Seasonal differences in the properties of gelatins extracted from skin of silver carp (Hypophthalmichthys molitrix)

The gelatins were extracted from skins of silver carp (Hypophthalmichthys molitrix) caught in winter and summer, respectively. The physicochemical (molecular weight distribution and melting point) and rheological characteristics (viscosity property), as well as functional properties (emulsifying capacity and stability) of the gelatin from winter silver carp skin were compared with those of the summer equivalent. The results showed the properties of the summer gelatin were superior to those of the winter one, showing higher viscosity, emulsion stability, melting point and lower concentration for gelling. The summer gelatin has slightly denser strands of the gel microstructure which was observed by scanning electron microscopy (SEM). Different properties of gelatins from skin of silver carp may be attributed to the big discrepancy of the environmental temperature in the two seasons.     

Participation of cathepsin L in modori phenomenon in carp (Cyprinus carpio) surimi gel

Cathepsin L (Cat L) in carp (Cyprinus carpio) dorsal muscles was purified and its molecular weight determined by SDS polyacrylamide gel electrophoresis (SDS–PAGE) was 36 kDa. Its optimal temperature and pH were 50 °C and 5.5, respectively. The results of the effects of specific substrates, activators and inhibitors on the enzymatic activity showed that Cat L belongs to the family of cysteine proteinases containing thiol. Compared to the control, the gel strength of surimi with the addition of purified Cat L decreased significantly by 24.33% while that of surimi with both purified Cat L and inhibitors increased by 13.7% and 21.6%, respectively, suggesting the participation of Cat L in the modori phenomenon occurring in carp surimi. Both the SDS–PAGE electrophoretic pattern and microstructure figure revealed that Cat L could hydrolyse the main protein in carp surimi and was one of the enzymes involved in the modori phenomenon.     

Production and refinement of oil from carp (Cyprinus carpio) viscera

Carp viscera oil can be obtained by both ensilage and fishmeal processes. This study examined the refinement of carp (Cyprinus carpio) oils obtained by both processes, and compared crude, neutralised, bleached, winterised and deodorised oils’ characteristics and lipid profiles. Refined oils obtained by the two processes did not present significant difference (p > 0.05) for Lovibond colour, free fatty acids, and thiobarbituric acid values. The major fatty acids identified in the carp crude, bleached and refined oils were oleic, palmitic, palmitoleic, linoleic and linolenic, constituting approximately 69.6% of the total fatty acids of the oils. The n − 3/n − 6 ratio was approximately 1.05 for refined oil. Therefore, carp viscera refined oil can be considered a rich source of essential fatty acids of the n − 3 and n − 6 series.     

Properties of collagen from skin,scale and bone of carp (Cyprinus carpio)

Carp (Cyprinus carpio) is one of the main species of freshwater fish produced in China. Acid-soluble collagens (ASC) were prepared from carp skin, scale and bone. The yields of skin ASC, scale ASC and bone ASC are 41.3%, 1.35% and 1.06% (on the dry weight basis), respectively. SDS–PAGE pattern showed that ASCs of carp skin, scale and bone were all type I collagen, which were composed of two α₁ and one α₂ chains. The molecular weight of α₂ chain is 116 KDa. The amino acid composition and peptide maps of ASCs were similar to each other, but they were totally different from those of cod skin ASC. Denaturation temperatures (T_d) of ASCs were around 28 °C. Fourier transform infrared spectroscopy proved that ASCs were integrate and native. The results suggest that carp skin, scale and bone collagens have potential as an alternative source of collagen for use in various fields.     

