Physical, mechanical, and barrier properties of carp and mammalian skin gelatin films

Films of 0.11 to 0.13 mm thickness were prepared using gelatins from the skins of cultured freshwater carp species and mammalian gelatins viz., porcine and bovine skin gelatin. A comparative study was made on the physical, mechanical, and barrier properties of these films. The amino acid composition, gel strength, clarity, and gel setting point of the gelatins were also determined. Carp skin gelatins had a lower imino acid content (19.16% to 20.86%) than mammalian skin gelatins (22.91% to 23.7%). Grass carp gelatin had gel strength of 230.2 B that is comparable to the reported value for bovine skin gelatin (227.2 B). The bloom values of rohu and common carp skin gelatins were 188.6 B and 181.3 B, respectively, which were significantly lower than mammalian gelatins. Mammalian gels have significantly higher (P < 0.05) setting temperatures (23.7 to 24.2 °C) than carp skin gelatins. Tensile strength (TS) was lowest for films from common carp and rohu skin gelatin (490 and 497 kg/cm(2), respectively) and highest for porcine skin gelatin film. The degree of transparency (L*) was significantly higher for films from grass carp, bovine hide, and pork skin gelatin films. Carp skin gelatin films had significantly lower water vapor permeability (WVP) and oxygen permeability (OP) than mammalian skin gelatin films, which indicated that carp skin gelatin based films have superior barrier properties than mammalian skin gelatin films.     

Mechanical properties of mammalian and fish gelatins based on their weight average molecular weight and molecular weight distribution

Acid porcine skin gelatins (type A), lime bone gelatins (type B) and gelatin from different cold water fish species were compared on the basis of low deformation mechanical properties, Bloom value, weight average molecular weight, molecular weight distribution and isoelectric point. The dynamic storage modulus and Bloom value for all types of gelatin increased with increasing weight average molecular weight. Type A and type B gelatins with similar weight average molecular weight exhibited different dynamic storage modulus (G′) and different Bloom values. This is most probably due to a different molecular weight distribution as well as the presence of different hydrolytic fragments. The present study suggests that it may be possible to improve the mechanical properties by removing low molecular weight molecules from a gelatin sample. The Bloom values for gelatin from haddock, saithe and cod were determined to be 200, 150 and 100 g from a linear correlation between G′ and Bloom.     

Water Vapor Permeability of Mammalian and Fish Gelatin Films

ABSTRACT:  Water vapor permeability of cold- and warm-water fish skin gelatins films was evaluated and compared with different types of mammalian gelatins. Alaskan pollock and salmon gelatins were extracted from frozen skins, others were obtained from commercial sources. Water vapor permeability of gelatin films was determined considering differences on percent relative humidity (%RH) at the film underside. Molecular weight distribution, amino acid composition, gel strength, viscoelastic properties, pH, and clarity were also determined for each gelatin. Water vapor permeability of cold-water fish gelatin films (0.93 gmm/m²hkPa) was significantly lower than warm-water fish and mammalian gelatin films (1.31 and 1.88 gmm/m²hkPa, respectively) at 25 °C, 0/80 %RH through 0.05-mm thickness films. This was related to increased hydrophobicity due to reduced amounts of proline and hydroxyproline in cold-water fish gelatins. As expected, gel strength and gel setting temperatures were lower for cold-water fish gelatin than either warm-water fish gelatins or mammalian gelatins. This study demonstrated significant differences in physical, chemical, and rheological properties between mammalian and fish gelatins. Lower water vapor permeability of fish gelatin films can be useful particularly for applications related to reducing water loss from encapsulated drugs and refrigerated or frozen food systems.     

