• Korean Journal of Agricultural Science
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• v.47 no.3
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• pp.459-470
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• 2020

This study investigated the effects of the proteolytic hydrolysates of α-lactalbumin (LA), β-lactoglobulin (LG) and bovine serum albumin (BSA) by alcalase on inflammatory cytokines. The proteolytic hydrolysates were separated into two fraction of peptides, ≤ 10,000 Da and > 10,000 Da, respectively, because various low molecular weight peptides were generated during the hydrolysis reaction time. Among the hydrolysate peptides, BSA (all types), β-LG (> 10,000 Da), and α-LA (> 10,000 Da) showed an inhibitory activity against thymic stromal lymphopoietin (TSLP) mRNA expression in lipopolysaccharide-induced RAW264.7 murine macrophages. α-LA (> 10,000 Da), β-LG (hydrolysates), and BSA (> 10,000 Da) showed an inhibitory activity against tumor necrosis factor (TNF)-α expression. α-LA (all types), β-LG (hydrolysates, > 10,000 Da), and BSA (> 10,000 Da) showed an inhibitory activity against interleukin-6 (IL-6) expression. α-LA (> 10,000 Da), β-LG (> 10,000 Da), and BSA (all types) showed an inhibitory activity against inducible nitric oxide synthase (iNOS) expression. α-LA (> 10,000 Da), β-LG (> 10,000 Da), and BSA (all types) showed an inhibitory activity against cyclooxygenase (COX)-2 expression. The lowest level of TNF-α production was measured with α-LA (> 10,000 Da) and β-LG (> 10,000 Da) for all types, and a similar low level was measured for all types of BSA. The highest level of IL- 6 production was measured with α-LA (≤ 10,000 Da) among α-LA, β-LG, and IL-6. The low level of NO production was similar with α-LA, β-LG, and BSA but not with α-LA (≤ 10,000 Da). These potential peptides from whey protein hydrolysates could be used for food, medicinal, and industrial applications.

• Korean Journal of Food Science and Technology
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• v.33 no.5
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• pp.603-610
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• 2001

The quality characteristics of soy yogurt prepared with different proteolytic enzymes and starter culture were evaluated. In order to facilitate the growth of lactic acid bacteria and subsequent production of lactic acid, soy protein isolate(SPI) was hydrolyzed using three kinds of proteases; one extracted from Aspergillus oryzae, bromelain and ${\alpha}-chymotrypsin$. The cultural systems employed thereafter for lactic fermentations were: 1) Bifidobacterium bifidum, 2) B. bifidum and Lactobacillus acidophilus, 3) B. bifidum and Lactobacillus bulgaricus. In soy yogurt, pH was more decreased by mixed culture method than single culture method with the accumulation of lactic acid. Viable cells of lactic acid bacteria in soy yogurts were measured $10^8$ CFU/g by the single culture method while $10^9$ CFU/g by the mixed culture method except ${\alpha}-chymotrypsin$ treatment. The amount of free amino acids in soy yogurts were substaintially increased by enzyme treatment. Viscosity was decreased by enzyme treatment, resulting in higher viscosity by ${\alpha}-chymotrypsin$ treatment. Water holding capacity was found to be higher in the single culture method in case of enzyme treatment. Among the various volatile flavor components isolated and identified after enzyme hydrolysis, acetaldehyde, ethanol, diacetyl, butyl alcohol contents tended to increase by lactic fermentation.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.23 no.2
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• pp.109-124
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• 1990

