Vibrational spectroscopy and biochemical changes in silver carp as related to quality of washed mince.

BACKGROUND: Biochemical and protein conformational changes in silver carp occurred during ice storage, affecting the physico-chemical and textural properties of its washed mince. Fourier-transform infrared spectroscopy (FTIR) and FT Raman could offer insightful molecular-level information that could be related to the freshness of fish and textural properties of washed mince. RESULTS: The K value increased from 15.8% to 85.0% after 14 days on ice. The surface hydrophobicity of silver carp muscle proteins increased during ice storage, and its thrice-washed mince showed the same trend. The yield and textural properties of washed mince continually decreased as the storage time was extended. Fourier-transform infrared spectroscopy revealed that, as storage time increased, the α-helix content of mince decreased, while the ß sheet content increased. Prolonged ice storage led to the exposure of buried aromatic amino acid residues and an increase in disulfide interchanges in mince and washed mince. Changes in the ∑ß sheet structure and Raman intensity at 828 cm-1 observed in mince correlated well with the K value. The α-helix content and Raman intensity of raw washed mince at 621 and 828 cm-1 showed a strong correlation with its textural properties. CONCLUSION: Silver carp should be processed to surimi within 7 days of ice storage to obtain a reasonably good yield and gel texture. Infrared and Raman spectroscopy can possibly be utilized to monitor freshness quality and protein conformational changes in silver carp and to estimate the textural properties of washed mince as affected by freshness. © 2019 Society of Chemical Industry.

Quantification of viable bacterial starter cultures of Virgibacillus sp. and Tetragenococcus halophilus in fish sauce fermentation by real-time quantitative PCR.

Real-time quantitative polymerase chain reaction (qPCR) methods were developed for the quantification of Virgibacillus sp. SK37 and Tetragenococcus halophilus MS33, which were added as starter cultures in fish sauce fermentation. The PCR assays were coupled with propidium monoazide (PMA) treatment of samples to selectively quantify viable cells and integrated with exogenous recombinant Escherichia coli cells to control variabilities in analysis procedures. The qPCR methods showed species-specificity for both Virgibacillus halodenitrificans and T. halophilus as evaluated using 6 reference strains and 28 strains of bacteria isolated from fish sauce fermentation. The qPCR efficiencies were 101.1% for V. halodenitrificans and 90.2% for T. halophilus. The quantification limits of the assays were 10(3) CFU/mL and 10(2) CFU/mL in fish sauce samples with linear correlations over 4 Logs for V. halodenitrificans and T. halophilus, respectively. The matrix effect was not observed when evaluated using fish sauce samples fermented for 1-6 months. The developed PMA-qPCR methods were successfully applied to monitor changes of Virgibacillus sp. SK37 and T. halophilus MS33 in a mackerel fish sauce fermentation model where culture-dependent techniques failed to quantify the starter cultures. The results demonstrated the usability of the methods as practical tools for monitoring the starter cultures in fish sauce fermentation.

Statistical Optimization of the Production of NaCl-Tolerant Proteases by a Moderate Halophile, Virgibacillus sp. SK37.

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 fish-based medium and to investigate the effects of the key factors (mass per volume ratios of dried anchovy, yeast extract and NaCl, and initial pH of the medium) on the secretion pattern 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 fingerprint 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.

Changes in volatile compounds and lipid oxidation in various tissues of Nile tilapia (Oreochromis niloticus) during ice storage.

Changes in the lipid oxidation and volatile compounds of a variety of tilapia tissues (Oreochromis niloticus) including the muscle, gills, and skin during ice storage were investigated by evaluating peroxide values (PVs), lipoxygenase (LOX) activity, fatty acid (FA) composition, and volatile substances. LOX activity and PV were determined in the gills, skin, and muscles throughout 9 days of storage in ascending order to the extended storage time. The highest level of LOX activity was found in the gills, whereas the highest PV was determined in the skin. FA content of all tissues decreased during the storage period. Oleic acid was the predominant monounsaturated fatty acid, whereas linoleic acid and docosahexaenoic acid were the main polyunsaturated fatty acids and omega-3 in all tissues. The fish gills were shown to have the highest level of volatile compounds followed by the skin and muscle, based on headspace solid-phase microextraction coupled with gas chromatography and mass spectrometry. Principal component analysis indicated gradual changes in the volatile compound composition with increasing storage time. 2-Butanone and nonanal in the muscle, 6-methyl-2-heptanone and 2-nonenal in the gills, and 1-heptanol, and 1-nonanol in the skin were found to be the potential freshness indicators. In addition, hexanal could be a general potential marker for measuring the degree of lipid oxidation in all tissues. PRACTICAL APPLICATION: Understanding the volatile compound formation related to lipid oxidation within storage time at various tissues of tilapia could be critical to the side-stream processing to yield the desired quality.

Effect of ultrasound and thermal pretreatments on antioxidant activity of egg white hydrolysate.

