Bone Gelatin Research Articles

Physical Properties of Edible Films from Pangasius catfish Bone Gelatin-Breadfruits Strach with Different Formulations

Abstract— This research aims to provide edible film made from fish bone gelatin of Pangasius catfish mixed breadfruit starch with different formulations. The study was carried out through three stages including breadfruit starch isolation, edible films fabrication and analysis of physical properties of resulted edible film such as thickness, water vapor transmission, tensile strength and percent of elongation as well as moisture content. Edible films which were formulated from fish bone gelatin of Pangasius catfish by breadfruit starch addition have thickness values ranged of ​​0.084-0.123 mm, 6.08 - 16.77% of moisture content, water vapor transmission of 1.07 - 1.60 g/ m²/hour, tensile strength of 0.245 - 1.186 MPa, and percent of elongation around 70 - 87.14%. The edible films from gelatin of Pangasius catfish bone with breadfruit starch addition have physical characteristic which fulfill the standard requirements issued by Japanese Industrial Standard (JIS)
 Keywords— edible film; biodegradable materials; fish gelatin;breadfruit starch; food packaging 

Effect Maltodextrin on Hedonic Attributes of Tray-Dried Fish Gelatin Powder from Bone of Pangasius sp.

Abstract—This research aims to drying liquid gelatin from bone of pangasius sp. through maltodexrtrin addition by using tray dehydrator. The stages of this study were including fish bone gelatin extraction and drying of liquid gelatin at 55 oC for 6 hours coupled with addition of maltodextrin (0%, 5%, 8%, 11%, and 14%). The analysis covering gelatin yield and organoleptic parameter i.e hedonic test toward color, aroma, texture, and overall preference. Then, proximate analysis was done also on the selected dried fish bone gelatin. The results shown that maltodextrin addition has significant effect to the hedonic attributes of resulting fish bone gelatin powder by tray drying process. The panelists preference toward fish bone gelatin powder with maltodextrin is like slightly whether to color, aroma, texture, and overall appearance. The yield of dried fish bone gelatin using tray dehydrator is about 16%. The moisture, ash, protein and fat compositions of dried fish bone gelatin in this study were 11.45%, 0.96%, 36.15%, and 0.045% respectively. 
 Keywords—drying; fish gelatin; halal gelatin; maltodextrin; organoleptic

Synthesis of Co/Mesoporous Silica-Alumina Catalyst from Sidoarjo Mud Templated by Bovine Bone's Gelatin for Hydrocracking of Waste Lubricant

The synthesis of Co/Mesoporous Silica-Alumina (MSA) catalyst from Sidoarjo mud templated by bovine bone’s gelatin for hydrocracking of waste lubricant has been done. The gelatin was obtained from bovine bone by the combination method of acids (CH 3 COOH 4% and HCl 1 M) and base (NaOH 0.1 M). The silica was obtained from Sidoarjo mud using the batch method (distilled water, HCl 6 M, and NaOH 6 M). The synthesis of MSA was carried out using silica from Sidoarjo mud with the variation of Si/Al of 25, 50, and 100, respectively, using gelatin as a template to produce MSA(25), MSA(50), and MSA(100); and was then analyzed by FTIR and acidity test. The MSA with the highest acidity and optimum thermal stability (MSA(x)) was analyzed by GSA and TEM. The cobalt metal from Co(NO 3 ) 2 ·6H 2 O as salt precursor was loaded to the MSA(x) using the Wet Impregnation (WI) and Ion Exchange (IE) method. The Co/SAM(x) was tested as a catalyst for hydrocracking of waste lubricant oil. The results showed that the MSA(100) has the highest acidity and optimum thermal stability with an average pore diameter of 3.37 nm, the pore volume of 0.87 cm 3 g –1 , and a surface area of 230.73 m 2 . The cobalt metal content loaded on the MSA(100) by WI and EI method was 0.50% and 0.15 wt.%, respectively. The hydrocracking of lubricant oil using Co/MSA(100)WI catalyst yielded a liquid fraction of 39.20 wt.% with selectivity for gasoline and diesel fraction of 29.30 and 9.48 wt.%. The Co/MSA(100)IE catalyst produced a liquid fraction of 28.10 wt.% with selectivity for gasoline and diesel fraction of 22.70 and 5.32 wt.%.

