Amino Acid Composition Research Articles

Putative MutS2 Homologs in Algae: More Goods in Shopping Bag?

MutS2 proteins are presumably involved in either control of recombination or translation quality control in bacteria. MutS2 homologs have been found in plants and some algae; however, their actual diversity in eukaryotes remains unknown. We found putative MutS2 homologs in various species of photosynthetic eukaryotes and performed a detailed analysis of the revealed amino acid sequences. Three groups of homologs were distinguished depending on their domain composition: MutS2 homologs with full set of specific domains, MutS2-like sequences without endonuclease Smr domain, and MutS2-like homologs lacking Smr and clamp in domain IV, the extreme form of which are proteins with only a complete ATPase domain. We clarified the information about amino acid composition and set of specific motifs in the conserved domains in MutS2 and MutS2-like sequences. The models of the predicted tertiary structure were obtained for each group of homologs. The phylogenetic analysis demonstrated that all eukaryotic sequences split into two large groups. The first group included homologs belonging to species of Archaeplastida and a subset of haptophyte homologs, while the second-sequences of organisms from CASH groups (cryptophytes, alveolates, stramenopiles, haptophytes) and chlorarachniophytes. The cyanobacterial MutS2 clustered together with the first group, and proteins belonging to Deltaproteobacteria (orders Myxococcales and Bradymonadales) showed phylogenetic affinity to the CASH-including group with strong support. The observed tree pattern did not support a clear differentiation of eukaryotes into lineages with red and green algae-derived plastids. The results are discussed in the context of current conceptions of serial endosymbioses and genetic mosaicism in algae with complex plastids.

B-165 The EXENT System reliably distinguishes daratumumab from endogenous monoclonal immunoglobulins in multiple myeloma patients

Abstract Background The use of therapeutic monoclonal antibodies (t-mAbs) in the treatment of multiple myeloma (MM) has created challenges in the clinical laboratory. Co-migration of t-mAbs (predominantly IgGK) and the endogenous M-protein during serum protein electrophoresis can result in inability to distinguish between a patient’s M protein and the t-mAb. This can hinder accurate assessment of therapeutic responses or may lead to extensive workup with immunofixation electrophoresis and gel-shift assays for accurate confirmation of a complete response, or to exclude the emergence of a new clone. There is a need for an analytically specific routine method to identify t-mAb interference. MALDI-TOF mass spectrometry is an especially promising tool, as due to its ability to identify M proteins’ molecular mass, it has the potential to recognize any t-mAbs with known amino acid composition. We have used the EXENT® Immunoglobulin Isotypes (GAM) Assay for the EXENT Analyzer (The Binding Site, part of Thermo Fisher Scientific) to assess the reproducibility of daratumumab (anti-CD38, IgGK) detection in an analytical study, and assessed the EXENT System’s ability to detect daratumumab in a cohort of MM patients. Methods The expected mass to charge ratio (m/z) of the +2-charge state of daratumumab’s kappa light chain was calculated based on published information. Daratumumab was spiked into normal human serum at 1.0 g/L, and this material was tested in 5 replicates a day over 5 days. In addition, 109 samples from 34 MM patients receiving daratumumab, carfilzomib, lenalidomide and dexamethasone (Dara-KRd) treatment were also tested. Samples were obtained after 14 or 28 days following daratumumab administration in various treatment cycles, ranging from cycle 4 to end-of-treatment. Results The expected m/z for the +2-charge state of daratumumab’s kappa light chain was calculated as 11,691.01 to 11,693.1. In the reproducibility experiment, the median m/z was 11691.3, within the expected range. The difference between the lowest and highest m/z was 1.3 across the 25 replicates. Daratumumab was detected in 103 out of 109 samples (97.3%): 51/57 (89.5%) in IgGK samples, and in all IgGL (23) and IgA (29) samples. In two IgGK patients, where daratumumab was not detected in 2 out of 4 samples and in 1 out of 3 samples, respectively, the patients’ own M proteins were in the proximity (24.8 and 27.1 m/z) of daratumumab’s m/z. In the other three cases where daratumumab was not modeled by the algorithm, the daratumumab peak overlapped with the endogenous M-protein's adduct peak (estimated m/z 11,688) in one sample; it was masked by the shoulder of a large (8.42 g/L) IgGK M-protein in another sample; and no obvious reason was identified in the third sample. There was no difference in the detection rate between 14 days and 28 days post administration. Conclusions The EXENT Immunoglobulin Isotypes (GAM) Assay has successfully identified the presence of daratumumab in approximately 90% of IgGK patient samples in addition to all other (non-IgGK) MM samples. These results demonstrate the potential of the EXENT Immunoglobulin Isotypes (GAM) Assay for recognizing known t-mAb interference.

