Characteristics and Select Functional Properties of Collagen from Golden Pompano (Trachinotus ovatus) Skins

Acid-solubilized collagen (ASC) and pepsin-solubilized collagen (PSC) from golden pompano skins were extracted and characterized. The molecular weight of ASC was about 130 kDa for α₁ and 115 kDa for α₂, which were slightly higher than those of PSC. Similar amino acid composition and Fourier transform infrared (FTIR) spectra were observed in both collagens, but slight differences were found in the peptide maps of collagen digested by V8 protease and trypsin. The denaturation temperatures (Tds) of ASC and PSC calculated from the reduced viscosity were 31.8 and 30.0°C, while the transition temperature (T_m) of ASC and PSC analyzed by DSC were 33.0 and 32.0°C, respectively. ASC has a lag phase, a growth phase, and a plateau phase in the turbidity–time curve, while PSC does not have similar phenomenon. It was found that the fibril gel of ASC could be formed at 25°C, leading to improved thermal stability.     

Isolation and characterization of collagens from the skin of giant grouper (Epinephelus lanceolatus)

ABSTRACT

Acid-solubilized collagen (ASC) and pepsin-solubilized collagen (PSC) were extracted from the skin of giant groupers (Epinephelus lanceolatus) with yields of 39.51 and 19.12%, respectively. ASC and PSC consisted of two different α chains (α1 and α2) and were characterized to be type I collagen with no disulfide bond. The imino acid contents of the ASC and PSC from giant grouper skin were 189 and 181 per 1,000 residues, respectively. The maximum endothermic temperatures (T_max) of ASC and PSC measured by differential scanning calorimetry (DSC) were 31.71 and 31.33°C, respectively. The denaturation temperatures of ASC and PSC measured by viscometry were 29.84 and 29.05°C, respectively. The maximum solubility in 0.5 M acetic acid was observed at pH 5 and pH 6 for ASC and PSC, respectively. A sharp decrease in solubility was observed for both ASC and PSC in the presence of NaCl above 3% (w/v).     

Isolation and Characterization of Pepsin-Soluble Collagen from the Skin of Peru Squid (Dosidicus gigas)

Pepsin-soluble collagen (PSC) was isolated from Peru squid (Dosidicus gigas) skin and physicochemical properties of the PSC were determined. The PSC exhibited a maximum absorbance at 220 nm. Sodium dodecyl sulfate polyacrylamide gel electrophoresis suggested the collagen containing α1 and α2 chain was classified as type I collagen. Amino acid composition indicated that the collagen had lower amino acid content than that of mammalian collagen. Denaturation temperature (T_d) of the PSC was 26.8°C. The PSC had relatively high solubility in alkaline condition or NaCl concentrations below 2%. Fourier transform infrared spectroscopy investigations showed the existence of helical arrangements of collagen. The lyophilized collagen had a uniform and regular network structure. These results suggested that Peru squid skin was a potential source of collagen for further research and application.     

Characterization of Acid-Soluble Collagen From Bone of Pacific Cod (Gadus macrocephalus)

Acid-soluble collagen (ASC) was isolated from Pacific cod (Gadus macrocephalus) bone. The ASC was rich in glycine. The amount of imino acid was lower than that of calf skin collagen, as was the transition temperature (48.6°C). Electrophoresis revealed two different α chains (α₁ and α₂), β-component, and γ-component. Fourier transform infrared spectroscopy measurement showed that ASC was in triple-helix structure. ASC had a solubility greater than 90% in a very acidic pH range (pH 1–4), and the solubility decreased with increasing NaCl concentrations up to 3%. Lyophilized ASC had a network ultrastructure with lace-like fibers, similar to calf skin collagen sponge.     

