• 한국미생물·생명공학회지
• /
• 제1권2호
• /
• pp.99-104
• /
• 1973

A study on the active center of the acid protease from Penicillium sp. was conducted, and also the milk clotting activity of acid prorease was measured. 1. PCMB failed to influence the proteolytic activity of acid protease, indicating that a reactive sulfhydryl group is not required for the enzymatic activity. 2. $\varepsilon$-amino caproic acid did not show any inhibitory effect on tile proteolytic activity of acid protease. 3. Also 2, 4-dinitro phenol did not show any inhibitory effect on the enzyme activity. 4. Acid protease from Penicillium sp. showed a strong milk clotting activity in the presence of Ca ion. 5. This enzyme had a strong proteolytic activity on various substrate, such as casein, denatured hemoglobin, ovalbumin, denatured bovine muscle protein, denatured percine muscle protein and denatured chicken muscle protein.

• 유기물자원화
• /
• 제6권1호
• /
• pp.1-12
• /
• 1998

피혁제조시 발생되는 크롬을 함유한 피혁 고형폐기물인 shaving scrap의 단백질 자원화 가능성을 검토하기 위하여 MgO를 기본으로 하여 alkaline inducing agents 및 alkaline proteolytic enzymes을 혼용처리하여 shaving scrap으로 부터 회수한 가수분해 단백질의 용해도, 무기성분 함량, 분자량분포 등을 비교 검토함으로서 최적 가수분해 조건 및 액체비료의 원료로 활용하기 위한 저분자 단백질의 회수방안을 조사한 결과는 다음과 같다. Alkaline inducing agents의 혼용처리에 의한 shaving scrap의 가수분해 실험결과 7% MgO를 기본으로 하여 alkaline inducing agents 종류에 따라 65~85% 범위로 용해도 차이가 뚜렷하였으며, 가수분해되는 정도는 NaOH>$Ca(OH)_2$>KOH순으로 나타났으며, 획득된 hydrolyzed protein의 평균분자량은 NaOH처리시 약 10 KD, $Ca(OH)_2$ 처리시 약 40 KD, KOH처리시 약 80 KD이었으며, 크롬함유량은 약 15 ppm이었다. Alkaline proteolytic enzymes의 혼용처리에 의한 shaving scrap의 가수분해 실험결과 alkaline proteolytic enzymes 종류에 따라 Alcalase>Esperase>Savinase순으로 용해도 차이를 보였으며, 0.5% Alcalase의 처리에 의해 용해도 85%수준, 평균분자량 1 KD 미만, 크롬 함유량 10ppm 이하인 저분자 형태의 hydrolyzed protein을 획득할 수 있었다.

• Animal Bioscience
• /
• 제34권11호
• /
• pp.1859-1869
• /
• 2021

Objective: The effects of a crude protease extracted from Cordyceps militaris (CM) mushrooms on the postmortem tenderization mechanism and quality improvement in spent hen breast were investigated. Methods: Different percentages of the crude protease extracted from CM mushrooms were introduced to spent hen breast via spray marination, and its effects on tenderness-related indexes and proteolytic enzymes were compared to papain. Results: The results indicated that there was a possible improvement by the protease extracted from CM mushroom through the upregulation of endogenous proteolytic enzymes involved in the calpain system, cathepsin-B, and caspase-3 coupled with its nucleotide-specific impact. However, the effect of the protease extracted from CM mushroom was likely dose-dependent, with significant improvements at a minimum level of 4%. Marination with the protease extracted from CM mushroom at this level led to increased protein solubility and an increased myofibrillar fragmentation index. The sarcoplasmic protein and collagen contents seemed to be less affected by the protease extracted from CM mushroom, indicating that substrate hydrolysis was limited to myofibrillar protein. Furthermore the protease extracted from CM mushroom intensified meat product taste due to increasing the inosinic acid content, a highly effective salt that provides umami taste. Conclusion: The synergistic results of the proteolytic activity and nucleotide-specific effects following treatments suggest that the exogenous protease derived from CM mushroom has the potential for improving the texture of spent hen breast.

