식육의 연도(Tenderness)는 풍미와 함께 식육의 품질을 결정하는 중요한 요소이다. 우리민족은 옛부터 식육의 연도증진을 위해 불고기, 육회 등의 양념에 배나 배즙을 첨가하여 고기를 연화시키는 방법이 이용되고 있는데, 이러한 고기의 연화는 배 속에 존재하는 고유한 단백질분해효소의 효과라고만 생각해왔을 뿐 그 ...
식육의 연도(Tenderness)는 풍미와 함께 식육의 품질을 결정하는 중요한 요소이다. 우리민족은 옛부터 식육의 연도증진을 위해 불고기, 육회 등의 양념에 배나 배즙을 첨가하여 고기를 연화시키는 방법이 이용되고 있는데, 이러한 고기의 연화는 배 속에 존재하는 고유한 단백질분해효소의 효과라고만 생각해왔을 뿐 그 분해효소에 관하여 생화학적 연구는 아직도 미약하다. 본 논문에서는 안성산 신고배 속에 함유된 것으로 알려진 단백질분해효소의 식육 연화제로서 최적의 조건과 특성을 생화학적으로 규명하고자 하였다. 한국산 신고배로부터 단백질분해효소를 추출하여 황산암모늄염석, DEAE-Sepharose ion exchange chromatography과 Mono Q Chromato- graphy 및 Mini Q column 을 이용하여 순차적으로 정제된 단백질분해효소를 전기영동상으로 확인 결과 분자량이 약 38 kDa 에서 단일 band가 나타났으며, specificity activity는 64,000 unit/㎎, 회수율은 38.79%, 정제도는 25배로 나타났다. 정제된 단백질을 MALDI-TOF MS/MS방법으로 Matrixscienceb (http://www.matrixscience) server의 Mascot MS Ion Search를 통하여 각각의 MS Spectrum과 일치하는 Peptide Sequence를 조사한 결과, 이 단백질이 미지의 단백질임을 밝혔다. 그리고 정제된 단백질분해효소는 pH 3.0∼7.5에서 활성을 보였으며 최적 pH는 6.5로 활성도가 90%로 나타났고, pH 8.0이상에서는 분해가 되지 않았으므로 활성이 없는 것으로 미루어 본 단백질분해효소는 알칼리성에서는 불활성되는 것으로 추정된다. 정제된 단백질분해효소는 4℃에서는 활성도가 25%로 나타났고 온도가 증가함에 따라 점차 분해활성이 증가하였으나 80℃에서 활성도가 20%로 낮아지는 경향을 확인하였으며 최적 온도는 70℃로 활성도가 85%로 측정되었다. 한국산 배에서 정제한 단백질분해효소를 actomyosin에 처리하여 시간이 경과함에 따라 냉전계방사형 주사전자현미경으로 관찰한 결과 acto- myosin의 구조가 변형된 것으로 확인 되였으며 이는 전기영동상으로 확인한 결과 band가 소편화 되는것과 일치한 경향이 나타났다. 본 효소는 고온에서도 상당 시간 비교적 안정한 특성을 보여 식품제조, 식육연화 등 식품산업 분야에서의 활용가능성이 높을 것으로 보이며, 나아가 단백질이 갖는 식품학적 기능성을 높이는 데에도 널리 활용 할 수 있을 것으로 판단된다.
