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Objective This study aimed to find global mechanisms related to carnosine synthesis in slow-growing Korat chickens (KRC) using a proteomic approach. Methods M. pectoralis major samples were collected from 10-week-old female KRC including low-carnosine (LC, 2,756.6±82.88 μg/g; n = 5) and high-carnosine (HC, 4,212.5 ±82.88 μg/g; n = 5). Results We identified 152 common proteins, and 8 of these proteins showed differential expression between the LC and HC groups (p

Wide-ranging bioactivities of enzymatically digested insect protein to produce peptides have been targeted for functional food development. In this study, fractionated peptides obtained from cricket (Acheta domesticus) protein hydrolysate by alcalase digestion were identified and evaluated for their bioactivities. Peptide fractions F44, F45, and F46, isolated through size exclusion chromatography, demonstrated strong cytoprotective effects on SH-SY5Y and HepG2 cells exposed to H2O2. This was evidenced by a 2-fold decrease in reactive oxygen species (ROS) accumulation in the cells and a 3-fold upregulation of genes encoding antioxidant enzymes. The F45 peptide fractions also showed chemical antioxidant activities ranging from approximately 290 to 393 mg trolox/g peptide, measured by DPPH, ABTS, and FRAP assays. Furthermore, F45 demonstrated the highest angiotensin-converting enzyme I (ACE) inhibitory activity, 57.93 %. F45 induced higher levels of Nrf2, SOD1, SOD2, CAT, GSR, and GPx4 gene expression in SH-SY5Y and HepG2 cells compared to cells treated with H2O2 and no peptides (p

ABSTRACT: This study aimed to differentiate the flavor characteristics of raw chicken breast meat from Thai slow-growing breeds (NC: native chicken, and KC: Korat/crossbred chicken) and fast-growing broilers (BR: broiler chicken) by using NMR-based metabolomic approaches along with multivariate data analysis. Chemical compounds related to chicken's flavor including free amino acids (FAA), ATP and its related compounds, sugars, as well as volatile compounds (VOC), were also investigated. BR had the highest total FAAs, followed by NC and KC (P < 0.05). In contrast, the accumulations of ATP degradation products, particularly ADP and IMP, were found at higher levels in the NC and KC (P < 0.05), while the highest total reducing sugars were noted in the KC (P < 0.05). Most VOCs found in the fresh breasts were products from the degradation of lipids, especially through lipid oxidation, which was found in varied types and proportions among samples. Not only chemical compounds but varying amounts of metabolites among samples were also detected. Apart from 21 identified metabolites, Glu, Gln, and betaine were the most prevalent in all samples with VIP > 1.00. Among 19 metabolic pathways, the most important pathways (P-value < 0.05, FDR < 0.05, impact > 0.05) were discovered to differentiate the flavor of raw chicken breast meat from various breeds. These metabolic pathways included (1) Ala, Asp and Glu metabolism; (2) D-Gln and D-Glu metabolism; (3) Purine metabolism; (4) β-Ala metabolism; (5) Aminoacyl-tRNA biosynthesis; (6) Nicotinate and nicotinamide metabolism; (7) Pyrimidine metabolism. Interestingly, based on the principal component analysis plot and partial least square-discriminant analysis (R2 = 0.9804; Q2 = 0.9782), NC and KC were clustered in the same area and discriminated from BR, indicating their similar flavor characteristics and metabolic profiles. Therefore, the findings could comprehend and distinguish the flavor of chicken breast meat of slow- from fast-growing chicken breeds based on their chemical characteristics and metabolite profiles.

ABSTRACT: Physico-chemical behaviors of natural actomyosin (NAM) from slow-growing Korat chicken (KC) breast and thigh at low (0.2 M) and high (0.6 M) NaCl concentrations were evaluated and compared to those from their corresponding muscles in fast-growing broiler chicken (BC). NAM from KC breast and thigh meat showed higher solubility than their corresponding in BC (p < 0.05). Breast NAM from both chickens showed the highest solubility at 0.4 M NaCl, while thigh NAMs showed the highest solubility at 0.8 M (p < 0.05). SDS-PAGE revealed higher protein extractability for breast NAMs at low ionic strength, regardless of breed and structural protein, particularly troponin, appeared to vary within muscle and breed. NAM from KC showed higher Ca2+–ATPase activity, surface hydrophobicity, but lower total sulfhydryl groups content (p < 0.05). Particle size of NAM solutions varied with ionic strength, in which KC-NAM showed larger size than at lower ionic strength, while BC-NAM appeared to be greater at higher ionic strength. NAM from KC breast showed higher thermal stability as higher initial (T0) and maximum (Tmax) denaturation temperatures of 48.4 and 54.8°C, respectively, recorded by microdifferential scanning calorimetry. The KC-NAM, particularly from breast, exhibited lower turbidity within 40-50°C, while turbidity increased in BC samples at low ionic strength when heated at 60°C. Dynamic rheology revealed different storage modulus (G′) depending on breed, muscle type and ionic strength. Breast BC-NAM formed more elastic gel with higher end point G′ at 80°C (Ge′; p < 0.05). Ionic strength showed reverse effects on different breeds as a stronger Ge′ value achieved in KC- NAM at low ionic strength, while high ionic strength induced stronger gel in BC samples. Aggregation of NAMs began at lower temperatures at higher ionic strength and actin dissociation probably occurred in breast NAM from KC as observed by a drop of G′ at around 70°C. The results of this study revealed differences between NAM of the two breeds, by which higher gel elasticity are expected in KC at lower ionic strength.

