Characteristics and chemical composition of skins gelatin from cobia (Rachycentron canadum)

Gelatin was obtained from cobia (Rachycentron canadum) skins, which is an important commercial species for marine fish aquaculture, and it was compared with gelatin from croaker (Micropogonias furnieri) skins, using the same extraction methodology (alkaline/acid pre-treatments). Cobia skins gelatin showed values of protein yield, gelatin yield, gel strength, melting point, gelling point and viscosity higher than the values found from croaker skins gelatin. The values of turbidity and Hue angle for cobia and croaker gelatins were 403 and 74 NTU, and 84.8° and 87.3°, respectively. Spectra in the infrared region had the major absorption band in the amide region for both gelatins, but it showed some differences in the spectra. The proline and hydroxyproline contents from cobia skins gelatin (205 residues/1000 residues) was higher than from croaker skins gelatin (188 residues/1000 residues). SDS-PAGE of both gelatins showed a similar molecular weight distribution to that of standard collagen type I. Therefore, cobia skins could be used as a potential marine source of gelatin obtainment for application in diversified industrial fields.     

罗非鱼皮明胶酶解物及其模拟消化产物的抗氧化活性

以罗非鱼皮为原料提取罗非鱼皮明胶,选用风味蛋白酶和胰蛋白酶制备罗非鱼皮明胶酶解物,采用ABTS自由基、DPPH自由基、羟基自由基及亚油酸过氧化体系,初步评价罗非鱼皮明胶酶解物的抗氧化活性,再通过模拟体外胃肠道消化实验,结合分子质量分布测定,进一步考察罗非鱼皮明胶酶解物的抗氧化活性。结果显示,在酶解过程中,风味蛋白酶及胰蛋白酶酶解的水解度逐渐升高,在3 h时达到最高,分别达到5.8%和25.36%。在酶解60 min时其TCA可溶性肽得率最高,风味蛋白酶、胰蛋白酶酶解物分别达56.82%和54.44%。通过比较半抑制浓度（IC₅0）,确定了酶解60 min时风味蛋白酶酶解物的清除DPPH自由基及抑制亚油酸过氧化能力较胰蛋白酶酶解物强。模拟体外胃肠道消化后,酶解物羟基自由基清除活性均显著提高（p<0.05）,亚油酸脂质过氧化活性明显降低,消化前后样品分子量分布范围均主要集中于3000⁵000 Da,消化后风味蛋白酶及胰蛋白酶酶解物3000⁵000 Da组分的含量分别提高了45%及13%。以上研究结果表明,罗非鱼皮明胶酶解后制备的明胶水解物具有一定的抗氧化能力,具有潜在的开发价值。      

Improvement of the antioxidant properties of squid skin gelatin films by the addition of hydrolysates from squid gelatin

Gelatin hydrolysates (HG1 and HG2) were obtained from giant squid (Dosidicus gigas) gelatins (G1 and G2) by hydrolysis with Alcalase. Antioxidant properties of both gelatins were highly increased by hydrolysis, especially ABTS radical scavenging capacity, whereas no significant differences were found between HG1 and HG2. The amino acid composition of HG1 and HG2 closely resembled the amino acid composition of the parent proteins, gelatins G1 and G2. Both, HG1 and HG2 were composed by peptides below 30 kDa, although no clear protein bands were observed in HG2. Edible gelatin films with increasing percentages of HG1 (0–10%) were made from G1, giving rise to increasing values of FRAP and ABTS, as well as changes in mechanical properties (decrease puncture force and increase puncture deformation) and water vapour permeability (increase). HG1 gelatin hydrolysate showed lower antioxidant capacity in the gelatin films than in the free form at the same amount added into the filmogenic solution, probably due to interactions with protein matrix.     

