Gelatin Peptides Research Articles

A Review on Fish Skin-Derived Gelatin: Elucidating the Gelatin Peptides-Preparation, Bioactivity, Mechanistic Insights, and Strategies for Stability Improvement.

Fish skin-derived gelatin has garnered significant attention recently due to its abundant availability and promising bioactive properties. This comprehensive review elucidates various intricacies concerning fish skin-derived gelatin peptides, including their preparation techniques, bioactive profiles, underlying mechanisms, and methods for stability enhancement. The review investigates diverse extraction methods and processing approaches for acquiring gelatin peptides from fish skin, emphasizing their impact on the peptide composition and functional characteristics. Furthermore, the review examines the manifold bioactivities demonstrated by fish skin-derived gelatin peptides, encompassing antioxidant, antimicrobial, anti-inflammatory, and anticancer properties, elucidating their potential roles in functional food products, pharmaceuticals, and nutraceuticals. Further, mechanistic insights into the functioning of gelatin peptides are explored, shedding light on their interactions with biological targets and pathways. Additionally, strategies aimed at improving the stability of gelatin peptides, such as encapsulation, modification, and integration into delivery systems, are discussed to extend the shelf life and preserve the bioactivity. Overall, this comprehensive review offers valuable insights into using fish skin-derived gelatin peptides as functional ingredients, providing perspectives for future research endeavors and industrial applications within food science, health, and biotechnology.

Feasibility of Enzymatic Protein Extraction from a Dehydrated Fish Biomass Obtained from Unsorted Canned Yellowfin Tuna Side Streams: Part II.

The enzymatic extraction of proteins from fish biomasses is being widely investigated. However, little or almost no research has paid attention to the exploitation of unsorted fishery biomasses. This work is part of a larger study, Part I of which has already been published, and focuses on an extensive characterization of two collagenous samples, namely gelatin (G) and hydrolyzed gelatin peptides (HGPs), extracted from a dehydrated fish biomass coming from unsorted canned yellowfin tuna side streams. The results indicate crude protein fractions of 90-93%, pH values between 3 and 5, white-yellow colors, collagen-like FTIR spectra, and 17% in terms of total amino acid content. Viscosity and the study of dynamic viscous-elastic behavior were analyzed. Thermo-gravimetric analysis was performed to assess the residual ashes. Both samples were investigated to determine their molecular weight distribution via size-exclusion chromatography, with a higher total average molecular weight for G compared to HGPs, with values of 17,265.5 Da and 2637.5 Da, respectively. G demonstrated technological properties similar to analogous marine gelatins. HGPs demonstrated antioxidant activity as per FRAP assay. All the results open up new perspectives for the potential use of these substances in biodegradable packaging, dietary supplements, and skin care cosmetics.

Cowhide gelatin peptide as a source of antioxidants for inhibiting the deterioration of pudding quality during storage

To investigate the effect of gelatin peptide on the inhibition of quality deterioration in stored pudding, gelatin peptide with antioxidant properties was added to pudding products. For this purpose, a pudding recipe containing gelatin peptides was created. The gelatin peptides were characterized based on their antioxidant activity and protein structure. It was found that gelatin peptides had better antioxidant properties, lower thermal stability and crystallinity, higher hydrophobic amino acid content, and greater surface hydrogen bond exposure than commercially available peptides. Properties such as the pH, colony growth, and sensory characteristics of the pudding were characterized at 4 °C and 25 °C. The results showed that the addition of 0.5–1.0 % gelatin peptide to pudding was capable of significantly (P< 0.05) slowing down the decline in pH and sensory scores of the pudding and significantly inhibiting colony growth. It could prolong its storage life by five days at 4 °C and three days at 25 °C.

Structural Characteristics and Antioxidant Mechanism of Donkey-Hide Gelatin Peptides by Molecular Dynamics Simulation.

This study aimed to explore the structural characteristics and antioxidant mechanism of donkey-hide gelatin peptides. After hydrolysis and ultrafiltration treatment, five gelatin peptides with different molecular weights (MWs) were obtained. Amino acid analysis showed that gelatin peptides with different MWs contained a large number of amino acids, including G, P, E, N, A, and R, and differences were noted in the content of various amino acids. Fourier transform infrared spectroscopy and circular dichroism revealed that these gelatin peptides differed in terms of the peak strength of functional groups and number of secondary structures. Moreover, 26 pentapeptides/hexapeptides were identified. Among them, we investigated by molecular docking how PGPAP, which has the best antioxidant activity, may interact with the Keap1 protein. The results showed that the PGPAP-Keap1 complex had a stable conformation, and Arg415, Gly462, Phe478, and Tyr572 were the key residues involved in the binding of the peptide PGPAP to Keap1. Our results demonstrated that PGPAP could serve as a bioactive peptide with antioxidant activity.

