Nutrients Amino Acids Research Articles

Enhancing the nutritional value of black soldier fly larvae through the nutrient amino acid transporter suppression in the excretion system

Abstract Larvae of the black soldier fly (Hermetia illucens, BSFL) are rich in valuable nutrients and offer a promising alternative protein source for animal feeds. Nonetheless, there is a pressing need to improve both the productivity and quality of BSFL to ensure the viability of BSFL products and facilitating the industrial production. To fulfil the needs of different animals, it is necessary to adjust the profile of essential amino acids (EAAs) in BSFL. Insects excrete surplus nutrients to maintain homeostasis; AAs are excreted by nutrient AA transporters (NATs) in the Malpighian tubules. We aimed to modify the composition of essential AAs by silencing the NAT in Malpighian tubules of BSFL (HiNATt). Silencing HiNATt resulted in a 77.3% increase in the total AA content while 56.2% decrease in body weight. Notably, the contents of some valuable essential AAs were strongly increased (histidine, 256.8%; valine, 198.1% in total compared to the intact larva). These results suggest that inhibiting the HiNATt function could modify the amount of accumulated EAAs. This finding opens a new avenue for producing BSFL with increased nutritional value as an alternative protein source.

Rapid determination for tyrosine isomers in food based on N-acetyl-L-cysteine/Upconversion nanomaterials target-induced quench by chiral Electrochemiluminescence sensor

Rapid determination of amino acid isomer is very important for the evaluation of the amino acid nutrition in different foods, so a fast and sensitive electrochemiluminescence (ECL) sensor was innovatively fabricated for the determination of tyrosine isomers in foods based on N-Acetyl-L-cysteine/upconversion nanomaterials possessed a good particular selectivity to L-tyrosine. Under the optimal conditions, for L-tyrosine, the limit of detection (LOD) of the sensor for L-tyrosine was 2.87 × 10−6 M, detection range of 5.5 × 10−5–5.5 × 10−3 M, for D-tyrosine, LOD was 2.56 × 10−5 M, detection range was from 5.5 × 10−4 to 5.5 × 10−3 M. The developed chiral sensor was used to determinate the tyrosine isomers in foods successfully, which provided a convenient method to quickly evaluate the nutritional value of amino acids in food.

Inhibition of Food-Borne Pathogen Growth and Biogenic Amine Synthesis by Spice Extracts

Food-borne pathogens and their toxins cause significant health problems in humans. Formation of biogenic amines (BAs) produced by microbial decarboxylation of amino acids in food is undesirable because it can induce toxic effects in consumers. Therefore, it is crucial to investigate the effects of natural additives with high bioactivity like spice extracts to inhibit the growth of these bacteria and the formation of BAs in food. In the present study, the antibacterial effects of diethyl ether spice (sumac, cumin, black pepper, and red pepper) extracts at doses of 1% (w/v) on Gram-positive (Staphylococcus aureus and Enterococcus faecalis) and Gram-negative (Klebsiella pneumoniae, Pseudomonas aeruginosa, Campylobacter jejuni, Aeromonas hydrophila, Salmonella Paratyphi A, and Yersinia enterocolitica) food-borne pathogen bacterial strains (FBP) were established. In addition, the accumulation of ammonia (AMN), trimethylamine (TMA), and biogenic amines (BAs) in tyrosine decarboxylase broth (TDB) was investigated by using high performance liquid chromatography (HPLC). Sumac extract exhibited the highest antibacterial potential against all FBPs, followed by cumin and peppers. AMN (570.71 mg/L) and TMA (53.66 mg/L) production were strongly inhibited by sumac extract in the levels of 55.10 mg/L for Y. enterocolitica and 2.76 mg/L for A. hydrophila, respectively. With the exception of S. aureus, black pepper dramatically reduced the synthesis of putrescine, serotonin, dopamine, and agmatine by FBP especially for Gram-negative ones. Furthermore, sumac extracts inhibited histamine and tyramine production by the majority of FBP. This research suggests the application of sumac extracts as natural preservatives for inhibiting the growth of FBPs and limiting the production of AMN, TMA, and BAs.

