• 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.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.29 no.6
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• pp.797-804
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• 1996

Two trypsins were purified from shrimp hepatopancreas through ammonium sulfate fractionation, Q-Sepharose ionic exchange, benzamidine Sepharose-6B affinity, and Sephacryl S-300 gel chromatography. Both enzymes had a single polypeptide chain with a molecular weight (M.W.) of 32 kDa by sodium dodecylsulfate polyacrylamide gel electrophoresis (SOS-PAGE), although trypsin A and B were estimated to be a molecular weight of 27.2 and 22.8 kDa, respectively, using Sephacryl S-300 gel filtration. Both trypsins had similar amino acid compositions and rich in glycine, valine, alanine, aspartic acid, and glutamic acid, but low in methionine and basic amino acids. Both enzymes were completely inactivated by soybean trypsin inhibitor (SBTI), phenylmethylsulfonyl fluoride (PMSF), tosyl-L-lysine chloromethyl ketone (TLCK), benzamidine, leupeptin, however, not affected by tosyl-L-phenylalanine chloromethyl ketone (TPCK) and pepstatin.

• 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.3 no.3_4
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• pp.188-194
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• 2000

A protease purified from hepatopancreas of shrimp, Penaeus japonicus, had maximum activity at $70^{\circ}C$ and in neutral and alkaline pH ranges. Specific activity at optimum reaction condition of the protease was estimated to be approximately 12 U/mg/min. The protease was stable in neutral and alkaline pH ranges and activity was retained after heat treatment at $50^{\circ}C$ for 30 min. Apparent $K_m$ and $V_{max}$ value against casein substrate were estimated to be $0.29\%$ and $7.8see^{-1}$, respectively, and those against N-CBZ-L-tyrosine p-nitropheny1 ester (CBZ­Tyr-NE) were 0.38 mM and $2,400 see^{-1}$, respectively. The N-termina1 sequence of the protease showed high homology to the trypsin from same species and the proteases from shrimp. Myosin heavy chain (MHC) from shrimp tail meat was the most susceptible to the protease and actin/tropomyosin were degraded progressively during 4 hr incubation, but to a lesser degree than MHC.

• Fisheries and Aquatic Sciences
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• v.2 no.2
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• pp.218-225
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• 1999

A protease without tryptic and chymotryptic activities was purified from the hepatopancreas of shrimp, Penaeus orientalis, using Q-Sepharose ionic exchange, benzamidine Sepharose-6B affinity, Mono-Q, and gel chromatography. Molecular weight (M.W.) of the protease was estimated to be 27kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS­PAGE). The amino acid composition of the protease was different from that of protease from P. japonicus or trypsin from P. orientalis. The protease was completely inhibited by benzamidine, $N\alpha-p-tosyl-L-lysine$ chloromethyl ketone (TLCK), and phenylmethylsulfonyl fluoride (PMSF) and was not affected by leupeptin, pepstatin, N-tosyl-L-phenylalanine chloromethyl ketone (TPCK), iodoacetate, and ethylenediamine tetra acetate (EDTA). The enzyme did not have any activity against Na-benzoyl-DL-arginine p-nitroanilide (BAPNA) or N-benzoyl-L-tyrosine ethyl ester (BTEE) which are specific substrates of trypsin and chymotrypsin, respectively. However, the protease showed hydrolytic activity for a carboxyl terminal of Tyr, Trp, Phe, Glu, and Cys.

• Fisheries and Aquatic Sciences
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• v.9 no.1
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• pp.7-13
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• 2006

Using biochemical methods, we determined the potential of local female shrimp populations as breeding stock to select the best adult prawns for improving larval production. As condition indexes, we selected total RNA, DNA, their ratio, and trypsin activity. The DNA content in the pleopods of each local population was similar, i.e., between $0.90{\pm}0.06\;and\;1.02{\pm}0.04(SE){\mu}g/mg$. In comparison, the RNA contents differed markedly between $2.00{\pm}0.09$ and $0.96{\pm}0.08\;{\mu}g/mg$. Therefore, the RNA/DNA (R/D) ratio in the pleopod could be used as a condition index because it represents a biochemical characteristic of the population. The mean pleopodal R/D ratio of the Goheung population was the highest at $2.52{\pm}0.19$, which indicated the best condition. Trypsin activity was influenced little by shrimp condition and more by the amount of food ingested. The gonadosomatic index (GSI) and R/D ratio in the gonads provided offsetting information about the instantaneous gonad maturity. The Goheung population had the highest instantaneous GSI, despite some spawning. Based on the condition indexes and time of gonad maturation, the Goheung shrimp population is suitable for use as breeding stock.

