• Journal of the Korean Chemical Society
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• v.21 no.6
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• pp.445-448
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• 1977

This study was carried out to understand the activity of ${\alpha}$-chymotrypsin, a proteolytic enzyme, to a oligopeptide in the presence of various coloring food additives. 1. The melting point of synthetic oligopeptide, Asp-Arg-Val-Tyr-Ile-His-Pro-D-Ala, ((8-D-Ala) angiotensin Ⅱ) was 210∼$212^{\circ}C$. Chemical formula and molecular weight were $C_{44}H_{67}N_{13}O_{12}{\cdot}2CH_3COOH{\cdot}H_2O$ and 970.08, respectively. 2.The amino acid rations by acid hydrolysis were Asp : 1.01, Arg : 1.03, Val : 1.00, Tyr :40.94, Ile : 1.00, His : 1.05, Pro : 1.04, D-Ala : 1.03. 3. ${\alpha}$-Chymotrypsin cleaved the oligopeptide bond between tyrosine and isoleucine (Tyr-Ile). 4. The addition of food coloring additives as determined by paper chromatogram, did not influence the inhibitory activity of ${\alpha}$-chymotrypsin on oligopeptide, (8-D-Ala) angiotensin II.

• Korean Journal of Environmental Agriculture
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• v.34 no.3
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• pp.244-251
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• 2015

BACKGROUND: Recent studies have described the importance of bacteria that can degrade polycyclic aromatic hydrocarbons (PAHs). Here we screened bacterial isolates from commercial gasoline for PAH degraders and characterized their ability to degrade PAHs, lipids and proteins as well as their enantioselective epoxide hydrolase activity, salt tolerance, and seawater survival. METHODS AND RESULTS: One hundred two bacteria isolates from commercial gasoline were screened for PAH degraders by adding selected PAHs on to the surface of agar plates by the sublimation method. A clear zone was found only around the colonies of PAH degraders, which accounted for 13 isolates. These were identified as belonging to Bacillus sp., Brevibacterium sp., Micrococcus sp., Corynebacterium sp., Arthrobacter sp., and Gordonia sp. based on 16S rRNA sequences. Six isolates belonging to Corynebacterium sp., 3 of Micrococcus sp., Arthrobacter sp. S49, and Gordonia sp. H37 were lipid degraders. Arthrobacter sp. S49 was the only isolate showing high proteolytic activity. Among the PAH-degrading bacteria, Arthrobacter sp. S49, Brevibacterium sp. S47, Corynebacterium sp. SK20, and Gordonia sp. H37 showed enantioselective epoxide hydrolase activity with biocatalytic resolution of racemic styrene oxide. Among these, highest enantioselective hydrolysis activity was seen in Gordonia sp. H37. An intrinsic resistance to kanamycin was observed in most of the isolates and Corynebacterium sp. SK20 showed resistance to additional antibiotics such as tetracycline, ampicillin, and penicillin. CONCLUSION: Of the 13 PAH-degraders isolated from commercial gasoline, Arthrobacter sp. S49 showed the highest lipid and protein degrading activity along with high active epoxide hydrolase activity, which was the highest in Gordonia sp. H37. Our results suggest that bacteria from commercial gasoline may have the potential to degrade PAHs, lipids, and proteins, and may possess enantioselective epoxide hydrolase activity, high salt tolerance, and growth potential in seawater.

• Journal of Life Science
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• v.21 no.2
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• pp.309-315
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• 2011

