• Title/Summary/Keyword: crude protease

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Persisting antibody reaction in paragonimiasis after praziquantel treatment is elicited mainly by egg antigens

  • Cho, Seung-Yull;Kong, Yoon;Yun, Doo-Hee;Kang, Shin-Yong;Kim, Lee-Soo;Chung, Young-Bae;Yang, Hyun-Jong
    • Parasites, Hosts and Diseases
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    • v.38 no.2
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    • pp.75-84
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    • 2000
  • Antibody responses in serum and cerebrospinal fluid (CSF) samples from patients with active and chronic paragonimiasis and in sera from patients on whom follow-up studies were done after praziquantel treatment were analyzed using antigens of Paragonimus westermani prepared from eggs, metacercariae, juveniles of 4-and 7-week old, adult worms and recombinant protein of 28 kDa cruzipain-like cysteine protease (rPw28CCP). The patient sera/CSFs of active and chronic paragonimiasis revealed strong antibody reactions against the crude extracts of 4-and 7-week old juveniles as well as against those from egg and adult. rPw28CCP also showed specific reaction to the sera with active paragonimiasis. After the treatment, levels of specific antibodies in the sera gradually decreased to negative range in most patients. In some cases with persisting high antibody levels, however, the reactions at 27 kDa egg Protein were sustained throughout the observation period of 34 months. The reactions at 35 and 32 kDa in adult extract and rPw28CCP disappeared rapidly after the treatment. Persistent antibody reactions even after successful treatment are provoked by continuous antigenic challenge from eggs which were not resolved by treatment.

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Effect of Lactic Acid Producing Bacteria on the Performance of Male Crossbred Calves Fed Roughage Based Diet

  • Ramaswami, N.;Chaudhary, L.C.;Agarwal, N.;Kamra, D.N.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.8
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    • pp.1110-1115
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    • 2005
  • To study the effect of feeding lactic acid producing bacteria on the performance of cattle calves, twenty four, day old male crossbred cattle calves (Bos indicus${\times}$Bos taurus), were distributed into two groups of 12 animals each. The animals were fed on calf starter containing wheat bran and green berseem ad libitum and milk as per requirement upto 8 weeks of age. The diet of calves of Group 2 was supplemented with 500 ml culture of Lactobacillus acidophilus-15. Total duration of the experiment was 31 weeks. There was no significant difference in intake and digestibility of dry matter (DM), organic matter (OM), neutral detergent fibre (NDF), acid detergent fibre (ADF) and crude protein (CP) between the groups. The rumen pH, protozoa numbers, concentration of volatile fatty acids (VFA), ammonia nitrogen ($NH_3-N$), trichloroacetic acid precipitable nitrogen (TCA-ppt N) and activity of microbial enzymes (carboxymethylcellulase, xylanase, amylase and protease) were not affected due to probiotic supplementation. Average live weight gain of the calves was improved (about 10%) and feed:gain ratio was reduced (about 5%) in the animals given Lactobacillus culture. The data indicated that crossbred calves could be reared on a diet devoid of cereal grain and addition of Lactobacillus culture in the diet resulted in an added advantage in growth performance of the animals.

Isolation and Culture of Meristotheca papulosa Protoplasts (갈래곰보, Meristotheca papulosa의 원형질체 분리와 배양)

  • 정규화;선상미;조용철;공용근;윤장택
    • Journal of Aquaculture
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    • v.12 no.1
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    • pp.7-14
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    • 1999
  • Protoplasts were isolated from the vegetative thalli of Meristotheca papulosa using several commercial and crude enzymes. The suitable enzyme combination for the protoplast isolation was 4% abalone acetone powder, 4% Macerozyme R-10 and 4% Hemicellulase in the filtered seawater buffered with 50mM MES (pH 6.0) containing 0.6M mannitol and 0.5% potassium dextran sulfate. Yield of protoplast was $107.6{\times}10^4$ cells per gram of fresh thallus. Protoplasts were whitish ovoid in shape and ranged between $7{\mu} m$~ $24{\mu} m$ in diameter. Division of protoplasts was first observed 9 days after culture in $ASP_{12}$ medium, and the germination occurred within 25 days. The addition of Guillard's antibiotics in culture media was harmful to the regeneration of protoplasts.