Physicochemical Properties of Pepsin-Solubilized Collagen Extracted from Nile Tilapia (Oreochromis niloticus) Scale

ABSTRACT

The aim was to investigate the physicochemical properties of pepsin-solubilized collagen (PSC) extracted from fish scales of Nile tilapia. The results indicated that Nile tilapia scales are rich in collagen. Therefore, the scales are a promising cost-effective collagen source. The conversion of hydroxyproline to collagen was 12.9. Based on the patterns of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), PSC comprised at least two different α chains, α1 and α2, and was classified to be type I with no disulfide. Fourier transform infrared spectroscopy spectrum showed that the bands of amide A, I, and II of PSC were found at 3,301, 1,631, and 1,239 cm^?1 wavelength. The content of imino acids was 187 residues per 1,000 total amino acid residues. Maximum solubility was observed at pH 4, and minimum was at pH 7. Almost no change in solubility was observed in the presence of NaCl up to 2% (w/v), and the decrease was more pronounced with increasing NaCl concentration. The temperature of denaturation was 35.4°C. Results show that PSC has physicochemical properties that can be applied in the cosmetics, biomedical, pharmaceutical, and food industries.     

Autolysis and Characterization of Sarcoplasmic and Myofibril Associated Proteinases of Oxeye Scad (Selar boops) Muscle

ABSTRACT

The autolytic profile of oxeye scad mince was characterized. Mince showed higher proteolytic activity than washed mince. The highest autolysis was observed at 65 and 60°C for mince and washed mince, respectively. Both mince and washed mince showed the optimum pH for autolysis at pH 9.0, and their activities decreased with increasing NaCl concentration (0–3.5%). Autolysis of washed mince was markedly inhibited by soybean trypsin inhibitor (SBTI), suggesting that myofibril-associated proteinase was serine proteinase. Sarcoplasmic proteinase was characterized to be heat-activated alkaline proteinase having the optimal pH and temperature of 9.0 and 60°C, respectively. The activities were stable at pH range of 8.0–11.0 at 20–40°C. The crude proteinase was inhibited by N-p-tosyl-L-lysine chloromethyl ketone, SBTI, and phenylmethanesulfonyl fluoride, suggesting the predominance of serine proteinases, especially trypsin. NaCl suppressed the activity while β-mercaptoethanol, dithiothreitol, and CaCl₂ activated the activity. Therefore, trypsin-like proteinase is a major endogenous proteinase responsible for autolysis in oxeye scad muscle. The present results can be used as scientific guidelines to predict the gel strength of surimi made from oxeye scad muscle.     

Elevated temperature affects growth and hemato‐biochemical parameters,inducing morphological abnormalities of erythrocytes in Nile tilapia Oreochromis niloticus

Temperature is considered as an important environmental factor, and the increasing water temperature resulting from global warming is a great concern. The present study was conducted to examine the effects of elevated water temperature on growth, hemato‐biochemical parameters in Nile tilapia, Oreochromis niloticus acclimatized to three temperatures (31°C, 34°C and 37°C) for 60 days. Additionally, erythrocytic cellular abnormalities (ECA) and erythrocytic nuclear abnormalities (ENA) tests were assayed using peripheral erythrocytes after exposure to the three temperatures. Fish were sacrificed on days 7, 15, 30 and 60 of exposure. Growth performances viz., weight gain, % weight gain and specific growth rate (SGR) showed decreasing tendency at 34°C but significantly declined at 37°C compared to 31°C. The abundance of haemoglobin (Hb) and red blood cells (RBCs) significantly decreased in response to temperature increases, while white blood cells (WBCs) displayed the opposite response. At days 7 and 15, blood glucose levels significantly increased in response to the temperature increase, while at days 30 and 60 glucose declined. Frequencies of ECA and ENA were significantly enhanced at the highest temperature throughout the experimental period. Dissolved oxygen decreased and free CO₂ increased significantly with increasing temperature throughout the study period. The present study revealed that temperatures higher than 34°C may be hazardous to O. niloticus.     

