Properties of protein-based film from round scad (Decapterus maruadsi) muscle as influenced by fish quality

The properties of film prepared from round scad (Decapterus maruadsi) stored in ice for different times were investigated. Degradation of myosin heavy chain (MHC) was more pronounced with the coincidental increase in total volatile base and trimethylamine contents as the storage time increased (P<0.05). Regardless of storage time, no changes in tensile strength (TS) and elongation at break (EAB) of resulting films prepared from unwashed mince were observed (P>0.05). For the films prepared from washed mince, TS decreased, whereas EAB increased when the storage time of fish increased (P<0.05). However, films prepared from washed mince showed the greater mechanical properties with the lower film solubility and protein solubility than did those from mince (P<0.05). Generally, films prepared from fish stored in ice for a longer time became less transparent, darker and more yellowish. The electrophoretic study revealed that similar protein patterns were observed between films, irrespective of storage time of fish and washing. Therefore, the quality of fish did not show the marked impact on the mechanical property of the resulting films, while washing likely affected the film forming ability.     

Roles of lipid oxidation and pH on properties and yellow discolouration during storage of film from red tilapia (Oreochromis niloticus) muscle protein

Protein-based films prepared from red tilapia (Oreochromis niloticus) washed and unwashed mince solubilised at pH 3 and 11 were prepared and characterised. Tensile strength (TS) of films from washed mince was greater than that of films prepared from unwashed mince for both pH used (P < 0.05). TS of films prepared at pH 3 was higher than that of films prepared at pH 11 for both of washed and unwashed mince (P < 0.05). Film from washed mince with pH 3 showed the highest TS, while that from unwashed mince with pH 11 had the lowest TS with the highest elongation at break (EAB) (P < 0.05). Films from washed mince had the lower value of thiobarbituric acid reactive substances (TBARS) than did those from unwashed counterpart, regardless of pH used. Nevertheless, TBARS was much higher in films prepared at acidic pH, compared with those prepared at alkaline pH. During storage of 20 days at room temperature, films became yellowish as evidenced by the increases in b^∗ and ΔE^∗-values. Films prepared at pH 11 showed the higher b^∗ and ΔE^∗-values than did those prepared at pH 3, especially for those from unwashed mince. However, films prepared from washed mince at pH 3 showed higher b^∗ and ΔE^∗-values than did those prepared at pH 11 (P < 0.05). Films generally had the increase in TS but the decreases in water vapour permeability (WVP), film solubility and protein solubility after 20 days of storage (P < 0.05). Therefore, lipid oxidation more likely played a role in yellow discolouration of fish muscle protein film, mainly by providing the carbonyl groups involved in Maillard reaction, while pH regulated the rate of reaction.     

Round scad protein-based film: Storage stability and its effectiveness for shelf-life extension of dried fish powder

Storage stability of different round scad (Decapterus maruadsi) protein-based films including (1) control film without palm oil and chitosan, (2) film with 25% palm oil (glycerol substitution) and (3) film with 25% palm oil and 40% chitosan (protein substitution) was investigated. During storage (54% relative humidity, 28-30 °C), tensile strength (TS) of the films without and with 25% palm oil or 25% palm oil in combination with 40% chitosan increased continuously, while elongation at break (EAB) decreased markedly when storage time increased up to 8 weeks (p < 0.05). No changes in water barrier properties were observed throughout the storage time up to 4 weeks (p > 0.05). Film solubility slightly increased but protein solubility decreased continuously during storage (p < 0.05). All films became darker and more yellowish as storage time increased. When round scad muscle protein-based films were used to cover dried fish powder, the samples covered with the film containing 25% palm oil and 40% chitosan showed lower thiobarbituric acid reactive substances (TBARS) and yellowness than other samples during the extended storage up to 21 days (p < 0.05). No differences in moisture content of all samples covered with different films were noticeable (p > 0.05), except those covered with HDPE, which had the lowest moisture content. Thus, round scad protein-based film incorporated with palm oil and chitosan could be a promising packaging material to prevent lipid oxidation in oil enriched foods.     

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.     