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Inhibitory effect of Ulmus davidiana Planch extracts on bone resorption mediated by processing of cathepsin K in cultured mouse osteoclasts

  • Park, Jun-Sung;Kim, Kyung-Ho;Jo, Hyun-Seog;Kim, Kap-Sung;Hwang, Min-Seob
    • Journal of Acupuncture Research
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    • v.22 no.2
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    • pp.55-70
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    • 2005
  • Objective: Ulmus davidiana Planch (Ulmaceae) has long been known to have anti-inflammnatory in the traditional Korean medicine. UD has been reported as a good enhancer for bone healing. Methods : In this experiment, we investigate the Inhibitory effects of UD on bone resorption using the bone cells culture. Different concentrations of crude extract of UD were added to mouse bone cells culture. The mitochondria activity of the bone cells after exposure was determined by colorimetric MIT assay. It was demonstrated that UD has potential effects on bone cells culture without any cytotoxicity. The most effective concentration of UD on bone cells were $100\;{\mu}g/ml$. Cathepsin K (Cat K) is the major cysteine protease expressed in osteoclasts and is thought to play a key role in matrix degradation during bone resorption. Results : When mouse long bone cells including osteoclasts and osteoblast were treated with the PI3-Kinase inhibitor, wortmannin (WT), WT prevented the osteoclast-mediated intracellular processing of Cat K. Similarly, treatment of osteoclasts-containing long bone cells with UD extracts prevented the intracellular maturation of Cat K, suggesting that UD may disrupt the intracellular trafficking of pro Cat K. This is similar to that of WT. Since secreted proenzymes have the potential to reenter the cell via mannose-6-phosphate (M6P) receptor, to prevent this possibility, we tested WT and UD in the absence or presence of M6P. Inhibition of Cat K processing by WT or UD was observed in a dose-dependent manner. Furthermore, the addition of M6P resulted in enhanced potency of WT and UD. Conclusion : UD dose-dependently inhibited in vitro bone resorption with a potency similar to that observed for inhibition of Cat K processing.

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Characteristics of Doenjang (Soybean Paste) Fermented with Multiple Starters Including Tetragenococcus halophilus

  • Ji Yeon Yoo;Dong Sin Kim;Tae Jin Kim;Yun Ji Kang;Min Jae Kim;Jeong Hwan Kim
    • Microbiology and Biotechnology Letters
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    • v.51 no.2
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    • pp.147-156
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    • 2023
  • Tetragenococcus halophilus CY54, an isolate from jeotgal, grows best in media with 5% NaCl and can grow at 18% and higher salt concentration. Three different doenjang samples were prepared with multiple starters including T. halophilus CY54. TBZA doenjang was prepared with T. halophilus, Bacillus subtilis, Zygosaccharomyces rouxii and Aspergillus oryzae. BZA doenjang was prepared with the same 3 starters except T. halophilus. KACC doenjang was prepared with a single starter, B. subtilis KACC16750. During 16 weeks of fermentation at 25℃, the viable counts were maintained in the range of 7-8 log CFU/g in all 3 samples. As fermentation progressed, pH decreased and titratable acidity (TA) gradually increased. Crude protein contents decreased slightly. TBZA doenjang showed higher amino-type nitrogen (ANN) and volatile basic nitrogen (VBN) contents, and KACC doenjang showed higher ammonia-type nitrogen (AMN) content. TBZA doenjang showed higher fibrinolytic and protease activity than other doenjang samples. Metabolites analyses by GC/MS showed that doenjang samples were separated from each other by partial least squares-discriminant analysis (PLS-DA) analysis. Seventeen major metabolites involved in the differences between samples were identified and they included organic acids, amino acids, sugars, fatty acids and alcohols. TBZA doenjang showed higher contents for most metabolites responsible for flavor and taste of fermented foods including doenjang. These results showed that T. halophilus could be useful as a starter for doenjang and can improve the product quality by accelerating the fermentation processes.