Standardized ileal amino acid digestibility of high-oleic full-fat soybean meal in broilers

High-oleic (HO) soybean may serve as a value-added feed ingredient to enrich poultry meat due to its fatty acid content. However, the amino acid (AA) nutrient digestibility of soybean meal (SBM) made from these soybeans has yet to be determined. The objective of this study was to determine apparent ileal AA digestibility (AID) and standardized ileal AA digestibility (SID) of high-oleic full-fat (HO-FF) SBM compared to normal oleic full-fat (NO-FF), normal oleic extruded expeller (NO-EE), and solvent-extracted SBM (SE-SBM) in broilers. A nitrogen-free basal diet (NFD) was fed to 1 treatment group with 10 chicks/cage to determine basal endogenous losses (BEL). Titanium dioxide was used as an inert marker. The test diets contained 57.5% of the basal NFD and 42.5% of 1 of the 4 soybean sources. A total of 272 Ross-708 male broilers were placed in 40 battery cages with 5 treatments and 8 replicates per treatment. A common starter diet was provided to all the chickens for 14 d. Experimental diets were provided as a mash for 9 d before sample collection. Chickens were euthanized with CO2 on d 23, and contents of the distal ileum were collected, frozen, and freeze-dried. The BEL were similar to the values found in the literature. At d 23, broilers fed the SE-SBM had the highest body weight gain and best FCR compared to chickens fed the HO-FF and NO-FF treatments (P < 0.001). Broilers fed the SE-SBM and NO-EE experimental diets had (P < 0.001) higher apparent ileal AA digestibility and AA SID than broilers fed the HO-FF and NO-FF treatments. In conclusion, the SID of AA from HO-FF is similar to the digestibilities of other full-fat soybeans found in the literature and is lower than that of NO-EE and SE-SBM.

Volatile Short-Chain Aliphatic Aldehydes Act as Taste Modulators through the Orally Expressed Calcium-Sensing Receptor CaSR.

Aldehydes are natural volatile aroma compounds generated by the Maillard reaction of sugars and amino acids in food and affect the flavor of food. They have been reported to exert taste-modifying effects, such as increases in taste intensity at concentrations below the odor detection threshold. The present study examined the taste-enhancing effects of short-chain aliphatic aldehydes, such as isovaleraldehyde (IVAH) and 2-methylbutyraldehyde, thus attempting to identify the taste receptors involved. The results obtained revealed that IVAH enhanced the taste intensity of taste solutions even under the condition of olfactory deprivation by a noseclip. Furthermore, IVAH activated the calcium-sensing receptor CaSR in vitro. Receptor assays on aldehyde analogues showed that C3-C6 aliphatic aldehydes and methional, a C4 sulfur aldehyde, activated CaSR. These aldehydes functioned as a positive allosteric modulator for CaSR. The relationship between the activation of CaSR and taste-modifying effects was investigated by a sensory evaluation. Taste-modifying effects were found to be dependent on the activation state of CaSR. Collectively, these results suggest that short-chain aliphatic aldehydes function as taste modulators that modify sensations by activating orally expressed CaSR. We propose that volatile aroma aldehydes may also partially contribute to the taste-modifying effect via the same molecular mechanism as kokumi substances.

Real time on-line amino acid analysis to explore amino acid trends and link to glycosylation outcomes of a model monoclonal antibody.

Bioreactor parameters can have significant effects on the quantity and quality of biotherapeutics. Monoclonal antibody products have one particularly important critical quality attribute being the distribution of product glycoforms. N-linked glycosylation affects the therapeutic properties of the antibody including effector function, immunogenicity, stability, and clearance rate. Our past work revealed that feeding different amino acids to bioreactors altered the productivity and glycan profiles. To facilitate real-time analysis of bioreactor parameters and the glycosylation of antibody products, we developed an on-line system to pull cell-free samples directly from the bioreactors, chemically process them, and deliver them to a chromatography-mass spectroscopy system for rapid identification and quantification. We were able to successfully monitor amino acid concentration on-line within multiple reactors, evaluate glycans off-line, and extract four principal components to assess the amino acid concentration and glycosylation profile relationship. We found that about a third of the variability in the glycosylation data can be predicted from the amino acid concentration. Additionally, we determined that the third and fourth principal component accounts for 72% of our model's predictive power, with the third component indicated to be positively correlated with latent metabolic processes related to galactosylation. Here we present our work on rapid online spent media amino acid analysis and use the determined trends to collate with glycan time progression, further elucidating the correlation between bioreactor parameters such as amino acid nutrient profiles, and product quality. We believe such approaches may be useful for maximizing efficiency and reducing production costs for biotherapeutics.