The effect of myofibrillar/sarcoplasmic protein ratios on the properties of round scad muscle protein based film

The effect of the ratios of myofibrillar protein (MP) to sarcoplasmic protein (SP) from round scad (Decapterus maruadsi) muscle on the properties of the resulting films was investigated. Tensile strength (TS) of films decreased with increasing SP content (p < 0.05). Films prepared from MP/SP ratio of 10:0 (w/w) exhibited the highest TS (p < 0.05). Elongation at break (EAB) of films prepared with SP content greater than 30% had the decreased EAB (p < 0.05). Water vapor permeability (WVP) of films increased when SP content increased up to 20% and decreased with increasing SP content up to 30% (p < 0.05). Solubility of films decreased but protein solubility increased with increasing SP contents (p < 0.05). The a*-value and ΔE* of film increased with increasing SP content. Films with all MP/SP ratios exhibited the negligible transmission to the light in UV range. Therefore, it is suggested that the type and ratio of proteins in fish muscle, both SP and MP, influenced the properties of film from round scad muscle. Results suggested that the removal of sarcoplasmic protein from fish muscle by thorough washing was an effective means to improve the mechanical properties as well as color of the fish muscle protein-based film.     

Compositions,functional properties and antioxidative activity of protein hydrolysates prepared from round scad (Decapterus maruadsi)

Composition, functional properties and antioxidative activity of a protein hydrolysate prepared from defatted round scad (Decapterus maruadsi) mince, using Flavourzyme, with a degree of hydrolysis (DH) of 60%, were determined. The protein hydrolysate had a high protein content (48.0%) and a high ash content (24.56%). It was brownish yellow in colour (L^∗ = 58.00, a^∗ = 8.38, b^∗ = 28.32). The protein hydrolysate contained a high amount of essential amino acids (48.04%) and had arginine and lysine as the dominant amino acids. Na⁺ was the predominant mineral in the hydrolysate. The protein hydrolysate had an excellent solubility (99%) and possessed interfacial properties, which were governed by their concentrations. The emulsifying activity index of the protein hydrolysate decreased with increasing concentration (p < 0.05). Conversely, the foaming abilities increased as the hydrolysate concentrations increased (p < 0.05). During storage at 25 °C and 4 °C for 6 weeks, the antioxidative activities and the solubility of round scad protein hydrolysate slightly decreased (p < 0.05). Yellowness (b^∗-value) of the protein hydrolysate became more intense as the storage time increased but the rate of increase was more pronounced at 25 °C than at 4 °C.     

Effects of washing with oxidising agents on the gel-forming ability and physicochemical properties of surimi produced from bigeye snapper (Priacanthus tayenus)

The effects of washing with hydrogen peroxide (H₂O₂) and sodium hypochlorite (NaOCl) solutions on the gel-forming ability and physicochemical properties of surimi produced from bigeye snapper (Priacanthus tayenus), stored in ice for up to 14 days, were investigated. Generally, pH and the trichloroacetic acid (TCA)-soluble peptide content of washed mince varied, depending on the type of oxidizing agent and storage time of the fish. With increasing time of storage, the pHs of water- and H₂O₂-washed mince were lower than that of NaOCl-washed mince (P < 0.05). However, no differences in the TCA-soluble peptide contents of the resulting mince washed with any media were observed (P > 0.05). Washing with 20 ppm NaOCl resulted in the highest increase in both the breaking force and the deformation of mince from fish stored in ice for all the times studied (P < 0.05). Natural actomyosin (NAM) extracted from NaOCl-washed mince had higher surface hydrophobicity and disulfide bond (SS) content than that of water-washed mince (P < 0.05). With no effect on Ca²⁺-, Mg²⁺-, or Mg²⁺–Ca²⁺-ATPase activities, NaOCl washing resulted in an increase in Mg²⁺–EGTA-ATPase activity of NAM (P < 0.05). The results suggested that washing mince with the appropriate type and concentration of oxidizing agent can improve the gelling ability of surimi, particularly from low quality fish.     