Effects of Long-Term Administration of Bovine Bone Gelatin Peptides on Myocardial Hypertrophy in Spontaneously Hypertensive Rats.

The research purpose was to investigate the effects and the underlying molecular mechanisms of bovine bone gelatin peptides (BGP) on myocardial hypertrophy in spontaneously hypertensive rats (SHR). BGP relieved myocardial hypertrophy and fibrosis in SHR rats in a dose-dependent manner by reducing the left ventricular mass index, myocardial cell diameter, myocardial fibrosis area, and levels of myocardial hypertrophy markers (atrial natriuretic and brain natriuretic peptide). Label-free quantitative proteomics analysis showed that long-term administration of BGP changed the left ventricle proteomes of SHR. The 37 differentially expressed proteins in the high-dose BGP group participated in multiple signaling pathways associated with cardiac hypertrophy and fibrosis indicating that BGP could play a cardioprotective effect on SHR rats by targeting multiple signaling pathways. Further validation experiments showed that a high dose of BGP inhibited the expression of phosphoinositide 3-kinase (Pi3k), phosphorylated protein kinase B (p-Akt), and transforming growth factor-beta 1 (TGF-β1) in the myocardial tissue of SHR rats. Together, BGP could be an effective candidate for functional nutritional supplements to inhibit myocardial hypertrophy and fibrosis by negatively regulating the TGF-β1 and Pi3k/Akt signaling pathways.

Optimization of a Novel Tyrosinase Inhibitory Peptide from Atrina pectinata Mantle and Its Molecular Inhibitory Mechanism.

In order to realize the multi-level utilization of marine shellfish resources and to develop the potential biological activity of processing by-products of Atrina pectinata, gelatin was extracted from the mantle and the potential whitening effect of its enzymatic peptides was explored. Taking tyrosinase inhibitory activity as the evaluation index, the enzyme hydrolysate process was optimized by response-surface methodology, and the optimal enzyme hydrolysate conditions were as follows: pH 5.82, 238 min enzyme hydrolysate time, and temperature of 54.5 °C. Under these conditions, the tyrosinase inhibition activity of Atrina pectinata mantle gelatin peptide (APGP) was 88.6% (IC50 of 3.268 ± 0.048 mg/mL). The peptides obtained from the identification were separated by ultrafiltration and LC-MS/MS, and then four new peptides were screened by molecular docking, among which the peptide Tyr-Tyr-Pro (YYP) had the strongest inhibitory effect on tyrosinase with an IC50 value of 1.764 ± 0.025 mM. The molecular-docking results indicated that hydrogen bonding is the main driving force for the interaction of the peptide YYP with tyrosinase. From the Lineweaver-Burk analysis, it could be concluded that YYP is inhibitory to tyrosinase and exhibits a mixed mechanism of inhibition. These results suggest that YYP could be widely used as a tyrosinase inhibitor in whitening foods and pharmaceuticals.

Feasibility of Enzymatic Protein Extraction from a Dehydrated Fish Biomass Obtained from Unsorted Canned Yellowfin Tuna Side Streams: Part I.

This study presents for the first time a scalable process for the extraction of valuable proteins starting from samples of unsorted mixed tuna scraps which were previously dehydrated by an industrial patented process. The aims of this work were both to avoid the onerous sorting step of tuna leftovers, which generally consists of isolating skin and bones for collagen/gelatin extraction, and to improve the logistic of managing highly perishable biomass thanks to the reduction in its volume and to its microbiological stabilization. In view of a zero-waste economy, all the protein fractions (namely, non-collagenous proteins NCs and ALKs, gelatin, and hydrolyzed gelatin peptides, HGPs) isolated in the proposed single cascade flowchart were stabilized and preliminarily characterized. The extraction flowchart proposed allows one to obtain the following most promising compounds: 1.7 g of gelatin, 3.2 g of HGPs, and 14.6 g of NCs per 100 g of dehydrated starting material. A focus on oven-dried gelatin was reported in terms of proximate analysis, amino acid composition, color parameters, FT-IR spectrum, pH, and viscoelastic properties (5 mPa·s of viscosity and 14.3 °C of gelling temperature). All the obtained extracts are intended to be exploited in food supplements, feed, fertilizers/plant bio-stimulants, packaging, and the cosmetic industry.

