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.     

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。     

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.     

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.     

酸/碱预处理对美洲鳗鲡（Anguilla rostrata）鱼骨明胶理化性质与凝胶特性的影响

以美洲鳗鲡（Anguilla rostrata）鱼骨为原料,采用酸或碱预处理结合热水浸提制备鱼骨明胶,并通过得率、凝胶强度测定、SDS-PAGE、紫外全波长扫描、红外光谱扫描、动态流变学测定以及扫描电镜等研究鱼骨明胶的理化性质和凝胶特性。结果表明:酸法预处理明胶（AG60）与碱法预处理明胶（BG60）得率分别为13.6%和6.88%,凝胶强度分别为101.95 g和78.74 g。AG60和BG60的羟脯氨酸含量为3.2 g/100 g和2.7 g/100 g。两种明胶均含有β和α₁、α₂链,其中AG60的α₁+α₂含量显著高于BG60。AG60与BG60均具有明胶的特征吸收峰,且无杂蛋白吸收峰。与BG60相比,AG60具有更高的凝胶温度与熔融温度,以及更短的胶凝时间。扫描电镜分析显示,AG60具有更致密、均一的凝胶网络结构。本研究表明,与碱法预处理相比,酸法预处理制备得到的鳗鱼骨明胶具有更高的得率与更好的凝胶特性。     

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.     

Use of lactic acid for extraction of fish skin gelatin

The ability of lactic acid compared to acetic acid for Dover sole (Solea vulgaris) skin swelling and the subsequent gelatin extraction was examined. The resultant gelatins were evaluated in terms of extraction yield, amino acid composition, molecular weight distribution, gel strength, viscoelastic properties, ability to refold into triple helical structures, and aggregation phenomena. Lactic acid (25 mM) proved to be an excellent substitute for acetic acid during the skin swelling process, as the gelatin preparation thus obtained presented quite similar properties to that prepared by using 50 mM acetic acid without the negative organoleptic properties of this acid. However, the application of 50 mM lactic acid gave rise to a highly hydrolysed gelatin, with lower folding ability, gel strength and viscoelastic properties than those obtained using 25 mM lactic acid or 50 mM acetic acid.     

Extracting Conditions for Megrim (Lepidorhombus boscii) Skin Collagen Affect Functional Properties of the Resulting Gelatin

Various procedures for extracting collagen and/or gelatin from megrim skins were compared on the basis of gelatin functional properties. It was possible to prepare a dry powder of soluble collagen with good viscoelastic and gelling properties, which can be converted into gelatin by dissolving at temperatures above 45 °C. Physical properties of gelatins are influenced more by extracting conditions than by imino acid composition. A high-quality, readily-dissolved gelatin was obtained from megrim skins using a pretreatment of the skins with NaCl and dilute NaOH, then swelling with 0.05 M acetic acid followed by an extraction step in water at 45 °C.     

Characterization of tilapia (Oreochromis niloticus) skin gelatin extracted with alkaline and different acid pretreatments

Tilapia production is growing worldwide and to better utilize wastes from the processing industry, one important application is production of high quality fish gelatin to meet the needs of markets that are not amenable to beef or porcine gelatin. The extraction process from tilapia skin gelatin was optimized through the use of a combination of alkali (0.3 M NaOH) with different types and concentrations of acids before thermal hydrolysis. The effects of acid pretreatments on the protein yields and the physicochemical properties of tilapia gelatin were investigated. Acid concentrations (0.01–0.20 M) influenced gelatin protein recovery: 10.52%–22.40% for citric acid, 1.92%–21.55% for acetic acid, and 4.47%–24.35% for HCl. It was possible to increase gelatin yield for each of the tested acids by adjusting the acid concentration. Gelatin viscosity and the molecular weight distribution of gelatin proteins were related to the acid concentration used. Gelatin prepared using too low a concentration (e.g. 0.01 M acetic acid or HCl) or too high a concentration (e.g. >0.05 M HCl or citric acid) yielded an extract with a smaller ratio of large molecule components, such as β-chains, and exhibited lower viscosity. The film forming properties of gelatins extracted from three acid-optimized pretreatments showed no significant difference in transparency, tensile strength and elongation at break; though the gelatin film made from 0.03 M citric acid pretreated gelatin had somewhat better water barrier property than those made with HCl or acetic acid.     