This study evaluated the long-term antihypertensive effect of corn gluten meal protein hydrolysate (CGM-H) on spontaneously hypertensive rats (SHRs). Plausible pathways governing blood pressure were also evaluated based on untargeted metabolomic analysis. The casein diet containing 1 % of CGM-H and peptides obtained from ultrafiltration membranes attenuated the development of hypertension in SHRs after a 6-week oral administration with similar efficacy, while the control group showed progressive hypertension. Plasma renin and angiotensin converting enzyme (ACE) activity and total peroxides level of the corn peptides-treated groups decreased when compared with the control (p

ABSTRACT: Korat chicken (KC) is a slow-growing crossbreed renowned for its excellent growth and firm texture. This study investigated the effect of various sous-vide (SV) conditions (60 and 70°C, 1–3 h) on their texture, protein structure and degradation, as well as consumer acceptability, with the traditional boiling served as control. Texture showed significant improvement under all SV conditions compared to the control, as demonstrated by increased water holding capacity (WHC), cooking loss, and decreased shear force, hardness, and chewiness (P < 0.05). These changes corresponded to the higher sensory scores (P < 0.05). Among the SV samples, increased temperatures and longer cooking times led to higher degradation of myofibrils and connective tissue, as evidenced by a decrease in water-, salt-soluble proteins, and soluble collagen (P < 0.05). These findings aligned with the scanning electron microscopy (SEM) results, which showed a looser muscle structure in the meat under more intense cooking conditions. Based on synchrotron radiation-based Fourier transform infrared (SR-FTIR) results, a gradual increase in antiparallel forms within the amide I bands (1,700–1,600 cm−1) of the total spectra with higher temperature and longer cooking times was observed (P < 0.05), while the fluctuations were observed in the changes of α-helix, β-sheet, and β-turn structures. This suggested that the antiparallel structure represented a looser configuration developing during intense SV cooking. Combined with the principal component analysis (PCA) results, the findings indicated that the suitable SV condition for KC breast meat was 70°C for varying durations (1–3 h), as it showed the strongest correlation with sensory scores, particularly in terms of tenderness. In summary, these findings provided a better understanding of molecular changes and discovered SV conditions to enhance the texture quality of the KC meat.

Changes in structural characteristics and antioxidant activity of tilapia hydrolysate glycated with glucose, fructose, or xylose at 90 °C for 12 h, and following in vitro gastrointestinal (GI) digestion were investigated. Fourier-transformed infrared (FTIR) band between 1,800 and 1,400 cm−1 confirmed the structural modifications of hydrolysate under glycations. Glycation drastically increased ATBS·+ and ONOO- scavenging activities (p

Wolffia is a protein-rich aquatic plant with the potential to help address food sustainability issues; however, a more efficient extraction process must be sought due to limited yield with conventional methods. This study aimed to investigate the effects of duckweed forms (fresh and dried), duckweed-to-aqueous solution ratio (DSR), as well as alkaline and ultrasonication extractions on yields, physicochemical and techno-functional properties of duckweed protein extracts (DPE). Three extraction methods were used: alkaline extraction (AE), ultrasound-assisted alkaline extraction (UAAE), and ultrasound-assisted water extraction (UAWE). The study showed fresh duckweed resulted in a 2.5-fold higher yield and protein recovery than oven-dried duckweed. UAE significantly enhanced extraction yield and protein recovery. The optimal extraction process was a DSR of 1:6 using UAAE at pH 8.5, resulting in a 16% yield and 34% protein recovery. Furthermore, the study found that UAE facilitated the extraction of non-polar/hydrophobic amino acids while AE proved to be efficient in extracting sulfur-containing amino acids. This study, for the first time, revealed the role of UAE and AE in promoting the extraction of different profiles of amino acids. The DPE produced using UAAE contained 65% protein, meeting the WHO/FAO/UNU recommended protein intake for preschool-aged children. Furthermore, the DPE exhibited excellent emulsifying properties and oil-holding capacity comparable to commercial soy protein isolates. Overall, UAAE was identified as a promising approach for producing techno-functional and nutritious protein ingredients from Wolffia. The resulting DPE proved to show great potential for functional plant-based food and feed applications.