Studies on the Chemical Compositions and Quality of Red Pepper Paste Brewed with Different Raw Materials (담금원료(原料)에 따른 고추장의 성분(成分)과 품질(品質)에 관(關)한 연구(硏究))

  • Park, Soo-Woong;Park, Yoon-Joong
    • Korean Journal of Agricultural Science
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    • v.6 no.2
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    • pp.205-212
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    • 1979
  • Red pepper paste were prepared by using various raw materials such as rice, glutinous rice, polished barley, polished wheat and corn powder, and their chemical compositions including reducing sugar, amino-N and ethanol were analyzed during the period of aging in order to elucidate effects of these starchy materials on the compositions and qualities of the products. The results obtained are summarized as follows: 1) Koji of these materials were manufactured respectively by inoculating a strain of Asporyzae and their activities of protease and saccharifying amylase were determined. Wheat koji was found to have the highest level of protease activity among the koji, while glutinous rice koji had the strongest amylase activity. 2) Contents of moisture, crude protein, crude fat, and sodium chloride in the red pepper pastes were not changed significantly, however total sugar content was decreased during the period of aging. 3) After 60 days of aging, the highest amount (160mg%) amino nitrogen was detected in the red pepper paste of polished wheat, but higher levels of reducing sugar and ethanol contents were detected in that of glutinous rice. 4) Amino acids in the products were analyzed after 60 days of aging: a) Total 17 amino acids were detected. b) Free amino acid ratio to total amino acid content was approximately 35 percent in average. c) Free glutamic acid was higher than any other free amino acid In the amount and free ratio, but free methionine was lower. d) Lysine content was relatively high, while histidine and glycine were detected in trace. 5) Sensory tests including color, odor and taste on the products resulted in the highest mark for the red pepper paste of glutinous rice but in the lowest mark for that of corn powder.

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Microbiological and Chemical Changes of Complete Feed during Spoilage (배합사료의 부패 동안 발생하는 미생물학적 및 영양학적 변화)

  • Yi, Kwonjung;Yeon, Jae-Sung;Kim, Juhyeon;Kim, Sam Churl;Moon, Hyung-In;Jeon, Che Ok;Lee, Sang Suk;Kim, Dong-Woon;Kim, Soo-Ki
    • Journal of Life Science
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    • v.25 no.10
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    • pp.1148-1155
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    • 2015
  • Commercial complete feeds contain enough nutrients to support animal growth and it is easy to be spoiled under proper temperature and humid conditions. The aim of this study was to investigate microbiological and chemical changes on complete feed for milking cow under open-air exposure with moisture 33% at 30℃ during 15 days. pH decreased 6.29 to 4.66 and water activity decreased gradually 0.99 to 0.95. Bacteria increased 6.2×106~1.6×107 to 2.1×109 CFU/g at 5 days and showed 108 CFU/g until 15 days. Fungi increased 103 CFU/g to 8.0×104 CFU/g. During the processing of spoilage, bacteria such as Acinetobacter oleivorans, Pediococcus acidilactici, Acinetobacter oleivorans, Weissella cibaria, and Methylobacterium komagatae were identified and fungi such as Fusarium sp. and Mucor sp. were also identified. Moisture content increased until 10 days (p<0.01). Crude protein was not changed so much whereas crude fat decreased 6.0% to 5.5% (p<0.01). Crude fiber and crude ash changed 2.0~ 3.0% and 4.5~ 4.8% levels with no significance, respectively. Gross energy was not almost changed at 4,400 kcal/g. During spoilage, lactate and propionate increased whereas acetate was not detected. Protease and lipase activities increased significantly during spoilage (p<0.01). Zearalenone content increased 59.2 μg/kg to 623.8 μg/kg, showing 10.5 times more production. During feed spoilage, pH decreased with microbial growth and various chemical changes were occurred.