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.     

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.     

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.     

基于鱼鳞贮藏方式改善鳙鱼鱼鳞明胶的功能特性

鱼鳞的贮藏方式会影响鱼鳞明胶的品质,本实验研究了太阳晒干后室温贮藏、4 ℃贮藏、－20 ℃贮藏和 60 ℃热风干燥后室温贮藏对鳙鱼鱼鳞明胶凝胶强度、乳化活性和起泡能力的影响,并通过傅里叶变换红外光谱、 十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和扫描电子显微镜表征经不同方式贮藏后制备得到的鱼鳞明胶的结构特性。 结果表明：4 种贮藏方式不会破坏明胶的分子质量分布（α链、β链和高分子聚合物）和傅里叶变换红外光谱的特征 性吸收峰（酰胺A带、酰胺I带、酰胺II带和酰胺III带）。4 种贮藏方式都能提升鱼鳞明胶的乳化活性和起泡能力, 其中,－20 ℃贮藏鱼鳞1 个月改善明胶乳化活性和起泡能力的效果最佳。除60 ℃热风干燥之外,其他贮藏方式均 能改善鱼鳞明胶的凝胶强度,这可能与其相应的结构特征变化有关。相比新鲜鱼鳞和60 ℃热风干燥鱼鳞,太阳晒 干和低温方式（4 ℃和－20 ℃）贮藏鱼鳞使得鱼鳞明胶α链、β链和高分子聚合物含量增加,酰胺A带吸收峰的波数 降低,鱼鳞明胶网络结构的致密性增加。     

氯化钠添加量对鱼皮明胶凝胶性能和结构的影响

以罗非鱼鱼皮为原料制备鱼皮明胶,通过单轴压缩实验、差示扫描量热实验(DSC)和红外光谱法(FT-IR)探讨了不同氯化钠添加量(0、0.5%、1.0%、1.5%、2.0%、3.0%,w/v)对不同浓度鱼胶凝胶样品(2%、4%、6.67%,w/v)的真实应力、杨氏模量、热焓值和结构的影响。结果表明,随着鱼胶浓度增加其真实应力和杨氏模量显著增加,表明鱼胶的凝胶强度和硬度显著增加,添加氯化钠后,鱼胶体系的凝胶强度和硬度随氯化钠添加量的增加呈现显著下降趋势,当氯化钠添加量达到3%时,鱼胶凝胶强度和硬度的值最小。差式扫描量热量热分析结果表明,随着氯化钠浓度的增加,热焓值下降,鱼胶的凝胶网络交联程度变弱。傅里叶变换红外光谱分析结果表明,随着氯化钠添加量增加,酰胺Ⅰ带吸收峰逐渐向低波数方向移动,氢键形成受到抑制,鱼胶凝胶强度降降低。研究表明氯化钠的存在破坏了鱼胶凝胶样品的空间结构,降低了鱼胶的凝胶性能。     

不同干燥方式对明胶特性的影响

系统研究了不同干燥方式(热风干燥,真空冷冻干燥和喷雾干燥)对明胶性质的影响,包括得率、色差、十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(sodium dodecyl sulfate-polyacrylamide gel electrophoresis,SDS-PAGE)、凝胶强度、凝胶持水力、疏水性以及傅里叶变换红外光谱(Fourier transform infrared spectroscopy,FTIR)。结果表明,由于贴壁行为和涡旋分离损失的影响,使得喷雾干燥明胶的得率明显较低(P<0.05)。喷雾干燥和真空冷冻干燥可减轻不良色泽的产生,并且降低干燥过程中明胶的氧化和降解,保留更多的高分子亚基,进而增加了明胶的凝胶强度和凝胶持水力。此外,喷雾干燥的明胶由于疏水基团的暴露以及较多的亚基组分可能含有更多的疏水基团,显示出最高的疏水性(P<0.05),冷冻干燥的明胶次之,热风干燥的明胶最低。红外结果表明,热风干燥和真空冷冻干燥破坏了明胶的构象,而喷雾干燥更有利于明胶在凝胶过程中,形成类三螺旋结构。该文可为绿色明胶的干燥方法和工业化应用提供理论依据和指导意义。     