• 한국미생물·생명공학회지
• /
• 제23권3호
• /
• pp.322-328
• /
• 1995

A thermophilic bacteria showing proteolytic activity against defatted soybean was isolated from soil. It was identified as Bacillus amyloliquefaciens based on its morphological and physiological characteristics. The Bacillus amyloliquefaciens NS 15-4 was cultivated at 50$\circ$C by rotary shaking in a medium containing defatted soybean. An extracellular protease from this strain was purified to homogeneity by ammonium sulfate precipitation, ion exchange, and hydrophobic interaction chromatographies. The molecular weight of the enzyme was estimated to be approximately 30,000 by SDS-PAGE and the N-terminal amino acid sequence of the enzyme was turned out to be AQSVPYGISQIKAPA. The optimum temperature and pH for the enzyme reaction were 60$\circ$C and 11, respectively, and its thermostability was increased by the addition of calcium ion. The enzyme was inactivated by phenylmethylsulfonylfluoride, suggesting it be a serine protease. Comparing with other commercial proteases, the enzyme showed relatively high proteolytic activity against defatted soybean, a water-insoluble protein substrate.

• 한국식품영양과학회지
• /
• 제23권5호
• /
• pp.856-862
• /
• 1994

In salt-preserved foods of every kinds, it was examined the growth condition of halophilic bacteria that induced a change of colour, taste, nutritive substance, a production condition of enzyme and a character of crude enzyme. Used bacteria is H. halobium a kind of extremely halophilic bacteria, and the required of optimum culture needed a quite long time of crude enzyme production is 168 hours. Optimum pH is about 7-7.5, so the traditional food of such neutrality pH as soybean paste and soy sauce particularly come into trouble because the growth can flourish in neutrality or alkaliescence, and the crude enzyme also appeared that best activation between pH 6 and pH 8. The optimum temperature is about 37$^{\circ}C$, the optimum temperature of enzyme is about 40 $^{\circ}C$ and the temperature stability is settled for 15 minutes and it is completely inactivated at 10 minutes. In the influence of each metal ion, Fe++ and Mn++ a stimulated the growth of H.halobium and the activation of enzyme, Cu++ and Zn++ were identified that made the growth and the activation of enzyme inhibit.

• Asian-Australasian Journal of Animal Sciences
• /
• 제16권6호
• /
• pp.884-888
• /
• 2003

Proteolytic enzymes are used for meat tenderization, an important process with regard to consumer preference. The proteolytic enzyme, IVRIN was isolated from the plant Cucumis pubescens W and its effect on physico-chemical properties and lipid profile of thigh and breast muscle of culled, desi and broiler birds was studied. Fifty-gram meat was treated with IVRIN containing 32.5 mg enzyme protein at $60^{\circ}C$ for 20 min. The pH of IVRIN treated meat was decreased significantly (p<0.01) and the effect was more pronounced in breast than thigh muscle. The water holding capacity (WHC) was increased significantly (p<0.01) in broiler as compared to desi and culled bird, and in breast compared to thigh muscle. IVRIN failed to produce any impact on muscle fiber diameter (MFD). The MFD of desi was significantly higher (p<0.01) than broiler and culled birds. The total lipid concentration in thigh and breast muscle of desi was lower (p<0.01) than broiler and culled birds, latter being similar in this respect. The cholesterol content was lower (p<0.01) in breast than thigh muscle, in broiler than desi and culled and in IVRIN treated than untreated meat samples. The phospholipid concentration was unaffected by IVRIN. Broiler and culled birds exhibited more phospholipid content than desi birds.

• 한국어병학회지
• /
• 제21권3호
• /
• pp.247-257
• /
• 2008

To understand the activity of non-specific defence factors in cultured Sebastes, the antibacterial effect of the serum, skin mucus and homogenate of various organs from cultured oblong rockfish (Sebastes oblongus) and rockfish(Sebastes schlegeli) against pathogenic bacteria, Aeromonas hydrophila, Edwardsiella tarda, Vibrio anguillarum, and Streptococcus sp. was compared with that of flounder(Paralichthys olivaceus) and seabass(Leteolabrax japonicus). And the activities of proteolytic enzyme, chitinolytic enzyme and haemolycin as non-specific defence factor were investigated on the oblong rockfish and rockfish. Samples from oblong rockfish showed the highest antibacterial activity by lysoplate assay on agar plate mixed with pathogens, followed in descending order by rockfish, seabass, and flounder. Turbidimetric assay was carried to evaluate the lysozyme activity of fish samples against lyophilized cells of Micrococcus lysodeiktikus. The serum, kidney, liver, stomach, intestine and eyeball of oblong rockfish and the mucus and gill of rockfish appeared to have the highest lysozyme activity among the fish strains investigated. All samples except skin mucus, liver, and eyeball of oblong rockfish and rockfish showed proteolytic enzyme activity. Chitinolytic enzyme activity was showed in random sampling and haemolytic activity was remarkable in oblong rockfish. Therefore, Sebastes strain was proved to have effective defense mechanisms based on the antibacterial activities, and lysozyme, proteolytic enzyme, chitinolytic enzyme, and haemolycin were considered to act as the non-specific defence factor of Sebastes.