The use of ultrasonic (US) treatment of egg white prior to enzymatic hydrolysis to produce hydrolysate with antioxidant activity was investigated. The state of egg white (raw vs. cooked form) along with two levels of Alcalase (1% and 10% (w/w) protein) was applied. Hydrolysis and antioxidant activity of hydrolysate increased by US pretreatment at intensity of 41.53 W/cm2 . The hydrolysate prepared from US treatment on raw egg white hydrolyzed by 1% Alcalase (US-R1%) showed the lowest degree of hydrolysis (DH); however, its 2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) radical scavenging and ferric reducing antioxidant power activities were the highest. In contrast, the highest cytoprotective effect and intracellular reactive oxygen species scavenging activity were more notable in the hydrolysate prepared from US treatment of boiled egg white hydrolyzed by 10% Alcalase (US-B10%), which also exhibited the highest DH and metal chelation ability. The hydrolysate possessing cellular antioxidant activity (CAA) showed the highest proportion of small molecular weight peptides (<200 Da). Fourier-transform infrared spectroscopy revealed an increase of N- and C-terminal ends at 1500 and 1400 cm-1 , respectively, in concomitant with a decrease of amide I. Principal component analysis showed clear differentiation of spectra from different levels of enzyme according to their DH, C-terminal ends, and antioxidant activity. Our findings suggested that cooked egg white followed by US pretreatment was beneficial to produce hydrolysate containing high CAA.

Glycation of tilapia protein hydrolysate decreases cellular antioxidant activity upon in vitro gastrointestinal digestion.

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 < 0.05) as well as ferric-reducing antioxidant power (FRAP). Xylose was the most effective sugar for glycation, yielding the highest chemical antioxidant activities (p < 0.05). However, glycated hydrolysates exhibited lower cellular antioxidant activity (CAA) on HepG2 cell when compared to hydrolysates. The extensive glycation of hydrolysates resulted in lower GI digestibility as confirmed by the FTIR spectra of C[bond, double bond]O, C-N, N-H, C-C, C-O, and C-H stretching vibrations. Glycation of tilapia hydrolysates only improved chemical antioxidant activities, but alleviated CAA, especially upon simulated GI digestion.

A comprehensive study of sous-vide cooked Korat chicken breast processed by various conditions: texture, compositional/structural changes, and consumer acceptance.

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, ß-sheet, and ß-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.

Reduction of red blood spots in cooked marinated chicken breast meat by combined microwave heating and steaming.

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.

Ultrasound-assisted extraction of collagen from broiler chicken trachea and its biochemical characterization.

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.

Heating temperatures affect meat quality and vibrational spectroscopic properties of slow- and fast-growing chickens.

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-5'-monophosphate (IMP) and guanosine-5'-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 ß-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; ß-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.

Investigating the Effect of Various Sous-Vide Cooking Conditions on Protein Structure and Texture Characteristics of Tilapia Fillet Using Synchrotron Radiation-Based FTIR.

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, ß-sheet, ß-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 ß-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.

Effects of collagen, chitosan and mixture on fibroblast responses and angiogenic activities in 2D and 3D in vitro models.

Despite accumulating evidences have demonstrated the potential of collagen and chitosan on tissue repair, it remains unclear on their combination effects. Here, we examined the regenerative effects of single collagen, chitosan and their mixture on fibroblasts and endothelial cells at cellular levels. The results showed that fibroblast responses, as indicated by high proliferative rate, increased spheroid diameter and migrated area existing from spheroid edge, and decreased wound area, were significantly promoted by either collagen or chitosan stimulation. Similarly, both collagen and chitosan resulted in increased endothelial cell proliferation and migration with accelerated tube-like network formation and upregulated VE-cadherin expression, although collagen strongly provided this effect. While the 1:1 mixture (100:100 µg/mL of chitosan to collagen) treatment caused a reduction in fibroblast viability, the lower ratio of chitosan (1:10 mixture; 10:100 µg/mL) did not produce any impact on both fibroblast and endothelial cell viabilities. The 1:10 mixture also significantly enhanced the additional effects on fibroblast responses and angiogenic activities as shown by higher endothelial growth, proliferation and migration with accelerated capillary-like network formation than those treated with the single substance. Further investigation of signaling proteins found that collagen significantly increased expressions of p-Fak, p-Akt and Cdk5 whereas chitosan upregulated p-Fak and Cdk5 expressions. Comparing to the single treatments, p-Fak, p-Akt and Cdk5 were higher expressed in the 1:10 mixture. These observations indicate that proper collagen-chitosan mixture provides the combination effects on fibroblast responses and angiogenic activities when a high concentration of collagen is used, possibly through Fak/Akt and Cdk5 signaling pathways. Therefore, this study helps to define the clinical use of collagen and chitosan as promising biomaterials for tissue repair.

