Bacillus polyfermenticus SCD가 생산하는 Protease의 정제와 특성 및 식육연화제로서의 이용 Partial Purification and Characterization of Protease Produced by Bacillus polyfermenticus SCD원문보기
본 연구는 Bacillus polyfermenticus SCD가 생산하는 protease의 정제와 특성 및 식육연화제로서의 이용을 조사하기 위하여 실시되었다. 결과를 요약하면 다음과 같다. 1. Bacillus polyfermenticus SCD가 생산하는 protease의 부분 정제와 특성 본 연구는 B. polyfermenticus SCD로부터 분리한 extracellular protease를 정제하고 그의 특성을 검토하였다. 48시간 배양했을 때 protease의 활성은 41.8 units/mL을 나타냈다. Ammonium sulfate 침전분리와 DEAE Sephadex-A50 column을 이용한 ion exchange chromatography를 거쳐 정제되면 specific activity가 2,973.1 units/mg로 58.4배 증가하였다. SDS-PAGE를 통해 확인한 결과 protease의 분자량은 약 44 kDa 또는 28 kDa이었다. Protease의 작용 최적온도는 40-50℃이다. Protease는 50℃에서 가장 안정하였으나 55℃이상 에서는 안정성이 급격히 저하하였다. Protease의 작용 최적 pH는 8.0 이었으며 pH 7.0-9.0에서 안정하였다. Protease는 계면활성제인 Triton X-100(0.1%)에 의해 활성이 2배 촉진되었으나 ...
본 연구는 Bacillus polyfermenticus SCD가 생산하는 protease의 정제와 특성 및 식육연화제로서의 이용을 조사하기 위하여 실시되었다. 결과를 요약하면 다음과 같다. 1. Bacillus polyfermenticus SCD가 생산하는 protease의 부분 정제와 특성 본 연구는 B. polyfermenticus SCD로부터 분리한 extracellular protease를 정제하고 그의 특성을 검토하였다. 48시간 배양했을 때 protease의 활성은 41.8 units/mL을 나타냈다. Ammonium sulfate 침전분리와 DEAE Sephadex-A50 column을 이용한 ion exchange chromatography를 거쳐 정제되면 specific activity가 2,973.1 units/mg로 58.4배 증가하였다. SDS-PAGE를 통해 확인한 결과 protease의 분자량은 약 44 kDa 또는 28 kDa이었다. Protease의 작용 최적온도는 40-50℃이다. Protease는 50℃에서 가장 안정하였으나 55℃이상 에서는 안정성이 급격히 저하하였다. Protease의 작용 최적 pH는 8.0 이었으며 pH 7.0-9.0에서 안정하였다. Protease는 계면활성제인 Triton X-100(0.1%)에 의해 활성이 2배 촉진되었으나 SDS(0.1%)에 의해서는 강한 저해를 받았다. 2. Bacillus polyfermenticus SCD가 생산하는 protease가 돈육의 연화에 미치는 영향 본 연구는 B. polyfermenticus SCD가 생산하는 protease가 돈육의 연화에 미치는 영향을 조사하고자 실시하였다. B. polyfermenticus SCD가 생산하는 protease의 특성을 조사하고 돈육을 각각의 효소(papain, Aspergillus oryzae와 B. polyfermenticus SCD가 생산하는 protease)가 포함된 용액으로 처리한 후 4℃에서 24, 72, 168시간 동안 저장하며 품질을 측정하였다. B. polyfermenticus SCD가 생산하는 protease는 다른 효소에 비해 활성이 낮았지만 육단백질을 쉽게 분해하였다. 소편화도는 대조구에 비해 처리구에서 높은 값을 나타내었다. 전자현미경으로 관찰한 결과 효소 처리된 육을 168시간 저장했을 때 근육 내의 결합조직이 많이 파괴되어 있었다. 가열감량은 저장기간 동안 증가하였다. 관능평가를 실시했을 때, 효소 처리구가 큰 차이를 나타내지는 않았지만 전체적으로 연도에서는 대조구에 비해 우수한 평가를 받았다. 3. Bacillus polyfermenticus SCD가 생산하는 protease가 육포의 품질에 미치는 영향 본 연구는 B. polyfermenticus SCD가 생산하는 protease가 육포의 품질에 미치는 영향을 조사하고자 실시하였다. 4개의 시료는 Streptomyces griseus와 B. polyfermenticus SCD가 생산하는 효소를 각각 0.005%, 0.01%로 첨가하였으며 텀블링 후 바로 건조한 것과 텀블링 후 24시간이 경과한 뒤 건조한 두 가지 시료로 구분하였다. 모든 처리구에서 돈육육포와 우육육포의 pH에서 유의적인 차이가 나타나지 않았으며, 효소를 처리하였을 때 우육육포의 색에 미치는 영향이 돈육 육포보다 컸다. 효소 처리구들이 대조구에 비해 유의적으로 낮은 경도를 나타내었으나 염지시간에 의해서는 유의적인 차이는 나타나지 않았다. 수분함량은 효소와 염지시간에 따라 영향을 받지 않았으나 수분활성도는 효소 처리구가 낮았다. 돈육육포는 효소 처리 육포가 높은 재수화율을 나타내었으며 우육육포는 텀블링 후 24시간 경과한 뒤 건조했을 때 높은 재수화율을 나타냈다. 관능평가를 실시했을 때, 효소 처리구에서 조직감, 다즙성, 전체적 기호도가 높게 나타났지만 염지시간에 의해서는 큰 차이를 나타내지는 않았다.