ABSTRACT: This study determined the effect of water bath cooking (70°C and 90°C for 40 min) and the extreme heat treatment by an autoclave (121°C for 40 min) on the quality of breast meat of a fast-growing chicken, commercial broiler (CB), and slow-growing chickens, Korat chicken (KC), and Thai native chicken (NC) (Leung Hang Khao), by vibrational spectroscopic techniques, including synchrotron radiation-based Fourier transform infrared (SR-FTIR) microspectroscopy and Fourier transform Raman (FT-Raman) spectroscopy. Taste-enhancing compounds, including inosine-5ˊ-monophosphate (IMP) and guanosine-5ˊ-monophosphate (GMP), were better retained in cooked KC and NC meats than in cooked CB meat (P < 0.05). The high heat treatment at 121°C depleted the amount of insoluble collagen in all breeds (P < 0.05). Shear force values of slow-growing chicken meat were not affected by high heating temperatures (P > 0.05). In addition, the high heat treatment increased protein carbonyl (P < 0.05), while no effect on in vitro protein digestibility (P > 0.05). SR-FTIR microspectroscopy performed better in differentiating the meat quality of different chicken breeds, whereas FT-Raman spectroscopy clearly revealed differences in meat qualities induced by heating temperature. Based on principal component analysis (PCA), distinct characteristics of chicken meat cooked at 70°C were high water-holding capacity, lightness (L*), moisture content, and predominant α-helix structure, correlating with Raman spectra at 3,217 cm−1 (O–H stretching of water) and 1,651 cm−1 (amide I; α-helix). The high heating temperature at 90°C and 121°C exposed protein structure to a greater extent, as evidenced by an increase in β-sheets, which was well correlated with the Raman spectra at 2,968 and 2,893 cm−1 (C–H stretching), tryptophan (880 cm−1), tyrosine (858 cm−1), and 1,042, 1,020, and 990 cm−1 (C–C stretching; β-sheet). SR-FTIR and FT-Raman spectroscopy show potential for differentiation of chicken meat quality with respect to breeds and cooking temperatures. The marked differences in wavenumbers would be beneficial as markers for determining the quality of cooked meats from slow- and fast-growing chickens.

ABSTRACT: The Korat chicken (KR) is a slow-growing Thai chicken breed with relatively poor feed efficiency (FE) but very tasty meat with high protein and low fat contents, and a unique texture. To enhance the competitiveness of KR, its FE should be improved. However, selecting for FE has an unknown effect on meat characteristics. Thus, understanding the genetic basis underlying FE traits and meat characteristics is needed. In this study, 75 male KR birds were raised up to 10 wk of age. For each bird, the feed conversion ratio (FCR), residual feed intake (RFI), and physicochemical properties, flavor precursors, and biological compounds in the thigh meat were evaluated. At 10 wk of age, thigh muscle samples from 6 birds (3 with high FCR and 3 with low FCR values) were selected, and their proteomes were investigated using a label-free proteomic method. Weighted gene coexpression network analysis (WGCNA) was used to screen the key protein modules and pathways. The WGCNA results revealed that FE and meat characteristics significantly correlated with the same protein module. However, the correlation was unfavorable; improving FE may result in a decrease in meat quality through the alteration in biological processes including glycolysis/gluconeogenesis, metabolic pathway, carbon metabolism, biosynthesis of amino acids, pyruvate metabolism, and protein processing in the endoplasmic reticulum. The hub proteins of the significant module (TNNT1, TNNT3, TNNI2, TNNC2, MYLPF, MYH10, GADPH, PGK1, LDHA, and GPI) were also identified to be associated with energy metabolism, and muscle growth and development. Given that the same proteins and pathways are present in FE and meat characteristics but in opposite directions, selection practices for KR should simultaneously consider both trait groups to maintain the high meat quality of KR while improving FE.

ABSTRACT: Egg yolk is a coproduct of egg white processing. The protein hydrolysis of egg yolks to exhibit antimicrobial activity is a strategy for its valorization. The objective of this study is to fractionate antibacterial peptides from pepsin-hydrolyzed egg yolks using flash chromatography. In addition, the mode of actions of the fractionated peptides were elucidated and plausible antibacterial peptides were reported. The fraction 6 (F6) obtained from a C18-flash column exhibited antibacterial activity against Staphylococcus aureus ATCC 29213 and Salmonella typhimurium TISTR 292 at minimal inhibitory concentration (MIC) values of 0.5 to 1 mmol/L (Leucine equivalent). The fractionated peptides induced DNA leakage as monitored by 260 nm. Propidium iodide and SYTO9 staining observed under a confocal microscope suggested the disintegration of cell membranes. Synchrotron-based Fourier-transform infrared spectroscopy analysis revealed that the egg yolk peptides at 1 × MIC induced an alteration of phospholipids at cell membranes and modified conformation of intracellular proteins and nucleic acids. Scanning electron microscopy revealed obvious cell ruptures when S. aureus was treated at 1 × MIC for 4 h, whereas damage of cell membranes and leakage of intracellular components were also observed for the transmission electron microscopy. Egg yolk peptides showed no hemolytic activity in human erythrocytes at concentrations up to 4 mmol/L. Peptide identification by LC-MS/MS revealed 3 cationic and 10 anionic peptides with 100% sequence similarity to apolipoprotein-B of Gallus gallus with hydrophobicity ranging from 27 to 75%. The identified peptide KGGDLGLFEPTL exhibited the highest antibacterial activity toward S. aureus at MIC of 2 mmol/L. Peptides derived from egg yolk hydrolysate present significant potential as antistaphylococcal agents for food and/or pharmaceutical application.

