSPE conditions, the combined HPLC-MS/MS method demonstrated good linearity (R2 ≥ 0.990) and a low detection limit (0.11-7.5 µg·kg-1) for AAAOPs. In addition, the method was applied to real sample and obtained satisfactory recoveries (86.8 % âˆ¼ 109.9 %). Especially, we applied this method to the detection of AAAOPs in meat samples and conducted a preliminarily study on its formation rules, which provides a reliable basis for assessing potential dietary risks.

A tunable LDI-MS platform assisted by metal-phenolic network-coated AuNPs for sensitive and customized detection of amino acids.

This study introduces a novel approach for the sensitive and accurate detection of small molecule metabolites, employing metal-phenolic network (MPN) functionalized AuNPs as both adsorbent and matrix to enhance laser desorption/ionization mass spectrometry (LDI-MS) performance. The MPN comprising tannic acid (TA) and transition metal ions (Fe3+, Co2+, Ni2+, Cu2+, or Zn2+) was coated on the surface of AuNPs, forming metal-TA network-coated AuNPs (M-TA@AuNPs). The M-TA@AuNPs provided a tunable surface for specific interactions with analytes, displaying distinct enrichment efficacies for different amino acids, especially for Cu-TA@AuNPs exhibiting the highest affinity for histidine (His). Under the optimized condition, the proposed method enabled ultrasensitive detection of His, with good linearity (R2 = 0.9627) in the low-concentration range (50 nM-1 µM) and a limit of detection (LOD) as low as 0.9 nM. Furthermore, the method was successfully applied to detect His from human urine samples, showcasing its practical applications in clinical diagnostics, particularly in the realm of amino acid-based targeted metabolomics.

Serum targeted metabolomics uncovering specific amino acid signature for diagnosis of intrahepatic cholangiocarcinoma.

Intrahepatic cholangiocarcinoma (iCCA) is a hepatobiliary malignancy which accounts for approximately 5-10 % of primary liver cancers and has a high mortality rate. The diagnosis of iCCA remains significant challenges owing to the lack of specific and sensitive diagnostic tests available. Hence, improved methods are needed to detect iCCA with high accuracy. In this study, we evaluated the efficacy of serum amino acid profiling combined with machine learning modeling for the diagnosis of iCCA. A comprehensive analysis of 28 circulating amino acids was conducted in a total of 140 blood samples from patients with iCCA and normal individuals. We screened out 6 differentially expressed amino acids with the criteria of |Log2(Fold Change, FC)| > 0.585, P-value < 0.05, variable importance in projection (VIP) > 1.0 and area under the curve (AUC) > 0.8, in which amino acids L-Asparagine and Kynurenine showed an increasing tendency as the disease progressed. Five frequently used machine learning algorithms (Logistic Regression, Random Forest, Supporting Vector Machine, Neural Network and Naïve Bayes) for diagnosis of iCCA based on the 6 circulating amino acids were established and validated with high sensitivity and good overall accuracy. The resulting models were further improved by introducing a clinical indicator, gamma-glutamyl transferase (GGT). This study introduces a new approach for identifying potential serum biomarkers for the diagnosis of iCCA with high accuracy.

Stereo-hindrance effect and oxidation cross-linking induced by ultrasound-assisted sodium alginate-glycation inhibit lysinoalanine formation in silkworm pupa protein.

Silkworm pupa protein isolate (SPPI) is rich in amino acids, making it chemically reactive, degradable, and easy to form lysinoalanine (LAL). We investigated how conformational cross-linking, induced by ultrasound-assisted sodium alginate, could inhibit the formation of LAL during the preparation of SPPI. Glycoconjugated SPPI (using 1 % sodium alginate under ultrasonication) showed the lowest LAL content i.e., 7.403 µg·mg-1, representing a 49.58 % decrease, with reference to the control. The ionic, hydrogen, and covalent bonds in the glycoconjugate increased by 171.79 %, 8.48 %, and 35.56 %, respectively. Glycation decreased arginine by 28.92 % and caused the oxidation of tyrosine, methionine and proline to form carbonyl groups. Some precursor amino acids, including lysine, serine, cysteine and threonine were not degraded during the combined treatment. The macromolecular aggregation caused by structural modifications strengthened the steric resistance of LAL cross-linking. The study outcomes provide a novel approach and theoretical basis for inhibition of LAL formation in SPPI.