Physico-Chemical Characteristics of Giant Featherback (Chitala lopis) and Mackerel (Scomberomorus commerson) Bone Gelatins and Their Potential as Gelling agents in Gel Mixtures for Roselle (Hibiscus sabdariffa L.) Soft Jelly Candies

The gel strength of gelatin depends on its raw material. Fish gelatin shows a broad range of gel strengths. Giant featherback and mackerel bone gelatins have the gel strengths 220 and 38 Bloom, respectively. Here, we prepared roselle soft jelly candy using the single gelatin and mixtures gelatins from bovine, giant featherback, and mackerel sources. Roselle calyces extract has been used as a food additive to improve taste, aroma, and antioxidant content. The soft jelly candies (1.5 % w/v roselle extract and 12 % gelling agent) were prepared using three types of gelatin (giant featherback (GF), mackerel (M), and bovine (B)) in four mixtures (GF:B = 1:3, GF:B = 1:1, M:B = 1:3, and M:B = 1:1). This experiment was arranged in a completely randomized design with three replications for each treatment. Physico-chemical and sensory characteristics of the jelly candies were determined. The data were analyzed by ANOVA continued by the LSD test. The results showed that water content, pH, gel strength, hardness, rigidity, and hedonic quality (taste, color, texture, and overall performance) of the roselle soft jelly candies were affected by choice of gelatin, but not total reducing sugar. Roselle soft jelly candy made with giant featherback and bovine gelatin (1:3) mix was the most preferred by the panelists.

Octenyl succinic anhydride modification of bovine bone and fish skin gelatins and their application for fish oil-loaded emulsions

Molecular modification of emulsifiers has attracted many attentions in the field of emulsions. In this work, an octenyl succinic anhydride (OSA) modification method is developed to modify bovine bone gelatins and fish skin gelatins, and then the effect of OSA modification on the gelatins and their application for fish oil-loaded emulsions are explored and compared. The results demonstrate that OSA can be successfully applied to modify gelatins and change their isoelectric points. Moreover, the degree of succinylation (DS) logarithmically increases with OSA-gelatin mass ratios in the chemical reaction process. Further, the OSA modification has no obvious effects on the initial emulsion droplet sizes of fish oil-loaded emulsions. Finally, the emulsion storage experiments suggest that the DS increase of OSA-modified bovine bone gelatins increases the droplet stability, phase transition time, and creaming index of fish oil-loaded emulsions, whereas the DS increase of OSA-modified fish skin gelatins only increases the droplet stability and has no obvious effects on the phase transition and creaming index of fish oil-loaded emulsions. This work will provide useful information to understand the relationship among molecular modification, molecular structure, and function of protein. It also will provide a potential route to optimize the preparation and application of fish oil-loaded emulsions in the field of food and drug industries.

Characteristics of Spanish Mackerel (Scomberomorus commerson) Bone Gelatin for Ice Cream Stabilizer

Ice cream is a milk-processed-based product that provides enough nutrients. The ice cream making process requires a stabilizer. The stabilizer used in this study was gelatin. This research used mackerel fish bones as the basic material for the gelatin, where the protein content is high enough to be made as gelatin. The purpose of this research was to determine the right amount of mackerel to make ice cream with good characteristics. The descriptive experimental method with an advanced regression analysis method was used to analyze the results of ice cream physical characteristics, while randomized block design (RBD) and descriptive method were used on the organoleptic results of the ice cream. The experiment consisted of four repetition sessions with different gelatin concentrations of 0%, 0.1%, 0.3%, and 0.5%, each treatment was repeated three times. The results showed additions of gelatin concentration on ice cream making had a very strong relationship with the characteristics of the ice cream namely overrun value, melting power, and viscosity with a correlation coefficient of 0.94, 0.911, and 0.995 respectively. While in terms of organoleptic, the higher the concentration gelatin, the softness of ice cream is higher, therefore more panelists preferred it.

