• Asian-Australasian Journal of Animal Sciences
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• v.29 no.11
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• pp.1632-1638
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• 2016

A study was conducted to compare the endogenous basal losses of phosphorus (EBLP) in pigs fed diets containing gelatin (GEL) or spray-dried porcine plasma (SDPP) as protein sources and to determine the standardized total tract digestibility (STTD) of phosphorus (P) in SDPP. The trial was carried out at the Federal University of Santa Maria, Brazil. Twelve castrated pigs with an initial body weight of 55 kg were individually allotted in metabolic crates during two 12-day periods, each with 7 days of adaptation and 5 days of total fecal collection. The beginning and the end of the collecting periods were determined according to the marker-to-marker approach, using ferric oxide as an indigestible marker. Pigs were submitted to four semi-purified diets, one being a P-free diet with 30% of GEL as the protein source and three were diets with 10%, 20%, and 30% inclusion of SDPP respectively. Data were subjected to analysis of variance and the model included the effects of period, animal and treatments; the results of the three diets with increased levels of SDPP were subjected to linear regression analysis. The intercept of the relation of between ingested P and absorbed P represented the EBLP, while the slope indicated the STTD of P in SDPP. The EBLP means obtained by P-free diet and regression method were compared with the Student t test. The EBLP were 128.95 mg/kg dry matter intake (DMI) and 153.63 mg/kg DMI (standard error = 77.0; p<0.06) using the P-free diet with GEL as the protein source and the regression method, obtained with diets containing increased levels of SDPP, respectively. The apparent digestibility of P was 87.9%, 94.2%, and 92.9% for the treatments containing 10%, 20%, and 30% inclusion of SDPP, respectively. The estimated STTD of P obtained with the linear regression was 97.4%. When the EBLP estimated by the P-free diet was used to corrected the apparent digestibility of P in diets containing SDPP, the STTD of P in SDPP was 96.9%, 98.8%, and 95.9% for 10%, 20%, and 30% SDPP, respectively. Therefore, it can be concluded that SDPP can replace GEL to estimate the endogenous losses of P. In addition, the STTD of P in SDPP estimated with the P-free diet was 97.2% and it was 97.4% by the regression method, utilizing SDPP.

• BMB Reports
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• v.34 no.3
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• pp.219-224
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• 2001

To identify the antioxidative peptides in the gelatin hydrolysate of bovine skin, the gelatin was hydrolyzed with serial digestions in the order of Alcalase, pronase E, and collagenase using a three-step recycling membrane reactor. The second enzymatic hydrolysate (hydrolyzed with pronase E) was composed of peptides ranging from 1.5 to 4.5 kDa, and showed the highest antioxidative activity, as determined by the thiobarbituric acid method. Three different peptides were purified from the second hydrolysate using consecutive chromatographic methods. This included gel filtration on a Sephadex G-25 column, ion-exchange chromatography on a SP-Sephadex C-25 column, and high-performance liquid chromatography on an octadecylsilane chloride column. The isolated peptides were composed of 9 or 10 amino acid residues. They are: Gly-Glu-Hyp-Gly-Pro-Hyp-Gly-Ala-Hyp (PI), Gly-ProHyp-Gly-Pro-Hyp-Gly-Pro-Hyp-Gly (PII), and Gly-ProHyp-Gly-Pro-Hyp-Gly-Pro-Hyp (PIII), as characterized by Edman degradation and fast-atom bombardment mass spectrometry. The antioxidative activities of the purified peptides were measured using the thiobarbituric acid method, and the cell viability with a methylthiazol tetrazolium assay The results showed that PII had potent antioxidative activity on peroxidation of linoleic acid. Moreover, the cell viability of cultured liver cells was significantly enhanced by the addition of the peptide. These results suggest that the purified peptide, PII, from the gelatin hydrolysate of bovine skin is a natural antioxidant, which has potent antioxidative activity.

• Journal of Microbiology
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• v.41 no.4
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• pp.289-294
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• 2003

Gelatinase is a proteolytic enzyme that hydrolyzes gelatin. Gelatinolytic activity was detected from culture broths of Streptomyces griseus IFO13350 and HH1 by paper disc assays on 0.5% agar plates containing 1% gelatin. The concentrated extracellular protein from the S. griseus was analyzed by SDS polyacrylamide gel, and two proteins, with molecular weights of 30 and 28 kDa, respectively, were identified to have gelatinase activity by gelatin zymography. The protein with a molecular weight of 28 kDa was confirmed to be S. griseus trypsin (SGT). The effects of metal ions and metal chelators on the protease activity of the SGT were studied. Of the metal ions tested, only manganese was found to enhance the protease activity, 2.6 times, however, $Co^{2+},\;Cu^{2+},\;and\;Zn^{2+}$, and metal chelators, such as EDTA and EGTA, inhibited the SGT activity. When the protease activity of the SGT was measured at various pHs, in the presence of 5 mM $MnCl_2$, its highest activity was at pH 11.0, whereas only 60% of the maximum activity was observed between pHs 4.0 and pH 6.0, and almost 80% activity between pHs 7.0 to pH 10.0. The protease activity was measured at various temperatures in the presence of 5 mM $MnCl_2$. The SGT was found to be stable up to $60^{\circ}C$ for 30 min, while only 16% of the enzyme activity remained at $60^{\circ}C$, and at $80^{\circ}C$ almost all the activity was lost. The optimal temperature for the protease activity was $50^{\circ}C$.