Isolation, identification, and mode of action of antibacterial peptides derived from egg yolk hydrolysate.

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.

Anti-Salmonella Activity of a Novel Peptide, KGGDLGLFEPTL, Derived from Egg Yolk Hydrolysate.

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.

Thigh muscle metabolic response is linked to feed efficiency and meat characteristics in slow-growing chicken.

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.

Efficient expression and purification of recombinant glutaminase from Bacillus licheniformis (GlsA) in Escherichia coli.

Glutaminase or L-glutamine aminohydrolase (EC 3.5.1.2) is an enzyme that catalyzes the formation of glutamic acid and ammonium ion from glutamine. This enzyme functions in cellular metabolism of every organism by supplying nitrogen required for the biosynthesis of a variety of metabolic intermediates, while glutamic acid plays a role in both sensory and nutritional properties of food. So far there have been only a few reports on cloning, expression and characterization of purified glutaminases. Microbial glutaminases are enzymes with emerging potential in both the food and the pharmaceutical industries. In this research a recombinant glutaminase from Bacillus licheniformis (GlsA) was expressed in Escherichia coli, under the control of a ptac promoter. The recombinant enzyme was tagged with decahistidine tag at its C-terminus and could be conveniently purified by one-step immobilized metal affinity chromatography (IMAC) to apparent homogeneity. The enzyme could be induced for efficient expression with IPTG, yielding approximately 26,000 units from 1-l shake flask cultures. The enzyme was stable at 30°C and pH 7.5 for up to 6h, and could be used efficiently to increase glutamic acid content when protein hydrolysates from soy and anchovy were used as substrates. The study demonstrates an efficient expression system for the production and purification of bacterial glutaminase. In addition, its potential application for bioconversion of glutamine to flavor-enhancing glutamic acid has been demonstrated.

Hydrolytic activity of Virgibacillus sp. SK37, a starter culture of fish sauce fermentation, and its cell-bound proteinases.

Fish sauce production relies on a natural fermentation process requiring 12-18 months for process completion. Virgibacillus sp. SK37 has been shown to be a potential strain for fish sauce acceleration. However, hydrolytic activity of proteinases bound at cell surface of this strain has not been well elucidated. Addition of 0.2 % CaCl(2) (w/w) in conjunction with starter cultures of Virgibacillus sp. SK 37 increased protein hydrolysis as measured by α-amino group content throughout fermentation (P < 0.05). Cell-bound proteinases from Virgibacillus sp. SK 37 were extracted into a free form by incubating the washed cells in Ca(2+)-free buffer at 37 °C for 2 h. Cell-bound proteinases revealed molecular mass of 19, 20, 22, 32, 34, and 44 kDa based on a synthetic peptide zymogram. The proteinases showed subtilisin-like serine characteristics with the highest activity at 50 °C and pH 8 and 11. Activity of the extracted proteinases increased ~4 times at ≥100 mM CaCl(2). In addition, CaCl(2) enhanced thermal stability of the extracted proteinases. Enzymes showed proteolytic activity in either the absence or presence of 10 and 25 % NaCl toward fish muscle, soy protein isolate, and casein substrates. Cell-bound proteinases were likely to play an important role in protein hydrolysis during fish sauce fermentation.

A comparative study of meat quality and vibrational spectroscopic properties of different chicken breeds.

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 ß-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.

Development of Flavor and Taste Components of Sous-Vide-Cooked Nile Tilapia (Oreochromis niloticus) Fillet as Affected by Various Conditions.

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).

Extremely halophilic strains of Halobacterium salinarum as a potential starter culture for fish sauce fermentation.

Proteinase-producing halophilic archaea were isolated from Thai fish sauce collected from industrial fermentation tanks at various periods of fermentation. Five isolates namely, J-1-S4, J-1-S13, J-1-S22, 2 m-40-15-R2, and P-1-S8, were identified as Halobacterium salinarum with slightly different colony and morphological characteristics among isolates. Starters of five isolates were prepared and added to the anchovy mixed with 25% solar salt and fermented for 180 days at 30-35°C. Halophilic bacteria/archaea counts of inoculated samples were decreased and undetected after 120 days of fermentation. At 180 days of fermentation, α-amino group contents of inoculated fish sauce samples (856-1010 mM) and total nitrogen content (2.2%-2.5%) were higher than the control without archaea inoculation (p < 0.05). All samples contained low amounts of biogenic amines, suggesting that all starters were not biogenic amine formers. The major volatile compound found in samples inoculated with H. salinarum P-1-S8 and H. salinarum 2 m-40-15-R2 was 3-methylbutanal, which contributes to the meaty note. Dimethyl disulfide, a compound that contributes to fecal note, was detected in all inoculated samples in a lower amount than in the commercial fish sauce (p < 0.05). Thus, H. salinarum accelerated protein hydrolysis and produced desirable volatile compounds during fish sauce fermentation in a strain-specific manner.