The characteristics of gelatin extracted from sturgeon (Acipenser baeri) skin using various pretreatments

The physicochemical characteristics of gelatin obtained by different pretreatments of sturgeon (Acipenser baeri) skin with alkaline and/or acidic solutions have been studied. Visual appearance, pH, gel strength, viscosity and amino acid profile of the gelatins were evaluated. Pretreatment with alkaline solutions of Ca(OH)₂ and/or acetic acid (HAC) provided gelatin with a favourable colour. Pretreatment with alkali removed noncollagenous proteins effectively, whilst acid induced some loss of collagenous proteins. Gel strength and viscosity of gelatin pretreated with HAC or alkali followed by HAC were as high as gelatin extracted in the presence of protease inhibitors. Amino acid composition had no significant effect on the gelatin characteristics. The total acid concentration for the highest gel strength was inversely proportional to ionisation strength, and the preferred pH for extracting gelatin with the optimum gel strength was approximately 5.0. The results showed that any available protons, regardless of the type or concentration of the acid, inhibit protease activity, which significantly affects the gelatin characteristics.     

斑点叉尾鮰鱼皮明胶的制备

目的以斑点叉尾鮰鱼皮为原料,采用酸碱法制备鱼皮明胶;方法选用NaOH溶液和H2SO4溶液进行明胶的提取,通过单因素试验和正交试验,确定斑点叉尾鮰鱼皮明胶最佳制备工艺;结果NaOH质量分数为0.3%,H2SO4质量分数为0.4%,处理时间均为120 min,提取温度45℃,提取时间6 h,此时,所得明胶的凝胶强度和黏度分别为672.2 g和9.46 mPa·s,明胶提取率为65.21%.     

Characteristics and functional properties of gelatin from zebra blenny (Salaria basilisca) skin

Gelatin was extracted from alkali-pretreated skin of zebra blenny (Salaria basilisca) using commercial pepsin with a yield of 18 g/100 g of skin sample. The polypeptides pattern, gel strength, viscosity, textural parameters and functional properties of the zebra blenny skin gelatin (ZBSG) were investigated. Amino acid analysis revealed that ZBSG contained almost all essential amino acids, with glycine being the most predominant one. ZBSG was identified as a type I gelatin, containing α₁ and α₂-chains as the major constituents. Its gel strength and viscosity were 170.2 g and 5.95 cP, respectively. Fourier transformed infrared spectroscopy (FT-IR) spectra showed helical arrangements in its structure. Its solubility and functional properties were concentration-dependent. While foam expansion (FE) and foam stability (FS) increased with the increase of concentration, emulsifying activity index (EAI) and emulsion stability index (ESI) were noted to decrease. ZBSG also showed strong clarification ability particularly for apple juice, without affecting nutritional value.     

Chemical composition and characteristics of skin gelatin from grey triggerfish (Balistes capriscus)

Gelatin was extracted from the skin of grey triggerfish (Balistes capriscus) by the acid extraction process with a yield of 5.67 g/100 g skin sample on the basis of wet weight. The chemical composition and functional properties of gelatin were investigated. The gelatin had high protein (89.94 g/100 g) but low fat (0.28 g/100 g) contents. Differences in the amino acid composition between grey triggerfish skin gelatin (GSG) and halal bovine gelatin (HBG) were observed. GSG contained a lower number of imino acids (hydroxyproline and proline) (176 residues per 1000 residues) than HBG (219 residues per 1000 residues), whereas the content of serine was higher (40 versus 29 residues per 1000 residues, respectively). The gel strength of the GSG (168.3 g) was lower than that of HBG (259 g) (p < 0.05) possibly due to lower hydroxyproline content. Grey triggerfish skin gelatin exhibited a slightly lower emulsifying activity and water-holding capacity but greater emulsifying and foam stability, foam formation ability and fat-binding capacity than the halal bovine gelatin (p < 0.05). SDS-PAGE of GSG showed high band intensity for the major protein components, especially, α- and β-components and a similar molecular weight distribution to that of standard calf skin collagen type I.     

