• Fisheries and Aquatic Sciences
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• 제1권2호
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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.

• 한국수산과학회지
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• 제29권6호
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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.

• 한국어업기술학회:학술대회논문집
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• 한국어업기술학회 1998년도 수산관련공동학술대회발표요지집
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• pp.266-267
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• 1998

• Fisheries and Aquatic Sciences
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• 제3권3_4호
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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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• 제2권2호
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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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• 제9권1호
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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.

• 환경생물
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• 제35권4호
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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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• 제1권2호
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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.

• 한국양식학회지
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• 제19권1호
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• pp.1-6
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• 2006

흰다리새우 양식기법의 최적화를 위하여 소화기간 동안의 간췌장, 전장 그리고 중장에서의 trypsin의 활성변화를 조사하였다. 체중에 대한 전장, 중장의 무게 그리고 그 무게 합의 비율은 섭취된 먹이의 이동 및 소화과정을 나타내는 지표로서 공급량과 잔류량에 의한 먹이섭취량을 측정하는 것 보다 더 정확한 지표로서 사용 가능하였다. 평균적으로 치대 먹이섭취량은 전장에서 먹이섭취 후 30분 이내에 체중에 0.3%로 나타났다. 또한 30분 이후부터 전장이 비워지기 시작되었으며 중장의 무게가 최대에 이르는 시각은 2시간째였다. $3{\sim}5$시간 후에는 먹이가 배설됨으로 인하여 중장의 체중에 대한 무게비가 감소하였으나 전장에서는 비교적 같은 비율을 유지하였다. 먹이섭취에 의한 trypsin활성변화는 간췌장에서 가장 커서 전장에서의 활성변화에 비하여 약 3배로 나타났다. 소화시간이 지날수록 간췌장에서의 trypsin 활성은 지속적으로 증가하였다. 전장에서의 trypsin 효소의 활성은 중장보다 약 $2{\sim}3$배정도 높았다 먹이섭취 후 2시간이 지났을 때 trypsin 활성은 303 n mol/mg/min였고, 4시간까지 이 활성이 유지($277{\sim}306$ n mol/mg/min)되었으며, 그 후에는 점차 감소하였다. 중장에서는 typsin 활성이 먹이를 섭취하여 한 시간이 지나면서 $65{\pm}29$ (SE) n mol/mg/min로 증가하였다. 그 이후에는 $80{\sim}97$ n mol/mg/min의 범위를 나타내었으며, 5시간이 경과하였을 때 $52{\pm}17$ (SE) n mol/mg/min로 감소하였고 소화관내에 잔류하고 있는 먹이 량은 최대 섭취량의 50%로 나타났다.

• Fisheries and Aquatic Sciences
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• 제27권7호
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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.