방어 普通肉과 血合肉 筋原纖維蛋白質의 熱安定性.II. 糖 및 아미노酸 變性 抑制.

卞在亨,鄭甫泳,崔映準 釜山水産大學校 1984 釜山水産大學 硏究報告 Vol.24 No.2

前報(下 等, 1984)에 이어 방어 普通肉과 血合肉에서 Trition X-100 界面活性劑를 含有하는 溶媒로 洗淨하여 筋原纖維蛋白質을 抽出하고 그 加熱變性에 대한 sorbitol, sucrose 等 糖類와 Na-glutamate, L-cysteine 等 아미노酸類의 抑制效果를 濃度, 加熱時間 및 混成添加의 各 條件別로 檢討하였다. 實驗의 結果를 要約하면 : 1. 糖類中 sorbitol와 sucrose, 아미노酸中 Na-glutamate와 L-cysteine을 各各 1∼10% 範圍로 添加하여 35℃에서 60分間 加溫한 結果, 普通肉과 血合肉의 各 筋原纖維蛋白質에 대하여 L-cysteine을 除外하면 모두 濃度의 增加와 더불어 變性抑制效果도 漸次 上昇하였다. 變性抑制效果는 sorbitol>Na-glutamate>sucrose의 順으로 컸으며, 特히 普通肉 筋原纖維蛋白質에서 더욱 效果的이었다. 2. 添加劑를 10%씩 添加하여 40℃에서 加溫하였을 때 ????-ATPase 活性의 半減에 所要되는 時間은 普通肉 筋原纖維蛋白質에 있어서는 sorbitol 54分, sucrose 35分, Na-glutamate 42分이었고 血合肉 筋原纖維蛋白質에 있어서는 sorbitol 31分, sucrose 30分, Na-glutamate 34分이었다. 3. sorbitol, sucrose, Na-glutamate를 各各 5%씩 서로 混合하여 添加하고 35℃에서 時間別로 加溫했을 때는 普通肉과 血合肉 筋原纖維蛋白質이 공통적으로 한 가지만을 添加했을 때 보다 變性抑制效果가 相乘的으로 나타났으며, 血合肉에서는 더욱 效果的이었다. 4. 混合添加物의 效果를 Arrhenius式을 適用하여 溫度에 대한 蛋白質의 變性速度常數로서 比較하여 보면, sorbitol과 Na-glutamate를 混合하여 添加했을 때는 血合肉 筋原纖維蛋白質이 普通肉 筋原纖維蛋白質에 比하여 1.38倍 더 安定하였으며, sorbitol과 sucrose를 混合하여 添加했을 때는 血合肉 筋原纖維蛋白質이 普通肉 筋原纖維蛋白質에 比하여 2.28倍 더 安定하였다. In the preceding paper, the authors have described on the differences between the ordinary muscle and dak muscle in specially referred to the thermal stability of the myofibrillar proteins, and the denaturation velocity and thermodynamic parameters of the proteins were discussed. The results provided a remarkable proof that the myofibrillar protein of the ordinary muscle is more stable than the protein of the dark muscle below 31℃. In this study, protective effect of some additives such as sorbitol and sucrose in saccharides and Na-glutamate and L-cysteine in amino acids on thermal denaturation of the two myofibrillar proteins has been investigated by the mixing of the additives and the heating time. When the additives were present in the proteins in the range of 1 to 10% and the temperature was maintained at 35℃ for 60 min., the protective effect was gradually risen with an increase of the concentration except L-cysteine. The protective effects of the additives against thermal denaturation of the protein of the ordinary muscle showed that the most effective is 10% sorbitol and follows 10% Na-glutamate and 10% sucrose in order, and in case of the dark muscle, 10% Na-glutamate is the most effective, and 10% sorbitol and 10% sucrose are followed to it in order. In the both of the muscles, 10% L-cysteine showed no protective effect. As the mixed additives of sorbitol, sucrose and Na-glutamate in 5% were used for the cooperative action against the thermal denaturation, a protective effect was confirmed in the both proteins. In the effects of the mixedadditives concerned, the denaturation constants of the protein in 5% Na-glutamate to 5% sorbitol st 30℃ was ???/sec in the ordinary muscle and ????/sec in the dark muscle, whereas those in ?? in ordinary muscle and ???/sec in the dark muscle, wheraw those in 5% sucrose to 5% sorbitol were ????/sec in the ordinary muscle and ????/sec in the dark muscle. The conclusion drawn from these experiments is that the mixed additive of Na-glutamate and sorbitol strongly enhances a protective effect in the both proteins of the muscles.

