• Fisheries and Aquatic Sciences
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• v.1 no.2
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• pp.209-215
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• 1998

Trypsin-like enzyme was purified from shrimp hepatopancreas through Q-Sepharose ionic exchange, benzamidine Sepharose-6B affinity, and Superdex 75 gel chromatography. Purity of trypsin-like enzyme was increased 69-fold with $44\%$ yield. The enzyme consisted of a single polypeptide chain with a molecular weight (M.W.) of 32 kDa judged by sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE). The enzyme was completely inactivated by serine enzyme inhibitors such as soybean trypsin inhibitor (SBTI), tosyl-L­lysine chloromethyl ketone (TLCK), and leupeptin. However, the enzyme was not affected by tosyl-L-phenylalanine chloromethyl ketone (TPCK) which is a chymotrypsin specific inhibitor. The enzyme had no activity against benzoyl-tyrosine ethyl ester (BTEE) which is a chymotrypsin specific substrate. The enzyme showed high activity on the carboxyl terminal of Phe, Tyr. Glu, Arg, and Asp. However. no activity was detected against the carboxyl terminal of Pro, Trp, Cys, Gly, Val, and Ala.

• The Korean Journal of Malacology
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• v.30 no.1
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• pp.41-49
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• 2014

This study was conducted to investigate the effects of cadmium (Cd) exposure on biochemical factors in the hemolymph and hepatopancreas of the abalone, Haliotis discus hannai. The abalone were exposed to 0, 5, 10, 20 and 40 ${\mu}g/L$ Cd for 4 weeks. The phenoloxidase (PO) activity was decreased in hemolymph of abalone exposed to 40 Cd ${\mu}g/L$ for 4 weeks compared to the control (P < 0.05). The hemolymph enzymes, alkaline phosphatase (ALP), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities were markedly elevated in 40 Cd ${\mu}g/L$ after 4 weeks. The hemolymph calcium concentrations were significantly decreased in 20 and 40 Cd ${\mu}g/L$ for 4 weeks. Hepatopancreas superoxide dismutase (SOD) and catalase (CAT) activities were significantly increased by Cd. SOD was increased in both 20 and 40 Cd ${\mu}g/L$ and CAT, in 40 Cd ${\mu}g/L$ after 2 weeks (P < 0.05). These results suggested that the abalone SOD and CAT including PO may serve as a protective mechanism against oxidative stress by Cd. We conclude that a Cd concentration, 40 ${\mu}g/L$ in water may curtail hemolymph homeostasis and anti-oxidative reactions in abalone hepatopancreas. From these results, these biochemical factors may represent a convenient method of monitoring heavy metal pollution in coastal areas.

• Proceedings of the Korean Society of Fisheries Technology Conference
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• 1998.06a
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• pp.264.2-265
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• 1998

• Proceedings of the Korean Society of Fisheries Technology Conference
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• 1998.06a
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• pp.266-267
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• 1998

• Fisheries and Aquatic Sciences
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• v.15 no.4
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• pp.283-291
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• 2012

This study examines the optimal fractionation method and conditions for the isolation of endoprotease- and exopeptidase-active fractions from crude extracts of cuttlefish hepatopancreas (HP) using four fractionation methods: ammonium sulfate fractionation (ASF), polyethylene glycol fractionation (PGF), ion exchange chromatography (IEC), and gel filtration chromatography (GFC). Total endoprotease activity highest in the fraction II (concentrate of fractions 34-42; 842.60 U) of GFC, followed by fraction III (40-60% ammonium sulfate fraction; 670.25 U) of ASF, fraction I (concentrate of fractions 8-12; 436.89 U) of IEC, and fraction II (10-20% polyethylene glycol; 307.31 U) of PGF. Total exopeptidase activity of these fractions was highest in fraction II (2,704.70 U) of GFC, fraction III (2,110.50 U) of ASF, fraction III (1,605.60 U) of PGF, and fraction II (concentrate of fractions 38-44; 1,196.22 U) of IEC. These results showed that fraction II of GFC had the highest activity toward both exopeptidase and endoprotease, with exopeptidase activity being 3.21 times higher than of endoprotease. These results suggest cuttlefish HP could be used as a potential source for the extraction of exopeptidase, an enzyme capable of catalyzing the cleavage of N- and C-terminal amino acids in polypeptides, Like endoprotease, the most efficient method for separating exopeptide-active fractions was GFC.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.36 no.3
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• pp.283-289
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• 2003

The effect on kidney, gill, and hepatopancreas of juvenile olive flounder, Paralichthys olivaceus were investigated by histological method under limit concentration of effulent of PCBs for 60 days. The free surface of epithelial cell layer in the renal tubules of kidney showed a strong positive reaction in alcian blue during all exposure period. There were also observed swelling of hemocyte in glomerulus and macrophage. At 60 days after exposure, epithelium of the renal tubules was fused and some of organism $(20\%)$ showed damage of glomerulus and eosinophlic cell in epithelium of the renal tubules. The phenomena such as the activation and increase of the chloride cell, and swelling of hemocyte in the capillary of gill were observed during all exposure period. At 60 days after exposure, swelling of the gill filament and mucous cell appeared in gill and some of organism $(30\%)$ showed swelling of the gill lamellar. Swelling of hemocyte in the capillary of hepatopancreas was observed and the free surface of epithelial layer of the bile duct showed positive reaction in alcian blue during all exposure period. Degenerated zymogen in the pancreas and swelling of the hepatocyte were occurred at 60 days after exposure.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.47 no.2
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• pp.135-143
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• 2014