Extraction and Physicochemical Characterization of Greater Lizardfish (Saurida tumbil) Skin and Bone Gelatin

ABSTRACT:  Type A gelatins were extracted from skins and bones of lizardfish and analyzed to determine their functional and chemical properties. Lizardfish skin gelatin had ash content of 2.2 ± 0.3% while bone gelatin had ash content of 12.2 ± 0.2%. Gel strength was 159.1 ± 14 and 135 ± 7.9 g, respectively, for skin and bone gelatins compared to 224.3 ± 7.7 g for porcine gelatin. Gelatin from skin exhibited higher viscosity and lower setting time than bone. Skin gelatin had higher imino acid content than bone gelatin. The total imino acid content was 21.71% and 19.83% for skin and bone, respectively. Both skin and bone gelatins contained more α chains than β and γ components. Both bone and skin gelatins also contained low molecular weight (< α) peptides. The differences in functional properties between the skin and bone gelatins appeared to be related to differences in amino acid composition and molecular weight distribution of the gelatins.     

Sea bream bones and scales as a source of gelatin and ACE inhibitory peptides

Sea bream scales and bones were used as sources of gelatin. Scales gave a higher gelatin yield than bones pretreated with HCl or Alcalase. Demineralization with EDTA was effective especially in the case of scale gelatin that showed the lowest ash content. The pretreatment of bones with HCl led to an increase in the removal of minerals. The gel strength and viscoelastic properties of sea bream scale gelatins were higher than those of bone gelatins, and only slight differences were found between gelatin extracted from bones pretreated with HCl or Alcalase, although the amino acid profile was similar in the three gelatins. The gel strength of scale gelatins was higher than that of a commercial bovine gelatin used for comparative purpose (Bloom 200–220). When the scales gelatin was hydrolyzed with Esperase, a high ACE-inhibitory activity was found in the peptide fraction below 3 kDa, and the amount of this peptide fraction required to inhibit 50% of the ACE activity (IC₅₀) was around 60 μg/mL.     

Mass spectrometric detection of marker peptides in tryptic digests of gelatin: A new method to differentiate between bovine and porcine gelatin

Gelatin is a mixture of polypeptides obtained by hydrolysis of collagen primarily from bovine and porcine skin and bones. The similarity between different gelatins makes it difficult to trace their species origin. In this work, a new method for differentiation between bovine and porcine gelatin was developed based on detection and identification of marker peptides in digested gelatins. Sequence alignment analysis indicates that bovine and porcine Type I collagen contain differential sequences. The gelatins were digested by trypsin, and the resulting peptides were analyzed by high performance liquid chromatography/tandem mass spectrometry (HPLC–MS/MS). The marker peptides specific for bovine and porcine were successfully detected in the digested bovine and porcine gelatin, respectively. Comparative analysis indicated that more marker peptides could be detected in gelatin with a higher mean molecular weight. It was found that proline hydroxylation was a key factor affecting the peptide identification. For peptides such as GPPGSAGSPGK and GPPGSAGAPGK detected in digested bovine and porcine gelatin, respectively, the sequence should be verified manually since the mass shift caused by proline hydroxylation can be confused with the mass difference between Ser and Ala residues. The results indicate that detection of marker peptides in the digested gelatin sample using HPLC–MS/MS is an effective method to differentiate between bovine and porcine gelatin.     

Determination of Total Protein Content in Gelatin Solutions with the Lowry or Biuret Assay

ABSTRACT:  Gelatins can be obtained from different sources and prepared using different processes, and the end product gelatin may vary in amino acid composition and molecular weight distribution. In the present study, the variation in "protein color" development among gelatins in colorimetric total protein content measurements was investigated at 540 nm using the Biuret assay and at 650 nm using the Lowry assay, with bovine serum albumin as the reference protein. In both the Biuret and Lowry assays, the color response varied significantly among gelatins. The difference in imino acid content was the major factor responsible for this variation, which probably influenced the gelatin helix → coil phase transition and resulted in the difference in gelatin associate state. Based on their "protein color" development abilities in both Biuret and Lowry, gelatins were classified into 2 major groups with the hierarchical cluster analysis: 1 group included all cold water fish gelatins, while the other included gelatins from warm water fish, avian, and mammalian species.     