A rapid processing method for fish sauce of high quality stability and favorable flavor was investigated using mackerel waste as starting material. The chopped waste was homogenized with water and hydrolyzed by commercial proteolytic enzymes such as Complex enzyme-2000($2.18\cdot10^4$ U/g solid, Pacific Chem. Co.) and Alcalase ($1.94\cdot10^4$ U/g solid, Novo) in a cylindrical vessel with 4 baffles and 6-bladed turbine impeller. Optimal pH and temperature for the hydrolysis with Complex enzyme-2000 were 8.0 and $50^{\circ}C$, and those with Alcalase were 9.0 and $55^{\circ}C$. In both cases, the reasonabe amount of added water and enzyme concentration based on the waste weight were $40\%,\;3\%$ and hydrolyzing time was 100 min. Thermal treatment of the hydrolysate with $6\%$ of invert sugar for 2 hours at $90^{\circ}C$ was adequated to inactivation of the enzymes and pasteurization of the hydrolysate. Flavor, taste and color of the hydrolysate were improved during the thermal treatment in which the browning reaction products might participate and result in antioxidative and bactericidal effects. Combined use of $0.005\%$ of Caryophylli flos with $6\%$ of invert sugar was also effective for the improvement of taste. Yield of the fish sauce based on the total nitrogen of the raw waste was $93.7\~94.9\%$, and $87.6\~87.9\%$ of the total nitrogen in the fish sauce was in the from of amino nitrogen. The pH, salinity and histamine content of the fish sauce prepared with $15\%$ of table salt were $6.1\~6.2$, $14.0\~14.5\%$ and less than $10mg\%$, respectively. The fish sauce was stable on bacterial growth during the storage of 60 days at $26\pm3^{\circ}C$ and the quality was also maintained.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.23 no.2
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• pp.125-136
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• 1990

To develope a rapid processing method for fish sauce, processing conditions of fish sauce from sardine waste was investigated. The chopped waste was homogenized and hydrolyzed by commercial proteolytic enzymes such as Complex enzyme-2000($2.18\cdot10^4$ U/g solid) and Alcalase($1.94\cdot10^4$ U/g solid) in a cylindrical vessel with 4 baffles and 6-bladed turbine impeller. Optimal temperature for the case of hydrolysis with Complex enzyme-2000 was 50 and that with Alcalase was $55^{\circ}C$. In both cases, the reasonable pH, amount of water for homo-genization, enzyme concentration and hydrolyzing time were 8.0, $40\%$ (W/W), $3\%$ and 100 min, respectively. Heating of the filtrated hydrolysate for 2 hours at $90^{\circ}C$ with $6\%$ of invert sugar was suitable for pasteurization of the hydrolysate and inactivation of enzymes. Flavor, taste and color of the hydrolysate was improved during the thermal treatment in which the browning reaction products might participate and result in antioxidative and bactericidal effects. Combined use of $0.005\%$ of Caryophylli flos with invert sugar was also effective for the improvement of taste. Yield of the fish sauce based on the total nitrogen in the raw sardine waste was $91.2\~92.3\%$ and $87.2\~87.8\%$ of the total nitrogen in the fish sauce was in the form of amino nitrogen. The pH, salinity and histamine content of the fish sauce prepared with $15\%$ of table salt were $6.1\~6.2$, $14.2\~14.4\%$ and less than $10mg\%$, respectively. The fish sauce was stable during the storage of 60 days at $26\pm3^{\circ}C$ on bacterial growth and its quality was also maintained.

• Food Science and Preservation
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• v.30 no.5
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• pp.885-895
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• 2023

This study aims to investigate the production and characteristics of protein hydrolysates derived from tuna byproducts (TP) using various proteolytic enzymes and to compare the antioxidant activity of the resulting hydrolysates. The TP were subjected to enzymatic hydrolysis using five different proteases: alcalase, bromelain, flavourzyme, neutrase, and papain, and the antioxidant activities of the hydrolysates were evaluated. Subsequent analysis of the available amino group contents and sodium dodecyl sulfate-polyacrylamide gel electrophoresis patterns indicated a high degree of hydrolysis in TP after treatment with all the enzymes, except for papain. Based on the RC₅₀ values obtained from four different antioxidant analyses, all the hydrolysates exhibited similar antioxidant activity, except for the flavourzyme hydrolysate, which showed significantly higher scavenging activity against ABTS radicals and hydrogen peroxide than the other hydrolysates. These findings suggest that protein hydrolysates derived from TP hold promise as potential sources of natural antioxidants.

