Acid Side Research Articles

Effects of Three Extraction Methods on Avocado Oil Lipid Compounds Analyzed via UPLC-TOF-MS/MS with OPLS-DA

Avocado oil is excellent functional oil. Effects of three extraction methods (squeezing extraction, supercritical carbon dioxide extraction, and aqueous extraction) on the species, composition, and contents of lipids in avocado oil were analyzed via ultra-performance liquid chromatography-time-of-flight tandem mass spectrometry (UPLC-TOF-MS/MS), and the differential components of lipids were revealed by OrthogonalPartialLeast Squares-DiscriminantAnalysis (OPLS-DA), S-plot combined with variable importance in the projection (VIP). The results showed that the fatty acid composition of avocado oil mainly consisted of oleic acid (36-42%), palmitic acid (25-26%), linoleic acid (14-18%), and palmitoleic acid (10-12%). A total of 134 lipids were identified first from avocado oil, including 122 glycerides and 12 phospholipids, and the total number of carbon atoms contained in the fatty acid side chains of the lipids was 32-68, and the number of double bonds was 0-9. Forty-eight differential lipid compounds with significant effects of the three extraction methods on the lipid composition of avocado oil were excavated, among which the differences in triglycerides (TG), phosphatidylethanol (PEtOH), and phosphatidylmethanol (PMeOH) contents were highly significant, which provided basic data to support the subsequent guidance of avocado oil processing, quality evaluation, and functional studies.

Comparison of In Vitro Antimelanoma and Antimicrobial Activity of 2,3-Indolo-betulinic Acid and Its Glycine Conjugates

Malignant melanoma is one of the most pressing problems in the developing world. New therapeutic agents that might be effective in treating malignancies that have developed resistance to conventional medications are urgently required. Semisynthesis is an essential method for improving the biological activity and the therapeutic efficacy of natural product precursors. Semisynthetic derivatives of natural compounds are valuable sources of new drug candidates with a variety of pharmacological actions, including anticancer ones. Two novel semisynthetic derivatives of betulinic acid-N-(2,3-indolo-betulinoyl)diglycylglycine (BA1) and N-(2,3-indolo-betulinoyl)glycylglycine (BA2)-were designed and their antiproliferative, cytotoxic, and anti-migratory activity against A375 human melanoma cells was determined in comparison with known N-(2,3-indolo-betulinoyl)glycine (BA3), 2,3-indolo-betulinic acid (BA4) and naturally occurring betulinic acid (BI). A dose-dependent antiproliferative effect with IC50 values that ranged from 5.7 to 19.6 µM was observed in the series of all five compounds including betulinic acid. The novel compounds BA1 (IC50 = 5.7 µM) and BA2 (IC50 = 10.0 µM) were three times and two times more active than the parent cyclic structure B4 and natural BI. Additionally, compounds BA2, BA3, and BA4 possess antibacterial activity against Streptococcus pyogenes ATCC 19615 and Staphylococcus aureus ATCC 25923 with MIC values in the range of 13-16 µg/mL and 26-32 µg/mL, respectively. On the other hand, antifungal activity toward Candida albicans ATCC 10231 and Candida parapsilosis ATCC 22019 was found for compound BA3 with MIC 29 µg/mL. This is the first report of antibacterial and antifungal activity of 2,3-indolo-betulinic acid derivatives and also the first extended report on their anti-melanoma activity, which among others includes data on anti-migratory activity and shows the significance of amino acid side chain on the observed activity. The obtained data justify further research on the anti-melanoma and antimicrobial activity of 2,3-indolo-betulinic acid derivatives.