Physicochemical and Reactive Oxygen Species Scavenging Properties of Collagen and Collagen Hydrolysates from Farmed Globefish (Fugu rubripes) Bone

Globefish (Fugu rubripes) bone collagen (GBC) was isolated and characterized. Electrophoresis and Fourier transform infrared (FTIR) spectra indicated that GBC was high-purity type I collagen with an intact triple-helical structure. Amino acid analysis revealed that GBC contained higher glycine content (364 residues/1,000 residues) than that of other fishery bones, and the imino acid content was 189 residues/1,000 residues. The denaturation temperature (T_d) of GBC was 27.1°C. GBC displayed a rapid fibril-forming rate and well-defined fibril morphology in vitro. Globefish bone collagen hydrolysates (GBCH) were optimized by neutral protease through response surface methodology (RSM) with the highest hydroxyl radical (?OH) scavenging capacity. The optimal hydrolysis condition was a reaction temperature of 48.4°C, time of 120 min, and enzyme/substrate ratio of 2520 U/g. Three fractions were collected by ultrafiltration, and the fraction GBCH-III (Mw<1 kDa) possessed the strongest ?OH scavenging ability. Reactive oxygen species (ROS) scavenging capacities were evaluated with two model systems: ?OH and nitric oxide (NO?). The ROS scavenging capacities increased in a dose-dependent manner. GBCH-III proved to be a more effective ?OH scavenger than reduced glutathione (GSH). These findings demonstrated that GBC and GBCH have potential as an alternative source of collagen-derived materials for use in pharmaceutical, nutraceutical, and cosmetic industries.     

The effects of temperature on respiration of Amur sturgeon,Acipenser schrenckii,at two acclimation temperatures

In order to clarify the respiratory responses strategy of Amur sturgeon Acipenser schrenckii exposed to water temperature changes, respiratory parameters of the fish were studied under two temperature regimes: fish acclimated at 13°C for Group I, temperature was increased to 16°C, 19°C, 22°C and 25°C and then returned stepwise to 22°C, 19°C, 16°C and 13°C; and fish acclimated at 25°C for Group II, the water temperature was reduced in steps to 22°C, 19°C, 16°C and 13°C, subsequently, returned to 16°C, 19°C, 22°C and 25°C. The results showed that the respiratory frequency (f_R), oxygen consumption rate (VO₂) and gill ventilation (V_G) of the fish were directly dependent on the acute temperature in both acclimation groups (p < .05). The initial 25°C VO₂ in Group II was significantly higher than the initial 13°C VO₂ in Group I (p < .05), but was significantly lower than that at 25°C in Group I (p < .05). In Group I, respiratory stroke volume (V_S.R) of fish significantly increased or decreased with the acute temperature increases or decreases, respectively (p < .05); oxygen consumption efficiencies (EO₂) of fish did not significantly show differences when temperature increased to 25°C from 13°C (p > .05), but the EO₂ significantly declined while returning to acclimation temperature (p < .05). In Group II, the V_S.R of the fish did not significantly change with acute temperature fluctuations between 25 and 13°C (p > .05), while the EO₂ increased with acute temperature increases (p < .05). The Q₁₀ values for f_R, VO₂, V_S.R, V_G and EO₂ were 1.53–1.72, 1.92–2.06, 1.07–1.60, 1.78–2.44 and 1.11–1.65 at 13–25°C of temperature interval respectively. Amur sturgeon showed partial metabolic compensation to temperature changes. The study results suggest that the ability of Amur sturgeon to regulate metabolism in response to acute temperature changes makes this species good adaptability in the aquaculture rearing.     

Extraction and Characterization of Pepsin Soluble Collagen from the Body Wall of Sea Cucumber Acaudina leucoprocta

Sea cucumber Acaudina leucoprocta is a potential alternative collagen source. However, the high levels of heavy metals contained in the body wall restricts its utilization. In this work, an efficient method was established to remove the heavy metals accumulated in the body wall of A.leucoprocta by demineralizing with 0.2 M ethylenediaminetetraacetic acid (EDTA). The resulting body wall of A.leucoprocta was then used for extracting pepsin-soluble collagen (PSC) with pepsin proteolysis. The PSC from the body wall of A.leucoprocta was obtained with a yield of 43.99 ± 0.65% (dry weight) and high purity. Maximum and minimum solubility for the isolated PSC in 0.5 M acetic acid was observed at pH 2.66 and 4.43, respectively. The solubility was remarkably decreased in the presence of NaCl. The denaturation temperature of PSC rehydrated in 0.5 M acetic acid was measured as 25.4°C. The PSC was characterized as type I collagen, which consists of three α₁ chains without α₂ chain. Interestingly, α₁ chain in PSC showed two isoforms with the pI values of 4.02 and 4.29. The heavy metals existing in PSC were all below the contaminant limit of edible gelatin. The PSC isolated from the body wall could be an alternative to mammalian collagens.     