Compositions and antioxidant properties of protein hydrolysates from the skins of four carp species

Compositions and antioxidant properties of protein hydrolysates prepared from four carp skins: black carp, grass carp, silver carp and bighead carp, using pepsin, with a degree of hydrolysis (DH) of 6–15%, were investigated. The yield of freeze‐dried hydrolysates was in the range of 54–62 g/100 g (dry skin). The content of protein and ash in four freeze‐dried hydrolysates was 72–81% and 8–17%, respectively. All hydrolysates contained high amount of hydrophobic amino acid residues (389–480 residues/1000 residues). Meanwhile, their antioxidant properties were evaluated by in vitro assays. The results revealed that all hydrolysates possessed potent antioxidant activities and showed dose dependency as the activity increased with sample concentration, capable of scavenging 72–88% of DPPH and 61–69% of hydroxyl radicals, respectively, at the highest tested concentration. The hydrolysates exhibited high reducing power and β‐carotene–linoleic acid oxidation inhibition. Among the four hydrolysates, the hydrolysate derived from bighead carp skin was superior to others in terms of yield, DH and antioxidant activities.     

The role of molecular size in antioxidant activity of peptide fractions from Pacific hake (Merluccius productus) hydrolysates

The antioxidative properties of Pacific hake hydrolysates and their peptidic fractions varying in molecular size were assessed. Hydrolysates produced by different proteases (Alcalase, bromelain, Flavourzyme, Protamex, Protease A“Amano”2, Protease N“Amano”K, Protin SD NY10, Umamizyme-K, Validase BNP-L, Validase FPexo) generally possessed good metal ion chelating (33–73% at 3 mg/ml), DPPH radical scavenging (18–30% at 1 mg/ml), ferric ion reducing power (abs_700nm 0.36–0.86 at 3 mg/ml) and ABTS radical scavenging (47–85% at 0.067 mg/ml) activity, as well as a good capability to suppress lipid peroxidation in a linoleic acid model system. Peptide size (<1.4 kDa) was important for ABTS radical scavenging activity, whereas specific peptide composition (which depended on the particular protease used) was the governing factor for effective lipid peroxidation. Validase BNP-L was the most promising enzyme for producing Pacific hake hydrolysates with good antioxidative activity in various assays and similar effectiveness as the synthetic antioxidant BHT to inhibit lipid peroxidation.     

Bioactive properties of enzymatic hydrolysates from abdominal skin gelatin of yellowfin tuna (Thunnus albacares)

Gelatin (90.6 ± 0.1%) was optimally prepared by response surface methodology from yellowfin tuna (Thunnus albacares, YT) abdominal skin. To investigate bioactive properties of enzymatic hydrolysates from the abdominal skin gelatin (ASG), ASG was hydrolysed with alcalase, protamex, neutrase and flavourzyme as affected by hydrolysis time. Antioxidant, nitrite scavenging and angiotensin‐I converting enzyme (ACE) inhibitory activities of the hydrolysates were determined. Antioxidant activities of the hydrolysates were found through linoleic acid peroxidation inhibitory effects. Alcalase‐derived hydrolysates (AHs) were more effective than others in metal ions chelating, superoxide anion scavenging and hydroxyl radical scavenging activities (P < 0.05). AHs showed significantly stronger nitrite scavenging activities (44.4–60.7%) than others (P < 0.05). Fraction A from AH showed strong ACE inhibitory activity (IC₅₀ of 0.75 mg mL^?1). These results suggest that YT ASG and its enzymatic hydrolysates could be functional food and/or pharmaceutical ingredients with potent antioxidant, anticarcinogenic and antihypertensive benefits.     

Biochemical properties and antioxidant activity of myofibrillar protein hydrolysates obtained from patin (Pangasius sutchi)

Antioxidant activities of myofibrillar protein hydrolysates (MPH) prepared from patin (Pangasius sutchi) using papain and Alcalase^® 2.4 L with different degrees of hydrolysis (DH) were investigated. With a DH of 65.83%, the hydrolysate prepared with papain exhibited the maximum of 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH) radical‐scavenging activity (71.14%) with a reducing power of 0.310. At a concentration of 1 mg mL^?1, the papain‐MPH exhibited a Trolox equivalent antioxidant capacity (TEAC) of 70.50 ± 1.22 μmol g^?1 protein. With a DH of 83.6%, the Alcalase‐MPH had the highest metal‐chelating activity. Low molecular weight peptides showed higher antioxidant activities than high molecular weight peptides. Both papain‐MPH and Alcalase‐MPH contained high amounts of the essential amino acids (48.71% and 48.10%, respectively) with glutamic acid, aspartic acid and lysine as the dominant amino acids. These results suggest that the protein hydrolysates derived from patin may be used as an antioxidative ingredient in both functional food and nutraceutical applications.     