Isolation, Identification, and Characterization of Bioactive Peptides in Human Bone Cells from Tortoiseshell and Deer Antler Gelatin.

Tortoiseshell and deer antler gelatin has been used to treat bone diseases in Chinese society. A pepsin-digested gelatin peptide with osteoblast-proliferation-stimulating properties was identified via LC-MS/MS. The resulting pentapeptide, TSKYR, was presumably subjected to further degradation into TSKY, TSK, and YR fragments in the small intestine. The above four peptides were chemically synthesized. Treatment of tripeptide TSK can lead to a significant 30- and 50-fold increase in the mineralized nodule area and density in osteoblast cells and a 47.5% increase in the number of chondrocyte cells. The calcium content in tortoiseshell was relatively higher than in human soft tissue. The synergistic effects of calcium ions and the peptides were observed for changes in osteoblast proliferation and differentiation. Moreover, these peptides can enhance the expression of RUNX2, OCN, FGFR2, and FRFR3 genes in osteoblasts, and aggrecan and collagen type II in chondrocyte (patent pending).

Determination of porcine derived components in gelatin and gelatin-containing foods by high performance liquid chromatography-tandem mass spectrometry

Detection and quantitation of porcine derived components in bovine and donkey gelatins and gelatin-containing foods is required for religious practices and consumer economic interests. In this study, shot-gun proteomics approaches were implemented to identify species-specific peptide biomarkers of three main gelatin origins, including porcine, bovine and donkey. Tryptic gelatin peptides were separated and analyzed with ultra-performance liquid chromatography-tandem high-resolution mass spectrometry (UPLC-HRMS), and 11 peptide biomarkers were selected for porcine, bovine and donkey. The gelatin pretreatment methods in three different food matrixes were optimized, and a label-free quantitative method using the MRM method of the most sensitive biomarker peptides was established. Among them, two porcine gelatin quantitative peptides were confirmed to have good linearity. The method has been successfully applied to the adulteration detection of porcine-derived components in gelatin and the quantification of porcine gelatin in candy and dairy products, with LODs of 0.07, 0.08 and 0.12 mg/kg, respectively. • A strategy for identification of peptide biomarkers of common gelatin species was established and improved with untargeted and targeted proteomics approaches. • Eleven species-specific peptide biomarkers were selected for porcine, bovine and donkey gelatin. • The pretreatment method of gelatin in different food matrixes has been optimized, especially for gelatin in dairy products. • The label-free MRM method is used for quantitative determination of adulterated porcine gelatin in mixed gelatin and gelatin-containing food, with good linearity, specificity, sensitivity and repeatability.

On the Variativity of Cell Adhesive Response under the Action of Related Short Peptides

Analysis of the participation of short peptides GER and FGER containing common tripeptide fragment in the regulation of adhesive response of CHO-K1 cells was conducted. Both peptides stimulated cell adhesion both to untreated plastic and to gelatin-coated plastic, but did not change cell attachment to poly-L-lysine-coated plastic. Tripeptide GER had larger stimulation effect on cell adhesion to untreated plastic. Peptide FGER increased the rate of cell attachment to gelatin in a wider range of concentrations as compared to adhesion to untreated plastic. Variativity of cell spreading to different substrates under peptide action was demonstrated. On untreated plastic both investigated peptides practically in equal extent stimulated cell spreading. On gelatin peptide FGER kept the stimulation effect on cell spreading, but peptide GER partly inhibited cell spreading as compared to cell spreading on untreated plastic. It was established that insertion of additional N-terminal hydrophobic amino acid residue Phe to tripeptide fragment GER changes the regulatory activity of peptide at the cell adhesion model depending on the stage of cell connection with substrate and/or on substrate properties. The structural-functional activity of investigated short peptides on the instance of different structural components of adhesive structures is discussed.