방어 普通肉과 血合肉 筋原纖維蛋白質의 熱安定性.I. 熱安定性比較

崔映準,卞在亨,鄭甫泳 釜山水産大學校 1984 釜山水産大學 硏究報告 Vol.24 No.2

방어 普通肉과 血合肉의 筋原纖維蛋白質의 熱安定性을 밝히기 위하여 背部 普通肉과 側部 血合肉에서 筋原纖維蛋白質을 各各 抽出하고 溫度의 變化가 筋原纖維蛋白質의 安定性에 미치는 影響을 ???-ATPase 活性度에 의하여 分析 檢討하였다. 그리고 이들 蛋白質의 變化速度와 熱力學的 狀態函數도 測定 比較하였다. 實驗結果를 要約하면 다음과 같다. 1. Trition X-100을 含有하는 溶媒로서 處理한 방어 普通肉과 血合肉의 筋原纖維蛋白質은 0.09% 1.70%의 脂質이 混入되어 있었으며, 이때의 ???-ATPase 活性은 各各 0.205∼0.225μM-Pi/min/mg-protein 및 0.231∼0.240μM-Pi/min/mg-protein 이었다. 2. ???-ATPase 活性은 普通肉과 血合肉의 筋原纖維蛋白質이 함께 溫度의 上昇과 더불어 1次 函數關係로 감소하였고, 31℃이하에서는 普通肉의 筋原纖維蛋白質이 血合肉의 筋原纖維蛋白質에 比하여 安定하였으나, 그以上의 溫度에서는 血合肉 筋原纖維蛋白質이 보다 安定하였다. 3. 25℃ 溫度條件에서의 普通肉 筋原纖維蛋白質의 活性化에너지는 5,500 cal/mole, 活性化엔탈피는 4,900cal/mole, 活性化 엔트로피는 -44.51 e.u. 자유에너지값은 18,100cal/mole 이었고, 血合肉의 筋原纖維蛋白質은 活性化에너지가 5,390 cal/mole, 活性化엔탈피가 4,800 cal/mole, 活性化엔트로피는 -44.17 e.u., 자유에너지 값은 17,900 cal/mole 이었다. 溫度의 上昇과 더불어 自由에너지값에 寄與하는 엔트로피의 影響은 증대하였다. 4. -30℃, -15℃ 및 0℃에서 貯藏하였을때의 各 筋原纖維蛋白質은 -30℃에서 가장 安定하였고 0℃와 -15℃의 順으로 安定性이 떨어졌다. This paper deals with the differences between the thermal stabilities of myofibrillar proteins of dorsal ordinary muscle and lateral dark muscle in yellowtail, Seriola quinqueradita. The myofibrillar proteins submitted to this experiment were extracted by the modified method using Triton X-100 detergent. Effects of temperature on the stabilities of the myofibrillar proteins were investigated by analysing ???-ATPase activity. The denaturation velocities and thermodynamic parameters of each of the proteins were compared. Lipid content and ???-ATPase activity of the myofibrillar protein of the ordinary muscle were found to be 0.09% and 0.205-0.225μM-Pi/min/mg-protein, respectively, while those of the dark muscle were 1.70% and 0.231-0.240μM-Pi/min/mg-protein. The relationship between the inactivation of ???-ATPase activity of the myofibrillar proteins and the increase in temperature showed a good first-order reaction. The half-life of ???-ATPase activity on the myofibrillar protein showed in general that the protein of the ordinary muscle is more stable than that of the dark muscle. Activation energy, activation enthalpy, activation entropy and free energy of the protein at 25℃ were 5,500 cal/mol, 4,900 cal/mol, -44.51 e.u. and 18,100 cal/mol, respectively for the ordinary muscle, and 5,390 cal/mol, 4,800 cal/mol, -44.17 e.u. and 17,900 cal/mol, respectively for the dark muscle. It may be concluded that an increase in temperature affects substantially to entropy of the proteins. The myofibrillar proteins of the both muscles found to be more stable at -30℃ than at 0℃ and -15℃.