Evaluation of the nutritional quality of yeast protein in comparison to animal and plant proteins using growing rats and INFOGEST model.

Yeast, identified as a microorganism, boasts a considerable protein content, positioning yeast protein as a highly promising alternative in the quest for sustainable protein sources. The primary aim of this study is to evaluate the protein quality of yeast protein and compare it with animal proteins (whey concentrate/isolate proteins) and plant proteins (soy, wheat, pea proteins). Notably, yeast protein exhibits the highest ratio of indispensable/dispensable amino acids (IAAs/DAAs, 0.91). However, in both in vivo and in vitro digestion experiments, yeast protein demonstrated lower true protein digestibility (TPD) and true ileal digestibility (TID) compared to other proteins. Despite this, the yeast protein's amino acid score (AAS, 1.37 for >3 years), protein digestibility-corrected amino acid score (PDCAAS, 100 % for >3 years), and digestibility-corrected amino acid score (DIAAS, 82.42 % for >3 years) of yeast protein surpassed those of plant proteins, yet remained lower than animal proteins primarily due to its lower digestibility.

Unraveling the abundance of vip3-type genes in Indian Bacillus thuringiensis across the agroclimatic landscape and impact of amino acid substitutions for safer agriculture.

Vegetative insecticidal protein (vip) genes of Bacillus thuringiensis (Bt) are candidates for gene pyramiding in the resistance management of pests. The prevalence of vip genes in Bt isolates is relatively under-explored. Bt isolates recovered from 29 diverse sources in nine agro-climatic zones of India were screened for the presence of vip3-type genes by PCR with 4 sets of oligonucleotide primers. Out of 155 Bt isolates, 70.32 % (109) and 44.52 % (69) isolates were positive for amplification of partial vip3-type genes with primer sets 1 and 4, respectively. The primer set-2 was found to be more efficient for amplifying full-length genes (29.03 % /45 isolates) as compared with primer set-3 (3.23 %/ 5 isolates), also corroborated in the amplification of full-length vip3 genes in ten Bt BGSC strains used as reference. Frequency analysis revealed presence of vip3 genes in Bt isolates across all agro-climatic zones. Thus, Indian Bt isolates from diverse sources have a rich repertoire of vip3-type genes. Our study reports the highest number (45) of full-length vip3-type genes detected in a native Bt isolates collection, demonstrating enrichment of Indian Bt isolates for vip3 genes. Twelve of these genes have been cloned, sequenced, and out of these, six were found to be effective against Helicoverpa armigera in our laboratory previously. Comparison of substitutions in deduced amino acids sequence of these genes and expression of Vip3 proteins in SDS-PAGE analysis of selected native Bt isolates positive for full-length vip3-type genes indicated their biopesticidal potential.

Dry-ageing impacts on meat quality, oxidative stability, and release of free amino acids in striploins from dairy crossbred yearling and 2-year-old steers.

This study determined the impacts of dry-ageing on meat quality, oxidative stability, and release of free amino acids (FAAs) in striploins from dairy-crossbred yearlings and 2-year-old steers (n = 12 each group) over 21 days of in-bag dry-ageing. Dry-ageing increased weight losses, with higher % drying rates in yearling meat during dry-ageing, likely due to the smaller loin size and lower intramuscular fat content (P < 0.05). Yearling meat showed greater (P < 0.05) decreases in moisture content, but both meats reached similar moisture levels by day 21. pH values increased with dry-ageing with variations at different ageing times. Dry-ageing reduced a*, b*, and chroma while increasing L* and hue angles on day 21 (P < 0.05), likely due to dehydration and lipid oxidation (higher TBARS, P < 0.05) after 14 days, especially in yearling meat. The decreased levels (P < 0.05) of some monounsaturated fatty acids and conjugated linoleic acid cis-9, trans-11 were likely linked with lipid oxidation. Total levels of FAAs and essential amino acids increased significantly, especially within the first 7 days, with distinct patterns between the two meats. Dry-aged yearling meat contained more FAAs associated with sweat taste (e.g., glutamine and glycine) and fewer FAAs associated with bitter taste (e.g., phenylalanine and tyrosine). Carnosine levels varied and significantly increased after 21 days. Dry-ageing demonstrated distinct effects on dehydration, lipid oxidation, and release of FAAs in meat from yearlings compared with 2-year-old steers, which can be tailored to develop high-quality beef products with unique flavours.

