Morphological, physiological, and biochemical responses of three different soybean (Glycine max L.) varieties under salinity stress conditions.

Salinity is one of the most detrimental factors for the growth performance and productivity of crops worldwide. Therefore, understanding crop responses or growth potentials and their effectiveness in salinity mitigation is highly important for the selection of salinity-tolerant plant varieties. In this study, the effects of salinity at various stress levels (0 mM, 50 mM, 100 mM, and 150 mM NaCl) on the morphological, physiological, and biochemical parameters of three soybean varieties ('Afigat', 'Gishama', and 'Pawi-2') were investigated. The results showed that salinity significantly reduced morphological traits including plant height, number of leaves per plant, stem thickness, shoot and root length, and fresh and dry weight. This reduction was more prominent in the 'Afigat' variety for all of these traits except shoot and root length. The concentrations of chlorophyll a and b decreased with increasing salinity. In addition, salinity significantly increased leaf electrolyte leakage (EL), lipid peroxidation, proline accumulation, and phenol and flavonoid content. The 'Pawi-2' variety was more tolerant than the other studied varieties in terms of membrane stability (less EL and a low malondialdehyde content) and proline, phenol, and flavonoid accumulation. Therefore, 'Pawi-2' may be considered as the most salt-tolerant variety in comparison with the other studied soybean varieties. Further complementary studies in field conditions including anatomical parameters are needed to confirm these findings.

Detection and validation of QTLs for green stem disorder of soybean (Glycine max (L.) Merr.).

In mechanically harvested soybean, green stem disorder (GSD) is an undesirable trait that causes green-stained seeds, which are graded lower in Japan. To obtain DNA markers for reduced GSD, we conducted a quantitative trait locus (QTL) analysis for 2 years using F4 and F5 lines from a cross between 'Suzuotome' (less GSD) and 'Fukuyutaka' (more GSD). We validated the effect of a detected QTL for GSD by first identifying F4 or F5 plants in which one or more markers in the QTL region were heterozygous. The F5 or F6 progeny of each plant was used to form a pair consisting of two groups in which the QTL region was homozygous for either the 'Suzuotome' or 'Fukuyutaka' allele in a similar genetic background, and the two groups within each pair were compared for GSD. Over 3 years of testing, the 'Suzuotome' allele of a QTL on chromosome 6 was found to reduce the level of GSD. This novel QTL was mapped to the region around DNA marker W06_0130, and was not closely linked to QTLs for important agronomic traits including yield components. Using this marker, the low level of GSD from 'Suzuotome' could be conferred to 'Fukuyutaka' or other high-GSD cultivars.

Electronic structure, global reactivity descriptors and nonlinear optical properties of glycine interacted with ZnO, MgO and CaO for bacterial detection.

Modern laboratory medicine relies on analytical instruments for bacterial detection, focusing on biosensors and optical sensors for early disease diagnosis and treatment. Thus, Density Functional Theory (DFT) was utilized to study the reactivity of glycine interacted with metal oxides (ZnO, MgO, and CaO) for bacterial detection. Total dipole moment (TDM), frontier molecular orbitals (FMOs), FTIR spectroscopic data, electronic transition states, chemical reactivity descriptors, nonlinear optical (NLO) characteristics, and molecular electrostatic potential (MESP) were all investigated at the B3LYP/6-31G(d, p) level using DFT and Time-Dependent DFT (TD-DFT). The Coulomb-attenuating approach (CAM-B3LYP) was utilized to obtain theoretical electronic absorption spectra with the 6-31G(d, p) basis set to be more accurate than alternative quantum chemical calculation approaches, showing good agreement with the experimental data. The TDM and FMO investigation showed that glycine/CaO model has the highest TDM (10.129Debye) and lowest band gap (1.643 eV). The DFT computed IR and the experimental FTIR are consistent. The calculated UV-vis spectra showed a red shift with an increase in polarity following an increase in the absorption wavelength due to the interaction with ZnO, MgO, and CaO. Among the five solvents of water, methanol, ethanol, DMSO and acetone, the water and DMSO enhances the UV-Vis absorption. Glycine/CaO model showed high linear polarizability (14.629 × 10-24esu) and first hyperpolarizability (23.117 × 10-30esu), indicating its potential for nonlinear optical applications. The results showed that all model molecules, particularly glycine/CaO, contribute significantly to the development of materials with potential NLO features for sensor and optoelectronic applications. Additionally, MESP confirmed the increased electronegativity of the studied structures. Additionally, glycine/ZnO nanocomposite was synthesized and characterized using IR and UV-visible spectroscopy to determine their structural and spectroscopic features. It was discovered that there was good agreement between the DFT computed findings and the related experimental data. The antibacterial activity of glycine/ZnO nanocomposites against Staphylococcus aureus (S. aureus) and Pseudomonas aeruginosa were studied in terms of concentration and time. The results showed that increasing the concentration of glycine/ZnO nanocomposite significantly enhanced its antibacterial efficacy by lowering optical density. Notably, Pseudomonas aeruginosa exhibited lower susceptibility to the nanocomposite compared to S. aureus, requiring higher concentrations for effective bactericidal action. In summary, this study contributes novel insights into the dual functionality of glycine-metal oxide complexes, with significant implications as optical biosensor for microbial detection.