The Change Rate of the Fbxl21 Gene and the Amino Acid Composition of Its Protein Correlate with the Species-Specific Lifespan in Placental Mammals

This article proposes a methodology for establishing a relationship between the change rate of a given gene (relative to a given taxon) together with the amino acid composition of the proteins encoded by this gene and the traits of the species containing this gene. The methodology is illustrated based on the mammalian genes responsible for regulating the circadian rhythms that underlie a number of human disorders, particularly those associated with aging. The methods used are statistical and bioinformatic ones. A systematic search for orthologues, pseudogenes, and gene losses was performed using our previously developed methods. It is demonstrated that the least conserved Fbxl21 gene in the Euarchontoglires superorder exhibits a statistically significant connection of genomic characteristics (the median of dN/dS for a gene relative to all the other orthologous genes of a taxon, as well as the preference or avoidance of certain amino acids in its protein) with species-specific lifespan and body weight. In contrast, no such connection is observed for Fbxl21 in the Laurasiatheria superorder. This study goes beyond the protein-coding genes, since the accumulation of amino acid substitutions in the course of evolution leads to pseudogenization and even gene loss, although the relationship between the genomic characteristics and the species traits is still preserved. The proposed methodology is illustrated using the examples of circadian rhythm genes and proteins in placental mammals, e.g., longevity is connected with the rate of Fbxl21 gene change, pseudogenization or gene loss, and specific amino acid substitutions (e.g., asparagine at the 19th position of the CRY-binding domain) in the protein encoded by this gene.

Ion Exchange Chromatography: A Versatile Technique for Comprehensive Characterization of Therapeutic Proteins and Peptides

Ion Exchange Chromatography (IEX) is an efficient and widely employed technique for the comprehensive characterization of therapeutic proteins and peptides. IEX separates biomolecules based on differences in their surface charge arising from amino acid composition and Post- Translational Modifications (PTMs). As a non-denaturing method, IEX is particularly valuable for resolving and quantifying charge variants like deamidation, oxidation, glycosylation, and terminal truncations in monoclonal antibodies, antibody-drug conjugates, and other biotherapeutics. The review provides insights into the principles, advantages, applications, recent advances, and limitations of using IEX (cation and anion exchange modes) for detailed analysis of proteins and peptides.

Biological active compounds from edible mushroom: Effect on hela cells

Background: Mushrooms are unique organisms rich in proteins, carbohydrates, vitamins, and minerals, which allows them to be used not only as a source of nutrition but also in cosmetics and medicine. The fungus P. ostreatus possesses two strong enzymatic complexes – lignin-decaying and cellulose-decaying. Objective: The research aimed to study the antitumor activity of treated mm-wave mycelial extracts of mushrooms on the HeLa cell line. Research on edible mushrooms against tumors is very actual and important for the field of functional food science and bioactive compounds field. Methods: The effect of the fungal culture extract on the cyto-photometric and morphometric indices of the HeLa cell line culture was studied according to the nature of proliferation and transcription of malignant tissues. Using high-resolution liquid chromatography, the amino acid composition of acid-soluble proteins in mycelial extracts of a mushroom culture treated with mm waves was studied. Results: Extracts of P. ostreatus mycelial culture significantly suppressed the growth of proliferative activity of human tumor cultures and completely suppressed the mitotic activity of continuous HeLa cell cultures to 72 hours. Our study revealed a significant increase in glutamic and aspartic acids in mycelial culture extracts treated by mm-waves. Conclusion: Based on HPLC analysis, we suggest polyfunctional peroxidase as the candidate for the role of biologically active compound in the composition of aqueous extracts of the mycelial culture. So, the polyfunctional peroxidase plays an active role in the therapeutic properties of the oyster mushroom. Keywords: anticancer activity, ant-inflammation activity, mm-waves, mushroom`s, mycelial extracts, HPLC of acid-soluble proteins, HeLa cell line.