Preparation and antioxidant activity of enzymatic hydrolysates from purple sea urchin (Strongylocentrotus nudus) gonad

Purple sea urchin (Strongylocentrotus nudus) gonad was treated separately with neutral protease, papain, pepsin and trypsin. The resultant hydrolysates were fractionated using a series of ultrafiltration membranes (molecular weight cut-offs of 10, 5, 3 and 1 kDa). Five fractions were prepared from each hydrolysate and the corresponding molecular weight ranges were below 10 kDa, 5-10 kDa, 3-5 kDa, 1-3 kDa and below 1 kDa. The peptide fractions were evaluated for antioxidant activity by using 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay and reducing power assay. Results indicated that all peptide fractions possessed DPPH radical scavenging capacity and reducing power in a dose-dependent manner. For all four hydrolysates, the below 1 kDa fractions exhibited the highest DPPH radical scavenging capacity. The below 1 kDa fractions prepared with neutral protease, papain and pepsin, and the 1-3 kDa fraction prepared with trypsin showed the highest reducing capacity among corresponding hydrolysates.     

Physicochemical and antioxidant properties of buckwheat (Fagopyrum esculentum Moench) protein hydrolysates

The enzymatic hydrolysis of common buckwheat (Fagopyrum esculentum Moench) protein isolate (BPI) by Alcalase and some physiochemical and antioxidant properties of the resulting hydrolysates were characterised. The hydrolysis resulted in remarkable decrease in the globulins or protein aggregates and concomitant increase in peptide fragments. The surface hydrophobicity of the hydrolysates decreased with increasing degree of hydrolysis (DH) and reached a minimum at DH 15%, but increased at further hydrolysis, whereas their amino acid compositions were unchanged. The polyphenol content of the hydrolysates gradually decreased with DH increasing from 0% to 15%, while it on the contrary increased upon further hydrolysis. The hydrolysates exhibited excellent antioxidant activities, including DPPH radical scavenging ability, reducing power and ability to inhibit linoleic acid peroxidation. The antioxidant activities of these hydrolysates were closely related to their polyphenol contents. The results indicated that polyphenol-rich buckwheat proteins are unique protein materials for the production of the hydrolysates with good nutritional and antioxidant properties.     

Antioxidant properties and protein compositions of porcine haemoglobin hydrolysates

Porcine haemoglobin hydrolysates were prepared through hydrolysis by Alcalase followed by Flavourzyme, and their protein compositions were analyzed using Sephadex G-50 gel filtration chromatography. The antioxidant activities, including reducing power, ferrous ion chelating ability, and DPPH radical scavenging activity, of the hydrolysates were evaluated. The results showed that the hydrolysates of haemoglobin exhibited low reducing powers, but high ferrous ion chelating abilities and DPPH radical scavenging activities. The hydrolysate, obtained through hydrolysis by 2% Alcalase for 4 h and followed by 1% Flavourzyme for 6 h, had the highest ferrous ion chelating ability of 63.54% at a concentration of 5.0 mg/mL. The hydrolysate, obtained through hydrolysis by 2% Alcalase for 4 hrs, had the highest DPPH radical scavenging activity of 41.94% at a concentration of 5.0 mg/mL. According to the results of protein composition analysis, we divided the hydrolysates into three groups, including high molecular weight (MW) group (Group I), medium MW group (Group II), and low MW group (Group III). The reducing power and ferrous ion chelating ability of the hydrolysates were significantly and positively correlated to the relative amount of Group I, and negatively correlated to the relative amount of Group III. This study revealed that the antioxidant activities of porcine haemoglobin hydrolysates were dependent on their protein compositions. The high MW protein fraction (Group I) was responsible for the high reducing power and ferrous ion chelating ability of the hydrolysate.     

Gelatin hydrolysates from sliver carp (Hypophthalmichthys molitrix) improve the antioxidant and cryoprotective properties of unwashed frozen fish mince

In this study, gelatin extracted from silver carp skin was hydrolysed by different enzymes (alcalase, papain and flavourzyme), aiming to improve the storage stability of unwashed fish mince during freeze-thaw cycles. Flavourzyme hydrolysates exhibited the highest DPPH• scavenging activity (1.2-fold of alcalase, 1.6-fold of papain) and reducing ability (1.2-fold of alcalase, 2.3-fold of papain), while the alcalase hydrolysates demonstrated the highest Fe²⁺ chelating ability. Moreover, fish mince treated with the mixture of flavourzyme hydrolysates and sucrose exhibited higher sulfhydryl and salt-soluble protein contents, greater Ca²⁺-ATPase activity and better water holding capacity. Hence, the gelatin hydrolysates of flavourzyme from silver carp skin are promising candidates as natural and high-performance antioxidant and cryoprotectants in fish mince processing.     