본 연구는 Bacillus polyfermenticus SCD가 생산하는 protease의 정제와 특성 및 식육연화제로서의 이용을 조사하기 위하여 실시되었다. 결과를 요약하면 다음과 같다. 1. Bacillus polyfermenticus SCD가 생산하는 protease의 부분 정제와 특성 본 연구는 B. polyfermenticus SCD로부터 분리한 extracellular protease를 정제하고 그의 특성을 검토하였다. 48시간 배양했을 때 protease의 활성은 41.8 units/mL을 나타냈다. Ammonium sulfate 침전분리와 DEAE Sephadex-A50 column을 이용한 ion exchange chromatography를 거쳐 정제되면 specific activity가 2,973.1 units/mg로 58.4배 증가하였다. SDS-PAGE를 통해 확인한 결과 protease의 분자량은 약 44 kDa 또는 28 kDa이었다. Protease의 작용 최적온도는 40-50℃이다. Protease는 50℃에서 가장 안정하였으나 55℃이상 에서는 안정성이 급격히 저하하였다. Protease의 작용 최적 pH는 8.0 이었으며 pH 7.0-9.0에서 안정하였다. Protease는 계면활성제인 Triton X-100(0.1%)에 의해 활성이 2배 촉진되었으나 SDS(0.1%)에 의해서는 강한 저해를 받았다. 2. Bacillus polyfermenticus SCD가 생산하는 protease가 돈육의 연화에 미치는 영향 본 연구는 B. polyfermenticus SCD가 생산하는 protease가 돈육의 연화에 미치는 영향을 조사하고자 실시하였다. B. polyfermenticus SCD가 생산하는 protease의 특성을 조사하고 돈육을 각각의 효소(papain, Aspergillus oryzae와 B. polyfermenticus SCD가 생산하는 protease)가 포함된 용액으로 처리한 후 4℃에서 24, 72, 168시간 동안 저장하며 품질을 측정하였다. B. polyfermenticus SCD가 생산하는 protease는 다른 효소에 비해 활성이 낮았지만 육단백질을 쉽게 분해하였다. 소편화도는 대조구에 비해 처리구에서 높은 값을 나타내었다. 전자현미경으로 관찰한 결과 효소 처리된 육을 168시간 저장했을 때 근육 내의 결합조직이 많이 파괴되어 있었다. 가열감량은 저장기간 동안 증가하였다. 관능평가를 실시했을 때, 효소 처리구가 큰 차이를 나타내지는 않았지만 전체적으로 연도에서는 대조구에 비해 우수한 평가를 받았다. 3. Bacillus polyfermenticus SCD가 생산하는 protease가 육포의 품질에 미치는 영향 본 연구는 B. polyfermenticus SCD가 생산하는 protease가 육포의 품질에 미치는 영향을 조사하고자 실시하였다. 4개의 시료는 Streptomyces griseus와 B. polyfermenticus SCD가 생산하는 효소를 각각 0.005%, 0.01%로 첨가하였으며 텀블링 후 바로 건조한 것과 텀블링 후 24시간이 경과한 뒤 건조한 두 가지 시료로 구분하였다. 모든 처리구에서 돈육육포와 우육육포의 pH에서 유의적인 차이가 나타나지 않았으며, 효소를 처리하였을 때 우육육포의 색에 미치는 영향이 돈육 육포보다 컸다. 효소 처리구들이 대조구에 비해 유의적으로 낮은 경도를 나타내었으나 염지시간에 의해서는 유의적인 차이는 나타나지 않았다. 수분함량은 효소와 염지시간에 따라 영향을 받지 않았으나 수분활성도는 효소 처리구가 낮았다. 돈육육포는 효소 처리 육포가 높은 재수화율을 나타내었으며 우육육포는 텀블링 후 24시간 경과한 뒤 건조했을 때 높은 재수화율을 나타냈다. 관능평가를 실시했을 때, 효소 처리구에서 조직감, 다즙성, 전체적 기호도가 높게 나타났지만 염지시간에 의해서는 큰 차이를 나타내지는 않았다.
This study was carried out to evaluate the characterization of protease produced by Bacillus polyfermenticus SCD and investigate the possibility as a new tenderizer of meat. The results obtained from experimental work were summarized as following; I. Partial Purification and Characterization of Prot...
This study was carried out to evaluate the characterization of protease produced by Bacillus polyfermenticus SCD and investigate the possibility as a new tenderizer of meat. The results obtained from experimental work were summarized as following; I. Partial Purification and Characterization of Protease Produced by Bacillus polyfermenticus SCD An extracellular protease from B. polyfermenticus SCD was purified and characterized. The activity of protease reached to 41.8 units/mL within 48 hr of the fermentation. The ammonium sulfate fractionation and ion exchange chromatography with DEAE Sephadex-A50 column enhanced the specific activity to 2,973.1 units/mg protein achieving 58.4-fold purification. The molecular mass was estimated to be 44 kDa or 28 kDa on SDS-PAGE. The optimum temperature for the activity of this protease was 40-50°C. The protease was highly stable at 50°C. However, the protease was quite unstable at temperature