어육단백질의 냉동변성에 관한 연구 : Ⅲ. 동결 저장중 방어 보통육과 혈합육 근원섬유 단백질의 단백질 상호작용, 용해도 점도 및 Ca-ATPase활성의 변화 Ⅲ. Changes of Protein-Protein Interaction, Solubility, Viscosity and Ca-ATPase Activity of Myofibrillar Protein from Ordinary and Dark Muscle in Yellowtail during Frozen Storage

김정한,최영준,변재형 國立統營水産專門大學 附設 水産科學硏究所 1989 수산과학연구소보고 Vol.1 No.-

The denaturation mechanism of the protein during freezing of myofibrillar protein extracted from the ordinary and dark muscle of yellowtail (Seriola quinqueradits) were investigated by analyzing the Protein-Protein interaction, Solubility, relative viscosity and Ca-ATPase activity. The rate of protein-protein interaction and inactivation of Ca-ATPase were the first reaction against the frozen storage time. The rate of protein-protein interaction in ordinary myofibrillar protein was 1.14-2.24 times as large as that of dark one, but Ca-ATPase was conversely. A similar relation-ship between ordinary and dark myofibrillar protein was recognized in relative viscosity and solubility, but the extent of decreased relative viscosity was large than solubility. The exposure of hydrophobic residures and protein-protein interaction were significantly related with the freezing denaturation of myofibrillar protein.