Decomposing Hofmeister effects on amino acid residues with symmetry adapted perturbation theory

Hofmeister effects, and more generally specific ion effects, are observed broadly in biological systems. However, there are many cases where the Hofmeister series might not be followed in complex biological systems, such as ion channels which can be highly specific to a particular ion. An understanding of how ions from the Hofmeister series interact with the proteinogenic amino acids will assist elucidation of why some binding interactions may be favoured over others. Using symmetry adapted perturbation theory (SAPT2 + 3), the interaction energies between a selection of anions and each amino acid have been investigated. The interaction strengths become more favourable in accordance with the Hofmeister series, and also with increasing polarity of the amino acids (with the exception of the negatively charged amino acid side chains). Furthermore, the interactions are generally most favourable when they simultaneously involve the side chain and both protic moieties of the backbone. The total interaction energy in these anion–amino acid complexes is also primarily determined by its electrostatic component, in a manner proportional to the þ (‘sho’) value of the anion.

Importance of aspartic acid side chain carboxylate-arginine interaction in substrate selection of arginine 2,3-aminomutase BlsG.

The fungicide nucleoside blasticidin S features a β-arginine, a moiety seldom revealed in the structure of natural products. BlsG, a radical SAM arginine-2,3-aminomutase from the blasticidin S biosynthetic pathway, displayed promiscuous activity to three basic amino acids. Here in this study, we demonstrated that BlsG showed high preference toward its natural substrate arginine. The combined structural modeling, steady-state kinetics, and mutational analyses lead to the detailed understanding of the substrate recognition of BlsG. A single mutation of T340D changed the substrate preference of BlsG leading to a little more preference to lysine than arginine. On the basis of our understanding of the substrate selection of BlsG and bioinformatic analysis, we propose that the D…D motif locationally corresponding to D293 and D330 of KAM is characteristic of lysine 2,3-aminomutase while the corresponding D…T motif is characteristic of arginine 2,3-aminomutase. The study may provide a simple way to discern the arginine 2,3-aminomutase and thus lead to the discovery of new natural compounds with β-arginine moiety.

The polar amino acid in the TatA transmembrane helix is not strictly necessary for protein function

The twin-arginine translocation (Tat) pathway utilizes the proton-motive force to transport folded proteins across cytoplasmic membranes in bacteria and archaea, as well as across the thylakoid membrane in plants and the inner membrane in mitochondria. In most species, the minimal components required for Tat activity consist of three subunits, TatA, TatB, and TatC. Previous studies have shown that a polar amino acid is present at the N terminus of the TatA transmembrane helix (TMH) across many different species. In order to systematically assess the functional importance of this polar amino acid in the TatA TMH in Escherichia coli, we examined a complete set of 19-amino-acid substitutions. Unexpectedly, although the polar amino acid is preferred overall, our experiments suggest that it is not necessary for a functional TatA. Hydrophilicity and helix-stabilizing properties of this polar amino acid were found to be highly correlated with the Tat activity. Specifically, change in charge status of the amino acid side chain due to pH resulted in a shift in hydrophilicity, which was demonstrated to impact the Tat transport activity. Furthermore, we identified a four-residue motif at the N terminus of the TatA TMH by sequence alignment. Using a biochemical approach, we found that the N-terminal motif was functionally significant, with evidence indicating a potential role in the preference for utilizing different proton-motive force components. Taken together, these findings yield new insights into the functionality of TatA and its potential role in the Tat transport mechanism.

Advances in Activity‐Based Protein Profiling of Functional Tyrosines in Proteomes

AbstractActivity‐based protein profiling (ABPP) is a chemical proteomic method for investigating functional states of proteins in native biological settings. By quantifying changes in probe binding states of active and regulatory protein sites, ABPP reveals functional information on protein regulation and can be configured in competitive settings to determine global selectivity profiles of tool compounds and drugs in lysates, cells, and animals. Chemical probes used for ABPP analyses can target protein families with conserved enzymatic or structural features or can broadly profile the proteome using electrophiles with reactivity towards functional groups on amino acid side chains. The latter approach has provided insights to protein sites involved in allosteric regulation and non‐enzymatic functions. This review introduces quantitative ABPP workflows and discusses electrophilic groups used for ABPP profiling of functional sites in the proteome with an emphasis on tyrosine residues.