Increasing concentrations of dietary threonine, tryptophan, and glycine improve growth performance and intestinal health with decreasing stress responses in broiler chickens raised under multiple stress conditions.

The current study aimed to compare the effects of increasing concentrations of dietary threonine (Thr), tryptophan (Trp), and glycine (Gly) on growth performance, stress biomarkers, and intestinal function in broiler chickens under multiple stress conditions. Five hundred sixty broiler chickens at 21 d old were randomly allotted to 5 treatments with 8 replicates. Birds in a positive control (PC) treatment were raised under low stock density (16.9 birds/m2 per cage) with recommended environmental conditions, whereas birds in 4 treatments were subjected to multiple stress conditions: a cyclic heat stress of 30 ± 0.3 °C for 10 h and 23 ± 0.2 °C for 14 h per day with high stock density (25.3 birds/m2 per cage). A basal diet was assigned to both PC and negative control (NC) treatments. Three additional diets were individually formulated to contain double concentrations of digestible Thr, Trp, or Gly + Ser compared with their concentrations in the basal diet. The experiment lasted for 14 d. Results showed that NC treatment had less growth performance (P < 0.001), jejunal goblet cell counts (P = 0.018), and trans-epithelial electrical resistance (TEER; P < 0.001), but greater (P = 0.026) feather corticosterone (CORT) concentrations than PC treatment. Thr treatment showed the least (P < 0.001) feed conversion ratio (FCR) among treatments under multiple stress conditions. Thr, Trp, and Gly treatments had less (P = 0.026) feather CORT concentrations, but had greater (P < 0.001) TEER than NC treatment. In conclusion, increasing concentrations of dietary Thr, Trp, or Gly improve the growth performance and intestinal health in broiler chickens with decreasing stress response under multiple stress conditions.

Hydrolyzed collagen supplementation prior to resistance exercise augments collagen synthesis in a dose-response manner in resistance-trained, middle-aged men.

BACKGROUND: Resistance exercise (RE) increases collagen synthesis in young and older men, while hydrolyzed collagen (HC) ingestion improves this response to RE in a dose-response manner in young men. However, the collagen synthesis response to RE with and without HC in middle-aged men is unknown. METHODS: Eight resistance-trained men (age: 49±8 years; height: 1.78±0.02m; mass: 90±4kg) took part in this double-blind, crossover design study, and undertook 4×10 repetitions of lower-limb RE at maximum load, after consuming 0g, 15g, or 30g vitamin C-enriched HC. We analyzed venous blood samples for N-terminal propeptide of type 1 pro-collagen (PINP), ß-isomerized C-terminal telopeptide of type 1 collagen (ß-CTx) and 18 collagen amino acids throughout all three interventions. RESULTS: The serum PINP concentration×time area-under-the-curve (AUC) was higher following 30g (169±28 µg/mL×h) than 15g (134±23 µg/mL×h, P<0.05) HC ingestion, and both 15g and 30g were higher than 0g HC (96±23 µg/mL×h, P<0.05). RE with 0g HC showed no change in serum PINP concentration. The AUCs for glycine, proline, hydroxyproline, alanine, arginine, lysine, serine, leucine, valine and isoleucine were greater with 30g than 15g and 0g HC ingestion (P<0.05), and greater with 15g than 0g HC ingestion (P<0.05). Plasma ß-CTx concentration decreased after RE independently of HC dose. CONCLUSIONS: Our study suggests connective tissue anabolic resistance to RE in middle-aged men but ingesting 15g HC rescues the collagen synthesis response, and 30g augments that response further. This dose-response is associated with the increased bioavailability of collagen amino acids in the blood, which stimulate collagen synthesis.