Broiler chicken tracheas are a co-product from chicken slaughterhouses which are normally turned into low value animal feed despite their high levels of collagen. Typical collagen extraction by acid and/or pepsin usually results in relatively low yield. Ultrasound-assisted extraction (UAE) could be a means to improve collagen yield. The objectives of this study were to investigate the effects of ultrasonic parameters on the yield and biochemical properties of trachea collagen (TC). Conventional extraction using acetic acid and pepsin for 48 h resulted in acid-soluble (AS) and pepsin-soluble (PS) collagen with a yield of 0.65% and 3.10%, respectively. When an ultrasound with an intensity of 17.46 W·cm−2 was applied for 20 min, followed by acid extraction for 42 h (U-AS), the collagen yield increased to 1.58%. A yield of 6.28% was obtained when the ultrasound treatment was followed by pepsin for 36 h (U-PS). PS and U-PS contained collagen of 82.84% and 85.70%, respectively. Scanning electron microscopy images revealed that the ultrasound did not affect the collagen microstructure. All collagen samples showed an obvious triple helix structure as measured by circular dichroism spectroscopy. Fourier transform infrared spectroscopy indicated that the ultrasound did not disturb the secondary structure of the protein in which approximately 30% of the α-helix content was a major structure for all collagen samples. Micro-differential scanning calorimetry demonstrated that the denaturation temperature of collagen in the presence of deionized water was higher than collagen solubilized in 0.5 M acetic acid, regardless of the extraction method. All collagen comprised of α1 and α2-units with molecular weights of approximately 135 and 116 kDa, respectively, corresponding to the type I characteristic. PS and U-PS collagen possessed higher imino acids than their AS and U-AS counterparts. Based on LC-MS/MS peptide mapping, PS and U-PS collagen showed a high similarity to type I collagen. These results suggest that chicken tracheas are an alternative source of type I collagen. UAE is a promising technique that could increase collagen yield without damaging its structure.

Prolyl oligopeptidase (POP) is known to be related to neurocognitive disorder as it degrades neuroactive peptides. POP inhibitory peptides from corn gluten meal hydrolysate (CGM-H) were identified and characterized. After a series of chromatographic separations, ALLTLSPLGPA was identified as the most effective POP inhibitor, with an IC50 value of 0.79 ± 0.004 mM. Its in silico gastrointestinal (GI)-digested peptide, SPLGPA, exhibited 3-fold lower POP inhibitory activity, while higher inhibition was observed in SH-SY5Y cells when compared to its parent peptide. In addition, 25 µM ALLTLSPLGPA notably reduced α-synuclein aggregation in KCl-treated SH-SY5Y cells. ALLTLSPLGPA and SPLGPA were characterized as mixed-type and uncompetitive inhibitors, respectively. They bound to POP via hydrogen bonds at the β-propeller domain. These results demonstrated that peptides derived from CGM could have potential for developing nutraceutical products targeting brain health.

The present study aimed to characterize the mode of action of a novel antimicrobial peptide isolated from egg yolk hydrolysate. The EYHp6, KGGDLGLFEPTL, exhibited inhibition against Salmonella enterica serovar Typhimurium TISTR 292 and S. enterica serovar Enteritidis DMST 15679 with a MIC value of 2 mM. In contrast, S. enterica serovar Newport ATCC 6962 and other strains of Typhimurium and Enteritidis were inhibited at 4 mM. EYHp6 increased the cell membrane permeability of S. Typhimurium TISTR 292, leading to DNA leakage. Membrane integrity determined by propidium iodide and SYTO9 staining visualized by confocal microscopy demonstrated that EYHp6 at 1 × MIC induced disruption of cell membranes. Electron microscopy revealed that treatment of S. Typhimurium with EYHp6 led to damage to the cell membrane, causing the leakage of intracellular contents. Synchrotron-based Fourier-transform infrared spectroscopy indicated that EYHp6 killed S. Typhimurium by targeting fatty acids and nucleic acids in the cell membrane. The peptide did not show hemolytic activity up to 4 mM. These findings suggest that EYHp6 could be a promising antibacterial agent for controlling the growth of S. enterica.

ABSTRACT: One of the defects commonly found in cooked marinated chicken breast products is a red blood spot (RBS), which is caused by undercooked blood in vessels. This problem was alleviated by microwave (MW) pre-heating for 6 to 7 min, followed by steaming. RBS formation decreased when samples were heated to a core temperature of 80°C and were completely eliminated at a core temperature of 82°C and 85°C when a MW pre-heating step was applied for 7 min. Based on synchrotron-based Fourier transform infrared spectroscopy (SR-FTIR), blood remaining in the blood vessel had a lower α-helical content when samples were cooked by the combination of MW heating and steaming as compared with those prepared by steaming alone (P < 0.05). MW pre-heating decreased cooking time by 28 to 48% as compared with steaming alone. Heating regimes had no effect on cooking loss, pH, water-holding capacity, and shear force. MW pre-heating for 7 min followed by steaming to a core temperature of 82°C appeared to be an effective heating regime to reduce the occurrence of RBS, with acceptable cooking loss.