Interaction of Positively Charged Oligopeptides with Blood Plasma Proteins.

In this project, we combine two areas of research, experimental characterization and molecular docking studies of the interaction of positively charged oligopeptides with crucial blood plasma proteins. The investigated peptides are rich in NH2 groups of amino acid side chains from Dap, Orn, Lys, and Arg residues, which are relevant in protein interaction. The peptides are 9- and 11-mer with the following sequences: (Lys-Dab-Dab-Gly-Orn-Pro-His-Lys-Arg-Lys-Dbt), (Lys-Dab-Ala-Gly-Orn-Pro-His-Lys-Arg), and (Lys-Dab-Dab-Gly-Orn-Pro-Phe(2-F)-Lys-Arg). The net charge of the compound strongly depends on the pH environment and it is an important aspect of protein binding. The studied oligopeptides exhibit therapeutic properties: anti-inflammatory activity and the capacity to diminish reactive oxygen species (ROS). Therefore, the mechanism of potential binding with blood plasma components is the next challenge. The binding interaction has been investigated under pseudo-physiological conditions with the main blood plasma proteins: albumin (BSA), α1-acid glycoprotein (AAG), and γ-globulin fraction (GGF). The biomolecular quenching constant (kq) and binding constant (Kb) were obtained by fluorescence spectroscopy at various temperatures. Simultaneously, the changes in the secondary structure of proteins were monitored by circular dichroism (CD) and infrared spectroscopy (IR) by quantity analysis. Moreover, molecular docking studies were conducted to estimate the binding affinity, the binding domain, and the chemical nature of these interactions. The results show that the investigated oligopeptides could be mainly transported by albumin, and the binding domain I is the most favored cavity. The BSA and GGF are able to form stable complexes with the studied compounds as opposed to AAG. The binding reactions are spontaneous processes. The highest binding constants were determined for Lys-Dab-Dab-Gly-Orn-Pro-His-Lys-Arg-Lys-Dbt peptide, in which the values of the binding constants Kb to BSA and GGF were 10.1 × 104 dm3mol-1 and 3.39 × 103 dm3mol-1, respectively. The positively charged surface of peptides participated in salt bridge interaction with proteins; however, hydrogen bonds were also formed. The secondary structure of BSA and GGF after contact with peptides was changed. A reduction in the α-helix structure was observed with an increase in the β-sheet and β-turn and random coil structures.

Impact of an Ionic Liquid on Amino Acid Side Chains: A Perspective from Molecular Simulation Studies.

Ionic liquids (ILs) are known to modify the structural stability of proteins. The modification of the protein conformation is associated with the accumulation of ILs around the amino acid (AA) side chains and the nature of interactions between them. To understand the microscopic picture of the structural arrangements of ILs around the AA side chains, room temperature molecular dynamics (MD) simulations have been carried out in this work with a series of hydrophobic, polar and charged AAs in aqueous solutions containing the IL 1-butyl-3-methylimidazolium tetrafluoroborate ([BMIM][BF4]) at 2 M concentration. The calculations revealed distinctly nonuniform distribution of the IL components around different AAs. In particular, it is demonstrated that the BMIM+ cations preferentially interact with the aromatic AAs through favorable stacking interactions between the cation imidazolium head groups and the aromatic AA side chains. This results in preferential parallel alignments and enhanced population of the cations around the aromatic AAs. The potential of mean force (PMF) calculations revealed that such favorable stacking interactions provide greater stability to the contact pairs (CPs) formed between the aromatic AAs and the IL cations as compared to the other AAs. It is further quantified that for most of the AAs (except the cationic ones), a favorable enthalpy contribution more than compensates for the entropy cost to form stable CPs with the IL cations. These findings are likely to provide valuable fundamental information toward understanding the effects of ILs on protein conformational stability.