Simultaneous quantification of alkanolamines and their dehydrogenation products in aqueous samples using high-performance liquid chromatography with pre-column derivatization with 2,4-dinitrofluorobenzene.

BACKGROUND: Diethanolamine, monoethanolamine, iminodiacetic acid, and glycine are important fine chemical intermediates, often requiring simultaneous quantitative analysis in various applications. This presents the challenge of accurately quantifying multiple substances within a single sample. The catalytic dehydrogenation of diethanolamine and monoethanolamine has garnered significant research interest, yet no analytical method has been reported for the simultaneous quantification of reactants and products in the dehydrogenation reaction mixtures of different alkanolamines. RESULTS: A high-performance liquid chromatography (HPLC) method has been developed for the simultaneous quantification of diethanolamine (DEA), iminodiacetic acid (IDA), glycine (Gly), and monoethanolamine (MEA) in aqueous solutions using 2,4-dinitrofluorobenzene (DNFB) for pre-column derivatization. The method demonstrated excellent linearity, with correlation coefficients (R2) of 0.9999, 0.9997, and 0.9998 for IDA, DEA, and Gly, respectively. The detection limits (LODs) were 0.02, 0.08, and 0.01 mg L-1, respectively. The quantification limits (LOQs) were 0.06, 0.24, and 0.03 mg L-1, respectively. Spiked recovery rates ranged from 96.99 % to 104.32 %, with relative standard deviations (RSDs) between 0.70 % and 3.03 %. For MEA and Gly, the R2 values were 0.9970 and 0.9990, the LODs were 0.12 and 0.01 mg L-1, the LOQs were 0.36 and 0.03 mg L-1, and spiked recoveries ranged from 96.24 % to 104.82 %, with RSDs between 1.22 % and 3.68 %. Compared to other methods, this HPLC approach offers superior sensitivity, accuracy, and precision. SIGNIFICANCE: This method provides a robust reference for the individual or simultaneous quantification of alkanolamines, glycine, and iminodiacetic acid in aqueous matrices. It offers new insights into the simultaneous analysis of alkanolamines with multiple organic acids in complex matrices. Additionally, the method can guide the optimization of catalytic dehydrogenation processes for alkanolamines, potentially extending the advantages of dehydrogenation catalysts to other reactions.

Investigating the electronic properties of edge glycine/biopolymer/graphene quantum dots.

This study systematically investigated four types of graphene quantum dots (GQDs) AHEX, ZTRI, ZHEX, and ATRI, and their interactions with glycine to form GQD-glycine complexes. Utilizing density functional theory (DFT) and the PM6 semiempirical method, the study analyzed electronic properties and structure-activity relationships. Global reactivity indices were calculated using Koopmans' theorem, and quantitative structure-activity relationship (QSAR) parameters were assessed via SCIGRESS 0.3. The study further explored interactions using density of states (DOS) and quantum theory of atoms in molecules (QTAIM) analyses. Key findings revealed that glycine interaction significantly increased the total dipole moment (TDM) and decreased the HOMO/LUMO energy gap (ΔE) for the GQD-glycine complexes. Notably, ZTRI/glycine showed a TDM of 4.535 Debye and a reduced ΔE of 0.323 eV, indicating enhanced reactivity. Further interactions with cellulose, chitosan, and sodium alginate identified the ZTRI/glycine/sodium alginate composite as the most reactive, with a TDM of 8.020 Debye and the lowest ΔE of 0.200 eV. This composite also exhibited the highest electrophilicity index (56.421) and lowest chemical hardness (0.145 eV), underscoring its superior reactivity and stability. DOS analysis revealed that biomolecules contributed the most to molecular orbitals, with carbon atoms contributing the least. QTAIM analysis confirmed the greater stability of the ZTRI/glycine/sodium alginate complex compared to other studied composites. These results highlight the enhanced reactivity and stability of GQDs when interacting with glycine and sodium alginate.

