• Title/Summary/Keyword: crude protease

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Changes in Microflora and Enzymes Activities of Traditional Kochujang Prepared with Various Raw Materials (담금원료에 따른 전통식 고추장의 숙성 중 미생물과 효소력의 변화)

  • Shin, Dong-Hwa;Kim, Dong-Han;Choi, Ung;Lim, Mi-Sun;An, Eun-Young
    • Korean Journal of Food Science and Technology
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    • v.29 no.5
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    • pp.901-906
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    • 1997
  • In order to reproduce and improve quality of traditional kochujang, various raw materials were added to prepare kochujang by replacing part of the glutinous rice. Chemical composition, microbial characteristics and enzyme activities were investigated during fermentation. Crude protein and salt contents of kochujang did not change significantly during fermentation, but moisture contents increased linearly. The pH and titratable acidity of kochujang changed little in garlic added group. The viable cell counts of aerobic bacteria and yeasts in the kochujang increased until 60 days of fermentation and then decreased slowly except for the garlic added group in which they increased during the last period of fermentation. Aerobic bacterial count did not show any remarkable differences among the samples and slowly decreased after 60 days of fermentation. The activities of liquefying and saccharifying amylases decreased until 45 days, but increased at 60th day. Acidic protease activities of each group were strong during the initial period, but neutral protease showed the highest activity from the 30 to 45 days of fermentation. Protease activities increased by addition of soy sauce, Chinese matrimony vine and purple sweet potato.

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Effect of Bacillus Strains on the Chungkook-jang Processing - II. Changes of the Components and Enzyme Activities During the Storage of Chungkook-jang - (균주(菌株)를 달리한 청국장 제조(製造)에 관한 연구(硏究) - 제 2 보 : 청국장의 저장중(貯藏中)의 성분(成分) 및 효소력(酵素力)의 변화(變化) -)

  • Suh, Jeong-Sook;Lee, Sang-Gun;Ryu, Myung-Ki
    • Korean Journal of Food Science and Technology
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    • v.14 no.4
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    • pp.309-314
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    • 1982
  • The growth process of the Chungkook-jang that occured by utilizing such traditional methods as Bacillus natto and Bacillus subtilis method has been examined. The results of the experiment in which the changing process of elements during the storage period had been measured are as follows: 1. During the growth period, concerning any change in pH, the storage period had been declined and after 18 days pH rose above 7.0. Salt content was between $5.28{\sim}6.40%$ and Bacillus subtilis fungus showed the highest titrable acidity. 2. Moisture content was between $50.94{\sim}56.74%$ and crude protein content range was $14.44{\sim}18.60%$ indicating irregularity in pattern resulting from the testing equipment groups, whereas crude fat and crude fiber tend to decrease in general. 3. During the storage, total sugar and ethyl alcohol content in all of groups tended to decrease and after 18 days Bacillus subtilis total sugar content was the lowest. 4. Amino nitrogen and water soluble nitrogen content increased with days, but no difference was found between groups. 5. Amylase and protease activity showed irregular pattern with time, but no significant difference between groups was found.

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Resource of Food Waste using Indigenous Bacteria Isolated from Soils (토양으로부터 분리한 토착유효미생물을 이용한 음식물쓰레기의 자원화)

  • Lee, Sang-Woo;Ham, Sun Nyeoo;Shin, Taek-Soo;Kim, Hye-Kyung;Yeon, Ik-Jun;Kim, Kawng-Yul
    • Journal of Korean Society of Environmental Engineers
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    • v.31 no.1
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    • pp.35-41
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    • 2009
  • This study was conducted to investigate feasibility of feedstuff for animal using food waste by fermentation mechanism of indigenous microorganism. To achieve this purpose, indigenous bacteria was isolated from soils to use as an inoculant. Enzyme test was performed to verify activity of amylase, protease and lipase using isolated bacteria. Bacteria(H1, D1), which vigorously express the enzyme activity, was selected and used in the fermentation experiments of food waste. From the analysis of 16s rDNA sequencing, H1 and D1 were identified as Bacillus subtilis and Paenibacillus polymyxa, respectively. In the fermentation experiment, food waste was mixed with rice bran and popped rice to control moisture and nutrient content. Isolated bacteria(B. subtilis and P. polymyxa) was used as an inoculant. From the measured data such as temperature, pH and ORP, it can be verified that food waste adding the indigenous bacteria was effectively fermented. From the nutritional analysis of manufactured feedstuff, it showed that the contents of crude protein, crude fat and crude fiber were enough to use as feedstuff for animal. In addition, harmful components such as Pb, Hg, Cd, aflatoxin and salmonella concentration were not exceeded permitted standards. Therefore, fermented food waste using indigenous bacteria can be used as feedstuff.