Extraction and functional properties of gelatin from the skin of cuttlefish (Sepia officinalis) using smooth hound crude acid protease-aided process

The characteristics and functional properties of gelatin from skin cuttlefish (Sepia officinalis) were investigated and compared to those of halal bovine gelatin (HBG). The gelatin extraction efficiency was improved by an acid-swelling process in the presence of smooth hound crude acid protease extract (SHCAP). The yields of gelatins from cuttlefish skin after 48 h with acid and with crude acid protease (15 units/g alkaline-treated skin) were 2.21% and 7.84%, respectively. The gelatin from skin cuttlefish had high protein (91.35%) but low fat (0.28%) contents. Compared to HBG, the cuttlefish-skin gelatin (CSG) has different amino acids composition than halal bovine gelatin. CSG contained slightly low hydroxyproline and proline (180‰) than HBG (219‰), whereas the content of serine was higher (49‰ versus 29‰). The gel strength of the gelatin gel from CSG (181 g) was lower than that of HBG (259 g) (p < 0.05) possibly due to lower hydroxyproline content. Cuttlefish-skin gelatin exhibited a similar emulsifying activity but greater emulsifying and foam stability than the halal bovine gelatin (p < 0.05). Foam formation ability, foam stability and water-holding capacity of CSG were slightly lower than those of the HBG, but fat-binding capacity was higher in the cuttlefish gelatin.     

Preparation and characterisation of gelatins from the skins of sin croaker (Johnius dussumieri) and shortfin scad (Decapterus macrosoma)

Gelatins were prepared from the skins of the tropical fish, sin croaker (Johnius dussumeiri) and shortfin scad (Decapterus macrosoma). Visual appearance, colour, pH, bloom strength, viscoelasticity, melting point and amino acid profiles of the fish gelatins were evaluated. Shortfin scad gelatin had higher melting and gelling temperatures than those of sin croaker gelatin. The bloom strengths of gelatins from sin croaker and from shortfin scad were 125 and 177 g, respectively, compared to 240 g for commercial bovine gelatin. The pH values were significantly different between the solutions of the two fish gelatins. The elastic modulus (G′) of the fish gelatin gels increased by more than 10-fold and the viscous modulus (G″) of fish gelatin solution increased sixfold after holding at 5 °C for 2 h. These viscoelastic properties of bovine gelatin only increased by less than twice.     

Purification and characterization of a leucine aminopeptidase from the skeletal muscle of common carp (Cyprinus carpio)

An aminopeptidase was purified from the skeletal muscle of common carp (Cyprinus carpio) to homogeneity, with 1160-fold purification and a yield of 4.3%. The purification procedure consisted of ammonium sulfate fractionation and sequential chromatographic steps, including DEAE-Sephacel, Sephacryl S-200, hydroxyapatite, Phenyl Sepharose and Macro-Prep High Q columns. The molecular mass of the enzyme was estimated to be 105 kDa and 100 kDa by SDS-PAGE under reducing conditions and gel filtration chromatography, respectively, suggesting it to be a monomer. The enzymatic activity was optimal at 35 °C and pH 7.0. The metal-chelating agents EDTA, EGTA and 1,10-phenanthroline specifically inhibited the enzyme activity while inhibitors of other proteinases did not show much effect, indicating that it was a metalloproteinase. Furthermore, bestatin, a specific aminopeptidase inhibitor strongly inhibited its activity. Divalent cations Mn²⁺, Mg²⁺ and Ba²⁺ slightly enhanced the enzymatic activity, while Co²⁺, Cu²⁺, Zn²⁺, Ca²⁺ and Fe²⁺ inhibited the activity to different extents. In addition, a sulfhydryl reagent was necessary to maintain its activity. Substrate specificity study revealed that the purified enzyme preferentially hydrolyzed Leu-MCA, followed by Arg-MCA, Ala-MCA and Tyr-MCA and it was thus regarded as a leucine aminopeptidase (LAP, EC 3.4.11.1). The apparent K_m and V_max values of the enzyme were 4.6 μM and 9.6 μmol min⁻¹ mg⁻¹, respectively for substrate Leu-MCA. This is the first report concerning purification and characterization of LAP from freshwater fish.     