Rice syrup meal (RSM) was enzymatically hydrolyzed using eight commercial proteases (Protamex, Neutrase, Flavourzyme, Alcalase, Protease M, Protease N, Protease A, Molsin F) for 4 hr at optimum pH and temperature. Proteolytic hydrolysates were examined in supernatant and precipitate using Lowry protein assay, semimicro Kjeldahl method and gravimetric method using weight difference before and after enzymatic hydrolysis. Although RSM contains a high amount of protein (71.2%), only a very small amount of protein was hydrolyzed. Two proteases (Protease M and Protease N) were found to be the most effective in the hydrolysis of RSM protein. In Lowry method, 57.5 and 59.0 mg protein/g RSM were hydrolyzed after Protease M and Protease N treatments, respectively. In gravimetric method, 80.0 and 85.4 mg protein/g RSM were hydrolyzed after Protease M and Protease N treatments. In Kjeldahl method, 67.43 and 70.43 mg protein/g RSM were hydrolyzed after Protamex and Protease N treatments, respectively. For synergistic effect, two or three effective commercial proteases (Protease M, Protease N and Protease A) were applied to RSM at one time. The highest hydrolysis of RSM protein was observed in both Lowry protein assay (80.3 mg protein/g RSM) and gravimetric methods (153.2 mg protein/g RSM) when three commercial proteases were applied at one time, suggesting the synergistic effect of those proteases.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.35 no.1
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• pp.97-104
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• 2002

Protein hydrolysate was prepared as a natural flavor stock from the sharp toothed eel (Muraenesox cinereus) mince using com-mercially available proteolytic enzymes, Alcalase, Neutrase, Protamex, and Flavourzyme. A 6 hr hydrolysis of mince, to which water of the equal weight of the mince was added, with $2\%$ (w/w, protein weight) Elavourzyme at $50^{\circ}$ yielded a hydrolysate of the highest acceptability. Removing the access lipid in liquified hydrolysate (not dehydrated) after enzyme hydrolysis, five times repetitive extraction using n-hexane (liquified hydrolysate : n-hexane=4 : 1, v/v) was effective, resulting in less than $1\%$ lipid content of the dehydrated-hydrolysate. The amino acid composition of the hydrolysate (prepared with Flavourzyme) was similar to that of the starting material. Hydrolysis led to an increase in concentration of not only total free amino acid but also free amino acid such as serine, glutamic acid, alanine, and methionine responsible for umami taste, especially up to about 40 times for methionine. Major free amino acids in amount were leucine, phenylalanine, valine, alanine, and isoleucine and they comprised about half of the total free amino acids, Moisture adsorption, fat adsorption, emulsifying capacity, and foaming capacity of the hydrolysate were 870.1 $\pm$ $7.9\%$, 352.0$\pm$ $5.3\%$, 50.3 $\pm$ $1.2\%$, and $87.5\pm$ $2.5\%$, respectively, and solubility was 83$\~$$84\%$ at acid pH range of 2$\~$4.

• Journal of the Korean Society of Food Science and Nutrition
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• v.13 no.1
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• pp.97-106
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• 1984

The Antarctic krill, Euphausia superba, is drawing attention over the world as the largest source of unutilized proteins in the ocean. For the use of krill as a human food, processing conditions of krill sauce by autolysis and/or commercial proteolytic enzyme digestion were examined. The krill was chopped and mixed with equal weight of water, and hydrolyzed by autolysis and/or commercial proteolytic enzyme digestion. The optimal conditions for hydrolysis of krill were $52.5^{\circ}C$, pH 7.0-7.5, 3 hours by autolysis, $52.5^{\circ}C$, pH 6.3, 3hours by bromelain (0.5 %) digestion, and $52.5^{\circ}C$, pH 7.0-7.5, 3 hours by commercial complex enzyme (5 %) digestion, respectively The maximum hydrolyzing rate of protein were 83.2 % by autolysis, 89.7 % by bromelain digestion, 92.7 % by commercial complex enzyme digestion. After krill meat hydrolyzed by autolysis at optimum condition, inactivated at $100^{\circ}C$ for 20 minutes and filtered with Buchner funnel. Two kinds of products were prepared with krill hydrolysate and preservatives: one contained 10 % of sodium chloride and 0.06 % of benzoic acid and the other 10 % of sodium chloride and 3 % of ethyl alcohol. These products were filled in the sterilized glass bottle and sealed. The pH, volatile basic nitrogen, amino nitrogen, color value (L, a and b values) and viable counts of bacteria were determined during storage at $37^{\circ}C$. The results showed that the products could be preserved in good condition during one month at $37^{\circ}C$. As a method to reduce the sodium level in krill sauce, it is convinced that sodium chloride could be replaced half in partially by potassium chloride. In the products prepared from krill by autolysis, bromelain or commercial complex enzyme digestion, hypoxanthine and 5'-IMP were abundant among the nucleotides and their related compounds as 15.3-20.4 ${\mu}mole/g$, dry solid, 2.2-2.5 ${\mu}mole/g$, dry solid, respectively. The abundant free amino acids were lysine, leucine, proline, alanine and valine. The contents of these amino acids were 67.4 %, 69.4 %, 69.8 % of the total free amino acids of each products. And TMAO, betaine and total creatinine were low in contents. The flavor of krill sauce prepared from krill by autolysis or enzyme digestion was not inferior to that of traditional Kerean soy sauce by sensory evaluation.