ABSTRACT: Chicken breed is one of the key factors that influence meat quality. The quality attributes of breast meat from commercial broiler (CB), Thai native chicken (NC, Leung Hang Khao), and the crossbred Korat chicken (KC) were investigated via synchrotron radiation-based Fourier transform infrared (SR-FTIR) microspectroscopy, Fourier transform Raman (FT-Raman) spectroscopy, and physicochemical analysis. The protein and carbonyl contents of KC and NC meats were higher than that of CB meat, but the lipid content was lower (P < 0.05). CB meat was characterized by high moisture, lightness (L*), and presence of taste-active nucleotides, namely, inosine 5′-monophosphate (IMP) and guanosine 5′-monophosphate (GMP). Moreover, NC meat had the highest insoluble collagen and inosine contents (P < 0.05). The predominant protein secondary structures of KC and NC meats were β-turns and random coils, whereas α-helices were mainly found in CB meat. Based on principal component analysis, the meat quality and spectra were clearly separated by breeds. The high moisture and lipid content of meat corresponded to O–H stretching (3,203 cm−1) and C–H stretching (2,854 cm−1) in the FT-Raman spectra, whereas PO2− stretching (1,240 cm−1), measured via SR-FTIR, was well correlated with the IMP content. In addition, the FT-Raman wavenumber of 934 cm−1, indicating C–C stretching, was correlated with high water-holding capacity (WHC) in KC meat. The quality of meat from slow- and fast-growing chickens significantly varies. Vibrational spectroscopy is a powerful technique that provides insightful molecular information correlated with various meat attributes.

ABSTRACT: The Korat chicken (KR), developed in Thailand, is a slow-growing breed developed as an alternative breed for Thai chicken producers. The growing interest in slow-growing chicken meat, due to its unique taste, distinct texture, health benefits, and higher broiler welfare have led to higher market demand for KR. However, its low feed efficiency (FE) has a significant negative impact on farm profitability. Understanding the molecular mechanism regulating FE allows for designing a suitable selection program and contributing to breeding more efficient chicken for poultry production. Thus, the objective of our study was to investigate the proteome differences and possible pathways associated with FE in male KR using a label-free quantitative proteomic approach. Seventy-five KR males were individually evaluated for FE, and duodenum samples from 6 animals (3 high-FE and 3 low-FE chickens) were collected at 10 wk of age for differential abundant proteins (DAPs), protein networks, functional enrichment, and pathway analyses. In this study, we found 40 DAPs significantly associated with FE pathways, including glycolysis/gluconeogenesis, peroxisome, oxidative phosphorylation, tight junction, and cysteine and methionine metabolism. Thus, variations in observed DAPs or genes related to DAPs could be interesting biomarker candidates for selection for higher feed utilization efficiency in chicken.

ABSTRACT: Carnosine enrichment of slow-growing Korat chicken (KRC) meat helps differentiate KRC from mainstream chicken. We aimed to investigate the effects of β-alanine and L-histidine supplementation on the carnosine synthesis in and quality and secondary structure of proteins in slow-growing KRC meat. Four hundred 21-day-old female KRC were used, and a completely randomized design was applied. The chickens were divided into 4 experimental groups: basal diet (A), basal diet supplemented with 1.0% β-alanine (B), 0.5% L-histidine (C), and 1.0% β-alanine combined with 0.5% L-histidine (D). Each group consisted of 5 replicates (20 chickens per replicate). On d 70, 2 chickens per replicate were slaughtered, and the levels of carnosine, anserine, and thiobarbituric acid reactive substances were analyzed. Biochemical changes were monitored using synchrotron radiation-based Fourier transform infrared microspectroscopy; 5 chickens per replicate were slaughtered, and the meat quality was analyzed. Statistical analysis was performed using ANOVA and principal component analysis (PCA). Group D chickens exhibited the highest carnosine meat content, followed by those in groups B and C. However, amino acid supplementation did not affect anserine content and growth performance. Higher carnosine levels correlated with increasing pH45 min and decreasing drip loss, cooking loss, shear force, and lipid oxidation. PCA revealed that supplementation with only β-alanine or L-histidine was related to increased content of β-sheets, β-turns, and aliphatic bending groups and decreased content of α-helix groups. This study is the first to report such findings in slow-growing chicken. Our findings suggest that KRC can synthesize the highest carnosine levels after both β-alanine and L-histidine supplementation. Higher carnosine contents do not adversely affect meat quality, improve meat texture, and alter the secondary structures of proteins. The molecular mechanism underlying carnosine synthesis in chickens needs further study to better understand and reveal markers that facilitate the development of nutrient selection programs.

Meat quality attributes vary with chicken age. Understanding the relationship between poultry age and the quality of the meat would be beneficial for efficient poultry farming to meet market needs. The Korat hybrid chicken (KC) is a new crossbred chicken whose meat quality is distinct from that of commercial broiler (CB) chickens and has not been well characterized. In this study, we characterized the physico-chemical properties of KC meat and correlate the findings with Raman spectral data. The protein content of KC breast and thigh meat increased with age. The pH of thigh meat decreased, while the water-holding capacity of breast meat increased as the age of the chickens increased. The amount of cholesterol in breast meat decreased as the rearing period was extended. Inosine 5′-monophosphate and guanosine 5′-monophosphate of breast meat decreased as KC grew older. The shear force values of meat from older birds increased concomitantly with an increase in total collagen. Principle component analysis revealed that the meat quality of CB was greatly different from that of KC meat. High shear force values of KC meat at 20 wk of age were well correlated with an increase in the β-sheet structure (amide I) and amide III of collagen. Raman spectra at 3,207 cm−1 and relative α-helical content were negatively correlated with shear force values of KC breast meat. These could be used as markers to evaluate KC meat quality.