Feeding behavior, life-history traits, and settling preference of Aulacorthum solani (Hemiptera: Aphididae) on resistant and susceptible soybeans.

Foxglove aphid, Aulacorthum solani Kaltenbach (Hemiptera: Aphididae), is a major pest worldwide. It can infest various crops, including soybean, and reduce yields. The use of insect-resistant cultivars can effectively manage pests. Dowling is a soybean cultivar that can control Aphis glycines through antibiosis and antixenosis. In this study, we investigated the feeding behavior of A. solani using electropenetrography (EPG), and its life-history traits using an age-stage, two-sex life table, and its settling preferences on Dowling (resistant) and Heinong 51 (HN51, susceptible) soybean cultivars. The Dowling cultivar showed strong antibiosis against A. solani. Aphids feeding on Dowling exhibited significantly reduced survival, fecundity, and longevity, and increased nymph duration. Moreover, Dowling had negative impacts on the demographic parameters of the aphids. The number of A. solani individuals was significantly less on Dowling than on HN51, indicating the antixenotic effects of Dowling. As shown using EPG, the mechanical blocking wave (F) of A. solani was significantly longer after feeding on Dowling than after feeding on HN51, indicating that Dowling has a higher mechanical resistance. In conclusion, Dowling exhibited strong resistance to A. solani. These results are beneficial for integrated pest management in soybean fields and breeding programs.

Lipidomics analysis of microalgal lipid production and heavy metal adsorption under glycine betaine-mediated alleviation of low-temperature stress.

Heavy metal pollution in the cold region is serious, affecting human health and aquatic ecology. This study investigated the ability of microalgae to remove heavy metals (HMs) and produce lipid at low temperature. The removal efficiency of different HMs (Cd2+, Cu2+, Cr3+ and Pb2+), cell growth and lipid synthesis of microalgae were analyzed at 15 °C. Moreover, addition of glycine betaine (GB) further enhanced the productivity of microalgae in treating HMs and lipid production, and simultaneously increased the antioxidant capacity of microalgae against environmental stresses. The results showed that the highest lipid productivity of 100.98 mg L-1 d-1 and the removal efficiency of 85.8 % were obtained under GB coupled with Cr3+. The highest glutathione content of 670.34 nmol g-1 fresh alga was achieved under GB coupled with Pb2+. In addition, lipidomics showed that GB was able to up-regulate the triglyceride and diglyceride content, influenced fatty acid composition to regulate the microalgal metabolism, and mediated lipid accumulation under 15 °C mainly through the regulation of glycerol ester metabolism. This study provided a new perspective on microalgal lipid production and the removal of HMs in cold regions and provided evidence for the use of phytohormones to improve the algal environmental resistance.

Beneficial or detrimental? How nickel application alters the ionome of soybean plants.

The use of nickel (Ni) in agriculture may represent one of the most significant cases of plant hormesis ever reported, as plants exhibit both positive and negative responses depending on the level of exposure to this element. For a more comprehensive understanding of this effect, the next step is to conduct studies on the dynamics of pre-existing chemical elements in the system (ionomic profile), especially when introducing Ni as a novel nutrient for the plants. This micronutrient is of particular interest to the fertilization of leguminous plants, such as the soybean, due to its additional effects on the biological nitrogen fixation process. This study thus evaluated the influence of five doses of Ni (0.0, 0.5, 1.0, 3.0, and 9.0 mg of Ni kg-1) on the ionomic profile of soybean genotypes using modern quantification techniques. The results revealed that the addition of Ni reduced the concentration of cationic micronutrients manganese (Mn), iron (Fe), zinc (Zn), and copper (Cu), while it increased the concentration of macronutrients nitrogen (N) and magnesium (Mg). The application of Ni also resulted in a reduction of the potentially toxic element aluminum (Al). Correlations were also observed for these elements, indicating that Ni could be a controlling agent in elemental absorption and translocation. The ionome of the leaf tissues exhibited the most significant alterations, followed by the grains, nodules, and roots. Exogenous agronomic doses of Ni proved beneficial for the growth and production of soybean plants, although a genotypic effect was observed. The treatment with 9.0 mg of Ni kg-1, resulted in a new ionomic profile related to toxicity, demonstrating suboptimal plant development. Thus, the application of Ni in appropriate doses had a significant impact on the ionomic profile of soybeans, improving plant development and implying resistance to potentially toxic elements such as Al.