beyond 55°C. The protease was active in the range of pH 7.0-9.0 with the optimum at pH 8.0. The protease was highly stable in the presence of the surfactants like Triton X-100 (0.1%), showing two-fold increases in the proteolytic activity. But, the protease activity was strongly inhibited by SDS (0.1%). II. The Effects of Protease Produced by Bacillus polyfermenticus SCD on the Tenderization of Pork Meat The purpose of this study was to examine the effect of protease from B. polyfermenticus SCD on the tenderization of pork meat. The characterization of B. polyfermenticus SCD protease was done, and pork loin samples were treated in solutions containing different enzymes (papain, proteases from Aspergillus oryzae, B. polyfermenticus SCD), then stored for 24, 72, 168 hr at 4°C. Each treated sample was subjected to quality assessment. B. polyfermenticus SCD protease activity was lower than the other enzymes tested, though it easily hydrolyzed the meat protein. The fragmentation index of the enzyme-treated meats was higher than those of the control ones. The remarkable disruption of the structure of intramuscular connective tissue with the enzyme-treated meat was observed at 168 hr of storage as shown by electron microscopy. Cooking loss increased with increasing storage time. Sensory evaluation did not show differently between meat treated with protease, but overall the tenderness of enzyme-treated meats was higher than those of the control ones. III. The Effects of Protease Produced by Bacillus polyfermenticus SCD on Quality of Jerky The objective of this study was to examine the effects of crude protease from B. polyfermenticus SCD on quality of jerky. Four test samples used various crude proteases. They were added with 0.005 or 0.01% crude protease from Streptomyces griseus or 0.005 or 0.01% crude protease from B. polyfermenticus SCD and two marination methods are: drying immediately after tumbling and drying after tumbling and then held for 24 hr. Neither pork nor beef jerky showed a significant difference in pH among all treatments, and each crude protease was found to have a greater effect on the color of beef jerky than on those of pork jerky. The hardness was significantly lower in pork and beef jerky treated with each crude protease than in the control, however the textural properties of jerky were not significantly different with regard to holding time after marinating. Moisture content was not affected by crude protease addition or holding time after marinating, however, the water activity was lower in jerky treated with crude protease. The rehydration capacity of pork jerky was higher in jerky treated with crude protease, and the rehydration capacity of beef jerky was higher in jerky dried after tumbling and held for 24 hr. Sensory characteristics such as tenderness, juiciness and overall acceptability were higher in jerky treated with crude protease, but were not affected by holding time after marinating.