Impact of microbial transglutaminase on gelling properties of Indian mackerel fish protein isolates

Impacts of microbial transglutaminase (MTGase) (0–0.6 units/g sample) on gel properties of Indian mackerel unwashed mince, surimi and protein isolates with and without prewashing were studied. Generally, lower myoglobin and lipid contents were found in protein isolate with and without prewashing, compared to those of unwashed mince and surimi (P < 0.05). Protein isolate had the decreased Ca²⁺-ATPase and protein solubility, indicating protein denaturation. When MTGase was incorporated, breaking force and deformation of all gels markedly increased, especially as MTGase levels increased (P < 0.05). At the same MTGase level, gel from protein isolate with prewashing exhibited the highest breaking force and deformation (P < 0.05). The addition of MTGase could lower the expressible moisture content of most gels. No change in whiteness of gel was observed with the addition of MTGase (P > 0.05), but gel from protein isolate gels had decreased whiteness as MTGase at high level was added. The microstructure of protein isolate gels without prewashing showed a similar network to unwashed mince gels, whilst a similar network was observed between surimi gel and gel from protein isolate with prewashing. Nevertheless, a larger void was noticeable in gels from protein isolates. All gels incorporated with MTGase (0.6 units/g) showed a slightly denser network than those without MTGase. Thus, gel with improved properties could be obtained from protein isolate from Indian mackerel with added MTGase.     

Characteristics and gel properties of muscles from sardine (Sardinella gibbosa) and mackerel (Rastrelliger kanagurta) caught in Thailand

Dark and ordinary muscle from sardine (Sardinella gibbosa) and mackerel (Rastrelliger kanagurta) were characterized. Lipid and myoglobin contents were higher in dark muscle than in ordinary muscle of both species, and higher contents of both constituents were found in sardine muscle than mackerel muscle. The extractable myoglobin contents in sardine dark and ordinary muscle were 14.27 and 2.18 mg/g, while mackerel dark and ordinary muscle contained 4.88 and 1.37 mg myoglobin/g sample, respectively. Alkali-soluble protein and stroma contents were greater in dark muscle than ordinary muscle. Mackerel muscle comprised a higher content of non-protein nitrogenous compounds than sardine muscle. The effect of washing conditions on the myoglobin extractability was investigated. A large amount of myoglobin was removed in the first washing cycle and only a small amount was removed in the second washing cycle. The highest removal of myoglobin from sardine (32.10–46.55%) and from mackerel muscle (103.20–313.66%) was achieved when the mince was washed with 0.2% NaCl and 0.5% NaCl, respectively. Washing media showed the marked effect on the color, expressible drip and textural properties of sardine and mackerel mince gels. The breaking force of directly heated and kamaboko gels from both sardine and mackerel mince washed with NaCl solution was higher than that of unwashed mince and water washed mince. However, no difference in deformation was observed. Washing also resulted in increased whiteness and lowered expressible moisture. In general, sardine surimi showed the superior gel-forming ability and whiteness to mackerel surimi.     

Gel-forming ability of small scale mud carp (Cirrhiana microlepis) unwashed and washed mince as related to endogenous proteinases and transglutaminase activities

Gel-forming ability of small scale mud carp (Cirrhiana microlepis) mince and washed mince was investigated with respect to their proteinase and transglutaminase (TGase) activities. Proteinases in sarcoplasmic fluid showed the optimum activity at 65 °C and pH 9, whereas autolytic activity was maximum at 70 °C and pH 10, indicating the presence of heat-stable alkaline proteinases. When mince was washed with three volumes of water twice, TGase and proteinases were mainly removed in the first washing cycle, resulting in a decreased autolytic activity of washed mince. Breaking force of the single washed mince gels was greater than the twice washed mince and the unwashed mince gels (p<0.05). Pre-incubation of mince pastes at 40 °C for 1 h prior to cooking (90 °C/30 min) increased breaking force of all samples, particularly the single washed mince (p<0.05). This coincided with an increase of higher molecular weight polymers observed on SDS-PAGE. Washing did not completely eliminate proteinases as it was evident by an increased trichloroacetic acid (TCA)-soluble oligopeptides of washed gels pre-incubated at 55 °C. Whiteness values of washed mince gels were greater than that of mince gels.     