Commercial cold-water fish skin gelatin and bovine bone gelatin: Structural, functional, and emulsion stability differences

Both fish gelatins and mammalian gelatins have been explored in the preparation and application of fish oil-loaded emulsion. However, the comprehensive comparisons and relationships of the structural, functional, and emulsions stability differences between these two gelatins remain unclear. In this work, structural, functional, and the emulsion application differences between commercial cold-water fish skin gelatin (CFG) and bovine bone gelatin (BBG) are explored. The results demonstrated the BBG-stabilized emulsion was more stable than the CFG-stabilized emulsion at 4 °C and higher temperatures (23 °C and 37 °C) decreased the creaming stability differences. The creaming index differences between CFG-stabilized and BBG-stabilized emulsions were mainly dependent on emulsion droplet size and interfacial layer thickness consisted of gelatins. Further, the possible reason for CFG's lower emulsion stability at 4 °C might be mainly dependent on its higher β-antiparallel percentage in the secondary structures and its lower thickness of gelatin film-like nanostructures. This work will provide useful information to understand the relationship between structure and function of protein. It also will provide a guide for the application of fish oil-loaded gelatin-stabilized emulsions in food beverages and other foods such as surimi.

Distinguishing tissue origin of bovine gelatin in processed products using LC/MS technique in combination with chemometrics tools

Gelatin, as a by-product of the meat industry, is extracted from bone and skin of mainly bovine and porcine origins. It is used widely in the food, drug, and cosmetic industries. Authenticity testing methods can be used to confirm whether labelled ingredients are present in the product. Generally, studies on gelatin are concerned mainly with determining species, but detecting tissue origin is also important from religious, health, and commercial perspectives. In the present study, for the first time, liquid chromatography/mass spectrometry (HPLC/MS) was used to differentiate bovine bone gelatin from gelatin derived from bovine skin. Tryptic-digested gelatins were measured using HPLC/MS and, subsequently, two powerful chemometrics approaches (i.e., PCA and PLS-DA) were used to classify samples as either skin or bone gelatins. Origin of bovine gelatins in different test samples were predicted accurately using this method. The results showed both the stability and reliability of the proposed procedure.

Dipeptidyl Peptidase IV (DPP-IV) Inhibitory Activity of Ultrafiltration and Gel Filtration Fraction of Gelatin Hydrolyaste Derived from Bone of Fish for Antidiabetes

Bioactive peptides have been investigated largely for many biofunctional properties. The aim of this research was to determine the inhibitory activity of ultrafiltration (UF) and gel filtration (GF) fractions of gelatin derived from bone of Pangasius catfish against dipeptidyl peptidase IV (DPP-IV). Previous studies have shown that gelatin from skins of salmon, hake, halibut, milkfish, tilapia and bone of pangasius catfish have the activity in DPP-IV inhibition. While, as inhibitor, most of previous and recent studies shown that separation and fractination of gelatin hydrolysate increase their activity. This research was conducted in three stages including gelatin hydrolysates fractination by ultrafiltration (UF), UF fraction loaded into gel filtration (GF) and DPP-IV inhibition measurement. The fish bone gelatin was hydrolyzed using flavourzyme at three enzyme/substrate ratios (E/S of 3%, 6% and 9%) with incubation times 4, 6, and 8 h. Then, the hydrolysates fractioned by ultrafiltration with 3 kDa cutoff membrane continued with Superfine G-25 sephadex column (65 cm x 3 cm, flow rate 1 mL/min). The result shown that both group of fractions i.e UF and GF have inhibitory activity regarding their capacity to inhibit DPP-IV. The UF fraction >3 kDa derived from 9% (E/S) ratio for hdyrolysis were superb as DPP-IV inhibitor than other fractions, the highest one also has bioactivity higher than other previous fish gelatin fractions.

Antihypertensive Effects in Vitro and in Vivo of Novel Angiotensin-Converting Enzyme Inhibitory Peptides from Bovine Bone Gelatin Hydrolysate.