Nutritional properties of commercially important cephalopods from the south-west coast of India

Cephalopods are widely regarded as healthy and nutritionally rich seafood resources. This study focuses on analysing the nutritional composition of six commercially important edible cephalopod species viz., Cistopus indicus, Amphioctopus marginatus, Amphioctopus aegina, Sepia pharaonis, Sepia elliptica and Sepia aculeata, from the Malabar coast of the Indian peninsula. The selected cephalopods were determined to be a rich source of protein ranging from 12.41 to 19.02 g 100 g-1. The amino acid profile demonstrated a balanced ratio of essential to non-essential amino acids (0.93-1.63) with a higher concentration of essential amino acids, attributing enhanced biological value of proteins in these cephalopods. S. pharaonis recorded the highest arginine-to-lysine ratio (~1.77), indicating excellent cholesterolemic index. Notably, octopus species had the lowest total cholesterol content (~100 mg 100 g-1) compared to cuttlefish species which recorded higher cholesterol levels (~160 mg 100 g-1). A. marginatus recorded more significant quantities of polyunsaturated fatty acids (PUFA) (36.85% total fatty acids) among all the species studied. Octopus species exhibited significantly greater Σn-3/Σn-6 ratio than cuttlefish species. The ideal atherogenicity/thrombogenicity indices (<1.0) in the edible tissues of the studied cephalopods qualify these species as potentially healthy food. A. aegina was found to exhibit greater vitamin E (α-tocopherol) contents (17.84 IU), whereas ascorbic acid was pronounced more in A. marginatus (47.06 μg 100 g-1). The higher vitamin D3 content in A. aegina (96.23 μg 100 g-1) among the species studied, highlights its potential role in preventing osteoporosis in adults. Among microelements such as iron and zinc were found in significant quantities in the cephalopod species studied. The findings from the current study provide valuable insights into the nutritional benefits of these species, positioning them as promising candidates for the seafood industry as possible healthy dietary options. Keywords: Amino acid composition, Atherogenicity index, Cephalopods, Fatty acid composition, Thrombogenicity index

Biochemical composition of biological substances extracted from red deer antlers

We aim to study biologically active agents in biological substances obtained from different parts of red deer antlers. Frozen red deer antlers were separated into skin and antler base, crushed, and subjected to ultrasonic enzymatic hydrolysis. The press cake from the extraction of antler base was dried in a vacuum unit. The content of protein, fat, ash, and moisture in the obtained biological substances was studied. In addition, the amino acid, fatty acid, and mineral compositions were analyzed, as well as the molecular weight distribution of peptide fractions. The concentrates from the skin and base of red deer antlers were found to contain the maximum of proteins (73.68–74.41%). The mineral biological substances comprise the maximum of macroand microelements (84.17%), in particular, a high level of 136,000 mg /kg of phosphorus, 29,230 mg /kg of sodium, and 5,140 mg /kg of calcium. All biological substances are characterized by a low-fat content in the range from 0.3 to 1.76%. Similar to the protein content, the total amount of amino acids prevails in concentrates from the skin and base of red deer antlers, equaling 40.90 and 43.66 g /100 g, respectively. Moreover, in both samples, the amount of essential amino acids is higher than that of non-essential ones. The amount of vitamins is low, being represented by B1, B2, B6 water-soluble vitamins. The fatty acid composition is most balanced in the concentrate from the red deer antler base, containing omega-3 and omega-6. The ratio of unsaturated to saturated fatty acids is equal to 0.96. In the concentrate from antler skin, the maximum content (54.7%) of peptides with a low molecular weight of less than 2.9 kDa and up to 6.4 kDa was found using the molecular-weight distribution of peptide fractions.

Study of the jellyfish Rhizostoma pulmo hydrolysis and characterization of the obtained product