Assembly, comparative analysis, and utilization of a single haplotype reference genome for soybean.

Cultivar Williams 82 has served as the reference genome for the soybean research community since 2008, but is known to have areas of genomic heterogeneity among different sub-lines. This work provides an updated assembly (version Wm82.a6) derived from a specific sub-line known as Wm82-ISU-01 (seeds available under USDA accession PI 704477). The genome was assembled using Pacific BioSciences HiFi reads and integrated into chromosomes using HiC. The 20 soybean chromosomes assembled into a genome of 1.01Gb, consisting of 36 contigs. The genome annotation identified 48 387 gene models, named in accordance with previous assembly versions Wm82.a2 and Wm82.a4. Comparisons of Wm82.a6 with other near-gapless assemblies of Williams 82 reveal large regions of genomic heterogeneity, including regions of differential introgression from the cultivar Kingwa within approximately 30 Mb and 25 Mb segments on chromosomes 03 and 07, respectively. Additionally, our analysis revealed a previously unknown large (>20 Mb) heterogeneous region in the pericentromeric region of chromosome 12, where Wm82.a6 matches the 'Williams' haplotype while the other two near-gapless assemblies do not match the haplotype of either parent of Williams 82. In addition to the Wm82.a6 assembly, we also assembled the genome of 'Fiskeby III,' a rich resource for abiotic stress resistance genes. A genome comparison of Wm82.a6 with Fiskeby III revealed the nucleotide and structural polymorphisms between the two genomes within a QTL region for iron deficiency chlorosis resistance. The Wm82.a6 and Fiskeby III genomes described here will enhance comparative and functional genomics capacities and applications in the soybean community.

Buffer system improves the removal of host cell protein impurities in monoclonal antibody purification.

Polysorbates (PS) are commonly used as stabilizers of biopharmaceuticals such as monoclonal antibodies (mAbs). However, they are prone to chemical and enzymatic degradation. The latter can be caused by residual host cell proteins (HCPs) in the drug substance. Degradation affects the functionality of the PS surfactant which can lead to formation of particles. An increasing number of publications describe enzymatic PS degradation. Significant efforts have been made to characterize HCP removal during Downstream Processing (DSP) of mAbs and to develop mitigation strategies. Here we describe the use of glycine buffer for acidic elution in Protein A affinity chromatography compared to acetate buffer, which is more commonly used in the biopharmaceutical industry. Increased turbidity was observed during pH re-adjustment after low pH virus inactivation when using glycine buffer. Analytical data suggests that this turbidity is caused by the formation of precipitates which include HCP and DNA impurities. Additionally, as a zwitterion, glycine does not contribute to conductivity; this further enhances HCP removal during anion-exchange flow-through chromatography. Although glycine is well known as a possible elution buffer for Protein A affinity chromatography, its positive impact on HCP removal and PS stability have not yet been described in literature.

Contextualized Metabolic Modelling Revealed Factors Affecting Isoflavone Accumulation in Soybean Seeds.