Angiotensin-I converting enzyme inhibitory and antioxidant activity of bioactive peptides produced by enzymatic hydrolysis of skin from grass carp (Ctenopharyngodon idella)

Grass carp skin pieces were homogenized in water and hydrolyzed by Alcalase®, collagenase, proteinase K, and/or trypsin at their optimum conditions. Samples were taken at various degrees of hydrolysis and were evaluated for antioxidant, antimicrobial, and angiotensin-converting enzyme inhibitory activities. Alcalase and collagenase completely hydrolyzed the skin with different rates, and released peptides with antioxidant and angiotensin-converting enzyme-inhibitory activity. These activities increased linearly with increasing degrees of hydrolysis. Subsequent incubation of the collagenase hydrolysates with trypsin slightly increased the antioxidant activity. Proteinase K, although only partially hydrolyzing the skin, also catalyzed the release of peptides with antioxidant and angiotensin-converting enzyme-inhibitory activities. These results show that skin by-products from grass carp can be a source of bioactive peptides produced by a one-step reaction. Such hydrolysates may be applied in food products to prolong shelf life and provide beneficial effects on blood pressure.     

The effect of enzymatic hydrolysis time and temperature on the properties of protein hydrolysates from Persian sturgeon (Acipenser persicus) viscera

Protein hydrolysate was prepared from the viscera of Persian sturgeon (Acipenser persicus), a major sturgeon species in the Caspian Sea. Hydrolysis was performed at three different temperatures (35, 45 and 55 °C), pH 8.5, using commercially available Alcalase^® and an enzyme to substrate ratio of 0.1 AU/g viscera protein over a 205 min incubation period. Protein and lipid content of the hydrolysate were 65.82%, and 0.18%, respectively. Protein recovery and degree of hydrolysis ranged from 34.97% to 61.96% and 13.32% to 46.13%, respectively. The highest degree of hydrolysis was observed at 55 °C after 205 min (p < 0.05). The amino acid score of the hydrolysates was similar to that of the FAO/WHO reference protein. It is revealed that Persian sturgeon visceral protein hydrolysate amino acid fulfils adult human requirements. Based on National Research Council guidelines, phenylalanine is the first limiting amino acid in the hydrolysate. However, the hydrolysate has the potential for application as an ingredient in formulated diets.     

Physical and chemical modification of SPI as a potential means to enhance small peptide contents and antioxidant activity found in hydrolysates

The hydrolytic behaviour of the thermal and chemical modified soy protein isolate (SPI) and the antioxidant activity of the resultant hydrolysates have been evaluated in the present study. Thermal treatment, especially in combination with the addition of sodium sulfite, increased the susceptibility of protein hydrolysis due to the unfolding of proteins and the cleavage of disulfide bonds. Succinylation and acylation resulted in significant decrease in amino acids content than that of phosphorylation due to steric hindrance effect as a result of introduction of longer and bulky side chains. Thermal and chemical modification resulted in a significant increase in small peptide content found in hydrolysates, which in turn enhanced significantly the 2,2-Diphenyl-1-picryhydrazyl (DPPH) radical scavenging activity of the hydrolysates, indicating that modification of proteins before hydrolysis could be an efficient way to enhance the small peptide content with desired antioxidant activity.Industrial relevanceEnzymatic hydrolysis of SPI using selected protease has been intensively studied, however, reports on optimization of the substrate by thermal and chemical modifications before hydrolysis to obtain desired hydrolysates are still limited. In this work, SPI was thermally treated or chemical modified and then hydrolyzed by Alcalase. The results indicated that physical and chemical modification of SPI could change the levels of formaldehyde nitrogen and small peptide nitrogen, and consequently enhanced significantly the DPPH radical scavenging activity of the hydrolysates. This work was helpful to provide a feasible and economic way to enhance the contents of small peptides with desired antioxidant properties.     