魚具類中에 分布하는 蛋白質分解酵素의 分離 및 精製에 關한 硏究

張東錫,金亨洛,趙鶴來,卞在亨 釜山水産大學校 1986 釜山水産大學 硏究報告 Vol.26 No.1-2

水産無脊椎動物중 낙지, 전복, 소라, 군소, 해삼, 개불과 水産脊椎動物인 말쥐치, 두툽상어, 먹장어, 고등어 그리고 정어리의 消化管 및 肉과 臟器組織을 區分하여 組織中에 分布하는 蛋白質分解組酵素를 抽出하여 그 活性을 比較·檢討하였다. 그리고 强한 活性을 보인 고등어 幽門垂에서 抽出한 粗酵素는 鹽析, DEAE-Sephadex A-50 column chromatography 및 Sephades G-100 겔여과에 의하여 3種類의 알칼리性 蛋白質分解酵素를 分離·精製하였으며 各 精製酵素에 대하여 活性最適條件, 基質親和度, 化學藥劑에 의한 影響 및 分子量 등을 究明하였다. 그리고 試料動物의 內臟에서 分離한 細菌中에서 가장 强한 蛋白質分解酵素를 生産하는 菌株를 選別하고 그 菌이 生産한 酵素를 精製하여 特性을 調査한 結果를 要約하면 다음과 같다. 1. 各 試料의 組織에서 抽出한 粗酵素中 活性이 强한 것으로는 정어리의 膵臟에서 抽出한 것이 活性最適條件인 pH 9.8, 45℃에서 固有活性은 360 이었고, 고등어 幽門??에서 抽出한 것은 pH 9.4, 45~50℃에서 固有活性은 275였다. 2. 고등어 幽門垂에서 抽出한 粗酵素에서는 3種의 알칼리性 蛋白質分解酵素가 分離·精製되었는데, 이 세 酵素를 SDS-PAG 電氣泳動法과 Se-phadex G-100겔 여과법에 의하여 分析·檢討한 結果 이들 酵素는 모두 monomeric polypeptide로서 구성되고, 分子量은 Enz. A가 27,500, Enz. B가 20,500, 그리고 Enz. C가 15,250 정도였다. 3. 酵素의 基質親和度를 測定한 結果 Km値는 casein 基質에 대하여 Enz. A는 5.0×??, Enz.B는 1.0×?? 그리고 Enz. C는 3.3×??였다. 4. 各 精製酵素는 soybean trypsin inhibitor에 의하여 活性이 沮害를 되었으며 Enz. A는 p-chloromercuribenzoate에 의해서도 沮害를 받았는데 Enz. B와 C는 serine系 蛋白質分解酵素로 判斷되었다. 5. 分離細菌中에서 제일 강한 蛋白質分解酵素를 生産하는 細菌은 Pseudomonas SPP. 였으며 酵素의 生産은 pH 7.0, 溫度 25℃, 食鹽 0.5% 염화칼슘 0.2%를 添加했을 때 제일 양호하였으며, 이 酵素는 分子量이 약 29,000되는 metal chelator sensitive neutral proteinase로 추정되었다. 6. 結論的으로 漁具類의 組織酵素는 계속적인 原料供給이 어려우므로 酵素의 産業的 利用을 위해서는 細菌이 生産한 菌體外 蛋白質分解酵素의 活用이 보다 有益할 것으로 생각된다. Proteolytic actvity of the tissue extracts from the octopus(Octopus variabilis), abalone (Haliotis discus hannai), top shell(Turbo cornutus), sea hare(Aplysia kurodai), sea cucumber(Stichopus japonicus), echiurid(Urechis unicinctus), file fish(Navodon modestus), cat shark(Scillion hinus tarazame), hag fish(Eptatretus burgeri), mackerel(Scomber japonicus) and sardine(Sardinops melanosticta) was compared to develope as an useful enzyme. The strongest proteolyticbacterium was selected among the isolated strains from the samples submitted, then the proteolytic exoenzyme produced by this strain was also characterized. Among the tissue enzyme extracts, the proteolytic enzymes from the pyloric caeca of mackerel and pancreas of sardine were estimated as strong alkaline proteinases. The optimum pH and temperature of the crude enzyme extracted with 1% NaCl from the pyloric caeca of mackerel and pancreas of sardine were pH 0.4, 50℃ and pH 9.8, 45℃, respectively. Specific activity of the former enzyme was 275 nM-Tyr. eq/mg-protein/min and that of latter one was 360 nM-Tyr. eq/mg-protein/min. Three kinds of alkaline proteinases were isolated from the pyloric caeca of mackerel, we named those as enzyme A, B and C. Molecular weight of enzyme A, B and C determined by SDS-polyacrylamide gel electrophoresis and Sephadex G-100 gel filtration method was to be 27,500, 20,500 and 15,250, respectively. Km value of enzyme A, B and C by the method of Lineweaver and Burk was determined to be 5.0×10??%, 1.0×10??% and 3.3×10??%, respectively, for 2% casein solution as a substrate. According to the analytical results, these enzymes were observed to be composed of monomeric polypeptide. The enzyme B and C were supposed to be a serine protease because these enzymes were remarkbly inhibited by soybean trypsin inhibitor. Pseudomonas spp. (named Pseudomonas FU 101) was identified as the strongest proteolytic bacterium among the isolated strains, which grew best at 25℃, pH 7.5. It is observed that the addition of 0.2% CaCl₂ and 0.5% NaCl in the culture medium was benefitted for the production of proteinase by Pseudomonas FU 101. The extracellular proteinase produced by the strain was supposed to be kind of metal chelator sensitive neutral protease, and it showed maximum activity at 35℃, pH 7.0. Molecular weight was estimated to be 29,000 by gel filtration. As a conclusion, both proteinases produced by Pseudomonas FU 101 and extracted from tissue of mackerel were pretty good in enzyme activity, but bacterial enzyme was more benefit for industrial use than the enzyme of mackerel tissue, because it is very difficult to supply continuously lots of pyloric caeca of mackerel as raw material for enzyme extraction.