Isoflavones, secondary metabolites with numerous health benefits, are predominantly found in legume seeds, especially soybean; however, their contents in domesticated soybean seeds are highly variable. Wild soybeans are known for higher seed isoflavone contents than cultivars. Here we used experimental and modelling approaches on wild soybean (W05) and cultivated soybean (C08) to delineate factors influencing isoflavone accumulation. We found imported nutrients were converted into storage compounds, with isoflavone accumulation in W05 seeds being faster than in C08 ones. The isoflavone accumulation during seed development was simulated using context-specific cotyledon metabolic models of four developmental stages on cultivar C08, and the metabolic burden imposed by increasing biomass was evaluated. Trade-off analyses between biomass and isoflavone suggest that high biomass requirement in cultivars could limit the reallocation of resources for secondary metabolite production. Isoflavone production in mature seeds was also influenced by biomass compositions. Seeds with higher carbohydrate contents favour isoflavone production, while those with highest protein and oil contents had lowest isoflavone contents. Although seeds could synthesize isoflavones on their own, the predicted fluxes from biosynthesis alone were lower than the empirical levels. Shadow price analyses indicated that isoflavone accumulation depended on both intrinsic biosynthesis and direct contribution from the plant.

Formulation of nutritious and functional meal-based biscuits from mixture of soybean, papaya fruit pulp, and baobab fruit pulp flours.

Background and objective: The current trend in the formulation of convenience foods like biscuits is directed towards using local ingredients endowed with health benefits effects. The present study aimed at valorizing local crops and fruits as substitutes for wheat flour (WF) and sugar in the formulation of healthy, nutritious and functional meal-based biscuits. Methods: Soybean (Glycine max L.), papaya fruit pulp (Carica papaya L.) and baobab fruit pulp (Adansonia digitata L.) flours were produced, characterized, and used to formulate biscuits following a simplex centroid mixing design. The physicochemical, microbiological, sensory and antioxidant properties of the biscuits were assessed. Results: The results showed that protein and lipid contents of the biscuits increased significantly (p < 0.05) with the proportion of soybean flour (SF) in the formulation. The significant increase (p < 0.05) in the mineral content of the biscuits was proportional to the incorporation of papaya fruit pulp (PFPF) and baobab fruit pulp (BFPF) flours in the formulation. The energetic value of the formulated biscuits was higher than those made with 100 % WF. The incorporation of SF and BFPF contributed to a significant increase (p < 0.05) in the crude fibres' content of biscuits. Biscuits made with SF, PFPF and BFPF were safe for human consumption. They were accepted by the panelists; rich in bioactive compounds (total phenolic, flavonoids), and displayed high antioxidant activities. The optimization procedure revealed that the optimum formulation (with the highest desirability of 0.81) was WF 25 g, SF 51.86 g, PFPF 8.06 g and BFPF 15.06 g. Conclusion: This study indicates that baobab fruit pulp, papaya fruit pulp and soybean flours can be used as a substitute for WF in the formulation of functional biscuits. Significance of the research: This study suggests that the formulated meal-based biscuits might have the potential to be used to fight/prevent malnutrition and cardiometabolic diseases, and to boost the immune system while reducing the dependence on wheat.

Case report: Unveiling genetic and phenotypic variability in Nonketotic hyperglycinemia: an atypical early onset case associated with a novel GLRX5 variant.

Nonketotic hyperglycinemia (NKH) is a rare, autosomal recessive metabolic disorder usually associated with mutations in genes AMT, GLDC or GCSH involved in the glycine cleavage complex. Other genes have been linked with less severe NKH, associated with deficiency of lipoate cofactor such as GLRX5, LIAS, BOLA3. We identified a new case of GLRX5-mediated NKH who presented at 2-month with severe developmental delay and seizures. The initial suspicion was raised by the MRI and then confirmed by glycine measurements in cerebrospinal fluid and blood. Genetic analysis revealed a previously undescribed homozygous variant in the GLRX5 gene [NM_016417.3:c.367G>C; p. (Asp123His)]. Despite medication and supportive care, he died at the age of 4 months after a sudden neurological deterioration. It was decided to limit therapeutic interventions due to the severity of the prognosis. The case was more severe than the previous GLRX5-mediated NKH described, regarding the early age at onset and the severity. Moreover, the genetic variant was located at a potentially crucial site for glutathione binding in the GLRX5 protein. This report, thereby, expands our understanding of NKH's genetic underpinnings and phenotypic variability, highlighting the crucial role of GLRX5 and other related genes in variant NKH.