In this study, we investigated the antihypertensive effects in vitro and in vivo of novel angiotensin-converting enzyme inhibitory (ACEI) peptides purified and identified from bovine bone gelatin hydrolysate (BGH). Thirteen ACEI peptides were identified from BGH, and among which, RGL-(Hyp)-GL and RGM-(Hyp)-GF exhibited high ACE inhibition with IC50 values of 1.44 and 10.23 μM. Molecular docking predicted that RGM-(Hyp)-GF and ACE residues of Glu384, His513, and Lys511 formed hydrogen-bonding interactions at distances of 2.57, 2.99, and 2.42 + 3.0 Å. RGL-(Hyp)-GL formed hydrogen bonds with Lys511 and Tyr523 and generated hydrogen-bonding interactions with His387 and Glu411 in the zinc(II) complexation motif at distances of 2.74 and 3.03 + 1.93 Å. The maximal decrements in systolic blood pressure in spontaneously hypertensive rats induced by one-time gavage of RGL-(Hyp)-GL and RGM-(Hyp)-GF at 30 mg/kg were 31.3 and 38.6 mmHg. RGL-(Hyp)-GL had higher enzyme degradation resistance than that of RGM-(Hyp)-GF in vitro incubation in rat plasma, and they were sequentially degraded into pentapeptides and tetrapeptides within 2 h. Our results indicate that BGH can serve as a nutritional candidate to control blood pressure.

Effect of phosphoric acid pretreatment on characterization of gelatin from broiler chicken (Gallus gallus domesticus L.) bones

Broiler chicken [Gallus gallus domesticus (Linnaeus, 1758)] bone is one of the alternative sources of collagen to form gelatin. This research was attempted to extract and characterize broiler chicken bone gelatin. Chicken bones were pretreated with phosphoric acid in various concentrations and curing times. The concentration of phosphoric acid was 8 %, 9 % and 10 %, while the curing time was 12 h and 24 h. Then, gelatin was extracted and physicochemical properties examined. Gelatin was pretreated by phosphoric acid 9 % for 24 h produced the best physicochemical properties with (15.4 ± 0.94) % of yield. Moisture, ash content, and pH of chicken bones gelatin were (3.96 ± 1.46) %, (24.0 ± 6.36) %, and (5.74 ± 0.003) %, respectively. Other physiochemical properties were (372.29 ± 4.62) g bloom of gel strength, (54.21 ± 1.24) % of protein content, and (60.21 ± 0.72) % of emulsion stability. Infrared spectra of chicken bones gelatin had the most with vibration peak at the wave number of 1 639 cm −1 to the amide I, of 1 545 cm −1 to the amide II, of 1 129 cm −1 to the amide III, of 2 292 cm −1 to the amide B, and 3 456 cm −1 to the amide A.

The proportion–ratio on dipeptidyl aminopeptidase-4 (DP-4) inhibition by gelatin compared to synthetic sitagliptin

ABSTRACTThe current study aims to determine the inhibition activity gelatin against dipeptidyl aminopeptidase 4 (DP-4). Two commercial gelatins, i.e., bovine and fish skin gelatin and one extracted (in our laboratory) gelatin, i.e., fish bone gelatin were selected for analysis. Each gelatin have same protein pattern (75–245 kDa) on sodium dodecyl sulfate polyacrylamide gel electrophoresis with mean of protein concentration of 1.72 mg/mL. The inhibition activity was measured on the capacity to inhibit DP-4 by using Gly-Pro-p-nitroanilide as their substrate. The sitagliptin was used as standard comparison. Based on the percent inhibition, gelatin has been shown to be the prospective DP-4 inhibitor.

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.

PENGARUH KONSENTRASI ASAM KLORIDA DAN LAMA PERENDAMAN TERHADAP SIFAT FISIK DAN KIMIA GELATIN TULANG IKAN TUNA (Thunnus albacare)

Type A gelatin was extracted from partial hydrolysis of collagen from tuna fish bone. The experiment was conducted to determine the effect of hydrochloride acid solution and soaking time on physical and chemical properties of tuna fish bone gelatin and to determine optimum conditions for extraction of tuna fish bone gelatin which is based on the highest yield and gel strength. The experiment used randomized complete block design (RCBD) with two factors. The first factor was concentration of hydrochloric acid consisting of 3 level (1, 3, and 5% v/v) and the second factor was soaking time (24, 48, and 72 hours). The results showed that concentration of hydrochloric acid and soaking time significantly affect at the physical and chemical properties of tuna fish bone gelatin. The optimum condition obtained the treatment of gelatin was concentration of hydrochloric acid 3% and soaking time 72 hours. Physical and chemical properties of gelatin from tuna fish bone, which is 120.37 gram Bloom gel strength; viscosity 4.75 cP; melting point 20.37 ° C; color was yellowish brown, moisture content 11.85%, protein content 80.90%.; fat content 2.73% and ash content 8.12%.