Novel multiplex PCR-RFLP assay discriminates bovine,porcine and fish gelatin substitution in Asian pharmaceuticals capsule shells

Gelatin is widely used in pharmaceuticals as a protective coating, such as soft and hard capsule shells. However, the animal source of gelatin is a sensitive issue because certain gelatins such as porcine and bovine gelatins are not welcome in Halal, Kosher and Hindus’ consumer goods. Recently, we have documented DNA barcoding and multiplex PCR platforms for discriminating porcine, bovine and fish gelatins in various fish and confectionary products; but those assays were not self-authenticating and also not tested in highly refined pharmaceutical products. To address this knowledge gap, here we report a self-authenticating multiplex PCR-restriction fragment length polymorphism (RFLP) assay to identify animal sources of various gelatin in pharmaceutical capsules. Three different restriction enzymes, BsaAI, Hpy188I and BcoDI were used to yield distinctive RFLP patterns for gelatin-based bovine (26, 94 bp), fish (97, 198 bp) and porcine (17, 70 bp) DNA in control experiments. The specificity was cross-tested against 16 non-target species and the optimised assay was used to screen gelatin sources in 30 halal-branded pharmaceuticals capsule shells. Bovine and porcine DNA was found in 27 and 3 of the 30 different capsules products. The assay was suitable for detecting 0.1 to 0.01 ng total DNA extracted from pure and mixed gelatins. The study might be useful to authenticate and monitor halal, kosher, vegetarian and Hindu compliant pharmaceuticals, foods and cosmetics.     

Rheological Properties of Fish Gelatins

ABSTRACT: The rheological properties of fish gelatins (cod, megrim, tuna, and tilapia) and conventional gelatin (bovine and porcine) were compared. The different fish gelatins had from low to high viscosity values. They also had from low to high gel strength values. However, they had lower melting and gelling temperatures than gels from conventional gelatins. Cold-water fish gelatins were more different from conventional gelatins than warm water fish gelatins reflecting the different amino acid composition, as cold-water fish gelatins are low in imino acids. Binary blends of cod and other fish or conventional gelatins seemed to be completely compatible.     

Rheological properties of channel catfish (Ictalurus punctaus) gelatine from fish skins preserved by different methods

Gelatins were extracted from channel catfish skins preserved by different methods using 50 mmol/l acetic acid. Molecular weight distribution, gel strength and viscoelastic properties of gelatin samples were studied. Compared to gelatins from fresh and frozen skins, gelatin from dried channel catfish skin exhibited higher gel strength. This can be explained by the large α-chains content of gelatin from the dried skins. The gelling point and melting point of dried channel catfish skin gelatin solution were similar to those of fresh skin gelatin solution, but distinctly different from those of frozen skin gelatin. After maturation at low temperature, melting points of gelatins increased. But the melting point of frozen skin gelatin was still the highest among the three gelatin samples studied.     

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.     

Comparative study on characteristics of gelatin from the skins of brownbanded bamboo shark and blacktip shark as affected by extraction conditions

Gelatins from the skins of brownbanded bamboo shark (BBS; Chiloscyllium punctatum) and blacktip shark (BTS; Carcharhinus limbatus) were extracted using the distilled water at different temperatures (45, 60 and 75 °C) and times (6 and 12 h). Yields of gelatin from the skins of BBS and BTS were 19.06–22.81% and 21.17–24.76% (based on wet weight), respectively. Gelatins from both species extracted at 45 °C for 6 h exhibited the highest bloom strength (206–214 g), which was higher than that of commercial bovine bone gelatin (197 g) (p < 0.05). Gelatin gels from BBS skin could set at room temperature (25–26 °C) within 24 min. However, gelatin gels from BTS skin was not able to set within 3 h at the same temperature. Scanning electron microscopic study showed that gelatin gel from BBS skin presented the thicker strand than those from BTS skin and bovine bone. Cross-linked components (β- and γ-chains) and α-chains were more degraded with increasing extraction temperatures, especially at 75 °C. Gelatin from BTS skin was more susceptible to hydrolysis than that from BBS skin. Fourier transform infrared (FTIR) study revealed that the major absorption bands of gelatin from the skins of both sharks shifted to a higher wavenumber, compared with their corresponding acid soluble collagen (ASC). Therefore, gelatins from the skin of BBS has a potential to replace mammalian for gelatin, due to its similarity in bloom strength and setting behavior to the commercial bovine bone gelatin.     