• Journal of Microbiology
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• v.41 no.3
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• pp.207-211
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• 2003

Extracellular protease, from Aeromonas hydrophila Ni 39, was purified 16.7-fold to electrophoretic homogeneity with an overall yield of 19.9％, through a purification procedure of acetone precipitation, and Q Sepharose and Sephacryl S-200 chromatographies. The isoelectric point of the enzyme was 6.0 and the molecular mass, as determined by Sephacryl S-200 HR chromatography, was found to be about 102 kDa. SDS/PAGE revealed that the enzyme consisted of two subunits, with molecular masses of 65.9 kDa. Under standard assay conditions, the apparent $K_{m}$ value of the enzyme toward casein was 0.32 mg/ml. About 90％ of the proteolytic activity remained after heating at 60$^{\circ}C$ for 30 min. The highest rate of azocasein hydrolysis for the enzyme was reached at 60$^{\circ}C$, and the optimum pH of the enzyme was 9.0. The enzyme was inhibited by the serine protease inhibitor, phenylmethylsulfonyl fluoride (PMSF), by about 87.9％, but not by E64, EDTA, pepstatin or 1,10-phenanthroline. The enzyme activity was inhibited slightly by Ca$\^$2＋/, Mg$\^$2＋/ and Zn/supb 2＋/ ions.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.40 no.4
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• pp.341-347
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• 2014

This study was aimed to investigate skin whitening effects of Nomura's jellyfish (Nemopilema nomurai) hydrolyzed extracts (NHE). The extracts were prepared through the hydrolysis of N. nomurai using commercial proteolytic enzymes such as Protamex, Alcalase, Flavourzyme and Neutrase with optimum pHs (pH 5-8) at $55^{\circ}C$. Their whitening activities were examined from the inhibition of melanin synthesis using B16-F1 cell lines. Among the examined samples, Neutrase-treated extract (N-NHE) showed the most significant inhibition effect on melanin synthesis by 89.9% at a concentration of $100{\mu}g/mL$. This sample decreased the expression of tyrosinase and TRP-1 (tyrosinase related protein-1) proteins as well. These results suggested the potential application of NHE as whitening ingredients in cosmetic preparation.

• Korean Journal of Food Science and Technology
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• v.20 no.3
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• pp.433-440
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• 1988

Dried anchovy (Engraulis japonica) was ground and treated with 0.3N NaOH solution and then hydrolyzed with proteolytic enzymes. Extracts obtained by centrifugation of alkali-enzyme treated anchovy slurry was compared with water extract for the yields of soluble solid, protein and ashes and organoleptic characteristics. The data for the yields of the soluble solids, protein and ash showed that a 2-3 folds increase in those yields was resulted by combined alkali-enzyme treatments when it was compared to water only extract. The organoleptic evaluation on the alkali-enzyme treated anchovy extracts also showed a 2-3 folds in flavor strength of all descriptions in odor and taste and a significant improvement in total odor or taste acceptability.

• Korean Journal of Agricultural Science
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• v.40 no.3
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• pp.221-226
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• 2013

Degrees of hydrolysis by alkaline protease produced from Serratia marcescens S3-R1 is 3.95-6.30% of whey proteins during 5, 15, 30, 60, 90, 120,180, 240 min incubation at $40^{\circ}C$. Proteolytic pattern of the whey proteins showed that various low molecular weight peptides were generated during the incubation periods. The biological function of in Raw 264.7 cells treated with whey protein hydrolytic peptides, anti-inflammatory effect showed exhibit in the expression of pro-inflammatory cytokines such as TNF-${\alpha}$, IL-6, COX-2 and iNOS by PCR analysis. COX-2 and iNOS gene expression inhibited in Raw 264.7 cells on whey protein hydrolysates below 3,000 dalton. The protease from Serratia marcescens S3-R1 showed a potential in production of low molecular weight whey protein hydrolysates which could be used for industrial application.

• Korean Journal of Food Science and Technology
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• v.22 no.5
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• pp.590-595
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• 1990

The destructive effect of peroxidized lipid on the amino acid in protein isolate and the proteolytic activity of protease were studied in the model system of rice bran. The content of amino acids in the protein isolate decreased significantly when they reacted with peroxidized lipid (pov. 1200 meq/kg). Most of amino acids were lost by more than 90% in salt soluble protein isolates when analyzed by the method of enzyme hydrolysis. Formaldehyde reduced the activity most severely among peroxidized products. Formic acid, peroxidized lipid and hydroperoxide were also found to reduce the protease activity. The damaging effect of the secondary products on the protease activity was more serious than that of the primary products of lipid peroxidation. The destruction of amino acids in the total protein and Inhibition of protease activity by the peroxidized lipid were apparent.