Characteristics of gelatin from the skin of unicorn leatherjacket (Aluterus monoceros) as influenced by acid pretreatment and extraction time

Gelatins from the skin of unicorn leatherjacket (Aluterus monoceros) pretreated with different acids (0.2 M acetic acid or 0.2 M phosphoric acid) and extracted with distilled water at 45 °C for various times (4 and 8 h) were characterized. Yields of 5.23–9.18 or 6.12–11.54% (wet weight basis) were obtained for gelatins extracted from the skin pretreated with 0.2 M acetic acid or 0.2 M phosphoric acid, respectively. Extracted gelatins contained α₁ and α₂ chains as the predominant components and some degradation peptides. The absorption bands of gelatins in FTIR spectra were mainly situated in the amide band region (amide I, amide II and amide ???) and showed the significant loss of molecular order of triple helix. Gelatin samples had a relative solubility greater than 90% in the wide pH ranges (1–10). The gel strength of gelatin from skin pretreated with phosphoric acid (GPA) was higher than that of gelatin from skin pretreated with acetic acid (GAA). Both GPA and GAA had the lower gel strength than that of commercial bovine gelatin (P < 0.05). Net charge of GAA and GPA became zero at pHs of 6.64–7.15 and 6.78–7.26, respectively, as determined by zeta potential titration. Emulsifying and foaming properties of GAA and GPA increased with increasing concentrations (1–3%, w/v). Those properties were governed by pretreatments and extraction time. Thus gelatin can be successfully extracted from unicorn leatherjacket skin using the appropriate acid pretreatment and extraction time.     

Chemical and sensory quality changes of fish fingers,made from mirror carp (Cyprinus carpio L., 1758), during frozen storage (−18 °C)

The effects of frozen storage at −18 °C on the chemical and sensory qualities of fish fingers produced from unwashed and washed mirror carp (Cyprinus carpio) mince were investigated. The amounts of moisture, crude protein, lipid, crude ash, ω3 polyunsaturated fatty acids (PUFA ω3), and ω6 polyunsaturated fatty acids (PUFA ω6) in fish fingers produced from unwashed mince (UWF) were found to be 68.50%, 15.5%, 6.00%, 2.20% 2.31%, and 55.2%, respectively, while they were found to be 70.23%, 10.8%, 2.14%, 1.80%, 2.28%, and 54.6%, respectively, in carp fingers produced from washed mince (WF). The thiobarbituric acid value (TBA, mg malonaldehyde/kg) was found to be significantly higher in mince of WF than in mince of UWF and increased significantly during frozen storage in both the mince of UWF and WF (p < 0.05). A significant decrease in pH was obtained throughout the washing treatment (p < 0.05). There were no significant differences of pH in either the mince of UWF or WF between the beginning and end of the storage periods (p > 0.05), whereas a sharp increase was observed in the fourth month in both groups. The protein solubilities of the mince of both UWF and WF decreased significantly throughout the storage periods (p < 0.05). Sensory parameters of colour, odour, flavour, and general acceptability for both groups decreased during the frozen storage period (p < 0.05) but were still within acceptable limits. It was also concluded that mirror carp was a good source for fish fingers and that product could be stored for five months in a frozen state without undesirable changes of sensory and chemical qualities.     

Effects of alkaline and acid pretreatment on the physical properties and nanostructures of the gelatin from channel catfish skins

The objective of this study was to illustrate the correlation between the physical properties and nanostructure of gelatins made of channel catfish (Ictalurus punctatus) skins. The gelatin samples were first pretreated with sodium hydroxide, acetic acid, or water, and then extracted with hot water before the measurement. Physical properties including the yield of protein, viscosity and textural properties were determined on gelatins obtained with different pretreatment conditions. The acid pretreatment group showed the highest gel strength and protein yield, and a reasonable viscosity. The water pretreatment group showed the lowest values for all of the physical properties. Four samples including water, 0.1 M acid and 0.25 and 1.0 M alkaline-pretreated groups’ nanostructures were then studied using atomic force microscopy (AFM). The AFM images showed that the acid-pretreated gelatin was composed of sponge-like aggregates, while the others showed separated individual aggregates. Annular pores were only found in the alkaline pretreatment group. There was no significant correlation between the diameters of the spherical aggregates and the physical properties; however, the different AFM patterns may relate to the gelatin's physical properties.     