The effects of various sous-vide (SV) cooking conditions (50-60℃, 30-60 min) on physicochemical properties related to the texture characteristics, protein structure/degradation, and sensory acceptability of tilapia fillet (Oreochromis niloticus) were investigated. With an increasing temperature and processing time of SV cooking, protein degradation (of both myofibrils and connective tissue) was more pronounced, as evaluated by the decrease in water- and salt-soluble proteins, total collagen, as well as the changes in the ratio of secondary protein structures (α-helix, β-sheet, β-turn, etc.), which were determined by synchrotron-FTIR (SR-FTIR). These degradations were associated with the improvement of meat tenderness, as estimated by shear force and texture profile analyzer (TPA) results. Among all SV conditions, using 60 ℃ for 45 min seems to be the optimal condition for tilapia meat, since it delivered the best results for texture characteristics and acceptability (p < 0.05). Moreover, principal component analysis (PCA) results clearly demonstrated that the highest texture-liking score of this condition was well associated with the intensity of β-sheets, which seem to be the crucial component that affected the texture of SV-cooked tilapia more so than other parameters. The findings demonstrated the potential of SR-FTIR to decipher the biomolecular structure, particularly the secondary protein structure, of SV-cooked tilapia. This technique provided essential information for a better understanding of the changes in biomolecules related to the textural characteristics of this product.

Nine novel peptides from tilapia hydrolysate were isolated and identified to be EKL, EKP, HKPA, ELSC, ALSC, ASLCH, SLCH, LPGYF, and LEVPGY. In addition, six fragments from in silico gastrointestinal digestion of the identified peptides, including EK, KPA, SC, CH, PGY, and EVPGY, were synthesized. The most effective parent and digested peptides showing ABTS·+ scavenging capacity were LPGYF and SC, respectively. All C- and Y-containing peptides were more effective than ascorbic acid (AsA). In contrast, K-containing peptides exhibited less antioxidant activity. All 15 peptides showed potent intracellular reactive oxygen species scavengers in the AAPH-induced HepG2 cell oxidative stress model. In addition, the digested peptides SC, CH, and PGY up-regulated the expression of CAT and SOD1 in HepG2 cells. The peptide PGY was the most effective cellular antioxidant. Thus, tilapia peptides could be potent nutraceutical products to reduce cellular oxidative stress.

Due to the overuse and abuse of antibiotics, several antibiotic resistant bacteria have emerged. Antimicrobial peptides (AMPs) have gained attention as alternative antimicrobial agents because of their unique mode of action that impedes bacterial resistance. Two novel antibacterial peptides were isolated from Alcalase-hydrolyzed chicken plasma by size exclusion and reverse-phase chromatography. They were identified by LC-MS/MS to be VSDH and CCCPKAF, which showed effective antibacterial activity toward Bacillus cereus DMST 5040, with varied modes of action. The peptide CCCPKAF caused cell membrane disintegration, as evidenced by propidium iodide (PI) uptake. In contrast, the peptide VSDH targeted intracellular molecules, including proteins and nucleic acids, as revealed by Synchrotron-based Fourier Transform Infrared (SR-FTIR). The secondary structure of intracellular proteins increased to a β-sheet structure concomitant with a decrease in the α-helix structure when exposed to 0.5 mM VSDH. Molecular docking analysis revealed that VSDH showed high binding affinity for the active sites of the various enzymes involved in DNA synthesis. In addition, it showed good affinity for a chaperone protein (Dnak), resulting in the misfolding of intracellular proteins. Nuclear magnetic resonance (NMR) and molecular dynamics simulations also indicated that VSDH chelated well with Mg2+, which could partly contribute to its antibacterial activity.

This study aims to shed light on the association between non-volatile and volatile compounds related to flavor/taste characteristics as well as sensory acceptability of Nile tilapia fillet (Oreochromis niloticus) cooked by various sous-vide (SV) conditions (50–60 ℃, 30–60 min), with fish cooked with boiling water used as control. Higher temperatures and longer processing times of SV cooking led to greater protein and lipid oxidation as indicated by the increase in total sulfhydryl (-SH), carbonyl, free fatty acid (FFA) contents as well as peroxide values (PV) and thiobarbituric acid reactive substance (TBARS) values. The differences in flavor/taste components including adenosine triphosphate (ATP)-related compounds, free amino acids (FAAs) and volatiles were also obtained, which directly affect sensory acceptability as evaluated by using the hedonic scale. Based on principal component analysis (PCA) results, the acceptability score was strongly correlated with inosine monophosphate (IMP) and acetoin, which seem to be the most crucial flavor enhancers for cooked tilapia. Among all samples, tilapia processed at 60 °C for 45 and 60 min, which contained significantly higher IMP and acetoin (p < 0.05) than others, had significantly higher flavor-liking and overall-liking scores, with a more than 7.5 meaning for high acceptability (p < 0.05), indicating the optimal SV conditions for tilapia fillet. Overall, the present finding indicated that the SV-cooking technique, at the optimal conditions, can improve the meat quality of cooked fish, in terms of flavor/taste characteristics, compared with traditional cooking (control).