Pharmacokinetics and tissue distribution of bronopol in grass carp,Ctenopharyngodon Idella,at 15 and 20°C

This study explored the pharmacokinetic characteristics of bronopol in grass carp (Ctenopharyngodon idella) under different temperatures. The concentrations of bronopol in the plasma and tissues (kidney, liver and muscle) of grass carp after soaking in bronopol (6.75 µg/ml) at 15 and 20°C were determined using liquid chromatography–mass spectrometry. The concentrations of bronopol in both plasma and tissue first increased and then decreased with soaking time at both tested temperatures. The peak bronopol concentrations at 15°C (plasma: 8.934 µg/ml; kidney: 9.23 µg/g; liver: 9.47 µg/g; and muscle: 7.98 µg/g) were slightly lower than those at 20°C (plasma: 9.654 µg/ml; kidney: 10.83 µg/g; liver: 13.40 µg/g; and muscle: 8.68 µg/g). In an analysis of pharmacokinetic parameters, the bronopol concentration–time profiles for plasma were best described by a two‐compartment, open pharmacokinetic model with first‐order absorption. The t_1/2β for plasma at 15 and 20°C was 82.933 and 92.041 hr, respectively, indicating that the elimination of bronopol was faster at 15°C than at 20°C. In addition, AUC_0‐t and AUC_0‐∞ values of bronopol were 477.892 and 495.809 µg/L hr at 15°C and 495.809 and 589.859 µg/L hr at 20°C, indicating that the content of bronopol in the organism was higher at higher temperature.     

Purification and characterization of three trypsin isoforms from viscera of sardinelle (<Emphasis Type="Italic">Sardinella aurita</Emphasis>)

Three trypsin isoforms A, B and C were purified to homogeneity from the viscera of sardinelle (Sardinella aurita). Purification was achieved by ammonium sulfate precipitation (20–70% (w/v)), Sephadex G-100 gel filtration and Mono Q-Sepharose anion-exchange chromatography. The molecular weights of these purified enzymes were estimated to be 28.8 kDa by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE). Based on the native PAGE and casein-zymography, each purified trypsin appeared as a single band. Trypsins A and C exhibited the maximal activity at 55°C, while trypsin B at 50°C. All isoforms showed the same optimal pH (pH 9.0) using Nα-benzoyl-dl-arginine-p-nitroanilide (BAPNA) as a substrate. The three trypsins were stable at temperatures below 40°C and over a broad pH range (7.0–11.0). The activities of the three isoforms were strongly inhibited by soybean trypsin inhibitor and phenylmethylsulfonyl fluoride, a serine protease inhibitor, and partially inhibited by ethylenediaminetetraacetic acid, a metalloenzyme inhibitor. Kinetic constants of trypsins A, B and C for BAPNA were evaluated at 25°C and pH 9.0. The values of K _m and k _cat were 0.125, 0.083 and 0.10 mM, and 2.24, 1.21 and 5.76 s⁻¹, respectively. The N-terminal sequences of the first 10 amino acids were “I V G G Y E C Q K Y” for trypsin A and “I V G G Y E A Q S Y” for trypsins B and C. These sequences showed highly homology to other fish trypsins.     