• 한국수산과학회지
• /
• 제16권2호
• /
• pp.147-153
• /
• 1983

To check the differences of the digestive enzymes by the bait habits and the proteolytic activities of the fissile extracts from the fish, omnivorous filefish (Navodon modestus), carnivorous cat shark (Scilliorhinus tarazame) and bloodsucking hag fish (Eptatretus burgeri) were sampled for this experiment. The activity of crude alkaline protease extracted from the muscle and the internal organs of the samples was determined with casein as substrate. The activity of the proteolytic enzymes showed remarkable differences by the organs of the fish. The optimum condition of the pretenses from the muscle revealed in range of pH 7.8-8.3, at $60-65^{\circ}C$, while those of the enzymes from the internal organs were at about pH 8.2, $45-55^{\circ}C$, but those of hag fish were at about pH 6.7, $45-55^{\circ}C$. The proteolytic activity of the enzyme of alimentary canal in filefish and in hag fish was 57 and 11 times stronger than that of muscle, respectively. The crude enzyme from the alimentary canal of file fish showed the strongest proteolytic activity in samples submitted and that of cat shark was the lowest. The activity of pancreatic alkaline protease in cat shark was 50 fold higher than that of muscle alkaline protease in the fish.

• BMB Reports
• /
• 제28권2호
• /
• pp.138-142
• /
• 1995

An anticoagulant/fibrinolytic protease was purified to homogeneity from the earthworm Lumbricus rubellus. The protein was a single chain glycoprotein of 32 kDa that exhibited strong proteolytic activity on human thrombin and fibrin clots. Proteolytic degradation of these plasma proteins by the purified enzyme occurred at a neutral pH range. Among several human plasma proteins tested as possible substrates for the protease reaction, the 32 kDa enzyme specifically hydrolyzed both thrombin and fibrin polymers without affecting other proteins, such as serum albumin, immunoglobulin, and hemoglobin. Treatment of the purified enzyme at neutral pH with either phenylmethylsulfonylfluoride or soybean trypsin inhibitor resulted in a loss of catalytic activity. The enzyme hydrolyzed the chromogenic substrate H-D-Phe-L-Pipecolyl-L-Arg-p-nitroanilide with a $K_m$ value of 1.1 ${\mu}M$ at a neutral pH. These results suggest that the anticoagulant/fibrinolytic enzyme from Lumbricus rubellus is a member of the serine protease family having a trypsin-like active site, and one of the potential clevage sites for the enzyme is the carbonyl side of arginine residues in polypeptide chains.

• Mycobiology
• /
• 제33권2호
• /
• pp.84-89
• /
• 2005

Five types of agricultural wastes were used for the production of xylanolytic enzyme by Aspergillus flavus K-03. All wastes materials supported high levels of xylanase and ${\beta}-xylosidase$ production. A high level of proteolytic activity was observed in barley and rice bran cultures, while only a weak proteolytic activity was detected in corn cob, barley and rice straw cultures. Maximum production of xylanase was achieved in basal liquid medium containing rice barn as carbon source for 5 days of culture at pH 6.5 and $25^{\circ}C$. The xylanolytic enzyme of A. flavus K-03 showed low thermostability. The times required for 50% reduction of the initial enzyme activity were 90 min at $40^{\circ}C$, 13 min at $50^{\circ}C$, and 3 min at $60^{\circ}C$. Xylanolytic activity showed the highest level at pH $5.5{\sim}10.5$ and more than 70% of the original activity was retained at pH 6.5 and 7.0. The higher stability of xylanolytic enzymes in the broad range of alkaline pH is useful for utilization of the enzymes in industrial process requiring in alkaline conditions. Moreover, the highest production of xylanolytic enzyme was obtained when 0.5% of rice bran was supplied in basal liquid medium. SDS-PAGE analysis revealed a single xylanase band of approximately 28.5 kDa from the culture filtrates.