The purpose of this work is to develop technological parameters for the hydrolysis of barrel jellyfish Rhizostoma pulmo involving the use of enzymatic agents «Enzy-mix» and alcalase, available through the domestic market, as well as to investigate the amino acid and mineral composition of the resultant hydrolysate.Methods used: the process of hydrolysis has been assessed based on the changes in peptide concentrations in the hydrolysate, the content of amino nitrogen, and the degree of hydrolysis. To investigate its chemical, amino acid, and mineral composition, the standard methods were used. Novelty: the use of jellyfish for hydrolysis after its thawing and the hydrolysis procedure not involving the addition of water into the raw material subjected to hydrolysis present a novel approach.Result: the optimal regime for hydrolysis of barrel jellyfish Rh. pulmo after its thawing has been identified, which was characterized by the use of 1% alcalase at the temperature 55±2 °C and duration of 1 h. It has been shown that alcalase is more effective in terms of amino nitrogen accumulation than enzymatic agent «Enzy-mix». The values of amino nitrogen after 1 h of hydrolysis involving the use of 1% alcalase were higher than the values of the same parameter obtained during the 3-hour hydrolysis involving 1.5% «Enzy-mix» at the temperature 40±2 °C. The hydrolysate, after its concentration to the dry matter exceeding 30%, contains no less than 15.1±0.59% of protein represented by 48% of essential amino acids, out of which leucine and isoleucine prevail (15 %). Among non-essential amino acids, the most prominent was glycine (17 %). For all essential amino acids, except for valine, the amino acid score exceeds 100%. In humans, consumption of around 20 g of the hydrolysate covers the daily physiological requirement of magnesium.Practical significance: the obtained data on the amino acid profile and mineral composition of the barrel jellyfish Rh. pulmo hydrolysate provide the basis for its consideration as a promising ingredient for the functional nutrition products.

Genomic insights into codon usage bias in Cannabis sativa and pathogenic interactions

This study delves into codon usage bias (CUB) across Cannabis sativa and its associated pathogens (Meloidogyne incognita, Alternaria alternata, Aspergillus flavus, Aspergillus niger, and Fusarium oxysporum). It finds an AT-bias in C. sativa and M. incognita, favouring AT ending codons, while fungal pathogens lean towards GC-ending codons. Analysis suggests weak codon bias overall, with host and nematode showing similar preferences, and fungal pathogens preferring GC-ending codons. In ENC/GC3 plot, parity plot analyses both natural selection and mutational pressure influence CUB, slightly favouring T-ending codons.Neutrality plot analysis demonstrated varying degrees of mutational pressure and selection across the organisms. Correspondence analysis revealed indistinguishable CUB effective in all pathogens, despite differences in evolutionary forces. Correlation analysis illustrated the impact of nucleotide composition, mutational pressure, and natural selection on CUB. Additionally, amino acid composition analysis indicated differential usage of amino acids across organisms. Codon context analysis unveiled common trends in codon pairing, emphasizing the prevalence of preferred codons in C. sativa and M. incognita. This comprehensive analysis provides insights into the factors shaping CUB and the evolutionary forces influencing the genomes of the host plant and its associated pathogens, contributing to a deeper understanding of molecular interactions in host-pathogen systems. This data informs conservation for Cannabis sativa by identifying pathogen-resistant genetic traits, guiding breeding and management for biodiversity preservation.

Variability of a number of Pm gene sequences in representatives of Triticeae

Aim. Comparison of the sequences of selected resistance genes deposited in GenBank to find types of differences in their nucleotide composition and to determine the significance of these differences for primer design. Methods. Sequence searches were performed in the GenBank biological database. Comparison of different sequences of the same gene was carried out in DnaSP6 by calculating the Tajima test for Pm2, Pm3 and the Kolmogorov-Smirnov test in GeneDoc for Pm21. Multiple alignment was performed in CLUSTALW before using the specified programs. Primers were designed in Primer3. Results. Variability of the compared sequences is represented by nucleotide substitutions, insertions/deletions, the presence of a transposon in one of the Pm3 sequences. The regions coding for leucine-rich repeats are the most variable. The sequences isolated from Triticum dicoccoides have two such regions, within each of which some of the changes in the nucleotide sequence will be reflected in the amino acid composition of the proteins. Conclusions. The differences between the compared sequences of Pm2, Pm3, Pm21 (within each of the genes) are random (p < 0.05). According to the results of the sequences analysis, combinations of primers were developed for the identification of sequences characteristic to resistance genes on the available plant material.

Prediction of protein-ligand binding sites modulating activity of MAST protein kinases

Aim. Identification of the protein-ligand binding sites, that may be the target of compounds, affecting individual human protein kinases of the MAST family (MAST1, 2, 3, 4 and MASTL / GWL). Methods. Literature and database search. Comparison of protein and ligand structures. Protein structure modeling, structural superimposition, etc. Results. The structural alignment demonstrates significant similarity of catalytic domains in MAST1, 2, 3, 4 and MASTL (GWL). It justifies transferring of reference ligands from PDB structures to human MASTs, discovering potential sites of ligand binding. 13 sites of ligand-binding were specified based on refrence ligands, transferred from RCSB Protein Data Bank structures, and differences in sites amino acid composition of MAST family members were discovered. Сonclusions. Based on the differences in the amino acid composition of the studied pockets in MAST1, 2, 3, 4 and MASTL (GWL), the sites B, C, D, E, F, were selected for further study and virtual screening for new selective inhibitors of individual members of MAST protein kinase family.