Characteristics of myoglobin and haemoglobin-mediated lipid oxidation in washed mince from bighead carp (Hypophthalmichthys nobilis)

Myoglobin (Mb) and haemoglobin (Hb) accounted for 61% and 39% of the total haem-protein in bighead carp (Hypophthalmichthys nobilis) dark muscle, respectively. Molecular weight of Mb and monomeric-Hb was ∼16 kDa. Haemin loss from metHb was more rapid, compared to metMb (pH 6.0, 4 °C). Pro-oxidative activities of oxyMb/Hb and metMb/Hb were examined in washed mince during 9 days of iced storage (pH 6.0). Soret measurements suggested the existence of holoMb throughout storage. For Hb, weakening of haem-globin linkage was observed, especially for metHb which had undetectable Soret after 3 days of storage. Loss of redness was more rapid and extensive in washed mince containing Hb, compared to Mb. During storage, Hb induced larger amounts of peroxides, thiobarbituric acid-reactive substances and hexanal than did Mb (p < 0.05), especially for met-form. Thus, Hb had lower haemin affinity and was a stronger pro-oxidant than Mb.     

Oxidation of lipid and protein in horse mackerel (Trachurus trachurus) mince and washed minces during processing and storage

Protein and lipid oxidation was followed during processing and storage of mince and washed minces prepared from horse mackerel (Trachurus trachurus). Briefly horse mackerel mince (M0) was washed with three volumes of water, mimicking the surimi production and different washed products were obtained: M1, M2 and M3, with one, two and three washing steps, respectively. The different products were characterised (i.e. lipid content, protein, water, iron, fatty acid profile and tocopherol content) and analysed for protein and lipid oxidation in order to investigate the impact of the washing steps on oxidation. Subsequently the different products were stored for up to 96 h at 5 °C and samples were taken out regularly for analysis. Lipid oxidation was investigated by measuring primary oxidation products (lipid hydroperoxides) and secondary oxidation products (volatiles). Protein oxidation was followed by determination of protein solubility, protein thiol groups and protein carbonyl groups using colorimetric methods as well as western blotting for protein carbonyl groups. Lipid and protein oxidation markers indicated that both lipid and protein oxidation took place during processing and the ranking for oxidation was as follows M0 < M1 < M2 ? M3 with M0 being significantly less oxidised than M3. Results indicated that washing creates an imbalance in the initial prooxidant-antioxidant equilibrium in the muscle tissue and contributes to the observed differences in the oxidative status of the four products obtained. In contrast, during storage of different products, lipid oxidation development was faster in M0 and the ranking was as follows M0 > M1 > M2 ? M3. Lipid and protein oxidation developed simultaneously in different minces during storage, but it was not possible to determine at which level these two reactions were coupled.     

Effect of pretreatment on lipid oxidation and fishy odour development in protein hydrolysates from the muscle of Indian mackerel

Impact of different pretreatments on chemical compositions of Indian mackerel mince was studied. Mince prepared using washing/membrane removal/alkaline solubilisation process (W–MR–Al) contained the lowest remaining myoglobin and haem iron content and also showed the lowest total lipid and phospholipid contents. When mince and W–MR–Al were hydrolysed using Alcalase for up to 120 min, a higher degree of hydrolysis (DH) was found in W–MR–Al after 30 min of hydrolysis. Furthermore, hydrolysate from W–MR–Al had lower peroxide value (PV), thiobarbituric acid reactive substances (TBARS) and non-haem iron content throughout hydrolysis period (P < 0.05). When hydrolysate powder produced from mince and W–MR–Al (0–0.3% w/v) were fortified in milk, the former resulted in the lower likeness score (P < 0.05) at all levels used. The addition of the latter, for up to 0.2%, had no effect on likeness of all attributes, compared with milk without fortification (P > 0.05). Therefore, the appropriate pretreatment of mince yielded hydrolysate with lower fishy odour.     