Amino Acid Composition, Antioxidant and Antihypertensive Activity in Extracts from Mexican Añejo Cheese.

Research background: Authentic Mexican cheeses have potential health benefits, although there are few studies on their bioactive components. In this study, we analysed soluble extracts from añejo cheese from Zacazonapan (Mexico), obtained from two dairies (A and B) that used milk from cows of different breeds and differed in processing. Experimental approach: The soluble extracts of Zacazonapan añejo cheese during ripening (0, 30, 95 and 180 days) were used to determine proximate composition, amino acid composition, peptide profile, molecular mass profile, antioxidant and antihypertensive activities. Results and conclusions: The molecular mass of the released protein fragments ranged from 0.10 to 22.43 kDa. During ripening, amino acids such as Pro, His, Tyr, Trp, Met, Cys, Ala, Gly, Leu and Val were present, which are associated with antioxidant activity. The inhibition of ß-carotene bleaching was below 50 % at all ripening times. A significant difference between the cheese samples was observed only after 180 days. Amino acids associated with angiotensin-I converting enzyme (ACE-I) inhibition were found in the extracts (Trp, Phe, Tyr, Pro, Lys and Arg). The highest activity was observed in cheese ripened for 180 days (IC50 of cheese A and B was 0.38 and 0.42 mg/mL, respectively). Novelty and scientific contribution: These results suggest that Zacazonapan añejo cheese is a potential source of antioxidant and antihypertensive peptides, which are influenced by the dairy factor (milk source and production technique) and ripening time. This is relevant as there are no reports of these bioactivities in this type of cheese.

N-Acetylcysteine Alleviates Depressive-Like Behaviors in Adolescent EAAC1-/- Mice and Early Life Stress Model Rats.

Exposure to adverse experiences during early life is associated with an increased risk of psychopathology during adolescence. In a previous study, we demonstrated that neonatal maternal separation (NMS) combined with social isolation led to impulsive and depressive-like behaviors in male adolescents. Additionally, it significantly reduced the expression of excitatory amino acid carrier 1 (EAAC1) in the hippocampus. Building upon this work, we investigated the effects of N-acetylcysteine (NAC), a precursor to glutathione, in early-life stress (ELS) model rats and in EAAC1-/- mice. EAAC1 plays a dual role in transporting both glutamate and cysteine into neurons. Our findings revealed that female adolescents subjected to in the ELS model also exhibited behavioral defects similar to those of males. NAC injection rescued depressive-like behaviors in both male and female NMS models, but it improved impulsive behavior only in males. Furthermore, we observed increased reactive oxidative stress (ROS) and neuroinflammation in the ventral hippocampus (vHPC) and prefrontal cortex of NMS model rats, which were mitigated by NAC treatment. Notably, NAC reversed the reduced expression of EAAC1 in the vHPC of NMS model rats. In EAAC1-/- mice, severe impulsive and depressive-like behaviors were evident, and the NAC intervention improved only depressive-like behaviors. Collectively, our results suggest that ELS contributes to depression and impulsive behaviors during adolescence. Moreover, the cysteine uptake function of EAAC1 in neurons may be specifically related to depression rather than impulsive behavior.

Vampire bats rapidly fuel running with essential or non-essential amino acids from a blood meal.