Key interactions convert amino acid side chains into strong acids and bases in the active sites of enzymes.

Enzymes catalyze reactions under mild conditions that might otherwise require extreme conditions such as high temperature or strong acid or base. To achieve this, amino acid side chains that are weak Brønsted acids or bases in the free amino acid become strong acids and bases in the active sites of many enzymes. Here we report on a set of 30 enzymes that represent all six major EC classes and a variety of different folds for which experimental studies of the mechanistic roles of the catalytic residues have been reported in the literature. Using macromolecular electrostatics techniques, it is shown that the catalytic aspartate and glutamate residues are strongly coupled to at least one other aspartate or glutamate residue, and often to multiple other carboxylate residues, with intrinsic pKa differences less than 1 pH unit. Sometimes these catalytic acidic residues are also coupled to a histidine residue, such that the intrinsic pKa of the acidic residue is higher than that of the histidine. Most catalytic lysine residues studied here are strongly coupled to tyrosine or cysteine residues, wherein the intrinsic pKa of the anion-forming residue is higher than that of the lysine. Some catalytic lysines are also coupled to other lysines with intrinsic pKa differences within 1 pH unit. Some general principles about the design of enzyme active sites are presented. These principles have significant implications for the engineering of novel enzymes. Supported by NSF CHE-1905214 (MJO).

Structural basis for severe pain caused by mutations in voltage gated sodium channels Nav1.7.

Gain-of-function mutations in voltage-gated sodium channel NaV1.7 cause severe inherited pain syndromes, including Inherited Erythromelalgia (IEM). We introduced IEM mutations into the bacterial sodium channel NaVAb and confirmed the negative shift in the voltage dependence of activation that recapitulates the gain-of-function of these mutants. Unexpectedly, our structures of NaVAb IEM mutations reveal two complementary pathogenic mechanisms at the atomic level. Mutations in the voltage sensor domain facilitate the outward movement of S4 gating charges by widening the pathway for gating charge movement and modifying hydrophobic interactions with surrounding amino acid side chains or membrane phospholipids. In contrast, IEM mutations in the S4-S5 linker stabilize the activated state by forming new hydrogen bonds with residues in the pore domain, including a conserved asparagine near the activation gate. These results provide key structural insights into the dual mechanisms by which these IEM mutations cause hyperexcitability and severe pain in this debilitating disease.

H+ flux of the voltage-gated proton channel Hv1 in giant unilamellar vesicle (GUV).

The voltage-sensitive proton selective channel HV1 is ubiquitously present in mammalian tissues. An aqueous pore provides the proton pathway. Intervening amino acid side chains render the channel impermeable to water [1]. Based on the hydrogen bonds formed between the permeating water molecules and the channel wall, HV1 would otherwise exhibit a water conductance comparable to aquaporins [2,3]. The proton wire involves Grotthuss-like proton hopping on top of intraluminal water molecules and titratable amino acids. Conceivably, HV1 exploits one or more of the arginines from the voltage sensor as part of the proton wire. Here we measured the unitary H+ transport rate by reconstituting the purified proton channel into giant unilamellar vesicles (GUV). We used the pHluorin tag to measure intravesicular pH and oxanol to probe the membrane potential. Fluorescence correlation spectroscopy served to measure channel density. The simultaneous assessment of diffusion limitations of H+ transport towards and away from the channel allows for evaluating the role of HV1's putative proton antenna. This project was supported by the Marie Skłodowska-Curie grant 860592 (Horizon 2020).