Retardation of myoglobin and haemoglobin-mediated lipid oxidation in washed bighead carp by phenolic compounds

Antioxidative activities of phenolic compounds (caffeic acid, gallic acid and tannic acid; 200 ppm) in washed mince (pH 6), with added myoglobin (Mb) and haemoglobin (Hb), from bighead carp (Hypophthalmichthys nobilis), during 9 days of iced storage, were studied. Tannic acid exhibited the preventive effect on discolouration of washed mince containing Mb or Hb during storage (P < 0.05). High peroxide value (PV) was found and large amount of, thiobarbituric acid-reactive substances (TBARS) and hexanal were formed in washed mince containing haem proteins, especially Hb. As determined by apo Streptococcal haem-associated protein, Hb had the lower haem affinity than Mb. Phenolic compounds, especially caffeic acid and gallic acid, could lower lipid oxidation induced by Mb or Hb throughout storage (P < 0.05). Prevention of haem release, as well as inhibition of lipid oxidation induced by haem proteins with selected phenolic compounds, should be an alternative means in lowering discolouration and lipid oxidation in fish muscle.     

The effects of pretreatments on antioxidative activities of protein hydrolysate from the muscle of brownstripe red snapper (Lutjanus vitta)

Different pretreatments of mince from brownstripe red snapper (Lutjanus vitta) including 1) washing; 2) membrane separation; 3) washing followed by membrane separation and 4) membrane separation followed by washing were conducted prior to hydrolysis. Among the resulting minces, that subjected to membrane separation with subsequent washing (MS/W) contained the lowest remaining myoglobin content, phospholipid content, heme iron and non-heme iron contents (p < 0.05) and showed the lowest TBARS values throughout 9 days of storage at 4 °C in the presence and absence of 0.15 mol L⁻¹ cupric acetate (p < 0.05). When hydrolysates from 1) mince, 2) MS/W and 3) protein isolate from MS/W (PI) with different degree of hydrolysis (DH) (20, 30 and 40%) were prepared using proteases from pyloric caeca of brownstripe red snapper, antioxidative activities determined by DPPH, ABTS radical scavenging activities, ferric reducing antioxidant power and metal chelating activity varied with hydrolysates and DH. Antioxidative activities increased with increasing DH up to 40% (p < 0.05). At all DH tested, hydrolysate prepared from MS/W exhibited the highest antioxidative activities determined by all assays, compared to those from mince and PI (p < 0.05). Hydrolysate from MS/W with 40% DH had the molecular weight lower than 6.5 kDa as determined by SDS-PAGE. In liposome oxidation system, the addition of hydrolysate from MS/W resulted in the lower TBARS, compared with the control throughout the incubation period of 48 h at room temperature (25-28 °C). Therefore, fish mince with membrane separation followed by washing was the most appropriate source for production of hydrolysate possessing antioxidative activity with the lowered amount of lipids and pro-oxidants.     

Autolysis of goatfish (Mulloidichthysmartinicus) mince: Characterisation and effect of washing and skin inclusion

Autolysis of goatfish mince and washed mince incubated at different temperatures (30–70 °C) was investigated. The highest autolytic activity was generally observed in mince and washed mince at 60 °C as evidenced by the highest trichloroacetic acid (TCA) soluble peptide content and the greatest disappearance of myosin heavy chains (MHCs). Autolysis of both mince and washed mince was maximised at pH 4, and lower autolytic activity was observed at pH 7. trans-epoxysuccinyl-l-leucyl-amido (4-guanidino) butane (E-64) showed the greatest inhibition of autolysis at pH 4, showing that at least one cysteine protease was active in goatfish muscle. Nevertheless, soybean trypsin inhibitor effectively inhibited the autolysis at neutral pH (pH 7), suggesting that goatfish muscle also contained at least one serine protease. Generally, autolysis of mince was more pronounced than that of washed mince, indicating that washing could lower the autolytic activity of mince. In the presence of skin, a higher autolysis was obtained with the goatfish mince. Therefore, both sarcoplasmic and myofibril-associated proteases in muscle as well as the contamination of skin likely contributed to the degradation of muscle proteins of goatfish.     