This study was to determine the effect of the organic raising system (OR) on growth performance, meat quality, and physicochemical properties of slow-growing chickens. Three hundred and sixty (one-day-old) Korat chickens (KRC) were randomly assigned to control (CO) and OR groups. The groups comprised six replicates of thirty chickens each. The chickens were housed in indoor pens (5 birds/m2), wherein those in OR had free access to Ruzi pasture (1 bird/4 m2) from d 21 to d 84 of age. In the CO group, chickens were fed with a mixed feed derived from commercial feedstuffs, while those in the OR group were fed with mixed feed derived from organic feedstuffs. The results revealed a lower feed intake (p < 0.0001) and feed conversion ratio (p = 0.004) in the OR. The OR increased total collagen, protein, shear force, color of skin and meat, and decreased abdominal fat (p < 0.05). The OR improved fatty acid with increased DHA, n-3 PUFA, and decreased the ratio of n-6 to n-3 PUFA in KRC meat (p < 0.05). The synchrotron radiation-based Fourier transform infrared spectroscopy and correlation loading analyses confirmed these results. In conclusion, our results proved that OR could improve growth performance and meat quality and suggested the raising system be adopted commercially. In addition, the observed differences in biochemical molecules could also serve as markers for monitoring meat quality.

Six-month-old (FS6) and 12-month-old (FS12) Thai fish sauces were separated by Sephadex-G25 desalting columns into 3 fractions, namely, F1 (peptides/Maillard reaction products), F2 (peptides/NaCl) and F3 (peptides/amino acids). The F3 fractions from both samples showed the highest chemical and cellular antioxidant activities and could inhibit ACE (P

Permeability of angiotensin I-converting enzyme (ACE) inhibitory peptides (KPLLCS, ELFTT, and KPLL) identified from the in vitro gastrointestinal digest of cooked chicken breast was investigated using the Caco-2 cell model system. KPLLCS was originally the most effective ACE inhibitor (IC50 0.37 μM), but it was degraded during permeation through Caco-2 cells. Among these peptides, KPLL showed the highest permeability in intact form, and partially degraded to KP and LL, which still showed ACE inhibitory activity after permeation. ACE inhibitory activity of permeated KPLL was highest of 56%. KPLL and KP possessed a mixed inhibitor characteristic, while LL was a non-competitive inhibitor. Based on molecular docking, K at N-terminus of KPLL is a key structure contributing to ACE inhibition as it can occupy the active site of ACE. Determining the structural stability and degree of peptide transport across epithelial cell is required to confirm their potential as ACE inhibitors.

The release of encrypted angiotensin converting enzyme (ACE) inhibitory peptides upon simulated in vitro gastrointestinal (GI) digestion of Korat crossbred (KC) and commercial broiler (BR) breast meat varied with cooking methods. Digestas of KC breast meat heated at 70 °C for 0.5 h (H-0.5) showed the highest ACE inhibitory activity, while digestas of KC chicken heated at 121 °C for 1 h showed the lowest inhibitory activity. ACE inhibitory activity varied with the protein digestibility. After a series of chromatographic techniques, we identified KPLLCS, ELFTT, and KPLL, novel potent ACE inhibitor peptides whose sequences were homologous to that of myosin and whose IC50 values were 0.37, 6.35 and 11.98 μM, respectively. ELFTT and KPLL were resistant to plasmin hydrolysis for up to 60 min. Therefore, KC chicken meat heated at H-0.5 could serve as a source of ACE inhibitory peptides upon GI digestion.

Changes in the lipid oxidation of silver carp (Hypophthalmichthys molitrix) stored in ice for 14 days and that of its respective washed mince were evaluated. Total lipid, phospholipid, polyunsaturated fatty acid (PUFA) and monounsaturated fatty acid (MUFA) contents of the skin, belly flap and mince decreased as the storage time in ice increased. The washing process decreased the lipid contents but concentrated their phospholipid counterparts. The fish belly flap exhibited the highest thio-barbituric acid reactive substances (TBARS) value, while the mince had the lowest. 1-Hexanol, 1-octen-3-ol, and 1-hexanal were key volatile compounds detected in the belly flaps of fish stored for 7–14 days. Hexanal was the only major volatile compound found in washed mince prepared from fish stored for an extended period in ice, but in a much lower amount compared with that in the belly flap. FTIR (Fourier transform infra-red) spectra revealed a decrease in the number of cis double bonds, methylene groups and phosphate groups in lipids extracted from fish stored in ice for 7–14 days as compared with those extracted from fresh fish. Principle component analysis (PCA) revealed that the FT-Raman band at 1747 cm−1 could be a potential marker for tracking the degree of lipid oxidation in the belly flap of silver carp stored in ice. In addition, IR bands indicating phosphate group (925, 825 cm−1) in oil extracted from washed mince were correlated with the extent of the lipid oxidation of the raw material.