Targeted metabolomics of muscle amino acid profles and hepatic transcriptomics analyses in grass carp (Ctenopharyngodon idellus) fed with broad beans

While tissue amino acid compositions reflect that of the dietary protein source, and the liver orchestrates amino acid metabolism. In this study, we investigated the muscle amino acid profiles in ordinary and crisp grass carp. The 22 amino acids were measured, and seventeen showed significant concentration differences. To understand the molecular mechanisms behind changes, we analyzed the liver transcriptome, and the 2519 differentially expressed genes (DEGs) were identified, with 1156 up-regulated and 1363 down-regulated genes. DEGs were enriched in ribosome-related biological processes. KEGG pathway analysis showed enrichment in tryptophan metabolism, lysine degradation, valine, leucine and isoleucine degradation, galactose metabolism, and glutathione metabolism with up-regulated genes, arginine and proline metabolism, arginine biosynthesis and alanine, aspartate, amino sugar and nucleotide sugar metabolism, N-Glycan biosynthesis and glutamate metabolism with down-regulated genes. A protein-protein interaction network with 260 nodes and 249 edges was constructed, and 3 modules were extracted. The top 10 hub genes with close connections to other nodes were ITM1, STT3B, SEL1L, UGGT1, MLEC, IL1B, ALG5, KRTCAP2, NFKB2, and IRAK3. In summary, this study identified candidate genes and focused on amino acid and glycan metabolism pathways, providing a reference for further investigation into liver amino acid metabolism in grass carp fed with broad beans.

Maternal dietary protein and amino acid intake is not associated with the amino acid composition of human milk in an affluent environment

Abstract Amino acids (AA) are essential nutrients in human milk (HM) and critical for infant growth and development. Several maternal lifestyle factors have been suggested to influence HM AA composition, with possible consequences for the breastfed infant. Whether maternal dietary protein and AA intake is associated with AA concentrations in HM is still largely unknown. Therefore, the aim of this study was to investigate the association between maternal dietary AA intake and AA concentrations in HM over the first month postpartum. Data from the observational longitudinal Amsterdam Mother’s Milk study were used, consisting of 123 lactating women in their first postpartum month. HM samples were collected three times, on day 10, 17 and 24 postpartum. Maternal dietary protein and AA intake on these collection days was assessed using three 24-h recalls. HM protein-bound and free AA (BAA and FAA, respectively) were analysed by liquid chromatography. Associations between maternal AA intake and AA concentrations in HM were assessed using linear mixed models. Maternal intake was negatively associated with milk concentrations of free arginine (–0·0003; P = 0·01) and free lysine (–0·0004; P = 0·03) and was positively associated with free glutamine (0·002; P = 0·03) and free threonine (0·0008; P = 0·03). However, these associations were attenuated after correction for multiple testing. Both the quality and quantity of dietary protein intake in lactating women do not seem to influence the amino composition of their breast milk when living in an affluent environment.

PeptiHub: a curated repository of precisely annotated cancer-related peptides with advanced utilities for peptide exploration and discovery.

Peptihub (https://bioinformaticscollege.ir/peptihub/) is a meticulously curated repository of cancer-related peptides (CRPs) that have been documented in scientific literature. A diverse collection of CRPs is included in the PeptiHub, showcasing a spectrum of effects and activities. While some peptides demonstrated significant anticancer efficacy, others exhibited no discernible impact, and some even possessed alternative non-drug functionalities, including drug carrier or carcinogenic attributes. Presently, Peptihub houses 874 CRPs, subjected to evaluation across 10 distinct organism categories, 26 organs, and 438 cell lines. Each entry in the database is accompanied by easily accessible 3D conformations, obtained either experimentally or through predictive methodology. Users are provided with three search frameworks offering basic, advanced, and BLAST sequence search options. Furthermore, precise annotations of peptides enable users to explore CRPs based on their specific activities (anticancer, no effect, insignificant effect, carcinogen, and others) and their effectiveness (rate and IC50) under cancer conditions, specifically within individual organs. This unique property facilitates the construction of robust training and testing datasets. Additionally, PeptiHub offers 1141 features with the convenience of selecting the most pertinent features to address their specific research questions. Features include aaindex1 (in six main subcategories: alpha propensities, beta propensity, composition indices, hydrophobicity, physicochemical properties, and other properties), amino acid composition (Amino acid Composition and Dipeptide Composition), and Grouped Amino Acid Composition (Grouped amino acid composition, Grouped dipeptide composition, and Conjoint triad) categories. These utilities not only speed up machine learning-based peptide design but also facilitate peptide classification. Database URL: https://bioinformaticscollege.ir/peptihub/.