Characteristic tryptic peptides and gelling properties of porcine skin gelatin affected by thermal action

Thermal action in extraction process had effects on characteristic tryptic peptides identification and gelling properties of porcine gelatin. SDS-PAGE, HPLC-LTQ/Orbitrap high-resolution mass spectrometry, texture analyser and rheometer were used to evaluate collagen depolymerisation degree, characteristic tryptic peptides and gelling properties of gelatins prepared in various thermal actions. Results showed that with increasing temperature and time, depolymerisation degree enlarged, while gel strength, gelling and melting temperature decreased. Mass spectra showed that 47 and 49 common characteristic tryptic peptides were identified in gelatins extracted at 50 °C and 100 °C with various times, respectively. Moreover, 34 common characteristic tryptic peptides were identified in all gelatin samples. Further comparison between this work and our previous investigations yielded 20 common characteristic tryptic peptides, which stably exist in various thermal actions. These common characteristic tryptic peptides may be very helpful for the accurate authentication of porcine gelatin.     

Rheological properties of gelatin from silver carp skin compared to commercially available gelatins from different sources

Gelatin is used as a functional ingredient in many foods, pharmaceuticals, and cosmetics as a stabilizing, thickening, and gelling agent. The rheological properties of gelatins are important in the potential functionality of gelatin. This study is designed to determine the rheological properties of gelatin extracted from the skins of silver carp (Hypophthalmichthys molitrix Valenciennes 1844). The extracted gelatin is compared with commercially available gelatins from different sources. The results indicate that the stress-strain relationship of gelatin gels remained in the linear region over a broad range of strains and stresses and gave similar elastic moduli at varying frequency, stress, and strain levels. One exception was a commercial high molecular weight fish skin gelatin that gave a lower elastic modulus indicating that its gel strength was low compared to the other gelatin samples studied. Gel strength varied between 220 and 1230 g while viscosity varied between 4.53 and 6.91 cP among the samples. Melting and gelling temperatures varied between 14.2 and 32.3 °C and 3.2 and 25.4 °C, respectively. Texture profile analysis was done at 2 deformation levels, 25% and 75%, and the results correlated well with gel strength. The correlations between hardness, cohesiveness, and gumminess and gel strength were 0.98, 0.82, and 0.99, respectively, at 25% deformation but lower at 75% deformation. The results suggest that rheological measurements might be used to quickly estimate gel strength using less material. In addition, the silver carp skin gelatin seemed to be of equal quality to some of the commercial gelatins.     

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.     

微波-快速冻融耦合鱼皮明胶理化性质分析

本实验采用微波-快速冻融耦合处理鱼皮获得鱼皮明胶，并通过等电点、透射比、凝冻强度测定以及十二烷基硫酸钠-聚丙烯酰胺凝胶电泳、流变学特性、氨基酸组成和傅里叶变换红外光谱分析对鱼皮明胶的理化性质进行表征。结果表明：与市场上的A型和B型明胶不同，微波-快速冻融耦合鱼皮明胶等电点为7，在450 nm波长处透射比为57.13%，在620 nm波长处透射比为78.07%，凝冻强度为524.40 Bloom g，满足GB 6783—2013《食品安全国家标准 食品添加剂 明胶》要求（在450 nm波长处透射比不低于30%，620 nm波长处透射比不低于50%，凝冻强度不低于50 Bloom g）。与传统酸法鱼皮明胶相比，微波-快速冻融耦合明胶氢键含量更高，三螺旋结构较为完整，亚氨基酸比例相近；但由于高分子亚基比例低，导致凝胶特性总体不及传统明胶。总体而言，微波-快速冻融耦合鱼皮明胶符合国标要求，该方法是明胶绿色清洁制备的良好探索。     

Repurposing fish waste into gelatin as a potential alternative for mammalian sources: A review