Effects of acid concentration and the UHP pretreatment on the gelatinisation of collagen and the properties of extracted gelatins

An ultra‐high‐pressure (UHP) transmission medium with HCl was applied as a pretreatment to extract gelatin. The effects of the acid concentration (0–1% (w/v)) on the gelatinisation of collagen and the properties of gelatin were investigated. An increase in the acid concentration decreased the thermostability of collagen and increased the yield of gelatin (from 64% increased to 80%). The content of the subunit components in the pretreated collagen and the gelatin declined slightly as the acid concentration was increased, resulted in slight decrease (P > 0.05) in the gel strength (from 435 g decreased to 408 g). The analysis of Fourier transform infrared spectroscopy (FTIR) spectra showed that the triple‐helical structure, secondary structures of the collagen and covalent cross‐linking in the collagenous fibres were damaged gradually as the acid concentration was increased. The scanning electron microscopy (SEM) images showed that the smooth surface of collagen was partly disrupted with microvoids after UHP/1% HCl pretreatment, which may be related to the damage on the covalent cross‐linking.     

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.     

Effect of Epigallocatechin Gallate on the Properties of Gelatin

Epigallocatechin gallate was added to gelatin, and the changes in the gelatin were characterized to determine the effects of epigallocatechin gallate modification. The microstructural changes in the samples were analyzed using Fourier transform infrared spectroscopy, X-ray diffraction, differential scanning calorimetry, sodium dodecyl sulfate polyacrylamide gel electrophoresis, and scanning electron microscopy. The results indicated that the gel strength and thermal stability of gelatin can be improved by appropriate epigallocatechin gallate addition. The optimal final concentration of epigallocatechin gallate was 1.0 g l^?1 in a gelatin solution (66.7 g l^?1). The concentration was also verified using Fourier transform infrared spectroscopy and X-ray diffraction. Covalent bonds were not observed in the epigallocatechin gallate-gelatin samples. Hydrogen bonds were the main molecular interactions observed in the epigallocatechin gallate-gelatin samples. The color of the epigallocatechin gallate-gelatin hydrogels or xerogels was darker because of the epigallocatechin gallate oxidation.     

Extraction of Gelatin from Megrim (Lepidorhombus boscii) Skins with Several Organic Acids

ABSTRACT: Light-colored, dry collagen was obtained and, after dissolving in warm water, turned into soluble gelatin. The type of acid used influenced the gelatin viscoelastic and gelling properties. Acetic- and propionic-acid extracts produced the gelatins with the highest elastic modulus, viscous modulus, melting temperature, and gel strength, especially when skins were previously treated with dilute NaOH. After such treatment, lactic acid was also shown to be suitable for collagen or gelatin extraction. The lowest degree of turbidity was achieved by using citric acid, whereas propionic acid led to the most turbid gelatin. No improvements of rheological properties were observed when acid concentration for extraction was increased above 0.05 M.     

胶原明胶化过程中分子结构的变化

本试验以猪皮胶原为原料,研究胶原明胶化过程微观结构的变化规律。研究表明,8 h的酸处理诱导下,胶原明胶化程度较高,热水提胶后明胶得率可达83.98%,随着酸处理时间进一步延长,明胶得率显著下降;差示量热扫描(DSC)、红外光谱(FTIR)和圆二色谱(CD)研究表明,随酸处理的进行,胶原的三螺旋结构逐渐松散,维系三螺旋结构稳定的价键逐渐被破坏,酸处理8 h后,胶原三螺旋结构被过度破坏,是导致明胶得率降低的主要原因。聚丙烯酰胺凝胶电泳(SDS-PAGE)分析显示,在酸处理的过程中,胶原肽链的降解始终存在,且明胶化胶原中所保留的亚基量直接影响明胶中亚基含量。因此,在明胶化过程中必须在肽链降解和三螺旋结构松散之间寻找平衡点,以获得高得率和高品质明胶。