Combinatorial Protein Engineering and Metabolic Engineering for Efficient Synthesis of l-Histidine in Corynebacterium glutamicum.

l-Histidine is an essential proteinogenic amino acid in food with extensive applications in the pharmaceutical field. Herein, we constructed a Corynebacterium glutamicum recombinant strain for efficient biosynthesis of l-histidine. First, to alleviate the l-histidine feedback inhibition, the ATP phosphoribosyltransferase mutant HisGT235P-Y56M was constructed based on molecular docking and high-throughput screening, resulting in the accumulation of 0.83 g/L of l-histidine. Next, we overexpressed rate-limiting enzymes including HisGT235P-Y56M and PRPP synthetase and knocked out the pgi gene in the competing pathway, which increased the l-histidine production to 1.21 g/L. Furthermore, the energy status was optimized by decreasing the reactive oxygen species level and enhancing the supply of adenosine triphosphate, reaching a titer of 3.10 g/L in a shake flask. The final recombinant strain produced 5.07 g/L of l-histidine in a 3 L bioreactor, without the addition of antibiotics and chemical inducers. Overall, this study developed an efficient cell factory for l-histidine biosynthesis by combinatorial protein engineering and metabolic engineering.

External and Internal Qualities of Chicken Eggs Early Production at Various Storage Times at Room Temperature

Eggs are one of the consumed foods that contain many essential amino acid nutrients such as lysine, tryptophan, and methionine. Egg quality can be seen from the storability of eggs after they are produced. This study aims to evaluate the effect of various storage lengths on the external and internal quality of chicken egg consumption of the Isa Brown strain at the beginning of production at room temperature. The research method is a Laboratories experiment using a Complete Randomized Design. The study treatment consisted of egg storage duration for 0 days (P0), 7 days (P1), 14 days (P2), and 21 days (P3) with 4 repeats and each test unit of 4 eggs. The research variables include external qualities: egg weight, egg index, shell weight, and eggshell thickness, then internal qualities: egg white index, yolk index, yolk color, and egg pH. The data obtained in this study were analyzed with ANOVA, if there are differences between treatments, then proceed with the the Least Significant Differenttest. The results showed that the duration of deviation had a very noticeable influence (P&lt;0.01) on the external quality of the egg including:egg weight, weightand thickness of the eggshell, and no influence (P&gt;0.01) on the egg index. The length of storage also has a very noticeable influence on the internal quality of eggs (P&lt;0.01). Based on the results of the study, it can be concluded that the initial chicken eggs of production at a storage duration of up to 21 days at room temperature experience a decrease in external and internal qualities but still meet the Standards National Indonesia 3926:2008

Taurine and Creatine Transporters as Potential Drug Targets in Cancer Therapy.

Cancer cells are characterized by uncontrolled growth, proliferation, and impaired apoptosis. Tumour progression could be related to poor prognosis and due to this fact, researchers have been working on novel therapeutic strategies and antineoplastic agents. It is known that altered expression and function of solute carrier proteins from the SLC6 family could be associated with severe diseases, including cancers. These proteins were noticed to play important physiological roles through transferring nutrient amino acids, osmolytes, neurotransmitters, and ions, and many of them are necessary for survival of the cells. Herein, we present the potential role of taurine (SLC6A6) and creatine (SLC6A8) transporters in cancer development as well as therapeutic potential of their inhibitors. Experimental data indicate that overexpression of analyzed proteins could be connected with colon or breast cancers, which are the most common types of cancers. The pool of known inhibitors of these transporters is limited; however, one ligand of SLC6A8 protein is currently tested in the first phase of clinical trials. Therefore, we also highlight structural aspects useful for ligand development. In this review, we discuss SLC6A6 and SLC6A8 transporters as potential biological targets for anticancer agents.

Bifidobacterium animalis A12, a Probiotic Strain That Promotes Glucose and Lipid Metabolism, Improved the Texture and Aroma of the Fermented Sausage.