• Korean Journal of Environmental Biology
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• v.35 no.4
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• pp.461-467
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• 2017

Shrimps infected with WSSV(White Spot Syndrome Virus) generally exhibit white spots in their inner space of carapaces as an acute clinical sign. In an effort to identify the correlation between this acute clinical sign and the condition, the index factors (RNA/DNA concentration and ratio, trypsin activity) were analyzed. A total 580 farmed Fenneropenaeus chinensis and 130 Lithopenaeus vannamei were collected from western and southern fifteen outdoor ponds in Korea. The status of the white spot pathology was divided into four stages (stage 0, stage I, stage II, and stage III), in accordance with the clinical signs as to the size and area of white spots. A significant decrease in RNA concentration and RNA/DNA ratio for multi-infected fleshy prawn (WSSV and vibrio sp.) occurred during the stage III (the whole carapace is covered with a white spot). In particular, RNA/DNA ratio was significantly lower as $1.47{\pm}0.04$ than other groups. A similar trend was also found in the single infection (WSSV), but the decrease was less than the multi-infection. In the species comparison, both species were vulnerable to the multi-infection, but L. vannamei was more sensitive than F. chinensis(ANOVA, p<0.05): A significant decrease in RNA concentration and RNA/DNA ratio was first found in stage II for the former species, while it was found in stage III for the latter species. Trypsin activity was also showed a similar tendency with nucleic acid variation. Multi-infected shrimp showed drastically decrease of trypsin activity. According to the results, clinical signs of the white spot under carapace have an only physiological effect on shrimp if they covered entirely with white spots.

• Fisheries and Aquatic Sciences
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• v.1 no.2
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• pp.201-208
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• 1998

A protease, which had no tryptic and chymotryptic activity, was purified from the hepatopancreas of shrimp, P. japonicus, through ammonium sulfate fractionation, Q­Sepharose ionic exchange, benzamidine Sepharose 6B affinity, and Sephacryl S-100 gel chromatography. Molecular weight (M.W.) of the protease was estimated to be 24 kDa by gel filtration and showed a single peptide band by sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE). The protease had a low ratio of acidic to basic amino acids, which is different with pro teases from marine animals. The enzyme was partially inhibited by benzamidine, tosyl-L-lysine chioromethyl ketone (TLCK), phenylmethylsulfonyl fluoride (PMSF), soybean trypsin inhibitor (SBTI), and pepstatin. The enzyme did not have any activity against benzoyl-D,L-arginine p-nitroanilide (BAPNA) or benzoyl-L-tyrosine ethyl ester (BTEE) which is a specific substrate of trypsin and chymotrypsin, respectively. However, the enzyme showed activity forward N-CBZ-L-tyrosine p-nitrophenyl ester (CBZ-Tyr-pNE), N­CBZ-L-tryptophan p-nitrophenyl ester (CBZ-Trp-pNE), and N-CBZ-L-proline p-nitrophenyl ester (CBZ-Pro-pNE). The protease did not showed tryptic and chymotryptic activity, which was not reported in shrimp hepatopancreas.

• Journal of Aquaculture
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• v.19 no.1
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• pp.1-6
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• 2006

The tryptic enzyme activities from hepatopancreas, foregut, midgut and feces were examined to optimize the feeding method in whiteleg shrimp, Litopenaeus vannamei. The highest tryptic enzyme activity was found in hepatopancreas. The enzyme activities of hepatopancreas were 4 times higher than those of foregut per mg dry weight at 30 minutes feeding. Post feeding period, the activities of hepatopancreas increased continuously up to 30 hours after feeding. Trypsin activities of foregut showed about 3 times higher than did those of midgut. Average activity of foregut reached the pick with $303{\pm}68\;(mean{\pm}SE)$ nmol/mg/min at two hours after feeding and kept the activity up to 4 hours after feeding and thereafter the activity decreased. Average tryptic enzyme activity of midgut increased to $96{\pm}26nmol/mg/min$ up to two hours after feeding and it decreased to $52{\pm}17nmol/mg/min$ at five hours after feeding eventhough the gastric evacuation rate was still 50% by then. Foregut clearance occurred in 30 minutes after feeding and midgut weight increased up to 2 hours after feeding. Also we found that the maximal food ingestion in foregut was equivalent to the average 0.3% of its body weight by 30 minutes after feeding. Up to 5 hours after feeding, the weight ratio of midgut to body weight reduced, but still the weight ratio of foregut to body weight kept the similarity until then. These indicated that the tryptic enzyme activity and the clearance rate are different among the digestive organs and between forgot and midgut during the post feeding period in whiteleg shrimp.

• Fisheries and Aquatic Sciences
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• v.27 no.7
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• pp.447-455
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• 2024

This study was conducted to investigate the effects of feeding Artemia nauplii enriched with poly-β-hydroxybutyrate (PHB) on Pacific white shrimp (Penaeus vannamei) post-larvae. A total 900 shrimp (initial average body weight: 2.50 ± 0.01 mg) were randomly distributed into 9 acrylic tanks (96 L) in triplicate groups. The Artemia reared in 0%, 0.25% and 0.50% PHB dissolved water (designated as Con, PHB25 and PHB50, respectively) were fed to the shrimp post-larvae for 14 days. The growth performance of shrimp was significantly higher in PHB25 and PHB50 groups than those in Con group. Survival was not significantly different among all the groups. Relative gene expressions of trypsin and amylase were significantly upregulated in PHB25 group than those of Con group. Relative gene expression of chymotrypsin was significantly upregulated in PHB25 and PHB50 groups compared to Con group. Relative gene expression of insulin-like growth factor-I was significantly upregulated in PHB25 group than in Con group. Relative gene expression of insulin-like growth factor-binding protein was significantly upregulated in both PHB25 and PHB50 groups than that of Con group. Relative expression of prophenoloxidase gene was significantly upregulated in PHB25 and PHB50 groups as compared to Con group. Relative expression of penaeidin-3a gene was significantly upregulated in PHB50 group as compared to Con group. This study demonstrated that PHB, as a functional nutritional enrichment for Artemia, improves the growth, digestibility and immunity of shrimp post-larvae.