Response boosting-based approach for absolute quantification of gelatin peptides using LC-MS/MS

Response-boosting of MS signal was observed in gelatin samples due to abundant Glycine residues produced by collagen enzymolysis. In this work, a new strategy utilizing response-boosting to enhance detection sensitivity was developed for absolute quantification of Asini Corii Colla, a kind of gelatin commonly used as food therapy products in Asia, by high performance liquid chromatography coupled to tandem mass spectrometry. Peptidomics analysis was used to evaluate the similarity between eight different protein matrices, and deer-hide gelatin was selected as the appropriate simulated matrix. Isotope-labelled internal standard was used to compensate the matrix effect and construct matrix-matched calibration curves. The established method showed reliability in absolute quantification of three species-specific gelatin peptides with good linearity (r2 > 0.997), precision (RSD < 8.5%), repeatability (RSD < 8.9%), accuracy (recovery 89.4%∼106.5%) and sensitivity (LOD 0.02 ∼ 0.98 ng/mL). Thus, the present response-boosting based protocol provides a promising application in quality control of food rich in gelatins.

The Anti-Inflammatory Effect of Bovine Bone-Gelatin-Derived Peptides in LPS-Induced RAW264.7 Macrophages Cells and Dextran Sulfate Sodium-Induced C57BL/6 Mice.

The bioactive peptides hydrolyzed from bone collagen have been found to possess health-promoting effects by regulating chronic diseases such as arthritis and hypertension. In the current study, the anti-inflammatory effect of bovine bone gelatin peptides (GP) was evaluated in 264.7 macrophages cells and followed by animal trials to investigate their interference on inflammatory cytokines and gut microbiota compositions in dextran sodium sulfate (DSS)-induced C57BL/6 mice. The GP was demonstrated to alleviate the extra secretion of interleukin-6 (IL-6), nitric oxide (NO) and tumor necrosis factor-α(TNF-α) in lipopolysaccharide (LPS)-induced RAW264.7 cells. In DSS-induced colitis mice, the gavage of GP was demonstrated to ameliorate the IBD symptoms of weight loss, hematochezia and inflammatory infiltration in intestinal tissues. In serum, the proinflammatory cytokines (TNF-α,IL-6, MCP-1, IL-1β) were suppressed along with the decreasing effect on toll-like receptor 4 and cyclooxygenase-2 by GP treatment. In the analysis of gut microbiota, the GP was checked to modulate the abundance of Akkermansia, Parasutterella, Peptococcus, Bifidobacterium and Saccharibacteria. The above results imply that GP could attenuate DSS-induced colitis by suppressing the inflammatory cytokines and regulating the gut microbiota.

Bioactive and Functional properties of gelatin peptide fractions obtained from sea bass (Dicentrarchus labrax) skin

Fish skin is one of the most common resources of gelatin, which can be hydrolyzed bioactive peptides. In this study, gelatin from sea bass skin (SBS) was hydrolyzed with flavourzyme® to obtain peptide fractions with different molecular weights and determined their bioactive and functional properties. All peptide fractions obtained showed antioxidant activity (DPPH and FRAP). The bass gelatin peptide fraction 1 (BF1) (≤5 kDa) showed the highest DPPH (44.9%) and FRAP (42.04 mmol Fe+2 /g and 22.98 mmol trolox/g) activities. Besides, the BF1 (≤5 kDa) peptide fraction showed the highest in vitro cytotoxic effect (16.58%) at 20 mg/mL concentration compared to the other peptide fractions. The highest emulsifying capacity (389.5 m2/g), emulsifying stability (53.2 min), foaming capacity (30.47%), and foaming stability (10.40%) were obtained from the control gelatin sample. Moreover, the BF3 (≥10 kDa) peptide fraction showed an excellent fat binding capacity (9.39 mL/g). Enzymatic hydrolysis decreased emulsifying and foaming capacity of gelatin while increasing its fat binding capacity. In particular, antioxidant and anticancer activities of peptide fractions with low molecular weight were found to be high. The results demonstrated that gelatin and hydrolysates from the SBS offer an important alternative as a functional food ingredient for food technology.

Cold setting of gelatin–antioxidant peptides composite hydrogels using a new psychrophilic recombinant transglutaminase (rTGase)