Comparison of Digestive Function Among Rabbits, Guinea-Pigs, Rats and Hamsters. II. Digestive Enzymes and Hindgut Fermentation

  • Yu, Bi;Chiou, Peter Wen-Shyg;Kuo, Chung-Yi
    • Asian-Australasian Journal of Animal Sciences
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    • v.13 no.11
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    • pp.1508-1513
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    • 2000
  • The aim of this trial was to study the response of laboratory animals including omnivores (rats) and herbivores (rabbits, guinea pigs and hamsters) to the same level of dietary fiber on their digestive enzymes and hindgut fermentation. Ten weanling animals of each of four species, rabbits, guinea-pigs, rats and Syrian hamster, were fed a basal diet of 18% crude protein and 10% crude fiber for six weeks. The digesta and tissue of each intestinal segment were collected to measure the activity of digestive enzymes. Rabbits contained the highest secreted pepsin activity in the stomach, whereas rats contained the highest protease and ${\alpha}-amylase$ activity in the small intestine, and lower fibrous hydrolases in the hindgut than rabbits, guinea pigs and hamsters. The total VFA productions in the caecum and colon were highest in rats, followed by hamsters and rabbits, while the guinea pigs contained the lowest VFA and a different pattern of VFA molar ratio from the other laboratory animals. The degree of hindgut fermentation in these laboratory animals was in reverse to the trend for their fiber digestion.

IgG antibody responses in early experimental sparganosis and IgG subclass responses in human sparganosis

  • Chung, Young-Bae;Kong, Yoon;Yang, Hyun-Jong;Cho, Seung-Yull
    • Parasites, Hosts and Diseases
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    • v.38 no.3
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    • pp.145-150
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    • 2000
  • Antigenic components in the crude extracts of Spirometra mansoni plerocercoid were analyzed in early experimental infections and in IgG subclass observed in clinical sparganosis. By IgG immunoblot, sera obtained serially from experimental mice, fed 5 spargana each, were reacted with the crude extracts. Protein bands at 36-26 kDa and 103 kDa showed positive reactions since two weeks after infection. In a differential immunoblot, in which a monospecific antibody against sparganum chymase at 36 kDa was pre-treated, the reactions at 36-26 kDa disappeared, indicating that the sparganum chymase and its degradation products invoked IgG antibody reactions. When 69 patients sera of human sparganosis were examined for their IgG subclass responses, IgG4 levels showed the highest reaction which was followed by IgG 1 The IgG4 antibody also reacted mainly with 36-31 kDa protease. These results indicate that 36 kDa chymase of 5. nansoni plerocercoid is the main antigenic component inducing Ige antibody response in early stage of experimental sparganosis and for specific IgG subclass reactions in human sparganosis.

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Development and Fractionation of Proteolytic Enzymes from an Inedible Seafood Product Distribution and fractionation of proteolytic enzymes (수산동물의 비가식 부산물을 이용한 단백질분해효소의 분획 및 효소제제의 개발 단백질분해효소의 분포 및 분획)