Structural basis of antibodies targeting neutralizing epitopes revealed by cryoempem

Engineered HIV-1 envelope glycoprotein immunogens can be designed to activate naïve B cells expressing germline antibody precursors that have the potential to evolve into broadly neutralizing antibodies. We applied cryo-EM based polyclonal epitope mapping (cryoEMPEM) to characterize the polyclonal antibody responses elicited by rhesus macaques immunized with a germline-targeting stabilized Env (BG505 SOSIP GT1.1). Antibodies against two neutralizing epitopes, CD4 binding site (CD4bs) and fusion peptide, were identified along with some non-neutralizing or strain-specific epitopes (V5 loop, glycans holes and base). A hybrid structural and bioinformatic approach was applied to predict monoclonal antibody sequences from the cryoEMPEM maps. Using IgDiscover and next-generation sequencing results, we identified candidate variable region sequences compatible with the amino acid side chain densities of CD4bs- or FP-directed cryoEMPEM responses. These monoclonal antibodies were then expressed and purified to confirm their binding to BG505 SOSIP GT1.1 antigens by biolayer interferometry and EM. Superimposition of the monoclonal antibody cryoEM models with the corresponding polyclonal cryoEMPEM model revealed high similarity, further demonstrating the efficacy of our structure-to-sequence method for antibody discovery. Overall, our work reveals that cryoEMPEM-identified antibodies targeting intended neutralizing epitopes were induced by BG505 SOSIP GT1.1 trimers in rhesus macaques and we successfully inferred monoclonal sequences from the structural data. Our structure-to-sequence pipeline offers an alternative to typical monoclonal antibody isolation strategies and is a promising complement to the future of iterative vaccine design.

Phosphatidyl serine preconditions the A2AAR for G-protein coupling via a conserved motif on TM6 and TM7.

19F NMR of A2AAR in nanodiscs of defined composition indicates that negative charge modulates activation of the receptor. The fully active state is obtained either when bound to G-protein, or without G-protein but in a membrane environment containing negatively charged headgroups. Mutagenesis indicates that a trio of residues on the intracellular ends of TM6 and TM7 are responsible for the lipid-dependent activity. In order to determine the mechanism, a series of molecular dynamics simulations were performed. Comparison of simulations of the inactive and the active, G-protein bound state shows that a glutamic acid on the helix of the G-protein coordinates interactions between these three residues. This suggested a hypothesis in which a negatively charged headgroup can stand-in for the glutamic acid side chain, coordinating the ends of TM6 and TM7 and preconditioning the receptor for G-protein coupling. To test this idea, simulations of the fully active state were performed with and without phosphatidyl serine headgroups, but without the G-protein. Consistent with our hypothesis, the TM6 -- TM7 interactions are similar to the G-protein coupled state in the presence of PS, but similar to the inactive state in the absence of PS. Moreover, the interaction of a PS headgroup with a key residue on TM6 recapitulates the same interaction with the glutamic acid, but only when the receptor is in the active state.

Total Synthesis of Taxol Enabled by Inter- and Intramolecular Radical Coupling Reactions.

Taxol is a clinically used drug for the treatment of various types of cancers. Its 6/8/6/4-membered ring (ABCD-ring) system is substituted by eight oxygen functional groups and flanked by four acyl groups, including a β-amino acid side chain. Here we report a 34-step total synthesis of this unusually oxygenated and intricately fused structure. Inter- and intramolecular radical coupling reactions connected the A- and C-ring fragments and cyclized the B-ring, respectively. Functional groups of the A- and C-rings were then efficiently decorated by employing newly developed chemo-, regio-, and stereoselective reactions. Finally, construction of the D-ring and conjugation with the β-amino acid delivered taxol. The powerful coupling reactions and functional group manipulations implemented in the present synthesis provide new valuable information for designing multistep target-oriented syntheses of diverse bioactive natural products.

Total Synthesis of Taxol Enabled by Inter‐ and Intramolecular Radical Coupling Reactions

AbstractTaxol is a clinically used drug for the treatment of various types of cancers. Its 6/8/6/4‐membered ring (ABCD‐ring) system is substituted by eight oxygen functional groups and flanked by four acyl groups, including a β‐amino acid side chain. Here we report a 34‐step total synthesis of this unusually oxygenated and intricately fused structure. Inter‐ and intramolecular radical coupling reactions connected the A‐ and C‐ring fragments and cyclized the B‐ring, respectively. Functional groups of the A‐ and C‐rings were then efficiently decorated by employing newly developed chemo‐, regio‐, and stereoselective reactions. Finally, construction of the D‐ring and conjugation with the β‐amino acid delivered taxol. The powerful coupling reactions and functional group manipulations implemented in the present synthesis provide new valuable information for designing multistep target‐oriented syntheses of diverse bioactive natural products.