Textural and biochemical properties of cobia (Rachycentron canadum) sashimi tenderised with the ultrasonic water bath

The present study investigated the tenderisation effects ultrasound processing (UT) on farmed cobia sashimi. Age-treated cobia trunk muscles (AT) were used as the control. The pH, total volatile base nitrogen, trimethylamine nitrogen, thiobarbituric acid reactive substances, ATP catabolism components, K₁ value, and texture were evaluated. The texture of AT sashimi reached the optimal firmness range with 8.53 N at day 7. However, AT samples could not be served raw after day 7 because of their poor freshness indexes, including a TVBN value of 18.53 g/100 g, a TMAN value of 3.25 mg/100 g, and a TBARS value 0.983 MDA mg/100 g. Moreover, the K₁ value of AT sashimi was 20.21% at day 5. UT was employed to efficiently tenderise cobia sashimi with an initial firmness of 9.70-7.82 N after 90 min of treatment. The results of this study indicate that UT accelerates the biochemical reaction rate, as evidenced by the increases in the TVBN, TMAN, and TBARS contents; however, these values were very low. The results of this study could provide basic information for the development of a novel ultrasonic tenderisation technique in raw seafood designed for restaurants and consumers.     

Purification and characterisation of a new antioxidant peptide from chickpea (Cicer arietium L.) protein hydrolysates

An antioxidant peptide was purified using consecutive chromatographic methods from chickpea protein hydrolysates (CPH). This peptide was designated as Fra.7. It had a molecular weight of 717.37 Da, and its amino acid sequence was identified as Asn-Arg-Tyr-His-Glu by an ABI 4700 proteomics analyser. This antioxidant peptide was identified for the first time from food-derived protein hydrolysates. The molar ratio of the five amino acids in the sequence was 1:1:1:1:1. This antioxidant peptide efficiently quenched the free radical sources 1,1-diphenyl-2-pycryl-hydrazyl (DPPH), hydroxyl, and superoxide free radicals. The Cu²⁺ and Fe²⁺ chelating activities were 76.92% and 63.08% at the peptide concentration of 50 μg mL⁻¹, respectively. Furthermore, the inhibition of the Fra.7 on lipid peroxidation was greater than that of α-tocopherol. The inhibition ratio of the linoleic acid autooxidation was 88.81% at the eighth day of analysis.     

Chemical compositions and characterisation of skin gelatin from farmed giant catfish (Pangasianodon gigas)

Gelatin was extracted from the skin of farmed giant catfish (Pangasianodon gigas) with a yield of 20.1 g/100 g skin sample on the basis of wet weight. The chemical composition and properties of gelatin were characterised. The gelatin had high protein (89.1 g/100 g) but low fat (0.75 g/100 g) content and contained a high number of imino acids (proline and hydroxyproline) (211 residues per 1000 residues). Giant catfish skin gelatin had a slightly different amino acid composition than calf skin gelatin. The bloom strength of the gelatin gel from giant catfish skin gelatin (153 g) was greater than that of calf skin gelatin (135 g) (P < 0.05). Viscosity, foam capacity and foam stability of gelatin from giant catfish skins were in general greater than those of the gelatin from calf skin tested. SDS-PAGE of giant catfish skin gelatin showed a high band intensity for the major protein components, especially, α-, β- and γ-components and was similar to that of standard calf skin collagen type I.     

Radical scavenging properties of protein hydrolysates from Jumbo flying squid (Dosidicus eschrichitii Steenstrup) skin gelatin

Gelatin extracted from Jumbo flying squid skin was hydrolyzed with serials of protease. The scavenging effects of gelatin hydrolysates on superoxide and hydroxyl radical were assessed by chemiluminescence analysis. Properase E and pepsin have shown efficient hydrolysis of squid skin gelatin to obtain high radical scavenging activity peptides. The conditions chosen for enzymic hydrolysis of squid skin gelatin with properase E were pH 9.0, 45 °C, 3 h reaction time, and enzyme‐to‐substrate ratio of 1:50. Hydrolysates combining properase E and pepsin showed the best radical scavenging activity. For fragmented hydrolysates by ultrafiltration, fractions UF‐2 (M_w < 2000 Da) had high yield and radical scavenging activity. Copyright © 2006 Society of Chemical Industry