• Journal of Conservation Science
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• v.34 no.3
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• pp.157-166
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• 2018

In this study, gelatin binding ability was increased by adding cross linking agent to improve adhesive characteristic of animal glue. Animal glue added genipin measured gel strength and viscosity, the structural analysis, the color retention degree, elution degree, and rupture strength. And the water resistance and ultraviolet light resistance with the addition of genipin were compared. As a result of the study, the gel strength and viscosity increased with the amount of genipin. As a result of the structural analysis, in gelatin, the absorption peak of the triple structure of collagen structurally stabilized was observed. As a result of the color retention degree, the film was observed because of the lowered brightness. The amount of elution glue was increased with addition of genipin at $50^{\circ}C$ distilled water condition and rupture strength has increased with the amount of genipin. In the water resistance and light fastness, there was no appearance before and after deterioration due to the addition of genipin. Based on the results of this study, it confirmed the adhesive characteristics of animal glue added genipin and examined the experimental method applicable for animal glue. After the addition of genipin, flexibility, re-solving, adhesive force, and curing speed, which are unique characteristics of glue, can be improved without disappearing, so it is expected that it will be applicable to production of animal glue and conservation of cultural heritage when homogeneous glue is secured.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.29 no.2
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• pp.246-255
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• 1996

To develop a natural antioxidative peptide, the gelatin was extracted from fish (Yellowfin sole) skin by hot $water(50^{\circ}C)$ extraction method and hydrolyzed with Alcalase, pronase and collagenase through a continuous 3-step membrane reactor. Each step enzymatic hydrolysates were determined the antioxidative activity and their synergistic effects, compared with $\alpha-tocopherol$ and butylated hydroxytoluene (BHT). Also, we tried to investigate the antioxidative disposition of peptide which was successfully separated by gel filtration, ion-exchange chromatography, and HPIC in cultured rat hepatocytes intoxicated with tert-butyl hydroperoxide (TBHP). Second step enzymatic hydrolysate (SSEH) among all hydrolysates and $\alpha-tocoperol$ was showed the strongest antioxidative activity. The optimum concentration of antioxidative activity for SSEH was $1\%(w/w)$ in linoleic acid. The synergistic effects were increased in using the hydrolysate with tocopherol and BHT. In the presence of the peptide isolated from SSEH, supplemented hepatocytes exposed to TBHP showed that delayed cell killing and decreased significantly the lipid peroxidation, compared with hepatocytes not cultured with isolated peptide.

• Microbiology and Biotechnology Letters
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• v.9 no.3
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• pp.159-164
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• 1981

Reinforced Calcium-alginate gel entrappment method for enzyme immobilization is described with an example of penicillin amidase from Bacillus megaterium KFCC 10029, a partially constitutive mutant of B. megaterium ATCC 14945. Penicillin amidase recovered from the fermentation broth by adsorption on celite is mixed with alginate and gelatin solution, and cast into a pellet or noodle form by coagulation in calcium salt solution followed by crosslinking with glutaraldehyde. Optimum pH and temperature of the immobilized enzyme preparation were 8.0 and 6$0^{\circ}C$, respectively. Kinetic constants such as Km value and the inhibition constant of 6-APA and phenylacetic acid were 2.6 mM, 7.4 mM and 21.2 mM, respectively. The enzyme leakage from the adsorbent during operation was successfully prevented owing to the increase of physical strength of gel coat. The half lives in a column reactor were 6 and 30 days at the respective temperature of 4$0^{\circ}C$ and 3$0^{\circ}C$, which were the 6-8 fold increased values as compared with those of without entrappment. The results highly recommended the use of reinforced Calcium-alginate gel entrappment method for the enhancement of physical strength and the operational stability of alginate gel entrapped enzyme.