Amino Acid Composition Determination From the Fractional Mass of Peptides

ABSTRACTA peptide's fractional mass is directly associated with its elemental composition and thus amino acid composition. Here it is demonstrated that a peptide's fractional mass alone can be a useful identifier or indicator of that composition for small to mid‐sized peptides (5–7 amino acids) and can significantly reduce the number of viable amino acid compositions for larger peptides (> 8 residues) to include or exclude certain possibilities. Separate consideration of the integer portion of the peptide's mass helps to reduce the number of possibilities where many duplicate fractional mass values are found. Adoption of this fractional mass strategy should aid approaches that are presently employed for peptide identification, including in the use of mass map data to search protein databases for proteomics applications.

Effects of fermented soybean meal and supplemental methionine and lysine on growth, digestibility, body composition and amino acid composition of Beluga (Huso huso)

Abstract There is an increasing concern about using limited high-quality protein sources in the aquafeed industry. The alternative solution to this problem is cost-effective plant proteins such as soybean meal (SBM). However, it is better to improve plant protein-containing diets through processing and supplemental amino acids. This study aimed to examine the effects of fermented soybean meal (FSBM) and supplemental methionine and lysine (ML) in diets on growth, digestibility, and body and amino acid composition in Beluga (Huso huso). Two basal diets (490 g fish meal Kg−1) of fish meal (FM) and fish meal with methionine and lysine (FM+ML) were replaced by 40%, 60% and 80% of FSBM and FSBM+ML (FM, FSBM40, FSBM60, FSBM80, FM+ML, FSBM40+ML, FSBM60+ML and FSBM80+ML; 444 g kg−1 crude protein and 19.77 MJ kg−1 gross energy). The diets were fed to triplicate groups of Beluga (394.6 ± 4.3 g) 3 times daily for 56 days. Results showed that Beluga fed 80% FSBM and FSBM+ML had lower growth and feed utilization. Moreover, Beluga fed FSBM+ML performed poorer than those fed FSBM (P < 0.05). No interactions between fermented SBM and ML were observed in apparent digestibility coefficients (ADC) and body proximate composition except for ADC of crude lipid. ADC in Beluga fed 80% were less than other diets. Substitution of FSBM and FSBM+ML in diets significantly reduced dry matter, ash and crude lipid; Crude lipid increased in groups fed FSBM+ML (P < 0.05). FSBM and FSBM+ML in 60% had higher and lower total amino acids in muscle and faeces (P < 0.05), respectively. Also, excreted total amino acids in faeces increased with ML (P < 0.05). Generally, in Beluga, FSBM-containing diets showed better effects on growth, feed utilization, whole-body lipid, and muscle and faecal amino acids than those with FSBM+ML. Fermented SBM could be replaced up to 60%.

Some aspects of the process of implementing circular economy principles in the dairy industry of Kyrgyzstan

The introduction of resource-saving milk processing technology based on the principles of a “green” economy is very significant and relevant, because at the same time, environmental protection is ensured, all the most valuable components of raw milk are used for food purposes, which will improve the health and material well-being of various categories of the population. Rational processing of non-traditional, environmentally friendly raw material – cheese whey from kyrgyz khaynak milk – is proposed. For the first time, it has been proven that it is a raw material on the basis of which a wide range of target products can be prepared with natural additives that greatly increase their functionality. To confirm the high biological value of whey proteins isolated from khainak milk, their amino acid composition was analyzed by capillary electrophoresis in the Research laboratory of the All-Russian Research Institute of the Dairy Industry (Moscow). The results of the analysis were the basis for the development of a series of new functional products, the recipe and technology of which are protected by patents of the Kyrgyz Republic.