This study was carried out to evaluate the characterization of protease produced by Bacillus polyfermenticus SCD and investigate the possibility as a new tenderizer of meat. The results obtained from experimental work were summarized as following; I. Partial Purification and Characterization of Protease Produced by Bacillus polyfermenticus SCD An extracellular protease from B. polyfermenticus SCD was purified and characterized. The activity of protease reached to 41.8 units/mL within 48 hr of the fermentation. The ammonium sulfate fractionation and ion exchange chromatography with DEAE Sephadex-A50 column enhanced the specific activity to 2,973.1 units/mg protein achieving 58.4-fold purification. The molecular mass was estimated to be 44 kDa or 28 kDa on SDS-PAGE. The optimum temperature for the activity of this protease was 40-50°C. The protease was highly stable at 50°C. However, the protease was quite unstable at temperature beyond 55°C. The protease was active in the range of pH 7.0-9.0 with the optimum at pH 8.0. The protease was highly stable in the presence of the surfactants like Triton X-100 (0.1%), showing two-fold increases in the proteolytic activity. But, the protease activity was strongly inhibited by SDS (0.1%). II. The Effects of Protease Produced by Bacillus polyfermenticus SCD on the Tenderization of Pork Meat The purpose of this study was to examine the effect of protease from B. polyfermenticus SCD on the tenderization of pork meat. The characterization of B. polyfermenticus SCD protease was done, and pork loin samples were treated in solutions containing different enzymes (papain, proteases from Aspergillus oryzae, B. polyfermenticus SCD), then stored for 24, 72, 168 hr at 4°C. Each treated sample was subjected to quality assessment. B. polyfermenticus SCD protease activity was lower than the other enzymes tested, though it easily hydrolyzed the meat protein. The fragmentation index of the enzyme-treated meats was higher than those of the control ones. The remarkable disruption of the structure of intramuscular connective tissue with the enzyme-treated meat was observed at 168 hr of storage as shown by electron microscopy. Cooking loss increased with increasing storage time. Sensory evaluation did not show differently between meat treated with protease, but overall the tenderness of enzyme-treated meats was higher than those of the control ones. III. The Effects of Protease Produced by Bacillus polyfermenticus SCD on Quality of Jerky The objective of this study was to examine the effects of crude protease from B. polyfermenticus SCD on quality of jerky. Four test samples used various crude proteases. They were added with 0.005 or 0.01% crude protease from Streptomyces griseus or 0.005 or 0.01% crude protease from B. polyfermenticus SCD and two marination methods are: drying immediately after tumbling and drying after tumbling and then held for 24 hr. Neither pork nor beef jerky showed a significant difference in pH among all treatments, and each crude protease was found to have a greater effect on the color of beef jerky than on those of pork jerky. The hardness was significantly lower in pork and beef jerky treated with each crude protease than in the control, however the textural properties of jerky were not significantly different with regard to holding time after marinating. Moisture content was not affected by crude protease addition or holding time after marinating, however, the water activity was lower in jerky treated with crude protease. The rehydration capacity of pork jerky was higher in jerky treated with crude protease, and the rehydration capacity of beef jerky was higher in jerky dried after tumbling and held for 24 hr. Sensory characteristics such as tenderness, juiciness and overall acceptability were higher in jerky treated with crude protease, but were not affected by holding time after marinating.
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