[Efficient biosynthesis of guanidoacetic acid by a recombinant strain of Bacillus subtilis].

Guanidinoacetic acid, as an energetic substance, has a wide range of applications in the food, pharmaceutical, and feed industries. However, the biosynthesis of guanidinoacetic acid has not been applied in industrial production. In this study, we designed the synthetic route of guanidinoacetic acid in a food-grade strain of Bacillus subtilis. By regulating the expression of key enzymes, lifting feedback inhibition, and increasing membrane permeability, we achieved the efficient synthesis of guanidinoacetic acid by whole-cell catalysis. Firstly, the optimal L-arginine:glycine amidinotransferase was screened based on the phylogenetic tree, and the expression of the key enzyme was enhanced by a strategy combining strong promoter and genome integration. Secondly, the ornithine cycle for L-arginine synthesis in Corynebacterium glutamicum was introduced to alleviate the feedback inhibition of the enzyme by the byproduct L-ornithine, and the L-arginine degradation pathway was knocked down to enhance substrate regeneration. Thirdly, the expression of N-acetylmuramoyl-L-alanine amidase (LytC) was up-regulated to increase the cell membrane permeability. Finally, after optimization of whole-cell production conditions, strain Bs-13 achieved guanidinoacetic acid production at a titer of 13.1 g/L after 24 h, with a proudction rate of 0.54 g/(L·h) and a glycine conversion rate of 92.7%. The above strategy improved the production of guanidinoacetic acid and provided a reference for the biosynthesis of guanidinoacetic acid.

Whole-genome comparison using complete genomes from Campylobacter fetus strains revealed single nucleotide polymorphisms on non-genomic islands for subspecies differentiation.

Introduction: Bovine Genital Campylobacteriosis (BGC), caused by Campylobacter fetus subsp. venerealis, is a sexually transmitted bacterium that significantly impacts cattle reproductive performance. However, current detection methods lack consistency and reliability due to the close genetic similarity between C. fetus subsp. venerealis and C. fetus subsp. fetus. Therefore, this study aimed to utilize complete genome analysis to distinguish genetic features between C. fetus subsp. venerealis and other subspecies, thereby enhancing BGC detection for routine screening and epidemiological studies. Methods and results: This study reported the complete genomes of four C. fetus subsp. fetus and five C. fetus subsp. venerealis, sequenced using long-read sequencing technologies. Comparative whole-genome analyses (n = 25) were conducted, incorporating an additional 16 complete C. fetus genomes from the NCBI database, to investigate the genomic differences between these two closely related C. fetus subspecies. Pan-genomic analyses revealed a core genome consisting of 1,561 genes and an accessory pangenome of 1,064 genes between the two C. fetus subspecies. However, no unique predicted genes were identified in either subspecies. Nonetheless, whole-genome single nucleotide polymorphisms (SNPs) analysis identified 289 SNPs unique to one or the C. fetus subspecies. After the removal of SNPs located on putative genomic islands, recombination sites, and those causing synonymous amino acid changes, the remaining 184 SNPs were functionally annotated. Candidate SNPs that were annotated with the KEGG "Peptidoglycan Biosynthesis" pathway were recruited for further analysis due to their potential association with the glycine intolerance characteristic of C. fetus subsp. venerealis and its biovar variant. Verification with 58 annotated C. fetus genomes, both complete and incomplete, from RefSeq, successfully classified these seven SNPs into two groups, aligning with their phenotypic identification as CFF (Campylobacter fetus subsp. fetus) or CFV/CFVi (Campylobacter fetus subsp. venerealis and its biovar variant). Furthermore, we demonstrated the application of mraY SNPs for detecting C. fetus subspecies using a quantitative PCR assay. Discussion: Our results highlighted the high genetic stability of C. fetus subspecies. Nevertheless, Campylobacter fetus subsp. venerealis and its biovar variants encoded common SNPs in genes related to glycine intolerance, which differentiates them from C. fetus subsp. fetus. This discovery highlights the potential of employing a multiple-SNP assay for the precise differentiation of C. fetus subspecies.

Optimal glycine allowance levels in low-protein diets and the dynamic requirement model for broilers.