Antioxidant activity and physicochemical composition of fermented Vigna angularis using Bacillus subtilis KCCM 11965P (Bacillus subtilis KCCM 11965P를 이용한 붉은팥 발효물의 항산화 활성 및 이화학성분 분석)

  • Jeong, Kyung Ok;Oh, Keun Su;Moon, Kwang Hyun;Kim, Dae Geun;Im, So Yeon;Lee, Eun Ji;Kim, Na Ri;Kim, Wook;Kim, Hae Jin;Lee, Jeong Ho
    • Food Science and Preservation
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    • v.24 no.7
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    • pp.975-982
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    • 2017
  • Health of human intestine has deteriorated due to excessive stress and western diet. In this study, Vigna angularis was fermented by Bacillus subtilis KCCM11965P in order to develop prebiotic resource for improving bowel movement. The contents of ash and crude protein were $3.35{\pm}0.04%$ and $21.1{\pm}0.19%$ respectively. Vigna angularis extract (1, 3, 5%) were incubated with 3% (v/v) Bacillus subtilis KCCM 11965P for 0, 24, 48, and 72 h. Total bacterial numbers showed that the combination of 3% powdered seeds and 72 h incubation time was optimum condition for this experiment. Total polyphenol content increased from $0.18{\pm}0.010mg/mL$ in pre-incubation to $0.23{\pm}0.007mg/mL$ in post-incubation with the condition mentioned above. DPPH radical scavenging activity also increased from $36.1{\pm}6.0%$ to $63.6{\pm}5.2%$. Analysis of protease activity showed $2.69{\pm}0.003unit/mL$ in combination of 5% powdered seeds and 72 h incubation time. Amylase activity increased from $1.0{\pm}0.1unit/mL$ in pre-incubation to $26.0{\pm}0.2unit/mL$ in post-incubation. The analysis of free amino acids after incubation with Bacillus subtilis KCCM 11965P showed that leucine increased from 5.22 mg/L to 67.59 mg/L and tyrosine, one of non-essential amino acid also increased 10.08 mg/L to 259.35 mg/L by incubation with 5% powdered seeds. Most of organic acid were reduced by incubation for 72 h. These results suggest that Vigna angularis could be utilized most as a prebiotic resources.

Activities of the Hydrolytic Enzymes Produced by Plant Pathogenic Fungi, Sclerotium rolfsii, Sclerotinia Sclerotinia and Sclerotiorum, and Helminthosporium sigmoideum var. irregulare (수종의 식물병원균(흰비단병균$\cdot$균핵병균 및 좀검은 균핵병균)이 생산하는 가수분해효소의 활성)