Xylan glucuronic acid side chains fix suberin-like aliphatic compounds to wood cell walls.

Wood is the most important repository of assimilated carbon in the biosphere, in the form of large polymers (cellulose, hemicelluloses including glucuronoxylan, and lignin) that interactively form a composite, together with soluble extractives including phenolic and aliphatic compounds. Molecular interactions among these compounds are not fully understood. We have targeted the expression of a fungal α-glucuronidase to the wood cell wall of aspen (Populus tremula L. × tremuloides Michx.) and Arabidopsis (Arabidopsis thaliana (L.) Heynh), to decrease contents of the 4-O-methyl glucuronopyranose acid (mGlcA) substituent of xylan, to elucidate mGlcA's functions. The enzyme affected the content of aliphatic insoluble cell wall components having composition similar to suberin, which required mGlcA for binding to cell walls. Such suberin-like compounds have been previously identified in decayed wood, but here, we show their presence in healthy wood of both hardwood and softwood species. By contrast, γ-ester bonds between mGlcA and lignin were insensitive to cell wall-localized α-glucuronidase, supporting the intracellular formation of these bonds. These findings challenge the current view of the wood cell wall composition and reveal a novel function of mGlcA substituent of xylan in fastening of suberin-like compounds to cell wall. They also suggest an intracellular initiation of lignin-carbohydrate complex assembly.

Analysis of riboflavin/ultraviolet a corneal cross-linking by molecular spectroscopy

Corneal cross-linking (CXL) with riboflavin and ultraviolet A light is a therapeutic procedure to restore the mechanical stability of corneal tissue. The treatment method is applied to pathological tissue, such as keratoconus and induces the formation of new cross-links. At present, the molecular mechanisms of induced cross-linking are still not known exactly. In this study, we investigated molecular alterations within porcine cornea tissue after treatment with riboflavin and ultraviolet A light by surface enhanced Raman spectroscopy (SERS). For that purpose, after CXL treatment a thin silver layer was vapor-deposited onto cornea flaps. To explore molecular alterations induced by the photochemical process hierarchical cluster analysis (HCA) was used. The detailed analysis of SERS spectra reveals that there is no general change in collagen secondary structure while modifications on amino acid side chains are the most dominant outcome. The formation of secondary and aromatic amine groups as well as methylene and carbonyl groups were observed. Even though successful cross-linking could not be registered in all treated samples, Raman signals of newly formed chemical groups are already present in riboflavin only treated corneas.

Novel terpestacin derivatives with l-amino acid residue as anticancer agents against U87MG-derived glioblastoma stem cells

Based on the natural product terpestacin, seventeen derivatives (1–17) with various l-amino acid side chains were designed and synthesized. Their anticancer activities against U87MG-derived glioblastoma stem cells (GSCs) were evaluated, and compounds 5, 11, 13 and 15 showed strong abilities to inhibit the proliferation (IC50 = 2.8–6.9 μM) and tumorsphere formation of GSCs. Besides, compounds 13 and 15 could effectively induce apoptosis and significantly inhibit the invasion of GSCs (95 and 97 % inhibition, respectively, at 2.5 μM). The levels of CD133 marker in GSCs also decreased in dose-dependent manners after the treatment of these active compounds. Compared to terpestacin and the positive control A1938, our derivatives showed stronger activities and compounds 13 and 15 are promising candidates for further development as anticancer agents by targeting GSCs.