Properties of biodegradable blend films based on fish myofibrillar protein and polyvinyl alcohol as influenced by blend composition and pH level

Effects of the ratios of fish myofibrillar protein (FMP) from bigeye snapper (Priacanthus tayenus) to polyvinyl alcohol (PVA) (FMP:PVA; 10:0, 8:2, 6:4, 5:5, 4:6, 2:8, 0:10) and pH levels (3 and 11) on the properties of resulting films were investigated. Both tensile strength (TS) and elongation at break (EAB) of films increased with increasing PVA content (p < 0.05). When PVA was incorporated up to 40%, films prepared at pH 11 had the higher TS than did those prepared at pH 3 (p < 0.05). However, as PVA content was greater than 40%, films prepared at pH 3 exhibited the higher TS than did those prepared at pH 11 (p < 0.05). Water vapor permeability (WVP) of the films prepared at pH 3 increased when PVA content increased up to 40% and decreased with further increases in PVA content (p < 0.05). PVA films had the higher TS, EAB and WVP than did FMP films and FMP/PVA blend films prepared at both pHs. Films exhibited the increased L∗ and a∗ values but decreased b∗ value with increasing PVA content at both pHs. Films prepared at pH 11 showed higher b∗ value than did those prepared at pH 3 when PVA content was greater than 40% (p < 0.05). FMP/PVA blend films exhibited the negligible transmission to the UV light. At pH 3, light transmission of the films increased as PVA content increased (p < 0.05). At all FMP/PVA ratios, films prepared at pH 11 were less transparent than those prepared at pH 3 (p < 0.05). Therefore, blend composition and pH level influenced the properties of FMP/PVA blend films.     

Thermal gelation properties of surimi-like material made fromsheep meat

Thermal gelation properties of surimi-like material made from sheep meat were investigated. The sheep meat was ground, washed 3 times at a meat to water ratio of 1:5 and dewatered by centrifugation. The effects of washing on the composition, functional properties and colour of the mince, were studied. The washing method resulted in a sharp reduction of the fat content and an increase of the water content and pH of the mince. Lightness (L*) and yellowness (b*) of the mince were improved by washing. A significant reduction (p<0.001) of a* (redness) value and a decrease in the a*/b* ratios and saturation index value of the washed mince were recorded. Gels were prepared from washed and unwashed mince after being blended with 2% NaCl and heated at 75°C for 20 min. The washed mince produced excellent gels as measured by the fold test, elasticity modulus and the percentage recovery. The gels made of washed mince had lower expressible fluid compared to that of unwashed mince. A fibrous protein network structure was evidenced in the gel made from washed mince while observed under a transmission electron microscope.     

Retardation of haemoglobin-mediated lipid oxidation of Asian sea bass muscle by tannic acid during iced storage

Lipid oxidation mediated by haemoglobin from tilapia was monitored in washed Asian sea bass mince with and without added tannic acid (200 and 400 ppm) during 10 days of iced storage. Control samples (without tannic acid) had the highest peroxide value (PV) within the first 2 days and possessed the greater amount of thiobarbituric acid-reactive substances (TBARS) throughout the storage of 10 days (p < 0.05). With addition of tannic acid, the lipid oxidation of washed Asian sea bass mince was retarded, especially when the higher level (400 ppm) was used, as evidenced by lowered PV and TBARS. The retarded formation of volatile lipid oxidation products in the samples with added 400 ppm tannic acid was found. Sensory analysis revealed that samples with added 400 ppm tannic acid possessed lower fishy odour score, compared with the control sample and that with added 200 ppm tannic acid (p < 0.05).     

Effect of heat treatment of film-forming solution on the properties of film from cuttlefish (Sepia pharaonis) skin gelatin

Effects of heat treatment at different temperatures (40–90 °C) of film-forming solution (FFS) containing 3% gelatin from cuttlefish (Sepia pharaonis) ventral skin and 25% glycerol (based on protein) on properties and molecular characteristics of resulting films were investigated. The film prepared from FFS heated at 60 and 70 °C showed the highest tensile strength (TS) with the highest melting transition temperature (T_max) (p < 0.05). Nevertheless, film from FFS heated at 90 °C had the highest elongation at break (EAB) with the highest glass transition temperature (T_g) (p < 0.05). With increasing heating temperatures, water vapor permeability (WVP) of films decreased (p < 0.05), but no differences in L*-value and transparency value were observed (p > 0.05). Based on FTIR spectra, the lower formation of hydrogen bonding was found in film prepared from FFS with heat treatment. Electrophoretic study revealed that degradation of gelatin was more pronounced in FFS and resulting film when heat treatment was conducted at temperature above 70 °C. Thus, heat treatment of FFS directly affected the properties of resulting films.