Exopeptidase active fractions from the hepatopancreas of the Argentina shortfin squid Illex argentinus, were obtained with acetone (AC 30-40%), ammonium sulfate (AS 60-70% saturation), anion exchange chromatography (AE-II, 0.2 M NaCl) and gel filtration chromatography (GF-I, 30-50 kDa) fractionation methods. A bitter peptide solution that has a bitterness equivalent to that of 2% glycylphenylalanine and prepared by tryptic hydrolysis of milk casein, was treated with the exopeptidase active fractions. The GF-I fraction was the best based on aminopeptidase activity (35.3 U/mg), percentage of recovery (30.7%) and a sensory evaluation (1.7). The amount of released amino acids increased as incubation time increased, and the bitterness of the enzyme reaction mixtures decreased. Incubation with the GF-I fraction for 24 h resulted in the hydrolysis of several peptides as revealed by the reverse-phase high performance liguid chromatography profile, with three peaks (3, 5 and 6) decreasing in area (%) and three peaks (1, 2 and 4) increasing in area (%). Therefore, the GF-I fraction appeared to be ideally suited to reduce bitterness in protein hydrolysates by catalyzing the hydrolysis of bitter peptides.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.53 no.1
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• pp.112-122
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• 2020

This study investigated some enzymatic properties and bitterness improvement of an aminopeptidase retentate fraction (ARF) from common squid Todarodes pacificus hepatopancreas extract (HPE), obtained by ultrafiltration with a 10 kDa molecular weight cut off membrane. Endoprotease and aminopeptidase (AP) activity, and the purity of the ARF (>10 kDa) increased by 6.69-18.11 U/mg and 1.5-2.6 fold, respectively, compared to HPE (2.63-9.37 U/mg). The AP activity toward LeuPNA was stable at 20-55℃ and pH 5-9, but decreased slightly with increasing concentration of NaCl in the reaction mixture. The ARF was the most active MetPNA and preferentially hydrolyzed Glu, Leu and AlaPNA. The bitterness tryptic casein hydrolysates (BTCHs) were treated with ARF, and the bitterness of ARF-BTCHs significantly decreased with increasing amounts of released amino acids Ala, Val, Met, Ile and Leu, which show strong correlations with bitterness. Therefore, the ARF of T. pacificus HPE obtained by ultrafiltration may have a considerable potential for application in protein hydrolysis and appears to be ideally suited to the purpose of lowing bitterness in protein hydrolysates.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.54 no.5
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• pp.724-732
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• 2021

This study used response surface methodology to investigate the optimal conditions to reduce the bitterness of alcalase-treated anchovy hydrolysate (AAH) by the aminopeptidase active fraction (AAF) derived from the common squid Todarodes pacificus hepatopancreas. The central composite design selected AAF/AAH ratio (X₁, %) and hydrolysis time (X₂, h) as independent variables, and the degree of hydrolysis (Y₁) and bitterness (Y₂) as dependent variables. The uncoded values of the multiple response optimization for independent variables were 3.4% for the AAF/AAH ratio and 9.2 h for the hydrolysis time. The predicted values of the yield and bitterness score of alcalase-AAF continuously treated anchovy hydrolysate (AAAH) under the optimized conditions were 68.9% and 4.6 points, respectively. Their measured values of 69.5% for yield and 4.6±0.5 points for bitterness were similar to the predicted values. The food components of AAAH were 91.4% (moisture), 7.5% (protein), 0.1% (lipid) and 0.6% (ash). The findings indicate the potential value for use as an anchovy seasoning base. The results also confirm that the bitterness of AAH was remarkably improved by AAF and implicates AAF derived from squid hepatopancreas as a good enzyme to catalyze reduced bitterness.

• Journal of the Korean Society for Marine Environment & Energy
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• v.14 no.3
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• pp.147-153
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• 2011

This study is to measure the differences of the trace-metals resulted from the different ecologies, such as nickel(Ni), copper(Cu), zinc(Zn), cadmium(Cd), plumbum(Pb), arsenic(As) and chrome(Cr) remaining in the parts of Korean Yeongdeok Crab (KYC) and Russian Snow Crab (RSC) based on ICP-MS. The recovery rate of each metal certified the reference materials (CRM) was in the average of 81~99%, which corresponded with the level required in Codex. The level of metals in the parts of KYC was in the order of Ni > As > Zn > Cu > Cr > Cd in the both male and female shell; the order of Zn > As > Cu > Cr > Ni > Cd in the leg flesh; the order of Zn > As > Cu > Cr > Cd > Ni in the body flesh; the order of Cu > Zn > As > Cd > Cr > Ni in the gill; the order of Cu > As > Zn > Cd > Ni > Cr in the male hepatopancreas; the order of Cu > Zn > As > Cd > Cr > Ni in the female hepatopancreas, thereby showing some differences. It was revealed that the levels of most metals (nickel, copper, zinc, arsenic and chrome) were similar between KYC and RSC except cadmium which was somewhat lower than that of KYC. However, the cadmium in RSC was discovered in high level in most of the parts, two times higher in the hepatopancreas, and four times in the gill. It was also revealed that the trace metal contents were changing according to the size of KYC; the metals with the highest level of Ni in shell, Zn in leg and body flesh, Cu in gill tended to decrease as growing, whereas the cadmium contents tended to increase overall and accumulated the most in hepatopancreas. The results showed there was a possibility that the phenomenon of bioaccumulation within hepatopancreas would increase as growing.