Bifidobacterium animalis A12 was used for the development of fermented sausage. The growth activity, tolerance, and enzyme activity of B. animalis A12 and its contribution to the texture and flavour of fermented sausages were evaluated. Additionally, the sensory texture, flavour components, and amino acid nutrients during the fermentation process were assessed. B. animalis had high tolerance to NaCl and nitrite, and B. animalis A12 had protease and lipase activities. The pH value of sausage fermented with B. animalis A12 was lower than that of sausage fermented without any fermentation strain. Hexanal, heptanal, decanal, cis-2-decanal, and 4-methoxy-benzaldehyde are the unique aldehydes flavour components of fermented sausages in the A12 group. The highest content of volatile flavour substances and amino acids, and the color and texture characteristics of fermented sausage in the experimental group at 18 h were better than those at other times. These results suggest that B. animalis A12 has the potential to be used as a starter culture for im-proving flavour and texture in fermented sausage.

Ninhydrin Loaded Microcapsules for Detection of Natural Free Amino Acid

Natural free amino acids present in plant extracts or tea infusions provide a unique flavor and potential effect on anxiety and blood pressure reduction. Accordingly, quantifying free amino acids in foods has been of interest to food science and analytical research fields. The ninhydrin solution-based assay is a colorimetric method based on the formation and detection of Ruhemann’s purple complex. Media-based colorimetric detection requires specialized facilities and personnel; moreover, it can suffer from the interference of the analyte color. In this study, we developed ninhydrin-loaded microcapsules and a simple free amino acids detection procedure, by simply dipping the microcapsules into the analyte solution for 3 min. Among the five tested natural free amino acids, theanine exhibited the highest colorimetric response to microcapsule-based detection, with a limit of detection of 0.826 mM.

Whole-Genome Resource of Lasiodiplodia pseudotheobromae BaA, the Causative Agent of Black Root Rot Morinda officinalis.

Whole-Genome Resource of Lasiodiplodia pseudotheobromae BaA, the Causative Agent of Black Root Rot Morinda officinalis.

Determination of amino acids in food and feed by microwave hydrolysis and UHPLC-MS/MS

The determination of amino acids in food and feed by chromatography has a long history and is described in several official methods, including standards from ISO, AOAC, and the European Commission (EC) regulation 152/2009. The procedure usually consists of labor- and time-consuming preparation techniques and ion-exchange chromatography with challenging chromatographic conditions. Consequently, several approaches have been published to overcome these drawbacks but the knowledge about their suitability for complex matrices such as food and feed is limited.In this paper, we describe the development of two new methods to determine amino acids in food and feed. These methods involve microwave hydrolysis and reversed-phase UHPLC-MS/MS with pre-column derivatization using 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC). Both methods provide streamlined sample preparations and a dramatic reduction in analysis time while offering a high degree of specificity and selectivity. Selectivity also enabled the simultaneous determination of the more uncommon substances hydroxyproline, hydroxylysine, taurine, ornithine, and γ-amino butyric acid (GABA) along with amino acids typically present in food and feed. The results were all satisfactory with regards to sensitivity, accuracy, precision, and comparability with laboratories that use other methods, for example from ISO, AOAC, or regulation (EC) 152/2009. We therefore concluded that both methods provide a reliable and modern approach to overcome many of the drawbacks that occur with the conventional standard methods.

Ablation of Tas1r1 Reduces Lipid Accumulation Through Reducing the de Novo Lipid Synthesis and Improving Lipid Catabolism in Mice.

Amino acid sensing plays an important role in regulating lipid metabolism by sensing amino acid nutrient disturbance. T1R1 (umami taste receptor, type 1, member 1) is a membrane G protein-coupled receptor that senses amino acids. Tas1r1-knockout (KO) mice were used to explore the function of umami receptors in lipid metabolism. Compared with wild-type (WT) mice, Tas1r1-KO mice showed decreased fat mass (P < 0.05) and adipocyte size, lower liver triglyceride (7.835 ± 0.809 vs 12.463 ± 0.916 mg/g WT, P = 0.013) and total cholesterol levels (0.542 ± 0.109 vs 1.472 ± 0.044 mmol/g WT, P < 0.001), and reduced lipogenesis gene expressions in adipose and liver tissues. Targeted liver amino acid metabolomics showed that the amino acid content of Tas1r1-KO mice was significantly decreased, which was consistent with the branched-chain ketoacid dehydrogenase protein levels. Proteomics analysis showed that the upregulated proteins were enriched in lipid and steroid metabolism pathways, and parallel reaction monitoring results illustrated that Tas1r1 ablation promoted lipid catabolism through oxysterol 7 α-hydroxylase and insulin-like growth factor binding protein 2. In summary, Tas1r1 disruption in mice could reduce lipid accumulation by reducing de novo lipid synthesis and improving lipid catabolism.