In most mammals, running is fuelled by oxidization of endogenous carbohydrates and lipids while amino acids contribute little (< 5-10%). Common vampire bats (Desmodus rotundus), however, specialize on a unique, protein-rich blood diet. Therefore, we hypothesized that (i) vampire bats would rapidly begin utilizing dietary amino acids to support running metabolism, and (ii) that relative reliance on essential and non-essential amino acids would be similar. We fed bats cow's blood enriched either with isotopically labelled glycine (non-essential amino acid) or leucine (essential amino acid). Bats were exercised at speeds of 10, 20 and 30 m min-1 on a respirometry treadmill, allowing us to assess metabolic rate (i.e. O2 consumption and CO2 production) and track the oxidation of labelled amino acids in exhaled CO2. Vampire bats oxidized amino acids as their primary fuel as indicated by a respiratory exchange ratio (RER = ratio of CO2 production to O2 consumption rates) of approximately 0.8-0.9 at all speeds, with the labelled meal accounting for as much as 60% of oxidized fuels at peak usage. Similar oxidation rates indicated bats did not discriminate between essential and non-essential amino acid use. These findings reiterate how strongly metabolism can be shaped by a specialized diet.

Effect of chickpea on the physicochemical, nutritional, antioxidant, and organoleptic characterization of corn extrudates.

BACKGROUND: Ready-to-eat snacks are very popular. However, they have a high glycemic index and lack proteins & micronutrients. This study prepared protein-enriched corn extrudates by adding chickpea grit supplements at varying concentrations (0-100 g kg-1). RESULTS: The protein contents of 100 g kg-1 supplemented extrudates increased by 66.66% and dietary fiber contents increased by 48.02% in comparison with the control. Bulk density increased by 1.46 times. However, the expansion ratio, porosity, and water absorption index decreased significantly (P < 0.05). The health-promoting characteristics of the extrudates increased in comparison with the control sample, i.e., total phenolic content increasing by 17.84%, 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) contents by 11.38%, and 2,2-azinobis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) content by 9.59%. Likewise, the potassium contents increased by 24.63% with the inclusion of 10% chickpea in corn extrudates. Sensory evaluation revealed that corn extrudates with up to 60 g kg-1 added chickpea achieved the highest acceptability among panelists. CONCLUSION: The addition of chickpea produced corn extrudates with higher protein and mineral content, which could mitigate malnutrition. © 2024 Society of Chemical Industry.

Enhanced photorespiratory and TCA pathways by elevated CO2 to manage ammonium nutrition in tomato leaves.

Plants grown under exclusive ammonium (NH4+) nutrition have high carbon (C) demand to sustain proper nitrogen (N) assimilation and energy required for plant growth, generally impaired when compared to nitrate (NO3-) nutrition. Thereby, the increment of the atmospheric carbon dioxide (CO2) concentration, in the context of climate change, will potentially allow plants to better face ammonium nutrition. In this work, tomato (Solanum lycopersicum L.) plants were grown under ammonium or nitrate nutrition in conditions of ambient (aCO2, 400 ppm) or elevated CO2 (eCO2, 800 ppm) atmosphere. Elevated CO2 increased photosynthesis rate and tomato shoot growth regardless of the N source. In the case of NH4+-fed leaves the positive effect of elevated CO2 occurred despite of the high tissue NH4+ accumulation. Under eCO2 ammonium nutrition triggered, among others, the modulation of genes related to C provision pathways (including carbonic anhydrase and glyoxylate cycle), antioxidant response and cell membranes protection. The enhanced photosynthate production at eCO2 facilitated C skeleton provision through the TCA cycle and anaplerotic pathways to promote amino acid synthesis. Moreover, photorespiratory activity was stimulated by eCO2 and contributed to yield serine as additional sink for NH4+ excess. Overall, these changes denote a connection between the respiratory and the photorespiratory pathways linked to ammonium nutrition. This metabolic strategy may allow crops to grow efficiently using ammonium as fertilizer in a future climate change scenario, while mitigating N losses.