  • HEU Min-Soo;AHN Sam-Hwan
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.32 no.4
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    • pp.458-465
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    • 1999
  • Distribution of the proteolytic activities of crude pretense extracted from the viscera of ten kinds of fish was examined. Their proteolytic activities on proteinous substrates (azocasein, hemoglobin, and casein) from the viscera of anchovy, bastard flatfish, mackerel and red sea bream were higher than those of other fishes, and the crude pretenses were further fractoinated with acetone or ammonium sulfate. Optimum concentrations for pretenses fractionation were $0\~55\%$ for acetone and $30\~70\%$ for ammonium sulfate. The fractionated viscera pretense of mackerel showed the highest proteolytic activity among four kinds of fishes. Activities of cathepsin D- and pepsin-like enzymes at pH 3.0, cathepsin L-, B-, H- and G-like enzyme at pH 6,0, and Hypsin- and chymotrypsin- like enzymes (pH 8.0) were detected in the fractionated viscera pretense, whereas activities of cathepsin L- and chymoeypsin-like enzyme were observed in commercial pretenses. Proteolytic activities of Alcalase, Protamex, and Aroase AP-10 for azocasein were slightly higher than the fractionated viscera pretenses, but their amidolytic activities at pH 6.0 and 8.0 toward synthetic substrates were lower than counterpart. The fractionated pretenses from fish viscera would be utilized as commercial pretenses.

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Production and Separation of Anti-hypertensive Peptide during Chunggugjang Fermentation with Bacillus subtilis CH-1023 (청국장 발효과정 중 항고혈압성 peptide의 생산 및 분리)

  • Cha, Woen-Suep;Bok, Su-Kyung;Kim, Myoung-Uk;Chun, Sung-Sook;Choi, Ung-Kyu;Cho, Young-Je
    • Applied Biological Chemistry
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    • v.43 no.4
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    • pp.247-252
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    • 2000
  • As functionality investigation of Korean traditional soybean fermentation foods, an antihypertensive peptide was separated during Chunggugjang fermentation by Bacillus subtilis CH-1023 and investigated inhibitory effect against angiotensin converting enzyme. After incubation at $20^{\circ}C,\;30^{\circ}C,\;40^{\circ}C,\;50^{\circ}C,\;60^{\circ}C$ for the $0{\sim}72$ hrs, protein content, protease activity and angiotensin converting enzyme inhibitory rate were determined. The protein content and protease activity were increased and reached maximum at 60 hrs fermentation with $40^{\circ}C$ and decreased after the 60 hrs fermentation. The optimum condition for antihypertensive peptide from Chunggugjang was appeared for 60 hrs at $40^{\circ}C$. Crude extract of Chunggugjang was partially purified by Amicon YM-3 membrane filtration and Sephadex G-10, G-25 gel filtration. The purified peptide showed inhibitory rate of 94.3% with 0.5 mg peptide content. The most prominent amino acid composition of the peptide from Chunggugjang was alanine, followed by phenylalanine, histidine.

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Production and Separation of Angiotension Converting Enzyme Inhibitor during Natto Fermentation (납두 발효과정 중 Angiotensin Converting Enzyme 저해물질의 생성 및 분리)

  • Cho, Young-Je;Cha, Woen-Suep;Bok, Su-Kyung;Kim, Myung-Uk;Chun, Sung-Sook;Choi, Ung-Kyu;Kim, Soon-Hee;Park, Kyung-Sook
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.29 no.4
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    • pp.737-742
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    • 2000
  • As functionality investigation of a soybean fermentation food, a angiotensin converting enzyme inhibitory peptide was separated during natto fermentation by Bacillus natto and inhibitory effect was investigated. After incubation at each 2$0^{\circ}C$, 3$0^{\circ}C$, 4$0^{\circ}C$, 5$0^{\circ}C$, 6$0^{\circ}C$ for the 0~72 hr, protein content, protease activity and angiotensin converting enzyme inhibition were determined. The protein content and protease activity were increased and reached maximum at 60 hr fermentation with 4$0^{\circ}C$ and decreased after the 60 hr fermentation during natto fermentation. The optimum condition for angiotensin converting enzyme inhibitors was appeared at fermentation for 60 hr at 4$0^{\circ}C$. Crude extract of natto was partially purified by Amicon membrane YM-3 and Sephadex G-10, G-25 gel filtration, stepwise. The inhibitory rate was increased in a concentration dependent manner, espcially the most potent activity about 74.74% at 1.0 mg peptide content. The most prominent amino acid of the peptide from natto was alanine, followed by phenylalnine, histidine.