KUALITAS FISIK ANTIMICROBIAL EDIBLE FILM (AmEF) DENGAN EKSTRAK DAUN TEH (Camellia sinensis) DARI GELATIN LIMBAH TULANG AYAM

This study intends to find out the impact of green tea leaves’ concentration and heating temperature on the physical quality of the antimicrobial edible film (AmEF) made from chicken bone gelatin. This study also utilizes chicken bone waste, in oder to increase both monetary value and the usefulness of that waste. The bone gelatin will then be utilized as AMeF and combined with green tea leaves’ extract (Camellia sinensis) that used as sausage wrapper. Several stages have been taken placed as follow: chicken bone gelatin production; green tea leaves extraction; and tea leaves’ extract-AMeF production. cCompletely randomized design with the factorial pattern was used in this study. Data was statistically analyzed using uni-variate analysis with 5% significance rate. Duncan’s Multiple Range Test (DMRT) will later be used to further test if there is a difference between each treatment. The physical qualities of AmEF that tested in this study include clarity, color, and thickness. The result of this study shows that a clarity value of AmEF was ranged between 0.62-2.12 abs, color was 0.87-2.60 abs, and 0.015-0.023 mm of thickness. The addition of tea extract and heating temperature have significant (P<0.05) effect on the clarity and color while have no effect on the thickness.

Characteristic and Functional Properties of Gelatin from the Bones of Alaska Pollock (Theragra chalcogramma) and Yellowfin Sole (Limanda aspera) with Papain-Aided Process

ABSTRACTFish gelatin was extracted from the bone of Alaska pollock (Theragra chalcogramma) and yellowfin sole (Limanda aspera) using papain-aided process and the characteristic and functional properties were analyzed. The bones of Alaska pollock and yellowfin sole yielded 9.07% and 7.15% gelatin, respectively. Protein was the main component (84.67–91.76 g/100 g) in both gelatins. There was no significant difference in whiteness between the two gelatins. The absorption peak of Alaska pollock bone gelatin (ABG) and yellowfin sole bone gelatin (YBG) in ultraviolet-visible (UV-vis) spectra was 209 nm and 211 nm, respectively. Scanning electron microscope (SEM) and Fourier transform infrared (FTIR) investigations indicated the existence of helical arrangements and uniform network structures in the two gelatins. The gel strength of ABG (105.64 g) was significantly lower than that of YBG (187.39 g), possibly due to the presence of shorter chains as indicated by the degradation of α-, β-, and γ-chains. ABG also had lower emulsibility and foamability than YBG, which was associated with relatively lower hydrophobic amino acids. Therefore, yellowfin sole would be a better potential marine source of gelatin obtained from fish processing by-products.

Purification and characterization of a novel calcium-biding decapeptide from Pacific cod (Gadus Macrocephalus) bone: Molecular properties and calcium chelating modes

The calcium-binding peptides were recognized as novel functional ingredients for improving calcium bioavailability. Pacific cod (Gadus Macrocephalus) bone gelatin hydrolysates (BGH) with considerable calcium binding activity (0.41 ± 0.08 μg/mg) were prepared and then separated by hydroxyapatite specific affinity column chromatography and reversed phase high performance liquid chromatography. A novel decapeptide was purified and identified as KGDPGLSPGK (designated Peptide-K, MW: 954.5 Da), which exhibited the highest calcium binding activity (2.53 ± 0.12 μg/mg) among BGH. The calcium-binding sites were investigated by circular dichroism (CD), Fourier transform infrared spectroscopy (FTIR), and mass spectrometry (MS). Results indicated that the carboxylate O atoms of Asp-3 and Lys-10 and the side chain amino N atom of Lys-10 in Peptide-K contributed to the calcium-chelating reaction. Possible chelating modes between Ca2+ and Peptide-K were constructed based on the obtained data. Results may contribute to a better understanding of the chelating process between Ca2+ and calcium-binding peptides.