Advances in separation and analysis of aromatic amino acids in food

苯丙酮尿症等代谢病患者因基因缺陷无法正常代谢食物中的芳香族氨基酸(aromatic amino acids, AAA),常规饮食可能会造成永久性生理损伤,低AAA肽是其重要的蛋白质等同物来源。AAA分离分析技术对低AAA肽的制备和检测至关重要。该领域研究者探索了多种高效的吸附、分离材料,从复杂的蛋白质水解液中选择性吸附脱除AAA以制备符合特定氨基酸限量的低AAA肽类食品,或者建立对AAA特异性提取富集的分离分析策略。该文分析了AAA的结构特点与理化性质,总结了近年来基于活性炭、树脂等吸附材料脱除AAA的技术进展,并从样品前处理、手性分离和吸附-传感3个维度综述了二维纳米材料、分子印迹、环糊精、金属有机骨架等材料在AAA分离分析中的应用。通过探讨各类技术的优缺点,为AAA吸附脱除和分离分析技术的进步提供支撑。

Analysis of amino acids in food using High Performance Liquid Chromatography with derivatization techniques: a review

Amino acids (AAs) are essential components needed for human health. AAs are typically present as free or bound into protein backbones. Determination of AAs is important to evaluate the nutritional value of protein-containing foods and to authenticate food products from any adulteration practice. Due to its capability to provide the separation capacity, chromatographic-based methods are considered the method of choice for the analysis of AAs. The common detectors used for the analysis of food components are specific detectors including UV-Vis and fluorescence. Unfortunately, most AAs are nonchromophoric or fluorescence compounds, as a consequence, some derivatization processes are needed using derivatizing agents to make AAs detectable using UV-Vis or fluorescence detectors. This review highlighted the use of HPLC for the analysis of AAs in food products by applying pre-column and post-column derivatization techniques to be detectable by UV-Vis or fluorescence detectors.

New Insight on Vitality Differences for the Penaeid Shrimp, Fenneropenaeus chinensis, in Low Salinity Environment Through Transcriptomics

Excessive rainfall changes salinity in shrimp farming ponds in short period and exerts low salinity stress on the outdoor breeding shrimp under global warming. Fenneropenaeus chinensis can have different performance on vitality in low salinity environments. To reveal mechanisms of vitality difference in shrimp living in low saline environments. This study based on the normal and moribund F. chinensis in 10 ppt salinity environment using high-throughput sequencing identifies 1,429 differentially expressed genes (DEGs), 586 of which are upregulated, while 843 of which are downregulated in the normal group (FCN10) as compared to the moribund group (FCM10). Meanwhile, another transcriptomic analysis is conducted on the normal and moribund shrimp from 25 ppt (FCN25 vs. FCM25) salinity environment as the control, in which 1,311 DEGs (upregulated: 327 genes, downregulated: 984 genes) are identified. In this study, intersective pathways, GO (Gene Ontology) categories and DEGs from the two groups of comparative transcriptome are investigated. The two intersective pathways (Metabolism of xenobiotics by cytochrome P450, Pentose, and glucuronate interconversions) significantly enriched by DEGs are related to detoxification. In these two pathways, there is one vitality regulation-related gene (VRRG), the Dhdh (dihydrodiol dehydrogenase), which is upregulated in both the groups of FCN10 and FCN25 as compared to the groups of FCM10 and FCM25, respectively. Similarly, in the 25 top intersective GO categories, four VRRGs are revealed. Three of them are upregulated (Itgbl, kielin/chordin-like protein, Slc2a8, solute carrier family 2, facilitated glucose transporter member 8-like protein and Cyp3a30, cytochrome P450 3A30-like protein); one of them is downregulated (Slc6a9, sodium-dependent nutrient amino acid transporter 1-like protein isoform X2). These GO categories are related to transmembrane transporter activity of substance, enzyme inhibitor activity, monooxygenase activity. RT-qPCR analysis further verifies the VRRGs. The study gives new insight into understanding the vitality differences for F. chinensis, in low salinity environment. The pathways and DEGs in response to low salinity stress in modulating the vitality of F. chinensis that could serve as tools in future genetic studies and molecular breeding.