  • Cho B. H.;Kim K.
    • Korean journal of applied entomology
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    • v.16 no.4 s.33
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    • pp.199-208
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    • 1977
  • Activities of various hydrolytic enzymes produced by three plant pathogenic fungi, Sclerotium rolfsii Sacc., Sclerotinia sclerotiorum (Lieb.) deBary and Helminthosporium sigmoideum var. irregulare Crallery et Tullius, were measured. Activties and amounts of the enzymes in mycelia, cultural filtrates, and sclerotia(except of sclerotia of H. sigmoideum var. irregulare) were estimated at various pH levels in order to find out optimal pH for their enzymatic activities. Enzymes such as cellulase (ex), invertase, xylanase, $\beta-amylase$, polymethylgalacturonase, polygalacturonase, phosphatase and protease were estimated. Culture solution for production of enzymes was prepared by adding of 10g, D-glucose, 1.3g $NH_4NO_3,\; 0.5g\; MgSO_4,\;7H_2O,\; and\; 1.0g\; KH_2PO_4$ into 1 liter of potato decoction plus 2ml of micro element solution consisting of 0.2mg. Fe, 0.2mg Zn, and 0.1mg Mn as the sulphates into 1 liter of distilled water. All tested mycelia and cultural filtrates were obtained from the cultures incubarted in previous solution for ten days at $25^{\circ}C$, and sclerotia were harvested from PDA plates of 3. days old, The crude enzyme solutions were prepared according to the method of Miyazaki etal. Ten days after incubation, activities of Cx produced by Scl. sclerotiorum were higher than those of the other fung and each of Cx from three fungi showed different pH optima, such as S. rolfsii and Scl. schlerotiorum in acid side (around pH 3.0), H. sigmoideum var. irregulare in neutral side (around pH 6.3). Invertase activities of S. rolfsii were 20 times higher than those of the other fungi in all samples. All tested fungi, however, showed no significant difference between the enzymatic activities of their cultural filtrate and mycelia and the activities in sclerotia of S. rolfsii and Scl. sclerotiorum were hardly recognized. There were multiple peaks on the xylanase activity curves of three fungi in terms of pH values. High activities of the xylanase were revealed in sclerotia of S. rolfsii and Scl. sclerotiorum, and in mycelia of H. sigmoideum var. irregulare. The highest activities of $\beta-amylase$ were shown both in mycelia and cultural filtrate of H. sigmoideum var. irregulae among the tested fungi, and their optimal pH was 6.2 in both mycelia and cultural filtrate. In the S. rofsii and Sel. sclerotiorum, however, the activities of cultural filtrates were higher than those of the other fungi, and optimal pH was 3.0 and 6.2 for cultural filtrate and both mycelia and sclerotia, respectively. Activities of PMG were high in cultural filtrates of all tested fungi, especially in Scl. sclerotiorum and H. sigmoideum var. irregulare. Mycelia of themalso showed the considerable activities. Optimal pH for enzymatic activities were variable with thekind of fungi or with the samples measured. The highest activities of PG were presented by mycelia of S. rolfsii and Scl. sclerotiorum. $9.l\mu /min.\; and\; 9.5\mu g/min.$, respectively. Optimal pH for activity of PG in mycelia was around 4.5 in S. rolfsii and around 3.0 in Scl. sclerotiorum. Phosphatase of S. rolfsii and Scl. sclerotiorum was more active in acid side (optimal PH3. 5) and that of H. sigmoideum var. irregulare showed one peak each in acid, neutral and alkaline side. But the highest peak was at pH 9.5. Protease of all tested fungi was more active at pH 10.0, especially that of the cultural filtrate of H. sigmoideum var. irregualre.

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A Study on the Preparation of Traditional Andong sikhe with Lactic Acid Bacteria (유산균을 이용한 전통안동식혜의 제조방법에 관한 연구)

  • Choi, Cheong;Son, Gyu-Mok
    • Journal of the Korean Society of Food Culture
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    • v.7 no.3
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    • pp.259-270
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    • 1992
  • The changes in life style today appear many ways. Many housewives turn away from home preparation of the time consuming traditional foods, such as 'Andong sikhe'. The importance, however, of succeeding the traditional cuisines is getting appreciated widely nowadays. This study aimed to investigate the preparation of Andong sikhe by use of pure culture inoculation and the improvement of storage stability by the addition of stabilizers to the product. Lactobacillus delbreuckii was selected for the pure culture inoculation in the fermentation. The changes in chemical composition such as total acidity, sugar content, amino acid and various forms of nitrogen during fermentation were determined. The changes in pH of the product, the enzyme activities and the population of lactic acid bacteria were also followed in the process of fermentation. The Lactobacillus dominated in the beginning of the fermentation but the Streptococcus out numbered the former as the fermentation proceeded. The crude protein content increased up to the 4th day of fermentation but slowly decreased there after. The pH of the product rapidly decreased to 4.2 by the 2nd day of fermentation. The total acidity reached to the 0.38% by the 2nd day of fermentation and kept on increasing slowly during the fermentation. The free sugar consisted of 6 kinds including maltose and one unknown sugar. The amino form nitrogen increased up to 38.5mg% at the 2nd day of fermentation and the product tasted best at this time. The ammonia form nitrogen, water soluble and salt soluble protein decreased during fermentation. Proline and aspartic acid were the two major free amino acids. The free methionine increased while the free lysine decreased in the process of fermentation. The major amino acids of water soluble and salt soluble protein were glutamic acid and aspartic acid. The arginine content of salt soluble protein increased as the fermentation proceeded. Linoleic, palmitic and oleic acid were the three major fatty acids and occupy 90% or more of the total fatty acids. The activities of acid protease and liquefying amylase reached to the maximum at the 4th day of fermentation while those of saccharogenic amylase and lipase reached to the peak at the 2nd day of fermentation.

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