Influence of Different Hydrolysis Processes by Trypsin on the Physicochemical,Antioxidant, and Functional Properties of Collagen Hydrolysates from Sphyrna lewini,Dasyatis akjei,and Raja porosa

ABSTRACT

Acid soluble collagen hydrolysates from the cartilages of Sphyrna lewini, Dasyatis akjei, and Raja porosa on different hydrolysis conditions by trypsin were prepared and named as S, D, and R, respectively. The hydrolysates (S1, D1, and R1) obtained from hydrolysis in pH 2.5, 3 h, 37°C presented wonderful foaming and emulsifying capacities, caused by their high average molecular weights (AMWs), high degree of Pro and Lys hydroxylation, and imino acid contents (17.1, 15.3, and 14.1%). At the concentration of 0.1% (w/v), the foaming capacities of S1, D1, and R1 were 104.75 ± 2.57, 63.87 ± 2.73, and 76.87 ± 2.02%; and the emulsifying activity indexes of them were 116.07 ± 1.89, 91.04 ± 1.79, and 123.85 ± 2.14 m²/g, respectively. Conversely, hydrolysates (S2, D2, and R2) obtained from hydrolysis in pH 7.8, 3 min, 37°C exhibited strong scavenging activity on 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical (EC₅₀ 0.568, 0.680, and 1.634 mg/mL), hydroxyl radical (EC₅₀ 0.253, 0.376, and 0.438 mg/mL), and moderate reducing power (0.0465–0.4702 at the concentration of 5 mg/mL), due to their low AMWs. The results indicated that S1, D1, and R1 could be used as emulsifiers and foaming agents, and S2, D2, and R2 could be used as natural antioxidants in food systems.     

Effects of manufacturing factors on the viscosity of a polysaccharide solution extracted from Gagome Kjellmaniella crassifolia

A highly viscous polysaccharide solution was extracted from Gagome Kjellmaniella crassifolia in 20°C water. The eluted sugar concentration was 0.16%, and the extracted carbohydrates consisted of fucoidan, laminaran, and alginate at an approximately ratio of 8.2:0.8:1.0. An increase in the extraction temperature resulted in a less viscous solution, even though the amount of eluted sugar was higher than that obtained at a lower temperature. The most viscous extraction solution was obtained at a neutral pH, with a more acidic or alkaline extraction solution being a less viscous. When the highly viscous polysaccharide solution was heated, the viscosity decreased markedly with increasing temperature. The viscosity of the polysaccharide solution increased after dialysis against water and decreased with the addition of either KCl or NaCl. However, the viscosity was recovered to previous levels by following re-dialysis against water. The removal of divalent cations by EDTA and the re-addition using CaCl₂ or MgCl₂ also caused reversible changes to the viscosity. These characterizations will be useful for widespread applications of viscous K. crassifolia polysaccharides.     

Thermal challenge of the Indian shrimp Penaeus indicus

This study investigated short‐term effects of increasing water temperature from 27 to 41°C on survival and feed consumption of Penaeus indicus at three different ages: PL₂₅ (postlarvae 25 days old), PL₅₀ and PL₉₀. For each age group, water temperature was maintained at 27°C in the control, but increased to 32, 35, 38 and 41°C at a rate of 1°C every eight hours. The temperature was then kept stable until the end of the 7‐day experiment. Results showed that increasing water temperature affected both survival and feed consumption of the experimental shrimps (p < .01). Survival was highest at 32 and 35°C ranging from 93.8% to 100%, but significantly reduced to 40.0%–81.6% at 38°C. No shrimp survived the 41°C treatment. PL₂₅ were more tolerant to 38–41°C than PL₅₀ and PL₉₀ in terms of survival. Increasing water temperature had no effects on feed consumption of PL₂₅ (p > .05). For PL₅₀ and PL_90, feed consumption significantly increased at 38 and 41°C (p < .01) and was similar within the range of 27–35°C. This study suggests that P. indicus in tropical areas can tolerate water temperatures of at least 35°C and should be considered for farming during the summer time.     