Altered gut metabolites and metabolic reprogramming involved in the pathogenesis of colitis-associated colorectal cancer and the transition of colon "inflammation to cancer".

Colitis-associated colorectal cancer (CAC) is fatal and can develop spontaneously or as a complication of inflammatory bowel diseases. Although co-administration of azoxymethane/dextran sulfate sodium (AOM/DSS) is a classic method for CAC modeling, its limitations need to be addressed. Accordingly, we aimed to optimize the AOM/DSS model to study CAC extensively and further investigate its pathogenic mechanisms relative to microbiota and metabolism. We optimized the CAC model via a single or enhanced injection of AOM combined with different administration modes and varying DSS concentrations. Subsequently, the fecal-microbiota composition was examined using 16S RNA sequencing, and fecal-colon-metabolome profiles were evaluated via ultra-high performance liquid chromatography-mass spectrometry. Two interval injections of AOM combined with 1.5 % DSS-free drinking resulted in a high tumor formation rate, uniform tumor formation, and low mortality. Based on this model, we innovatively divided the pathogenesis of CAC into three stages, namely inflammation induction, proliferation initiation, and tumorigenesis, and examined the pathological characteristics in each stage. Gut microbial dysbiosis and metabolic alteration drove colorectal tumorigenesis by aggravating inflammation while promoting cell proliferation and carcinogenesis in mice. For the first time, we dynamically demonstrated the process of colon "inflammation to cancer" transformation and provided novel insights to clarify the role of amino acid metabolism in the formation of CAC.

Over-expression of SiADCL1 in Arabidopsis modulates folate and amino acid metabolism to impact on flowering time.

Foxtail millet is a C4 crop rich in folate (FA). This study explores the roles of the 4-amino-4-deoxychorismate lyase (ADCL) - a member of the transaminase IV group of enzymes - in FA metabolism and conferred phenotypes. Phylogenetic comparisons identified diversity in the transaminase IV/ADCL gene family in the foxtail millet genome which was associated with genomic duplications. Molecular docking studies suggested that SiADCL1 bound most strongly to aminodeoxychorismate (ADC) and most likely had the highest catalytic activities. SiADCL1 which was highly expressed in roots, peduncles and flag leaves. Over-expression of SiADCL1 in Arabidopsis significantly increased total FA content (1.14-1.84 fold) and this was linked to a delayed flowering time. Metabolomic and transcriptomic characterization of the derived over-expression lines, found that FA promotes the change of methylation-related genes, ethylene synthesis, amino acid metabolism and flowering-related genes. This study revealed a potential gene coexpression network linked with FA and targeted key genes that could be exploited in foxtail millet breeding programs.

Analysis of protein-protein interaction of the mitochondrial translocase at work by using technically effective BPA photo-crosslinking method.

The BPA photo-crosslinking method exploits the property of p-benzoyl-L-phenylalanine (pBpa), an amino acid containing a photoreactive side chain, and allows for the crosslinking with nearby proteins upon Ultraviolet irradiation. This feature enables the capture of two proteins within a close proximity with high spatial resolution at the level of amino acid residues. In this chapter, we introduce an example of the employment of the BPA photo-crosslinking method to the Translocase of the Outer Mitochondrial membrane complex of mitochondria in Saccharomyces cerevisiae as a model protein translocase. Here in, we provide three procedures (i) the introduction of pBpa into proteins of interest in living yeast cells by in vivo suppressor tRNA system; (ii) analysis of in vivo subunit-subunit interactions intra-complex; and (iii) analysis of translocase channel-substrate interactions in organello. The use of in vivo and in organello crosslinking tools enable the robust analysis of translocases in a near-to physiological condition.

Waste Nitrogen Upcycling to Amino Acids during Anaerobic Fermentation on Biochar: An Active Strategy for Regulating Metabolic Reducing Power.