This study used a new psychrophilic transglutaminase (TGase) to catalyze the formation of protein and peptide hydrogels by forming isopeptide bonds. The psychrophilic recombinant TGase (rTGase) was overexpressed in E. coli and purified using a Ni-Sepharose column followed by stepwise dialysis to recover the cold-active enzyme (specific activity of 1.63 U/mg). The rTGase catalyzed the formation of cold-set hydrogels at 4 °C, which were comprised of high molecular weight gelatin and antioxidant peptides. The antioxidant bioactivity of the cold-set gelatin–peptides composite hydrogels was significantly improved with rTGase at 4 °C for 10 days. To understand the mechanism of the psychrophilic rTGase on the enhanced bioactivity of cross-linked hydrogels, in silico molecular docking studies were performed to assess the binding of antioxidant peptides into the rTGase active pocket, as well as to analyze the interactions of Gln and Lys residues in the antioxidant peptides with amino acid residues in the rTGase catalytic site. The formation of covalent cross-linkages in the gelatin–gelatin, gelatin–peptide and peptide–peptide matrices, and possible structures of composite hydrogels formed by rTGase catalysis in cold conditions, were deduced. The new psychrophilic rTGase enabled stabilization of the antioxidant bioactivity of protein- and peptide-containing cold-set hydrogels formed at low temperatures.

Highly transparent, writable and photoluminescent foldable polymer film: When fluorescent dyes or pigments join cellulose-based microgel

We prepared a self-dispersed cellulose-based microgel via chemically bonding hydrophilic gelatin peptide chain onto cellulose glucose chain. Following, a variety of highly transparent, foldable, and writable photoluminescent polymer films was obtained through loading organic dyes and inorganic pigments onto cellulose-based microgel matrix, respectively. Benefiting from the coupling sites and network effect of microgel as well as the abundant hydroxyl, amino, and imino groups in its structure, the microgel containing organic dyes and inorganic pigments exhibit good dispersion and stability, and the resultant photoluminescent films emit bright yellow, orange, yellow-green, and blue-green fluorescence under UV light, respectively, especially the cellulose-based microgel stabilized inorganic alkaline earth aluminate hybrids exhibit continuous luminescence for a long time in the dark. Compared with the existing regenerated cellulose or CNCs-based fluorescent films, the cellulose-based microgel fluorescent films present higher transmittance and good biodegradability. This study can bring new ideas for the development of flexible luminescent devices.

Gelatin-based films activated with red propolis ethanolic extract and essential oils

Red propolis ethanolic extract (RPEE) and essential oils are well-known for their antimicrobial and antioxidant activities. The aim of this study was to develop gelatin-based active films loaded with RPEE in combinations with clove essential oil and/or basil essential oil and to study the effects of these components on the antioxidant, antimicrobial and physical properties of films. The active films were produced by a spreading technique, with RPEE and essential oils in different concentrations. These compounds were evaluated for antioxidant and antimicrobial activities, as well as for their minimal inhibitory concentration and minimal bactericidal concentration. The films were characterized through physical, antimicrobial and antioxidant properties. RPEE and essential oils both presented high antioxidant activity. All essential oils showed antimicrobial activity against all evaluated bacteria, while the RPEE presented activity only against the Gram(+) bacteria. All films showed antioxidant activity. Films with clove essential oil presented the highest total contents of phenol and antioxidant activity. The RPEE in films increased their UV/visible light barrier and modified their thermal properties. Changes in infrared spectra suggested possible interactions between phenols of the active compounds and the gelatin peptides. Overall, physical properties of active films were typical of biopolymer-based films.

Antioxidant mechanism, antibacterial activity, and functional characterization of peptide fractions obtained from barred mackerel gelatin with a focus on application in carbonated beverages

The present study aimed to use fish by-products to generate gelatin peptides with potential applications in carbonated beverages. After ultrafiltration, the F < 3 kDa (fraction < 3 kDa) showed the highest peptide concentration (227.22 mg/g) as well as antibacterial (MIC of ≤ 0.5 mg/mL) and antioxidant activities, including hydroxyl and superoxide radical scavenging, ferrous chelation, and ferric reduction (with IC50 values of 0.88, 1.04, 0.50 mg/mL, and 0.58, respectively). 2,2-diphenyl-1-picrylhydrazyl scavenging was the highest in the 3 < F < 10 kDa (IC50 of 0.64 mg/mL). In vitro gastrointestinal digestion significantly decreased all biological activities. Solubility, water holding capacity, and emulsifying activity of the F < 3 kDa were the highest while foaming properties and overfoaming were reversibly related to the molecular weight. All abovementioned properties, in addition to in vitro cytotoxicity analysis in different cell lines and better flavor characteristics, indicated that the F < 3 kDa could be safely and properly used as an ingredient for the fortification of carbonated beverages.

Characterization and Antioxidant Activity of Collagen, Gelatin, and the Derived Peptides from Yellowfin Tuna (Thunnus albacares) Skin.