Chemical Quality of Chicken Bone Waste Gelatin Extracted using Chloride Acid

This study aims to isolate and extract the chicken bone gelatin through hydrolysis process and was analyzed chemically the extracted gelatin product. Research activities included several stages: (1) chicken bone waste preparation (2) ossein extraction (3) gelatin production (4) gelatin chemical analysis. The extraction time was 60 minutes with 5 treatment levels of HCl concentration ie K1 = 2%, K2 = 3%, K3 = 4%, K4 = 5% and K5 = 6%. Each treatment was repeated three times. Gelatin analysis consists of water content, ash content, protein content, pH value, viscosity, and gel strength. The highest water content was 15,76% at 2% HCl concentration, the highest ash content was 44,40% at 2% HCl concentration, and the highest protein content was 23,23% at 3% HCl concentration. The highest pH value was 4.14 produced by gelatin at 2% HCl concentration, viscosity was 5,13 cP and gel strength of 252,43, the best bloom was produced by gelatin at 6% concentration level. The conclusion of this research is the best quality of gelatin is obtained by using chicken bones and the use of chloride acid for 24 hours and 6% HCl concentration level.

A simple and eco-friendly method of gelatin production from bone: One-step biocatalysis

Conventional gelatin production based on liming is time-consuming and leads to significant land pollution and wastewater. Thus, an efficient and ecologically compatible method of gelatin production from bone has to be developed. In this study, a novel one-step enzymatic method was proposed, and the new method showed excellent results, with time reduction for ossein hydrolysis from 3–8 weeks to 3 h and without the need for liming and deliming, thereby reducing alkali usage substantially and saving nearly 60% water. In addition, 4 steps with low yield involved in conventional gelatin extraction were replaced by 1 step with relatively high yield in the new method. The gelatin obtained from the one-step enzymatic method possesses similar physicochemical properties except for the isoelectric point. Moreover, environmental assessment and financial analysis demonstrated the environmental benefit and economically feasible of the enzymatic method, suggesting the potential of one-step biocatalysis as alternative route to sustainable development for bone gelatin industry.

The hydroxyproline content of fish bone gelatin from Indonesian Pangasius catfish by enzymatic hydrolysis for producing the bioactive peptide

Abstract. Atma Y, Lioe HN, Prangdimurti E, Seftiono H, Tahufik M, Fitriani D, Mustopa AZ. 2018. The hydroxyproline content on fish bone gelatin from Indonesian Pangasius catfish by enzymatic hydrolysis for producing of bioactive peptide. Biofarmasi J Nat Prod Biochem 16: 64-68. Gelatins has been widely used in food, medicines, cosmetics, photography industries. In the food industry, gelatin used as food additives and functional foods. The applications of gelatin as a functional food due to their bioactivity in a form of peptides. Bioactive peptides from gelatin are mostly obtained through enzymatic hydrolysis processes. This study was conducted to measure the hydroxyproline content of gelatin bone of Indonesian Pangasius catfish (Pangasius sutchi) before and after enzymatic hydrolysis. The hydroxyproline is one of the dominant amino acids in gelatin. Gelatin hydrolysis was carried out using a flavourzyme at concentration of 6% and the incubation series of 0, 4, 6 and 8 hours. The standard concentration of hydroxyproline was used in range of 0-1 μg. The results showed that the linear curve of the hydroxyproline standard solution was y=0.0554x+0.0406, with the coefficient of determination (R²) = 0.9435. The incubation time of enzymatic hydrolysis (6% enzyme concentration) affected the hydroxyproline content. The hydroxyproline from fish bone gelatin was 18.91±2.87 mg/mL, 63.81±1.28 mg/mL, 46.21±1.28 mg/mL and 37.64 ± 0.64 mg/mL respectively during 0, 4, 6 dan 8 h incubation time. This hydroxyproline content was significantly different at 95% confidence level from each treatment time.