• Korean Journal of Food Science and Technology
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• v.29 no.1
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• pp.77-81
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• 1997

In order to develop a new type of natural seasoning material combining fish meat with seaweed, a processing method of the mixture of enzyme treated mackerel meat and Laminaria powder was studied. Mackerel meat previously boiled and deboned was treated with proteolytic enzyme to enhance taste of meat by proper hydrolysis. The enzyme-treated meat was dried at $100{\pm}2^{\circ}C$ for 4 hrs, and finally mixed with kelp power, moistened in advance, plus binding agents (0.02% calcium carbonate) to aid the formation of pellets by extrusion. Boiled mackerel meat of enzyme treated (0.03% Protease-A) at $50^{\circ}C$ for 90 min was adequate to result an increase in 6 times of total free amino acid content and about 10% increase of taste-enhancing amino acids such as glutamic acid, glycine, arginine, lysine.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.23 no.5
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• pp.361-372
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• 1990

Processing conditions of whole sardine into modified fish sauce were investigated. Thawed and chopped sardine was homogenized and hydrolyzed using commercial proteolytic enzymes such as complex enzyme-2000($2.18{\cdot}10^4U/g solid$) and alcalase($1.94{\cdot}10^4\;U/g solid$) in a cylindrical vessel with 4 baffles and 6-bladed impeller. Optimal pH, enzyme concentration and temperature for the hydrolysis with complex enzyme-2000 were 7.0, $7\%$ (W/W) and $52^{\circ}C$, and-those with alcalase were 8.0, $6\%$ (W/W) and $60^{\circ}C$. In both cases, the reasonable amount of water for homogenization, agitation speed and hydrolyzing time were $100\%$ (W/W), 100 rpm and 210 minutes. Thermal treatment of the filtered hydrolysate at $90^{\circ}C$ for 2 hours with $6\%$ of invert sugar was adequated to inactivation of the enzymes and pasteurization of the hydrolysate. Flavor, taste and color of the hydrolysate were improved during the heating process in which the browning products might participate. The content of free amino nitrogen in the fish sauce seasoned with $15\%$ of table salt was ca. $1,640 mg\%$. Yield of the fish sauce based on the contents of proteinous and free amino nitrogen in the raw whole sardine was ca. $86\%$, and ca. $96\%$ of these compounds of the fish sauce was in the form of free amino nitrogen. The pH, salinity and histamine content of the fish sauce were $6.1\~6.3,\;14.2\~14.3\%$ and less than $10\;mg\%$.

• Journal of Nutrition and Health
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• v.45 no.5
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• pp.429-436
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• 2012

This study was performed to examine the characteristics of protein of red crab (Chionoecetes japonicus) shell powder hydrolyzed by commercial proteases. Red crab shell was digested by commercial proteases, such as Protamex (P), Neutrase (N), Flavourzyme (F), Alcalase (A), Protease M (PM) and Protease A (PA). Protein yield analyzed by Biuret assay, absorbance at 280 nm and brix revealed that PA was the enzyme having the highest proteolytic activity. SDS PAGE showed that molecular weight of proteins produced by protease treatments was various and below 150 kDa. Combinational treatment of proteases (PA + P, PA + PM, PA + F, PA + A) was tried whether these increase protein hydrolysis from red crab shell powder compared to a PA single treatment. Soluble protein content was similar, but amino acid concentration by combinational treatments was higher than PA single treatment [PA + P 247.4 mg/g > PA + F (206.4 mg/g) > PA + A (133.4 mg/g) > PA + PM (59.1 mg/g) > PA (54.9 mg/g)]. Amino acid composition by combinational treatments was slightly different. Most abundant essential amino acids were phenylalanine, glycine, alanine, and leucine, whereas tyrosine and cystine were not detected.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.12 no.4
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• pp.235-240
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• 1979