Mammalian gelatin is extensively utilized in the food industry because of its physicochemical properties. However, its usage is restricted and essentially prohibited for religious people. Fish gelatin is a promising alternative with no religious and social restrictions. The desirable properties of fish gelatin can be significantly improved by various methods, such as the addition of active compounds, enzymes, and natural crosslinking agents (e.g., plant phenolics and genipin), and nonthermal physical treatments (e.g., ionizing radiation and high pressure). The aim of this study was to explore whether the properties of fish gelatin (gel strength, melting or gelling temperature, odor, viscosity, sensory properties, film-forming ability, etc.) could be improved to make it comparable to mammalian gelatin. The structure and properties of gelatins obtained from mammalian and fish sources are summarized. Moreover, the modification methods used to ameliorate the properties of fish gelatin, including rheological (gelling temperature from 13–19°C to 23–25°C), physicochemical (gel strengths from ∼200 to 250 g), and thermal properties (melting points from ∼25 to 30°C), are comprehensively discussed. The relevant literature reviewed and the technological advancements in the industry can propel the development of fish gelatin as a potential alternative to mammalian gelatin, thereby expanding its competitive market share with increasing utility.     

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.     

Physicochemical and Enzymatic Properties of Five Kiwifruit Cultivars during Cold Storage

Samples of Abbot, Alison, Bruno, Monty, and Hayward cultivars of kiwifruit (Actinidia deliciosa) were obtained from the Iran Research Center of Citrus (Tonekabon, located in north of Iran) and their physicochemical properties were studied during cold storage (at T = 1 ± 1 °C, RH = 80 ± 5%) at 0-, 9-, and 18-week intervals. The mean chemical composition of the fruits were as follows: ash = 0.66–0.96%, moisture = 75.2–84.7%, starch = 0.3–7.0%, and ascorbic acid = 54.8–261.0; K = 125.0–372.0 mg 100 g⁻¹ fresh weight, Mg = 18.0–32.0 mg 100 g⁻¹ fresh weight, Na = 1.4–3.1 mg 100 g⁻¹ fresh weight, Fe = 0.17–0.52 mg 100 g⁻¹ fresh weight, Cu = 0.04–0.24 mg 100 g⁻¹ fresh weight, Zn = 0.16–0.49 mg 100 g⁻¹ fresh weight, Mn = 0.04–0.10 mg 100 g⁻¹ fresh weight, and P = 25.2–49.3 mg 100 g⁻¹ fresh weight; glucose = 0.7–2.39%, fructose = 1.20–3.13%, and sucrose = 0.0–5.8%. At the same time, the values of the parameters °Brix = 6.5–14.8% and acidity = 1.8–2.5% of the studied cultivars (mutual effects of cultivar and storage time) were investigated. The increase in peroxidase (POX = 0.0–6.65 U ml⁻¹) and the decrease in pectinesterase (PE; poor activity to 0) activities were also determined. The statistical analysis showed that the Bruno cultivar had the highest content of ascorbic acid (115.0–261.0 mg 100 g⁻¹ fresh weight), which is an important compound in fruits during storage, while Hayward had the best overall quality particularly with regards to its resistance to softening. This study confirms that long-term cold storage at 1 ± 1 °C and 80 ± 5% RH is suitable for maintaining the highest quality of Iranian grown cultivars of kiwifruit.     

Re-extraction,Recovery, and Characteristics of Skin Gelatin from Farmed Giant Catfish

This study was aimed to investigate the re-extraction process for gelatin recovery from the skin of farmed giant catfish. The first extraction was done by incubating the acid-treated fish skin at 45 °C for 12 h. The remnant was re-extracted at temperatures of 60–90 °C for 1–12 h. The gelatin yield of the first extraction was 10.14%, while the re-extraction at high temperature provided higher recovery (19.5%). Low band intensity of α₁ and α₂ chains of gelatin was observed when it was re-extracted at high temperature for a longer time. The absorption bands of amide I and II from both extracted gelatins were similar. Low-transition temperature with high transition enthalpy of gelatin extracted at 90 °C was observed. The obtained results suggested that the re-extraction process could be applied as a supplemental step for other sources to obtain high recovery with the desired properties.