This study proposes a novel strategy that utilizes biochar (BC) during anaerobic fermentation (AF) to generate amino acids (AAs) toward nitrogen upcycling. The BC, pyrolyzed at 800 °C (BC800) to enhance graphite structures and electron-accepting sites, effectively addresses issues related to biosynthetic reducing power nicotinamide adenine dinucleotide phosphate insufficiency by altering cellular conditions and alleviates feedback inhibition through the immobilization of end products. This process establishes unique microbial signaling and energy networks, with Escherichia coli becoming dominant in the biofilm. The conversion rate of ammonia-N to AAs-N within the biofilm reached 67.4% in BC800-AF, which was significantly higher compared to the levels in other AF reactors with BC pyrolyzed at 600 and 400 °C (45.9 and 22.5%, respectively), as well as a control AF reactor (<5%). Furthermore, in BC800-AF, the aromatic AAs (Aro-AAs) were as high as 70.8% of the AAs within the biofilm. The activities of key enzymes for Aro-AAs biosynthesis uniquely positively correlated with the electron-accepting capacity on BC800 (R2 ≥ 0.95). These findings hold promise for transforming existing AF reactors into factories that produce BC-based AAs, providing a more sustainable fertilizing agent than chemical fertilizers.

HSP70 promotes amino acid-dependent mTORC1 signaling by mediating CHIP-induced NPRL2 ubiquitination and degradation.

Mechanistic target of rapamycin complex 1 (mTORC1) is a master regulator of cell growth and its dysregulation leads to a variety of human diseases. Although NPRL2, an essential component of the GATOR1 complex, is reported to effectively suppress amino acid-induced mTORC1 activation, the regulation of NPRL2 protein stability is unclear. In this study, we show that chaperon-associated ubiquitin ligase CHIP interacts with NPRL2 and promotes its polyubiquitination and proteasomal degradation. Moreover, HSP70 mediates CHIP-induced ubiquitination and degradation of NPRL2. Consistently, overexpression of HSP70 enhances whereas HSP70 depletion inhibits amino acid-induced mTORC1 activation. Accordingly, knockdown of HSP70 promotes basal autophagic flux, and inhibits cell growth and proliferation. Taken together, these results demonstrated that HSP70 is a novel activator of mTORC1 through mediating CHIP-induced ubiquitination and degradation of NPRL2.

Glycation of sunnhemp protein with dextran via dry heating: Thermal, micro-structural characterization, and amino acid profiling.

This study aims to obtain sunnhemp protein isolate (SHPI) and dextran conjugates by dry heating method of Maillard conjugation. The effects of different incubation time (0, 1, 3, 5, 7, and 9 days) on the molecular flexibility, available lysine content, antioxidant properties, molecular structure, and thermal and micro-structural properties of conjugates were compared with SHPI (no conjugation) at 60°C and 79% relative humidity. The results indicated the formation of SHPI-dextran conjugates as confirmed by the change in molecular flexibility, lysine content, antioxidant activities, color, and water activity values. The molecular structure revealed the confirmation of covalent bonding between SHPI and dextran. Differential scanning calorimetry and thermo-gravimetric analysis results exhibited improvement in the thermal stability of SHPI when conjugated with dextran. The microstructural characterization showed that Maillard conjugation changed the surface structure of SHPI. The analysis of amino acid composition displayed that lysine, arginine, and phenylalanine were the dominant Maillard reaction sites of SHPI and dextran. Among all the conjugated samples, 5 days of incubation time was selected as an optimum condition for the development of SHPI-dextran conjugates on the basis of the aforementioned characterization. Overall, it was concluded that Maillard conjugation of sunnhemp protein with dextran via dry-heating technique could modify and improve its various attributes. PRACTICAL APPLICATION: The conjugation of plant proteins with polysaccharide through the Maillard reaction under dry heating conditions represents a natural and green technique for improving the techno-functional properties of proteins. The study has the potential to establish framework for the utilization of Sunnhemp protein isolate-dextran conjugates. This approach offers the potential for cost-effective production of emulsifiers and development of effective encapsulating matrices. The investigation expands on an underutilized plant protein source facilitating an alternative to animal-based proteins and contributing to the development of a sustainable circular bioeconomy.