• Journal of Applied Biological Chemistry
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• v.48 no.2
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• pp.75-78
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• 2005

Halotolerant protease produced by Halomonas marisflava KCCM 10457 was partially purified through ammonium sulfate precipitation and Sephacryl S-200HR gel permeation chromatography. Optimal pH and temperature of protease were 11.0 and $45^{\circ}C$, respectively. Enzyme activity was inhibited by $Cu^{2+}$, $Hg^{2+}$, $Fe^{2+}$, and $Fe^{3+}$, and selectively inhibited by p-chloromercuribenzoic acid (PCMB), suggesting this enzyme is cysteine protease. The enzyme is halotolerant, because it retained 77% of original activity in presence of 3.33 M NaCl. The protease showed broad substrate specificity to various natural proteins; BSA, casein, egg albumin, gelatin, and hemoglobin.

• Journal of Food Hygiene and Safety
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• v.10 no.4
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• pp.213-217
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• 1995

A screening method has been developed for detecting sulfamethazine(SMZ) contamination of meat or feeds by using horseradish peroxidase (HRP) labeled protein A (Prot AHRP)and an indirect competitve enzyme-linked immunosorbent assay(ELISA). The assay is based on competitve binding of guinea pig anti-SMZ with SMZ in smaple and SMZ-gelatin conjugate(SMZ.GEL). Percent binding (B.Bo$\times$100) was calculated from the absorbance in the absence (B0) and presence (B) of SMZ. By the sandard curve prepared by plotting log(SMZ) vs percent binding of each known reference solution, the detection limit was 1.0ppb or less. Cross reacton with sulfadimethoxine, sulfaguaniding, sulfamerazine, sulfamthoxpyridazine, sulfanilamide, sulfisomidine and sufisoxazole were not observed. But sulfamerazine crossreacted in the test. The EC-50 value (concentration causing 50% inhibition of color development compared with blank) of sulfamerazine was 2.0 ppm. Further quality control will make the ELISA system ideal for the detection of SMZ in meat or feeds.

• Journal of the Korean Society of Food Culture
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• v.20 no.3
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• pp.384-390
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• 2005

This study was designed to establish Korean food culture by considering traditional fruit preserve. A historical approach of traditional fruit preserve were reviewed by the cooking book published from 1670 to 1988 in Korea. The results obtained from this study are as follows. Cooking method of fruit preserve was firstly published to Omshikdimibang in 1670 and published moderate cooking book in 1988 but fruit preserve has not been cooked yet. Cooking method of fruit juice and starch had been started in Samkuk dynasty and was completed in Lee Dynasty. Compared with the western fruit jelly, mung bean starch was used as a gel forming material in Korean traditional fruit preserve while gelatin was used as a gelling agent in western fruit jelly. Western fruit jelly was succeed in innovation, but traditional fruit jelly was failed to innovation.

• Journal of Pharmacopuncture
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• v.16 no.2
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• pp.46-54
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• 2013

Objective: This study was undertaken to isolate a fibrin(ogen)olytic enzyme from the snake venom of Gloydius blomhoffii siniticus and to investigate the enzymatic characteristics and hemorrhagic activity of the isolated enzyme as a potential pharmacopuncture agent. Methods: The fibrinolytic enzyme was isolated by using chromatography, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and fibrin plate assay. The characteristics of the enzyme were determined by using fibrin plate assay, protein hydrolysis analysis, and hemorrhage assay. Its amino acid composition was determined. Results: The fibrin(ogen)olytic enzyme with the molecular weight of 27 kDa (FE-27kDa) isolated from G. b. siniticus venom consisted of two heterogenous disulfide bond-linked polypeptides with the molecular weights of 15 kDa and 18 kDa. When more than $20{\mu}g$ of FE-27kDa was applied on the fibrin plate, fibrinolysis zone was formed as indicating its fibrinolytic activity. The fibrinolytic activity was inhibited completely by phenylmethanesulfonylfluoride (PMSF) and ethylenediaminetetraacetic acid (EDTA) and partially by thiothreitol and cysteine. Metal ions such as $Hg^{2+}$ and $Fe^{2+}$ inhibited the fibrinolytic activity completely, but $Mn^{2+}$ did not. FE-27kDa preferentially hydrolyzed ${\alpha}$-chain of fibrinogen and slowly hydrolyzed ${\beta}$-chain, but did not hydrolyze ${\gamma}$-chain. High-molecular-weight polypeptides of gelatin were hydrolyzed partially into polypeptides with molecular weights of more than 45 kDa. A dosage of more than $10{\mu}g$ of FE-27kDa per mouse was required to induce hemorrhage beneath the skin. Conclusion: FE-27kDa was a serine proteinase consisting of two heterogeneous polypeptides, hydrolyzed fibrin, fibrinogen, and gelatin, and caused hemorrhage beneath the skin of mouse. This study suggests that the potential of FE-27kDa as pharmacopuncture agent should be limited due to low fibrinolytic activity and a possible side effect of hemorrhage.