식육의 연도(Tenderness)는 풍미와 함께 식육의 품질을 결정하는 중요한 요소이다. 우리민족은 옛부터 식육의 연도증진을 위해 불고기, 육회 등의 양념에 배나 배즙을 첨가하여 고기를 연화시키는 방법이 이용되고 있는데, 이러한 고기의 연화는 배 속에 존재하는 고유한 단백질분해효소의 효과라고만 생각해왔을 뿐 그 분해효소에 관하여 생화학적 연구는 아직도 미약하다. 본 논문에서는 안성산 신고배 속에 함유된 것으로 알려진 단백질분해효소의 식육 연화제로서 최적의 조건과 특성을 생화학적으로 규명하고자 하였다. 한국산 신고배로부터 단백질분해효소를 추출하여 황산암모늄염석, DEAE-Sepharose ion exchange chromatography과 Mono Q Chromato- graphy 및 Mini Q column 을 이용하여 순차적으로 정제된 단백질분해효소를 전기영동상으로 확인 결과 분자량이 약 38 kDa 에서 단일 band가 나타났으며, specificity activity는 64,000 unit/㎎, 회수율은 38.79%, 정제도는 25배로 나타났다. 정제된 단백질을 MALDI-TOF MS/MS방법으로 Matrixscienceb (http://www.matrixscience) server의 Mascot MS Ion Search를 통하여 각각의 MS Spectrum과 일치하는 Peptide Sequence를 조사한 결과, 이 단백질이 미지의 단백질임을 밝혔다. 그리고 정제된 단백질분해효소는 pH 3.0∼7.5에서 활성을 보였으며 최적 pH는 6.5로 활성도가 90%로 나타났고, pH 8.0이상에서는 분해가 되지 않았으므로 활성이 없는 것으로 미루어 본 단백질분해효소는 알칼리성에서는 불활성되는 것으로 추정된다. 정제된 단백질분해효소는 4℃에서는 활성도가 25%로 나타났고 온도가 증가함에 따라 점차 분해활성이 증가하였으나 80℃에서 활성도가 20%로 낮아지는 경향을 확인하였으며 최적 온도는 70℃로 활성도가 85%로 측정되었다. 한국산 배에서 정제한 단백질분해효소를 actomyosin에 처리하여 시간이 경과함에 따라 냉전계방사형 주사전자현미경으로 관찰한 결과 acto- myosin의 구조가 변형된 것으로 확인 되였으며 이는 전기영동상으로 확인한 결과 band가 소편화 되는것과 일치한 경향이 나타났다. 본 효소는 고온에서도 상당 시간 비교적 안정한 특성을 보여 식품제조, 식육연화 등 식품산업 분야에서의 활용가능성이 높을 것으로 보이며, 나아가 단백질이 갖는 식품학적 기능성을 높이는 데에도 널리 활용 할 수 있을 것으로 판단된다.
Meat tenderness is one of the major palatabilities and has a major influence on the quality evaluation. It must be maintained and improved in most of the meat producing species. The tough meat is primarily due to the formation of permanent cross bridges in muscle and between the actin and myosin fil...
Meat tenderness is one of the major palatabilities and has a major influence on the quality evaluation. It must be maintained and improved in most of the meat producing species. The tough meat is primarily due to the formation of permanent cross bridges in muscle and between the actin and myosin filaments and secondly insoluble collagen in muscle. Meat tenderness is affected greatly by the age of the animal. The proteolytic enzymes of plant origin papain, bromelin and ficin are effective in increasing tenderness of meat. However, action of these enzymes can result in extensive degradation of the skeletal muscle. The proteolytic properties of the protease, from Korean pear (Ansung Shingo) were investigated and evaluated for its bio-chemical and cathartic abilities as a meat tenderizer in degrading and solubility of actomyosin myofibrils and collagen of connective tissue without extensive degradation of the skeletal muscle. For the purification of protease were the analysis of salting out with ammonium sulfate, DEAE-Sepharose chromatography, Mono Q chromatography and Mini Q Membrane column were used. And purification of protease was confirmed by using SDS- PAGE and the enzymatic properties of protease were surveyed. Optimum pH and temperature of protease were 6.5 and 60℃, respectively. The hydrolysis of the actomyosin, identified by of the SDS-PAGE, indicated the activity of protease of muscle protein. The purification of protease was confirmed by the detection of single bands using SDS-PAGE. Molecular weight (M.W.) of the protease was 38,000. Specificity activity was 64,000 unit/mg and yield was 38.79%, degree of purification was 25 folds. The quantitative analysis of purified protein was carried out with MALDI-TOF MS/MS Method. The obtained precursor value was 1647.6 and with this precursor point was the peptide sequence analyzed. However identical peptide was not detected by using the Mascot MS Ion search of the Matrix sciences server. Purified protease was stable in the range of pH 3.0~7.5 and the optimum pH were 6.5 and enzyme reaction were active in 90%. The hydrolysis of purified protease could not found in alkali pH range above the pH 8.0. These results indicated that the protease isolated from Korean pear are active in neutral pH range, and not in alkali. And the activity