This study aimed to investigate the effects of different glycine levels in low-protein diets on the growth, nitrogen deposition, and expression of intestinal amino acid and glucose transporters in broilers from 29 to 42 d of age, in order to determine the optimal glycine supplementation level. A total of 240 male broilers at 29 days old were randomly assigned to 5 groups: the control group with a crude protein level of 20%, and experimental groups with low-protein diets (LP130) containing 18% crude protein, supplemented with glycine to achieve standardized ileal digestible (SID) glycine + serine to lysine ratios of 134% (LP134), 140% (LP140), and 145% (LP145). The results showed that the LP134 group had similar growth performance and slaughter performance compared to the control group (P > 0.05), whereas other low-protein diet groups had significantly lower growth performance (P < 0.05). Regression analysis determined that the optimal ratio for SID glycine + serine to lysine was 137%. A dynamic model for glycine + serine requirements was established through binary regression analysis: y = 599.051 × BW^0.75 + 8.381 × ADG (R2 = 0.998, P < 0.001). Feeding LP134, LP140, and LP145 diets significantly improved nitrogen deposition rates in broilers (P < 0.05). Low-protein diets significantly upregulated mRNA levels of b0,+AT, EAAT3, and SGLT1 genes in the duodenum (P < 0.05). In conclusion, appropriate glycine supplementation in low-protein diets can enhance growth performance, and nitrogen deposition efficiency, and regulate the expression of intestinal amino acid and glucose transporters. The optimal ratio of SID glycine + serine to lysine in low-protein diets for broilers aged 29 to 42 d is 137%.

A New Mechanism for Formation of Glycine from Glyoxylic acid: the Aza-Cannizzaro Reaction.

Glyoxylic acid and glycine are widely considered to have been important prebiotic building blocks. Several mechanistic routes have been previously examined for conversion of glyoxylic acid to glycine. Here we provide evidence for a new mechanistic path. Glycine is spontaneously formed from glyoxylic acid in ammonium-rich aqueous solutions at neutral pH; oxamic acid is generated as well. Hydride transfer from the glyoxylate-derived hemiaminal to the corresponding iminium ion appears to underlie this transformation. This proposed mechanism parallels the well-known Cannizzaro reaction mechanism, which leads us to suggest the designation "aza-Cannizzaro reaction." This discovery offers a new perspective on prebiotic nitrogen incorporation because glycine can be a source of nitrogen for more complex molecules, including other α-amino acids.

Fermented Fish Collagen Attenuates Melanogenesis via Decreasing UV-Induced Oxidative Stress.

Excessive melanogenesis leads to hyperpigmentation-related cosmetic problems. UV exposure increases oxidative stress, which promotes melanogenesis-related signal pathways such as the PKA, microphthalmia-associated transcription factor (MITF), tyrosinase (TYR), tyrosinase-related protein-1 (TRP1), and tyrosinase-related protein-2 (TRP2) pathways. Glycine is a source of endogenous antioxidants, including glutathione. Fermented fish collagen (FC) contains glycine; thus, we evaluated the effect of FC on decreasing melanogenesis via decreasing oxidative stress. The glycine receptor (GlyR) and glycine transporter-1 (GlyT1) levels were decreased in UV-irradiated keratinocytes; however, the expression levels of these proteins increased upon treatment with FC. The FC decreased oxidative stress, as indicated by the decreasing expression of NOX1/2/4, increased expression of GSH/GSSG, increased SOD activity, and decreased 8-OHdG expression in UV-irradiated keratinocytes. Administration of conditioned media from FC-treated keratinocytes to melanocytes led to decreased p38, PKC, MITF, TRP1, and TRP2 expression. These changes induced by the FC were also observed in UV-irradiated animal skin. FC treatment increased the expression of GlyR and GlyT, which was accompanied by decreased oxidative stress in the UV-irradiated skin. Moreover, the FC negatively regulated the melanogenesis signaling pathways, leading to decreased melanin content in the UV-irradiated skin. In conclusion, FC decreased UV-induced oxidative stress and melanogenesis in melanocytes and animal skin. FC could be used in the treatment of UV-induced hyperpigmentation problems.