A study on the amino acid composition of raw frozen krill, and krill solubles manufactured in forms of paste and powder has been carried out. The raw frozen krill was thawed, chopped, mixed and homogenized with same amount of water. The mixture was autolyzed or hydrolyzed by tile addition of $0.2\%$ pronase-p, a commercial proteolytic enzyme, to the weight of the raw frozen krill at $45^{\circ}C$ for 4 hours. After a thermal inactivation of enzymes at $95^{\circ}C$ for 15 minutes, the autolysate and the hydrolysate were centrifuged and filtered through gauzes, respectively, and then tile lipid layer in the supernatant was removed, The autolysate and the hydrolysate were finally concentrated under reduced atmospheric pressure in a rotary vacuum evaporator at $45^{\circ}C$ for 1 hour to produce the krill solubles in form of paste. The powdered krill solubles were prepared by the addition of $5\%$ starch to the autolysate and hydrolysate and by means of concentration in the rotary vacuum evaporator at $45^{\circ}C$ for 30 minutes and a forced air drying at $58^{\circ}C$ for 3 hours with a air velocity of 3m/sec. Among the amino acids in raw frozen krill, glutamic acid, lysine, and aspartic acid showed high values in quantity and then followed leucine, alanine, arginine, glycine and proline. The qnantity of histidine was very small and that of cystine was only in trace. The krill solubles in forms of paste and powder prepared by autolysis and hydrolysis with pronase-p revealed almost the same patterns in amino acid composition as in raw frozen krill. In case of free amino acids, a large quantity of it in raw frozen krill consisted of lysine, arginine, proline, alanine and leucine. The quantities of cystine, histidine and glutamic acid were, in contrast, very small. In the soluble krill paste prepared by autolysis, lysine, leucine, threonine and alanine existed in large quantities among the free amino acids and cystine, aspartic acid and histidine existed in small quantities. The contents of almost all of the free amino acids ill soluble krill paste perpared by hydrolysis with pronase-p were increased slightly as compared with those in soluble krill paste prepared by autolysis. In this product, the contents of cystine, histidine and serine were very low and lysine, leucine, arginine and proline were the dominant group in quantities among the free amino acids. The krill solubles in forms of paste and powder were not inferior to whole egg in the view point of its essential amino acid composition.

• Journal of the Korean Society of Food Science and Nutrition
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• v.40 no.1
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• pp.89-93
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• 2011

This study was conducted to investigate the proteolysis and extraction conditions of deer antler for application of food materials. ProteAX (A) was the most effective enzyme for proteolysis of deer antler and the proteolysis condition was 0.5% (w/w) for enzyme concentration and 5 hr for proteolysis time. The effect of mixing enzyme ProteAX (A)+KFEN 2 (C) treatment in $60^{\circ}C$, 5 hr was investigated; soluble solid and protein content were the highest with A 0.5% (w/w) and B 0.5% (w/w) concentration. Result for DAH (deer antler hydrolysate) and DA (deer antler) prepared with extraction in $95^{\circ}C$ atmospheric pressure (AP, 6~18 hr) and extraction under $120^{\circ}C$ pressure condition (UP, 15~60 min) after hydrolysis on preceding established condition descriptions indicated that difference in pH according to enzyme treatment and extraction conditions was not significant. Sugar content of DA was $1.5^{\circ}Brix$, DA-UP (under pressure) and DAH-AP (atmospheric pressure) were $2.2^{\circ}Brix$; the highest sugar content of $2.7^{\circ}Brix$ was observed in DAH-UP for 60 min extraction. Also total free sugar, crude protein and collagen content were the highest in DAH-UP for 60 min recording at 1.97%, 742.7 mg/100 g and 498.8 mg/100 g, respectively. From these results, deer antler hydrolysate prepared with extraction under pressure was the most effective for functional characteristics enhancement. Hereafter, various practical uses of materials with enhanced characteristics of antler is expected.