Production of protein hydrolysate as nutraceuticals is typically based on the activity of the hydrolysate, which might not yield the optimal activity under physiological condition due to structural modification of peptides upon gastrointestinal (GI) digestion. This study systematically compared the chemical and cellular antioxidant activities of the in vitro digesta of tilapia protein and its hydrolysates prepared with various degree of hydrolysis (DH) by Alcalase. The enzymes used in the in vitro GI digestion analysis significantly contributed to the peptide content, Trolox equivalent antioxidant capacity (TEAC), and oxygen radical absorbance capacity (ORAC). Proteins and all hydrolysates were slightly digested by pepsin but hydrolyzed extensively by pancreatin. Both hydrolysate and digesta predominantly scavenged free radicals via hydrogen atom transfer (HAT). The antioxidant activities of the hydrolysates increased with the increasing DH up to 16 h of hydrolysis. However, the digesta of 10-h hydrolysate displayed the highest chemical and HepG2 cellular antioxidant activities, while the protein digesta displayed the lowest. Principal component analysis (PCA) showed that the TEAC of the digesta was positively correlated with the cellular antioxidant activity (CAA). Therefore, the production of protein hydrolysate should be optimized based on the activity of the hydrolysate digesta rather than that of hydrolysates.

Angiotensin I-converting enzyme (ACE) inhibitory activity of tilapia mince (TM) hydrolyzed by Virgibacillus halodenitrificans SK1-3-7 proteinases with a 48% degree of hydrolysis showed the highest ACE inhibitory activity with an IC50 of 0.54 mg/mL. After ultrafiltration (UF) and chromatographic separation via anion exchange, cation exchange and size exclusion chromatography, the fraction with the highest ACE inhibitory activity with an IC50 of 0.15 mg/mL was obtained. The peptides showed uncompetitive inhibition characteristics with an inhibition constant (Ki) of 0.18 mg/mL. The peptides showed high thermostability at 100 and 121 °C, as well as pH stability at a wide pH range of 2–10, and also maintained ACE inhibitory activity after in vitro gastrointestinal digestion. The most potent novel ACE inhibitory peptide identified was MILLLFR with an IC50 of 0.12 µM. TM hydrolysate prepared by V. halodenitrificans SK1-3-7 proteinases could serve as a source of peptides with ACE inhibitory activity.

The objectives of this study are to optimize the conditions for providing high yield of NaCl-tolerant extracellular protease from Virgibacillus sp. SK37 based on a fi sh-based medium and to investigate the eff ects of the key factors (mass per volume ratios of dried anchovy, yeast extract and NaCl, and initial pH of the medium) on the secretion patt ern of proteases. Based on the predicted response model, the optimized medium contained 1.81 % of dried anchovy, 0.33 % of yeast extract and 1.25 % of NaCl at pH=7.8. Under these conditions, a 5.3-fold increase in protease production was achieved, compared with the broth containing only 1.2 % of dried anchovy (5 % of NaCl at pH=7). The cubic regression adequately described the protease production. Protease activity was determined using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) on the synthetic substrate (Suc-Ala-Ala-Pro-Phe-AMC). Proteases of molecular masses of 19, 34, 35 and 44 kDa were secreted in the presence of NaCl, whereas those of 22 and 42 kDa were the main proteases detected in the absence of NaCl. In addition, no secreted proteases were detected when initial pH of the medium was pH=6. The peptide mass fi ngerprint of the medium cultured with 10 % NaCl showed a higher abundance of peptides with lower mass of 500– 1000 m/z compared with the medium containing 0 % NaCl, indicating the higher proteolytic activity of the high-salt medium. The Virgibacillus sp. SK37 proteases showed a marked preference towards Lys, Arg and Tyr in the presence of NaCl and towards Lys and Arg in the absence of NaCl.

Objective: This study aimed to find global mechanisms related to carnosine synthesis in slow-growing Korat chickens (KRC) using a proteomic approach. Methods: M. pectoralis major samples were collected from 10-week-old female KRC including low-carnosine (LC, 2,756.6±82.88 μg/g; n = 5) and high-carnosine (HC, 4,212.5 ±82.88 μg/g; n = 5). Results: We identified 152 common proteins, and 8 of these proteins showed differential expression between the LC and HC groups (p<0.05). Heat shock 70 kDa protein 8, Heat shock 70 kDa protein 2, protein disulfide isomerase family A, member 6, and endoplasmic reticulum resident protein 29 were significantly involved in protein processing in the endoplasmic reticulum pathway (false discovery rate<0.05), suggesting that the pathway is related to differential carnosine concentration in the M. pectoralis major of KRC. A high concentration of carnosine in the meat is mainly involved in low abundances of Titin isoform Ch12 and Connectin and high abundances of M-protein to maintain homeostasis during muscle contraction. These consequences improve meat characteristics, which were confirmed by the principal component analysis. Conclusion: Carnosine synthesis may occur when muscle cells need to recover homeostasis after being interfered with carnosine synthesis precursors, leading to improved muscle function. To the best of our knowledge, this is the first study to describe in detail the global molecular mechanisms in divergent carnosine contents in meat based on the proteomic approach. [ABSTRACT FROM AUTHOR]