Determination of Amino Acids in Food and Feed by Microwave Hydrolysis and Uhplc-Ms/Ms

Determination of Amino Acids in Food and Feed by Microwave Hydrolysis and Uhplc-Ms/Ms

​Response of In ovo Supplementation of Amino Acid and Minerals on Egg Hatchability of Broiler Chicken

Background: In ovo supplementation of nutrients either amino acids or minerals in broiler chicken has been common more often individually or few nutrients with positive results. In view of the above, in ovo supplementation with combination of amino acids and minerals as a complete nutrient capsule for the gut and immune system development was tested for hatchability since the modern commercial broilers are very fragile to any changes in the internal environment. Methods: Four trials were conducted to study the response of in ovo supplementation of combinations of Lys, Met, Arg, Thr, Glu, Zn, Se and Cu on hatchability of broiler eggs. In trial I, the fertile eggs on day 18 were divided into five groups of 32 eggs in each group. Group I served as control without any amino acids and mineral supplementation. Group II, III, IV and V supplemented with Lys, Met, Arg, Thr, Glu, Zn, Se and Cu @ 2.2, 1, 2.5, 1.6, 2.5 mg, 80, 0.3, 16 µg; 4.4, 2, 5, 3.2, 5 mg, 80, 0.3, 16 µg; 11, 5, 12.5, 8, 12.5 mg, 80, 0.3, 16 µg and 22, 10, 25, 16, 25 mg, 80, 0.3, 18 µg respectively. In trial II, seventy fertile eggs were divided into seven groups, Group I served as control. Group II, III, IV, V, VI and VII were supplemented with graded levels (1x to 6x) of amino acids. The 1x consisted of Lys, Met, Arg, Thr and Glu @ 2.2, 1, 2.5, 1.6 and 2.5 mg, respectively. On day 18, 10 eggs per group were supplemented with graded levels of AA combination into the amniotic fluid. In trail III and IV it was similar to trial II but was without Lys and Met, respectively. Result: Results indicated that hatchability (Trial 1) in Group V, IV, III, II and I were 0, 18, 21.8, 71 and 90.6%, respectively. The Combination of five amino acids (Trial II) (Lys, Met, Thr, Arg, Glu) for in ovo supplementation reduced the hatchability, whereas, combination of four amino acids (Trial III) (Met, Thr, Arg, Glu) without Lys at lower doses resulted in good hatchability (86.6%). In ovo supplementation of amino acid or mineral-amino acid combinations in commercial broiler eggs should be restricted to few amino acids.

Studies of the content of optical isomers of amino acids in food

Food products undergo a wide range of chemical changes during their processing and storage. As a result of such reactions, both new chemical compounds and optical isomerization of compounds already present in the composition can be formed. The second case concerns the formation of D-enantiomers of amino acids from their L-forms. D-forms of amino acids not only have no biological value for the body, but also often have a negative effect on the human body due to the impossibility of metabolizing them and, as a consequence, their accumulation in the body. The aim of the work was to study the quantitative content of D-isomers of amino acids in milk that passed the ultra-pasteurization process and dairy products based on bacterial starter culture. The research results showed that in both cases of the considered technological methods, amino acid isomerization occurs. The highest degree of isomerization was observed in kefir samples relative to other samples. However, from the results obtained, it is not possible to estimate which amino acid is most susceptible to the racemization process, since different samples contained different D-isomers of amino acids. The smallest amount of D-isomers is found in milk that has not undergone any industrial processing. Studies have shown that technological processing of milk inevitably leads to the formation of D-isomers of amino acids, and this, in turn, at least reduces the nutritional and biological value of the product, which makes it necessary to conduct deeper studies in this direction to establish the most important factors in the process of racemization of amino acids in food products.