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Isolation and Purification of Fibrinolytic Enzyme of Edible Mushroom, Sarcodon aspratus(Berk.)S. Ito (능이버섯으로부터 Fibrin 분해활성이 있는 단백질의 분리 및 정제)

  • 이종호;양정례;정청송;김희숙;조재선
    • Journal of Life Science
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    • v.11 no.6
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    • pp.561-567
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    • 2001
  • To isolate and purify fibrinolytic active substance from Sarcodon aspratus(N $H_4$)$_2$S $O_4$ precipitation, DE52 anion exchange column chromatography, Sephacryl-S 200gel filtration chromatography and Mono S cation FPLC were carried out and the characterizations of the purified enzyme were investigated. The bound active fraction on DE52 anion exchange column chromatography were eluted with 0.2 M NaCI and the fibrionlytic enzyme was purified after following Sephacryl-S200 gel fitration chromatography and Mono S cation EPLC. The specific activity of purified enzyme was 55.2 U/mg protein and increased 11.3 fold comparing crude extract and the yield was 49.5%. 12% SDS-PAGE electrophoresis and gel filtration chromatography revealed that Sarcodon aspratus fibrionloytic enzyme was highly purified and had 29.300 Da molecular weight. Enzyme activity of the purified fibrinolytic enzyme from Sarcodon aspratus was increased on higher pH and was stable until pH 10.5. On temperature dependent stability, the enzyme activity was decrease sharply but remained 25% relative activity on 8$0^{\circ}C$. This enzyme activity was inhibited by heavy metal ion, C $U^{2+}$ and $Co^{3+}$ with 68% and 38%, respectively. And also, the enzyme activity was inhibited with $Ca^{2+}$ chelator EDTA and serine protease inhibitor PMSF. These results from this study suggested that the fibrinolycit enzyme from Sarcodon aspratus is a serine protease and the enzyme activity was increased by $Ca^{2+}$ or $Mg^{2+}$ ion.n.ion.n.

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Changes of RuBisCO Content and Protease Activity during the Life Span of Tobacco Leaf (담배잎의 일생에 있어서 RuBisCO 함량과 Protease활성의 변동)

  • 이학수
    • Journal of the Korean Society of Tobacco Science
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    • v.15 no.1
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    • pp.63-74
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    • 1993
  • Changes in the amount of ribulose 1, 5-bisphosphate carboxylase/oygenase(=RuBisCO) protein, namely fraction I protein, and the protease activity were determined in the 10th leaf of tobacco(Nicotiana tabacum, var. Ky-57) from 10 days after emergence through senescence at 5 days interval. The amount of RuBisCO per deveined leaf rapidly increased during the early growing season, reached a maximal quantity at the around 20 days after leaf emergence, when the leaf has gone through its most rapid expansion, and began gradually to decrease till 30 days after leaf emergence, thereafter significantly declined to 45 days that the leaf has been dried up partly. The pattern of the ratio of RuBisCO protein to soluble protein in quantity changed similar to that of RuBisCO contents in a leaf, that was 43%, 60%, and 21% at the around 10 days, 20 days, and 45 days, respectively. And RuBisCO contents was linearly correlated with the concentration of chlorophyll(r=0.98) throughout the life span of the leaf. So, it was assumed that the leaf color can be a useful indicator for judging whether RuBisCO contents higher or not in tobacco leaves without homogenization. On the other hand, the protease activities for degradation of casein were assayed at pH 5.5. 7.0. and 8.5 with crude extracts desalted on Sephadex G-25. The highest caseolytic activity was found at pH 5.5 throughout the life sawn of the leaf. Also, the activity at 5.5 became gradually to increase from 30 days after leaf emergence, when RuBisCO protein had became to disappear and remarkably increased in the last stage of senescence, although nitrogen contents of the leaf had reached low levels. The caseolytic activity at pH 5.5 was in negative correlation with RuBisCO contents throughout the life span of the leaf, but not in lineality between them. In other words, the caseolytic activity increased in a rapid exponential manner when RuBisCO contents had reached some low levels. These results showed that the leaf age, namely harvesting time, is a very important factor for the production of the tobacco leaf containing higher RuBisCO protein. It was concluded that the practical harvesting time is between 20 days and 30 days after the leaf emergence from the present results.

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