Skin waste from tuna processing needs to be utilized, such as extraction of its collagen and gelatin. Their functional properties can be improved by enzymatic hydrolysis for conversion to peptides. Thus, the research objectives were to examine the characteristics and antioxidant activity of collagen, gelatin, and the derived peptide from yellowfin tuna skin. Collagen was extracted using 0.75 M acetic acid at 4 °C, while gelatin was prepared using 0.25% citric acid and extracted at 65 °C. Hydrolysis was carried out with 2% Alcalase, followed by fractionation with a molecular weight cut off sieve for both collagen and gelatin. Collagen yield was 22.6% with pH value of 6.63 and whiteness of 96.7%. Gelatin yield was 20.0% with pH value of 4.94 and whiteness of 51.0%. Hydrolysis for three hours resulted in 52.7% and 45.2% degree of hydrolysis for collagen and gelatin, respectively. The molecular weights of collagen peptides ranged from 2.94 to 11.93 kDa, while those of gelatin peptides ranged from 3.54 to 16,620 kDa. Antioxidant activities of these peptides were higher than those before hydrolysis. The high antioxidant activity (IC50) of collagen peptides were found in <3, 3–10, and 10–30 kDa fractions as well as in the gelatin peptides.

Identification and characterization of the peptides with calcium-binding capacity from tilapia (Oreochromis niloticus) skin gelatin enzymatic hydrolysates.

The aim of this study was to isolate and identify the peptides with calcium-binding capacity from the different tilapia skin gelatin enzymatic hydrolysates. The complex protease was selected and its hydrolysates were further separated using gel filtration chromatography (Sephadex G-25) and reverse phase high-performance liquid chromatography. Two purified peptides with strong calcium-binding capacity were identified as Tyr-Gly-Thr-Gly-Leu (YGTGL, 509.25 Da) and Leu-Val-Phe-Leu (LVFL, 490.32 Da). The calcium-binding capacities of YGTGL and LVFL reached 76.03 and 79.50 µg/mg, respectively. The structures of the complex of purified peptides and calcium (YGTGL-Ca and LVFL-Ca) were characterized by ultraviolet-visible spectroscopy (UV-VIS), scanning electron microscopy (SEM), X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), and mass spectrometry (LC-MS/MS). The results of UV-VIS, SEM, and XRD indicated that YGTGL-Ca and LVFL-Ca were formed as new compounds. The results of FTIR and LC-MS/MS indicated the nitrogen atom of the amino group and the oxygen atom of the carboxyl group in terminates of the peptides provided primary binding sites. Moreover, the hydrophobic amino acids in purified peptides could provide more chelating spaces. This study was of great significance for the development of calcium supplement foods. PRACTICAL APPLICATION: Compared with inorganic calcium and organic calcium, the bioactive gelatin peptide chelated calcium has the characteristics of high utilization rate, high solubility, and high absorption rate. The raw materials are extracted from the tilapia processed waste, which reduce the cost, make full use of resources, and improve the bioavailability. The tilapia skin gelatin peptide calcium chelate can be directly absorbed by the human body, and the absorption efficiency is high, further improving the resource utilization rate and having high economic benefits, which is a comprehensive supplement that can also be used as a functional food.

Infrared spectroscopic study of the coil-helix transition of highly concentrated gelatin formulations

The aim of this study was to investigate the applicability of ATR-FTIR spectroscopy as an analytical tool to monitor the gel formation of highly concentrated gelatin formulations. Spectral changes induced by the coil-helix transition have been studied and related to the elasticity parameter G′ obtained by oscillatory rheology in simultaneous measurements. A principal component analysis of the amide I band allowed the evaluation of triple helix formation kinetics. It was found that the key frequencies of the amide I band at 1657 and 1612 cm−1 represent the transition of the gelatin molecules from the random coil to the triple helical conformation in the emerging gel. A direct correlation between the conformation of the gelatin molecules and the gel elasticity was obtained for a commercially available pharmaceutical grade limed bone gelatin in concentrations between 20 and 40% w/w. The same was valid upon addition of small gelatin peptides or a helix inhibitor. No such correlation between triple helix content and G′ was found for limed bone gelatins of the same Bloom value but an asymmetric molecular weight distribution with extremely high fractions of high or low molecular weight components. This suggests that early gel elasticity is not solely linked to the triple helix nucleation. Hence, our results indicate that FTIR spectroscopy can be applied to gain a better understanding of the relationship between triple helix content and elastic gel properties of pharmaceutical gelatin capsule shell formulations.