Partial characterization of pyloric-duodenal lipase of gilthead seabream (<Emphasis Type="Italic">Sparus aurata</Emphasis>)

In the present study, we report the isolation and characterization of seabream Sparus aurata pyloric caeca-duodenal lipase. Optimum activity was found at pH 8.5 and salinity of 50 mM NaCl. Lipase activity was sensitive to divalent ions, and extreme pH values (4, 5, and 12), being more stable at alkaline than acid pH. Optimum temperature was found at 50°C, but lipase was stable at temperatures below 40°C. Lipase has a bile salt sodium taurocholate requirement for increased activity. Gradient PAGE electrophoresis revealed the presence of four isoforms with apparent molecular masses of 34, 50, 68, and 84 KDa, respectively. Pyloric-duodenal lipase was able to hydrolyze emulsified alimentary oils. Results confirm the presence of true lipases in Sparus aurata digestive tract.     

Effect of temperature on excess post-exercise oxygen consumption in juvenile southern catfish (<Emphasis Type="Italic">Silurus meridionalis</Emphasis> Chen) following exhaustive exercise

The effects of temperature on resting oxygen consumption rate (MO_2rest) and excess post-exercise oxygen consumption (EPOC) after exhaustive exercise (chasing) were measured in juvenile southern catfish (Silurus meridionalis) (8.40 ± 0.30 g, n = 40) to test whether temperature has a significant influence on MO_2rest, maximum post-exercise oxygen consumption rate (MO_2peak) and EPOC and to investigate how metabolic scope (MS: MO_2peak − MO_2rest) varies with acclimation temperature. The MO_2rest increased from 64.7 (10°C) to 160.3 mg O₂ h⁻¹ kg⁻¹ (25°C) (P < 0.05) and reached a plateau between 25 and 30°C. The post-exercise MO₂ in all temperature groups increased immediately to the peak values and then decreased slowly to a steady state that was higher than the pre-exercise MO₂. The MO_2peak did not significantly differ among the 20, 25 and 30°C groups, though these values were much higher than those of the lower temperature groups (10 and 15°C) (P < 0.05). The duration of EPOC varied from 32.9 min at 10°C to 345 min at 20°C, depending on the acclimation temperatures. The MS values of the lower temperature groups (10 and 15°C) were significantly smaller than those of the higher temperature groups (20, 25 and 30°C) (P < 0.05). The magnitude of EPOC varied ninefold among all of the temperature groups and was the largest for the 20°C temperature group (about 422.4 mg O₂ kg⁻¹). These results suggested that (1) the acclimation temperature had a significant effect on maintenance metabolism (as indicated by MO_2rest) and the post-exercise metabolic recovery process (as indicated by MO_2peak, duration and magnitude of EPOC), and (2) the change of the MS as a function of acclimation temperature in juvenile southern catfish might be related to their high degree of physiological flexibility, which allows them to adapt to changes in environmental conditions in their habitat in the Yangtze River and the Jialing River.     

Determination of optimal temperature(s) in juvenile red‐spotted grouper Epinephelus akaara (Temminck & Schlegel) based on growth performance and stress responses

This study sought to determine the optimal temperature(s) for aquaculture of juvenile red‐spotted grouper Epinephelus akaara (Temminck & Schlegel) (mean initial BW: 3.1 g). Growth performance, insulin‐like growth factor 1 (IGF‐1) expression and thermal stress responses (plasma cortisol, glucose, and hepatic heat shock protein 60 expression) were evaluated at three constant temperatures (24°C, 26°C and 28°C) in a 2‐week trial. At the end of the trial, final BW was significantly higher at 26°C and 28°C than at 24°C (p < 0.05); a quadratic regression analysis of final BW showed the optimum temperature for growth was 27.5°C (p < 0.05, R² = 0.806). The highest hepatic IGF‐1 expression was observed at 26°C (p < 0.05). On the other hand, hepatic heat shock protein 60 expression was highest at 28°C (p < 0.05), suggesting thermal stress. In conclusion, temperature optima, which support excellent growth but induce minimal thermal stress, was 26°C. This fine information within a narrow temperature range is expected to give empirical information for red‐spotted grouper farmers to sustain maximal production efficiency with avoiding thermal stress and to determine the future location of production, especially in consideration of arising seawater temperatures.