Comparative Biochemical Properties of Proteinases from the Hepatopancreas of Shrimp. : 1. Purification of Protease from the Hepatopancreas of Penaeus japonicus

Pyeun, Jae Hyeung,KIM, Hyeung Rak,KIM, Doo Sang,AHN, Chang Bum,Choi, Sung Mi,Oh, Eun Sil,Cho, Deuk Moon 한국수산학회 1998 Fisheries and Aquatic Sciences Vol.1 No.2

A protease, which had no tryptic and chymotryptic activity, was purified from the hepatopancreas of shrimp, P. japonicus, through ammonium sulfate fractionation, Q-Sepharose 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 proteases from marine animals. The enzyme was partially inhibited by benzamidine, tosyl-L-lysine chloromethyl 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), N-CBZ-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.

Purification and Characterization of Trypsins Affecting on the Autolysis of Shrimp , Penaeus japonicus

Pyeun, Jae Hyeung,KIM, Hyeung Rak,KIM, Doo Sang,AHN, Chang Bum 한국수산학회 1996 한국수산과학회지 Vol.29 No.6

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 32kDa by sodium dodecylsulfate polyacrylamide gel electrophoresis(SDS-PAGE), although trypsin A and B were estimated to be a molecular weight of 27.2 and 22.8kDa, 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 fluorid(PMSF), tosyl-L-lysine chloromethyl ketone(TLCK), benzamidine, leupeptin, however, not affected by tosyl-L-phenylalanine chloromethyl ketone(TPCK) and pepstatin.

Variability of Current and Sea Level Difference in the Western Channel of the Korea Strait in Winter 1995 ~ 96

Pyeun, Jae Hyeung,Kim, Jeong Sook,Yoon, Hong Joo,Lee, Sang Ryong,Byun, Sang Kyoung,Park, Moon Jin 한국수산학회 1998 Fisheries and Aquatic Sciences Vol.1 No.2

As a part of the long-term ADCP mooring program to measure the mass flux through the Korea Strait, current velocity data were obtained for 39 days in the deepest point of the strait. Near-surface velocity of this abservation was compared with Izuhara-Pusan sea level difference (SLD) to investigate the geostrophic relationship. Principal direction of the Tsushima Current at the mooring station is 44.6 degrees to the north from the east. Variability of the tidal current is greater than the nontidal current by a factor of two. Correlation coefficient of tidal current a against SLD is 0.46 but the nontidal current is not correlated. The current velocity (U in ㎝/s) can be estimated from the demeaned SLD (in. ㎝) by the relation U=23.63+0.64SLD where the maximum range of SLD is 52.9 ㎝. Current is coherent with SLD at semidiurnal, diurnal and 42.7-hour periods. A dominant nontidal variability with about 5-day period is not coherent with SLD.

혈합육어(멸치, 고등어, 황다랭이 및 날개다랭이)의 Trypsin - 1. 정제와 반응조건

변재형(Jae-Hyeung Pyeun),조득문(Deuk-Moon Cho),허민수(Min-Soo Heu) 한국식품영양과학회 1993 한국식품영양과학회지 Vol.22 No.4