of purified protease showed in 25% at 4℃ and the activity of purified protease was detected at the level of 20% at 80℃. the activity is dependent on the increased temperature. The optimum temperature was 70℃, and the optimum activity was at the level of 85%. The fragmentation of actomyosin was investigated by using the Cyro-FE-SEM(cyro filed emission scanning electron microscope). These results were correspondences to the fragmentation band with SDS-PAGE. In conclusion, this study has demonstrated that the tenderization of meat was achieved through the fragmentation of myofibrils by using the protease from korean pear (Ansung shingo) without an undesirable mushy texture, and revealed that this protease from korean pear was an effective tenderizer of meat.
Meat tenderness is one of the major palatabilities and has a major influence on the quality evaluation. It must be maintained and improved in most of the meat producing species. The tough meat is primarily due to the formation of permanent cross bridges in muscle and between the actin and myosin filaments and secondly insoluble collagen in muscle. Meat tenderness is affected greatly by the age of the animal. The proteolytic enzymes of plant origin papain, bromelin and ficin are effective in increasing tenderness of meat. However, action of these enzymes can result in extensive degradation of the skeletal muscle. The proteolytic properties of the protease, from Korean pear (Ansung Shingo) were investigated and evaluated for its bio-chemical and cathartic abilities as a meat tenderizer in degrading and solubility of actomyosin myofibrils and collagen of connective tissue without extensive degradation of the skeletal muscle. For the purification of protease were the analysis of salting out with ammonium sulfate, DEAE-Sepharose chromatography, Mono Q chromatography and Mini Q Membrane column were used. And purification of protease was confirmed by using SDS- PAGE and the enzymatic properties of protease were surveyed. Optimum pH and temperature of protease were 6.5 and 60℃, respectively. The hydrolysis of the actomyosin, identified by of the SDS-PAGE, indicated the activity of protease of muscle protein. The purification of protease was confirmed by the detection of single bands using SDS-PAGE. Molecular weight (M.W.) of the protease was 38,000. Specificity activity was 64,000 unit/mg and yield was 38.79%, degree of purification was 25 folds. The quantitative analysis of purified protein was carried out with MALDI-TOF MS/MS Method. The obtained precursor value was 1647.6 and with this precursor point was the peptide sequence analyzed. However identical peptide was not detected by using the Mascot MS Ion search of the Matrix sciences server. Purified protease was stable in the range of pH 3.0~7.5 and the optimum pH were 6.5 and enzyme reaction were active in 90%. The hydrolysis of purified protease could not found in alkali pH range above the pH 8.0. These results indicated that the protease isolated from Korean pear are active in neutral pH range, and not in alkali. And the activity of purified protease showed in 25% at 4℃ and the activity of purified protease was detected at the level of 20% at 80℃. the activity is dependent on the increased temperature. The optimum temperature was 70℃, and the optimum activity was at the level of 85%. The fragmentation of actomyosin was investigated by using the Cyro-FE-SEM(cyro filed emission scanning electron microscope). These results were correspondences to the fragmentation band with SDS-PAGE. In conclusion, this study has demonstrated that the tenderization of meat was achieved through the fragmentation of myofibrils by using the protease from korean pear (Ansung shingo) without an undesirable mushy texture, and revealed that this protease from korean pear was an effective tenderizer of meat.
※ AI-Helper는 부적절한 답변을 할 수 있습니다.