Studying the protein complex of flax cake obtained from the seeds of a new variety of kazakh selection

In the Republic of Kazakhstan, pressing of flax seeds (squeezing) is most often used, therefore cake is more common than meal and its resulting physical reserves exceed meal several times. The biochemical characteristics of the protein complex of flaxseed have not been sufficiently studied, which, in our opinion, limits the use of protein products obtained from flax seeds such as flaxseed cake in the production of compound feed for farm animals and poultry. The article presents studies of the protein complex and amino acid composition of flaxseed cake of the Kazakhstan variety “Kostanay 11”. Flaxseed cake contains 36,5% protein, the fat content is 7,5%. The fractional composition of flaxseed cake was compared with the flax varieties “Northern” and “Golden” grown in the Republic of Kazakhstan. The amino acid composition of flaxseed cake was compared with other cakes used in feeding farm animals and poultry. The purpose of the study is to study the fractional composition of the protein complex of flaxseed cake from flax variety "Kostanay 11". A characteristic feature of the protein complex of flax seeds is the high total content of water- and salt-soluble proteins – albumins 29,2% and globulins 16,8%, which are considered well digestible. Flaxseed cake from flax variety "Kostanay 11" in its amino acid value is equal to the known cakes common in feed production and is second only to soybean cake. This allows us to assert that flaxseed cake obtained from the seeds of the “Kostanay 11” variety remained a highly digestible and highly nutritious product in terms of protein absorption, suitable for its inclusion in the diets of all types of farm animals and poultry, starting with feed for young animals.

Utility of Arabidopsis KASII Promoter in Development of an Effective CRISPR/Cas9 System for Soybean Genome Editing and Its Application in Engineering of Soybean Seeds Producing Super-High Oleic and Low Saturated Oils.

This study reports the use of the Arabidopsis KASII promoter (AtKASII) to develop an efficient CRISPR/Cas9 system for soybean genome editing. When this promoter was paired with Arabidopsis U6 promoters to drive Cas9 and single guide RNA expression, respectively, simultaneous editing of the three fatty acid desaturase genes GmFAD2-1A, GmFAD2-1B, and GmFAD3A occurred in more than 60% of transgenic soybean lines at T2 generation, and all the triple mutants possessed desirable high-oleic traits. In sharp contrast, not a single line underwent simultaneous editing of the three target genes when AtKASII was replaced by the widely used AtEC1.2 promoter. Furthermore, our study showed that the stable and inheritable mutations in the high-oleic lines did not alter the overall contents of oil and protein or amino acid composition while increasing the oleic acid content up to 87.6% from approximately 23.8% for wild-type seeds, concomitant with 34.4- and 3.7-fold reductions in linoleic and linolenic acid, respectively. Collectively, this study demonstrates that the AtKASII promoter is highly promising for optimization of the CRISPR/Cas9 system for genome editing in soybean and possibly beyond.

Effect of Low-Proportion Replacement of Dietary Fishmeal with Neanthes japonica Meal on Growth Performance, Body Composition, Muscle Texture, Serum Biochemistry, Digestive Enzymes and Gene Expression in Juvenile Tiger Puffer Takifugu rubripes

The polychaeta Neanthes japonica is a common by-product in mariculture ponds. It is rich in essential nutrients, but has not been well-explored. Therefore, this 56-day experiment investigated the effects of substituting N. japonica meal for dietary fishmeal on juvenile tiger puffer (15.49 ± 0.02 g, n = 450 fish). The control diet (CON) contained 40% fishmeal. Freeze-dried N. japonica meal (FNM) was supplemented into CON at the levels of 3% (FNM3), 6% (FNM6), and 9% (FNM9), replacing fishmeal. The fifth diet contained 6% oven-dried N. japonica meal (ONM6), replacing fishmeal. The results indicated that no significant difference was observed in growth, feed efficiency, and somatic index among all the treatment groups. The feed intake of the FNM6 group was significantly higher compared to CON. No significant difference was detected in fish proximate composition, as well as the fatty acid composition, amino acid composition, and muscle texture. The supplementation of N. japonica meal decreased the activities of intestinal lipase and α-amylase. The addition of freeze-dried N. japonica meal significantly up-regulated the expression of the intestinal amino-glycine transporter pat1. It was concluded that adding 9% N. japonica meal to the feed had no significant effect on the growth performance and body composition of juvenile tiger puffer.