Effects of superchilling and cryoprotectants on the quality of common carp (Cyprinus carpio) surimi: Microbial growth,oxidation, and physiochemical properties

The objective of the present study was to investigate the effect of superchilling with cryoprotectants (a mixture of sucrose and sorbitol) on microbial growth, lipid oxidation, and proteolytic degradation in common carp (Cyprinus carpio) surimi. With increasing storage time, the microbial count of surimi increased from 4.4 log₁₀ CFU/g (0 days) to 7.2, 6.2, 5.9, and 5.5 log₁₀ CFU/g (35 days; P < 0.05) in the samples superchilled at −1 °C, superchilled at −3 °C, superchilled at −3 °C with cryoprotectants, and frozen at −18 °C, respectively. The total volatile basic nitrogen and thiobarbituric acid-reactive substances exhibited an increasing trend (P < 0.05) similar to that obtained with the microbial growth, whereas the whiteness and lightness stability decreased (P < 0.05) with increasing storage time. The SDS-PAGE analysis revealed that the degree of protein degradation increased with increasing storage time. Compared with the sample frozen at −18 °C, superchilling at −1 °C and −3 °C resulted in a marked reduction in the microstructure deterioration of the myofibrillar protein gels, and the addition of cryoprotectants further reduced this deterioration. These results suggest that superchilling with cryoprotectants offers an effective approach for reducing microbial growth and lipid oxidation and limiting proteolytic degradation in common carp surimi.     

Effect of microwave heating on the low-salt gel from silver carp (Hypophthalmichthys molitrix) surimi

Gels from silver carp (Hypophthalmichthys molitrix) surimi were obtained using microwave (MW) heating (15 W/g power intensity for 20–80 s) at different levels of salt (0 g/100 g, 1 g/100 g, or 2 g/100 g). And the gel heated by MW was compared with the gel obtained by conventional water-bath heating (85 °C for 30 min). The gel strength increased when the salt level was increased. The mechanical and functional properties of non-salted, low-salt and regular-salt products were improved by MW heating for 60 s and 80 s, significantly (p < 0.05), except for the cook loss. The content of TCA-soluble peptides indicated that the MW heating inhibited the autolysis of proteins significantly (p < 0.05) during gelling. The SDS-PAGE and total content of –SH group proved that MW enhanced the cross-linking of proteins effectively through disulphide bonds and non-disulphide covalent bonds. The microstructure of the samples revealed that a fine compact network, with particles of protein aggregates, was formed in the low-salt gels (1 g/100 g) heated by MW for 60 s. All of these properties might be responsible for the formation of a superior textural low-salt gel induced by MW.     

Extraction and properties of gelatin from channel catfish (Ietalurus punetaus) skin

Response surface method was used to determine the optimum operating conditions for extracting the gelatin from channel catfish skin. The optimal conditions for maximum gel strength are 68.8 h for the time of treatment with calcium hydroxide solution, 43.2 °C for the extraction temperature, 5.73 h for the extraction time with hot water. The gelatin from channel catfish skin showed a high gel strength, 276±5 g. Compare to porcine skin gelatin, the gelatin from channel catfish skin has different amino acids composition and a lower thermo-stability.     

Suppression of thermal denaturation of myosin and salt-induced denaturation of actin by sodium citrate in carp (Cyprinus carpio)

The effect of sodium citrate (Na-citrate) on myosin and actin denaturation in myofibrils was investigated. Na-citrate significantly suppressed the thermal inactivation of Ca²⁺-ATPase of carp myosin in a concentration-dependent manner. The effect was greater than that of sorbitol. A similar effect was observed with myofibrils in which myosin is stabilized by F-actin binding. Na-citrate dissolved myofibrils at lower concentration than NaCl. Nevertheless, Na-citrate at 1 M failed to denature F-actin in myofibrils, while 1 M NaCl denatured F-actin almost completely. Na-citrate suppressed the NaCl-induced F-actin denaturation. Sorbitol did not show such protective effect on F-actin denaturation. Moreover, Na-citrate suppressed the freeze denaturation of myofibrils at lower concentration than sorbitol. Thus, Na-citrate was proved to be superior to sorbitol. It was suggested that Na-citrate alone could substitute sorbitol as cryoprotectant in surimi and NaCl as dissolving reagent of myofibril in thermal gel production.     