혈합육어 중 연근해 온대산 멸치의 내장과 고등어의 유문수, 그리고 원양 열대산 황다랭이 및 날개다랭이의 유문수를 각각 시료로 하여 trypsin을 분리 정제하였으며, 그 분자량과 아미노산 조성 및 효소의 반응 최적조건에 관하여 비교분석하였다. 실험결과를 요약하면 다음과 같다. 1. 멸치의 내장과 고등어, 황다랭이 및 날개다랭이의 유문수에서 각각 황산암모늄염석, benzamidine-Sepharose 6B 친화성 크로마토그래피, DEAE-Sephadex A-50 크로마토그래피, Sephadex G-75 겔 여과 크로마토그래피 등의 방법을 혼용하여 고등어부터는 2종의 trypsin(고등어 trypsin A와 고등어 trypsin B로 명명함)을, 그리고 다른 어류로부터는 각각 1종의 trypsin을 분리 정제하였다. 2. 멸치 trypsin과 고등어 trypsin B는 고등어 trypsin A와 황다랭이 trypsin 및 날개다랭이 trypsin에 비하여 그 고유활성이 월등히 높았다. 3. 이들 혈합육어 trypsin의 분자량은 22kDa~26kDa 범위였다. 4. 이들 trypsin은 공통적으로 glycine, serine, aspartic acid를 많이 함유하고 있었으며, tryptophan, methionine, lysine, tyrosine의 함유량은 적었다. 5. BA-p-NA기질에 대한 반응 최적조건은 멸치 trypsin은 PH 8.0에서 45℃, 고등어 trypsin A와 B는 PH 8.0에서 50℃, 그리고 황다랭이 trypsin은 PH 9.0에서 55℃, 날개다랭이 trypsin은 PH 9.0에서 50℃였다. 6. 위에 든 실험 결과들은 혈합육 어류의 서식온도가 이들 어류의 trypsin의 반응 최적 온도와는 어느정도 관계가 있을 것으로 추측되었다. Deterioration of fish muscle is known to occur more quickly in the dark fleshed fish than in the white fleshed fish, causing by their high intestinal proteolytic activity. Muscle degradation which suffer post-mortem autoproteolysis is affected by trypsin with its unique activation function towards other enzymes. To compare physicochemical and enzymatic properties for the trypsins of the dark fleshed fish, trypsins from the viscera of anchovy (Engraulis japonica), and the pyloric caeca of mackerel(Scomber japonicus), yellowfin tuna(Thunnus albacores) and albacore (Thunnus alalunga) were purified through ammonium sulfate fractionation, benzamidine-Sepharose 6B, DEAE-Sephadex A-50, and Sephadex G-75 chromatography. Two trypsins from mackerel (designated mackerel trypsin A and mackerel trypsin B), and one each from anchovy, yellowfin tuna and albacore were isolated as electrophoretical homogeneity. The purities of anchovy trypsin, mackerel trypsin A and H, yellowfin tuna trypsin, and albacore trypsin increased to 78.1, 4.8, 9.3, 120, and 16o-fold, respectively, compared to crude enzyme solutions. Molecular weights of the trypsins from the dark fleshed fish estimated by SDS-polyacrylamide electrophoresis were ranged from 22kDa to 26kDa. The trypsins contained higher amount of glycine, serine and aspartic acid, and less amount of tryptophan, methionine, lysine and tyrosine. Optimal conditions for amidolytic reactions of the enzymes were pH 8.0 and 45℃ for anchovy trypsin, pH 8.0 and 50℃ for mackerel trypsin A and B, pH 9.0 and 55℃ for yellowfin tuna trypsin, and pH 9.0 and 50℃ for albacore trypsin. It was supposed that the habitat temperature of the dark fleshed fish is slightly connected with the optimal reaction temperature of the trypsins of the fish.