Iron nutrition in agriculture: From synthetic chelates to biochelates

Iron (Fe) is an essential micronutrient for plant growth and productivity and, among other micronutrients, is the one required in higher amounts. In most soils, Fe occurs at relatively high concentrations; nevertheless, in calcareous soils, Fe bioavailability can be very limited, thus leading plants to develop Fe deficiency symptoms. At present, Fe fertilization still represents the most frequent approach adopted in agriculture to prevent or remediate Fe chlorosis. For soil applications, Fe fertilizers are based on Fe chelated with aminocarboxylate synthetic ligands (e.g. HEDTA, EDTA, DTPA, EDDHA), which are effective in maintaining Fe in the soil solution, even in alkaline soils, and thus in increasing its bioavailability for plant uptake. Nevertheless, Fe-aminocarboxylate complexes presents some limitations, related for instance to pH-dependent effectiveness, stability, persistence in the environment and ligand exchange reactions. In this context, vegetal-derived protein hydrolysates might represent good sources of biochelating ligands for developing innovative Fe fertilizers for sustainable agriculture. Protein hydrolysates are composed by small peptides, which, on one hand, can chelate Fe via amino acid side chains, carboxylate groups of C-termini, amine groups of N-termini and N atoms of amide groups of the peptide backbone, whilst, on the other hand, small peptides can also play a signaling role, triggering the acquisition of nutrient and morphogenetic processes in plants. The present paper reviews the current state of knowledge on traditional Fe fertilizers and, on the other side, explores the possible advantages in the application of biochelates as innovative Fe fertilizers. To further corroborate the hypothesis, three experimental trials have been carried out on three horticultural crops (cucumber, tomato, and strawberry) using a Fe-biochelate as Fe source and comparing it with the widely used traditional synthetic chelate Fe-EDDHA. The results in the three crops clearly show that Fe-biochelate is, at least, as efficient as Fe-EDDHA as source of micronutrient, even under circumneutral (pH 6.0) and alkaline conditions (pH 8.0), thus proving that Fe-biochelates can be promising alternatives to synthetic Fe chelates for the Fe nutrition management of crops. Considering the potential drawbacks of synthetic chelates (e.g., leachability, persistence, remobilization of toxic metals in soil), these findings might contribute enhancing the agriculture sustainability. In addition, Fe-biochelate could also find an application in soilless cultivation systems as an alternative to synthetic Fe chelates for Fe enrichment of edible plant tissues (biofortification), increasing their nutritional value.

Evaluation of Urea and Molasses Treatment on The Nutritional Composition of Guinea Corn Stalk

The majority of the world’s population relies on livestock as source of protein whereas the success of livestock production is reliant on availability of quality feeds which has been challenged by competition impose by geometric growth in human population. This study assessed the effect of molasses and urea treatment on the physical quality, proximate and amino acid composition of guinea corn silage as a means to augment protein content in guinea corn silage feed stocks. Guinea corn stalks samples collected randomly on farmlands around Federal University Wukari, Nigeria. After pulverization was divided into two portions and one portion subjected to molasses and urea treatment while the other served as control. Effect of treatment was assessed by estimating and comparing the physical quality, proximate and amino acid composition of the treated and untreated sample using standard methods. The result showed that the mean pH value became more acidic after treatment from 6.12 to 5.24. While the moisture, lipid and crude protein content were significantly increased due to treatment (P <0.05). There was also a significant elevation in amino acid content due to urea and molasses treatment in 14 out of the 18 amino acids determined at 95% confidence limit (P<0.05). And the total amino acid content of the silage sample was improved from 43.80 g/100g to 62.27 g/100g. The nature of amino acids in the treated silage (test sample) were in the order of prevalence total amino acid with acidic side chain (TAAA)>total amino acid with basic side chain (TBAA)>total aromatic amino acid (TArAA)>total sulphur containing amino acid (TSAA). The most concentrated amino acid was glutamic acid, an important amino acid responsible in building the immune system.