Effects of concentration on nanostructural images and physical properties of gelatin from channel catfish skins

Physical properties are crucial to gelatin utilization and the physical properties are determined by structure. Therefore, it is important to investigate the nanostructure and physical properties of gelatin over the full range of concentrations which are widely applied in research and industry. Nanostructure of gelatin can be investigated by atomic force microscopy (AFM). However, it is hard to obtain reliable AFM images of gelatin with high concentrations (1–6.67%). In this study, methods for imaging gelatin with high concentration were explored and developed, which mainly included six steps. Then the relationships among concentration, nanostructure and physical property of gelatin extracted from channel catfish skins (Ictalurus punctatus) were studied. The high-resolution AFM images show fibril structure in gelatins with concentrations from 1% to 6.67%. However, in low concentrations (<1%), most nanostructures of gelatin were spherical aggregates and fibril structure only existed occasionally. Correspondingly, there were no significant differences of gel strength, texture profile and viscosity among several groups of gelatin when the concentration was lower than 1%, in contrast, these properties changed dramatically when the concentration was greater than 1%. It indicates that there must be some close relationships among concentration, nanostructure and physical property of gelatin. The illustration of nanoscale transition would help us understand the macroscale changes of physical properties.     

Bacterial microflora of carp (Cyprinus carpio) and its shelf-life extension by essential oil compounds

The microflora of common carp (Cyprinus carpio) skin, gill and intestine were analysed and the antimicrobial activities of garlic oil and nine constituents of essential oils (allyl isothiocyanate, carvacrol, cinnamaldehyde, citral, cuminnaldehyde, eugenol, isoeugenol, linalool and thymol) against the carp isolates were studied to identify compounds that might extend the shelf-life of carp fillet. A total of 90 isolated strains were identified to belong to seven genera: Acinetobacter (6), Alcaligenes (2), Bacillus (2), Flavobacterium (20), Micrococcus (2), Moraxella (6) and Pseudomonas (4), and two families Enterobacteriaceae (14) and Vibrionaceae (34). The dominant micro-organisms of carp were found to be Flavobacterium (37%) and Vibrionaceae (33%) in skin, Flavobacterium (33%) in gill and Vibrionaceae (63%) and Flavobacterium (37%) in intestine. Against these isolates, thymol, carvcarol and cinnamaldehyde had the strongest antimicrobial activities, followed by isoeugenol, eugenol, garlic oil, and then citral. The antimicrobial properties of the other constituents tested (cuminnaldehyde, linalool and allyl isothiocyanate) were low. In tests of mixed compounds, a combination of carvacrol and thymol had the highest antimicrobial activity. Moraxella, Flavobacterium and Vibrionaceae were more sensitive to the compounds, whereas Alcaligenes strains were resistant. Dipping carp fillets in a solution of 0.5% carvacrol and 0.5% thymol before storage at 5°C and 10°C reduced both the total microbial load by about 100-fold and the Volatile Bases Nitrogen (VB-N), as compared with controls. In addition, dipping treatment delayed bacterial growth and extended the shelf-life of the fillets from 4 to 12 days at low temperature (5°C). However, the treated and control fillets showed little difference during storage at 10°C. Data from sensory evaluation showed that dipped fillets in 1% (carvacrol+thymol) extended the shelf-life of carp fillets by 8 and 4 days at 5° and 10°C, respectively. Thus, carvacrol and thymol dipping can improve the microbial stability of fish fillets by removing bacteria and by inhibiting bacterial growth.