海産腹足類의 Paramyosin의 分離 및 그 特性에 關한 硏究

변재형(Jae-Hyeung Pyeun) 한국식품영양과학회 1973 한국식품영양과학회지 Vol.2 No.1

The muscle of abalone, Notohaliotis discus (REEVE), and top-shell, Turbo cornutus Solander, were examined for protein composition.<br/> Then paramyosins which are known as one of the important structural protein of the muscle fibrils were isolated from the both muscle and their physico-chemical properties such as solubility, salting-out behaviour, intrinsic viscosity, ATPase activity, etc. involving amino acid composition and N-terminal amino acid residues were investigated to elucidate phylogenie characteristics more intensively from the viewpoint of comparative biochemistry.<br/> The analysis of protein composition resulted in the following estimations: abalone muscle; water-soluble protein of 22 %, salt-soluble protein, 34%, alkali-soluble protein, 20%, and stroma protein, 24%, and top-shell muscle; water-soluble protein of 16%, salt-soluble protein, 30%, alkali-soluble protein, 29%, and stroma protein, 25%, respectively.<br/> It is demonstrated in sedimentation analysis that paramyosin and myosin-actomyosin account for approximately 65% and 35% of the salt-soluble protein of abalone, and that the composition of both sediments in top-shell was approximately 70% and 30%, respectively.<br/> The ultracentrifugally homogenous paramyosins isolated essentially according to Bailey's ethanol-dried method from both of the muscle showed a S°_(20,w) of 3. 14s for abalone and a S°_(20,w) of 3.50s for top-shell.<br/> The both paramyosins were commonly rich in arginine, aspartic acid, and glutamic acid, while scarcely contained proline and tryptophan, in rough accord with the other paramyosins thus far reported.<br/> It is clear that these gastropod paramyosins showed of having the characteristic N-terminal amino acid residues such as N-aspartic acid, N-valine, N-serine, and N-threonine in common.<br/> The abalone paramyosin completely salted in with KCl beyond 0.35μ and the top-shell paramyosin beyond 0.30μ. The abalone paramyosin was salted-out between 18% and 30% saturation of ammonium sulphate and the top-shell paramyosin between 22% and 29% saturation.<br/> The intrinsic viscosities at abalone and top~shell paramyosins at 25℃ were estimated respectively to be 3.1 dl/g and 2.6 dl/g showing somewhat higher than the values for some other paramyosins from lamellibranchs.<br/> In regard with the ATPase activity, the para myosin specimens did not exhibit any significant activity over through the pH conditions of 5 to 9.5. irrespective of the presence of Ca^(++) or Mg^(++). So was the case with the abalone paramyosin prepared by a slightly modified Bailey's wet-extraction method.

멸치 육과 내장으로부터 분리한 Cathepsin L, Chymotrypsin 및 Trypsin의 단백질분해 특성

번재형 ( Jae Hyeung Pyeun ),허민수 ( Min Soo Heu ),조득문 ( Deuk Moon Cho ),김형락 ( Hyeung Rak Kim ) 한국수산학회 1995 한국수산과학회지 Vol.28 No.5

어류의 사후 초기의 변화를 육 및 장기조직중에 분포하는 단백질분해효소의 작용과 관련하여 검토할 목적으로 멸치의 육 및 장기에서 분리한 cathepsin L과 chymotrypsin 및 trypsin의 단백질 기질에 대한 특성과 근원섬유단백질에 대한 분해능을 전기영동적으로 분석하여 다음의 결론을 얻었다. 이들 세 효소의 casein에 대한 친화도는 유사하였고, 근원섬유단백질에 대한 친화도는 casein에 대한 친화도보다 높았다. 멸치와 방어의 근원섬유단백질에 대한 cathepsin L과 chymotrypsin의 활성은 trypsin보다 훨씬 높게 나타났다. 0~25%까지의 식염농도에서 세 효소의 단백질분해활성은 식염의 농도에 반비례하였으며, 식염의 공존상태에서 세 효소는 casein 보다 근원섬유단백질에 대하여 높은 활성을 나타내었다. 근원섬유단백질의 효소 분해시에 cathepsin L은 chymotrypsin과 trypsin에 비하여 염농도와 온도에 의한 영향이 적었다. 따라서, 멸치의 사후변화와 젓갈 숙성중의 자가소화는 trypsin보다는 cathepsin L과 chymotrypsin의 단백질분해활성이 더욱 깊이 관여할 것으로 판단된다. Proteolytic properties of enzymes from the muscle and viscera of anchovy have been examined. Cathepsin L, chymotrypsin, and trypsin showed similar Km values for casein. However, they had higher Km values for myofibrillar proteins than those for casein. The k(cat) of cathepsin L and chymotrypsin for myofibrillar proteins were higher than that of trypsin, and also cathepsin L and chymotrypsin caused higher hydrolysis in myofibrillar proteins of anchovy and yellowtail. In the presence of sodium chloride (0~25%), proteolytic activity for myofibrillar proteins from yellowtail was higher than that for casein. Proteolytic activity was decreased with the increase of sodium chloride concentration. Cathepsin L had been less affected by NaCl concentration and temperature on the hydrolysis of myofibrillar proteins than chymotrypsin and trypsin. Cathepsin L and chymotrypsin were more responsible to the autolysis of muscle proteins from fish than trypsin.