• Title/Summary/Keyword: protease treatment

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A Study on the Thermal Characterization of Barley ${\beta}-Glucan$ [mixed-linked $(1-3),(1-4)-{\beta}-D-Glucan$] by Differential Scanning Calorimetry (DSC에 의한 보리 ${\beta}-Glucan$ [mixed-linked$(1-3),(1-4)-{\beta}-D-Glucan$의 열적 특성에 관한 연구)

  • Cha, Hee-Sook;Kim, Mi-Ok;Koo, Sung-Ja
    • Korean Journal of Food Science and Technology
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    • v.25 no.1
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    • pp.22-27
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    • 1993
  • Crude ${\beta}-glucan$ extracted from Barley was purified by stepwide enzyme treatment with thermostable ${\alpha}-amylase$, amyloglucosidase and protease. The thermal properties of Barley ${\beta}-glucan$ were investigated by Differential Scanning Calorimetry. Three endotherms have been observed on DSC thermograms of Barley ${\beta}-glucan$. The first endotherm which produced the gelatinization phenomena commonly observed in Barley ${\beta}-glucan$ became the focus of this study. The temperature range and the enthalpy of gelation exhibited maximum values with increasing concentration of Barley ${\beta}-glucan$. Gelating Barley ${\beta}-glucan$ registered an enthalpy of approximately 0.23 cal/g and exhibited onset temperature (To), peak temperature (Tp) and conclusion temperature (Tc) of $48.8^{\circ}C,\;61.2^{\circ}C\;and\;78.5^{\circ}C$ respectively. The temperature and enthalpy of gelatinizing Barley ${\beta}-glucan$ at both alkali and acid conditions were lower than those at pH 7. With salt present, the Tp and Tc of gelating Barley ${\beta}-glucan$ produced lower temperatures than in conditions where salt was absent, and the enthalpy abruptly decreased. However, increasing salt concentrations did not affect the gelation temperature and the enthalpy of Barley ${\beta}-glucan$. The 'true melting' temperature of Barley ${\beta}-glucan$ was near $184^{\circ}C$ and the melting enthalpy was approximately 34.6 cal/g. The Barley ${\beta}-glucan$ decomposition temperature was in the range of $316^{\circ}C{\sim}346^{\circ}C$.

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Survey of Genes Responsive to Long-Term Heat Stress Using a cDNA Microarray Analysis in Mud Loach (Misgurnus mizolepis) Liver (장기 고온 스트레스에 대한 미꾸라지(Misgurnus mizolepis) 간 조직 내 유전자 발현 반응의 cDNA microarray 분석)

  • Cho, Young Sun;Lee, Sang Yoon;Noh, Choong Hwan;Nam, Yoon Kwon;Kim, Dong Soo
    • Korean Journal of Ichthyology
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    • v.18 no.2
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    • pp.65-77
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    • 2006
  • Gene transcripts potentially responsive to the heat stress were surveyed by cDNA microarray analysis in mud loach (Misgurnus mizolepis). Transcriptional profiles of hepatic tissue in the fish exposed to either $23^{\circ}C$ or $32^{\circ}C$ for 4 weeks were compared each other by 3 replicated hybridization assays using 1,124 unigene clones selected from mud loach liver expressed sequence tags (ESTs). A total of 93 clones showed the substantially increased mRNA levels (>2-fold) in $32^{\circ}C$-exposed group when compared in $23^{\circ}C$control group. It includes various enzymes and proteins involved in energy pathway, protease/protein metabolisms, immune/antioxidant functions, cytoskeleton/cell structure, transport and/or signal transduction. Maximum level of increase was up to 15-fold relative to $23^{\circ}C$ treatment. Heat exposure also resulted in the significant decrease (less than 50% relative to $23^{\circ}C$-exposed fish) of the transcriptional activities in 85 genes. Besides the above categories, yolk protein (vitellogenin) and ribosomal proteins were notably down regulated in the fish exposed to heat stress. A number of novel gene transcripts were also detected in both up-regulated and down-regulated groups.

Production of Phenyl Lactic Acid (PLA) by Lactic Acid Bacteria and its Antifungal Effect

  • Song, June-Seob;Jang, Joo-Yeon;Han, Chang-Hoon;Yoon, Min-Ho
    • Korean Journal of Soil Science and Fertilizer
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    • v.48 no.2
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    • pp.125-131
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    • 2015
  • Phenyllactic acid (PLA) which is known as antimicrobial compound can be synthesized through the reduction of phenylpyruvic acid (PPA) by lactate dehydrogenase (LDH) of lactic acid bacteria (LAB). LAB producing PLA was isolated from Korea Kimchi and identified to Lactobacillus plantarum SJ21 by 16 rRNA gene sequence analysis. Cell-free supernatant (CFS) from L. plantarum SJ21 was assessed for both the capability to produce the antimicrobial compound PLA and the antifungal activity against four fungal pathogens (Rhizoctonia solani, Aspergillus oryzae, Botrytis cinerea, and Collectotricum aculatum). PLA concentration was investigated to be 3.23mM in CFS when L. plantarum SJ21 was grown in MRS broth containing 5mM PPA for 16 h. PLA production also could be promoted by the supplement of PPA and phenylalanine in MRS broth, but inhibited by the supplement of 4-hydroxyphenylpyruvic acid and tyrosine as precursors. Antifungal activity demonstrated that all fungal pathogens were sensitive to 5% CFS (v/v) of L. plantarum SJ21 with average growth inhibitions ranging from 27.32% to 69.05% (p<0.005), in which R. solani was the most sensitive to 69.05% and followed by B. cinerea, C. aculatum, and A. oryzae. The minimum inhibitory concentration (MIC) for commercial PLA was also investigated to show the same trend in the range from $0.35mg\;mL^{-1}$ (2.11 mM) to $0.7mg\;mL^{-1}$ (4.21 mM) at pH 4.0. The inhibition ability of CFS against the pathogens was not affected by heating or protease treatment. However, pH modification in CFS to 6.5 caused an extreme reduction in their antifungal activity. These results may indicate that antifungal activities in CFS were caused by acidic compounds like PLA or organic acids rather than proteins or peptides molecules.

Gentiopicroside Ameliorates the Progression from Hepatic Steatosis to Fibrosis Induced by Chronic Alcohol Intake

  • Yang, Hong-Xu;Shang, Yue;Jin, Quan;Wu, Yan-Ling;Liu, Jian;Qiao, Chun-Ying;Zhan, Zi-Ying;Ye, Huan;Nan, Ji-Xing;Lian, Li-Hua
    • Biomolecules & Therapeutics
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    • v.28 no.4
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    • pp.320-327
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    • 2020
  • In current study, we aimed to investigate whether the gentiopicroside (GPS) derived from Gentiana manshurica Kitagawa could block the progression of alcoholic hepatic steatosis to fibrosis induced by chronic ethanol intake. C57BL/6 mice were fed an ethanol-containing Lieber-DeCarli diet for 4 weeks. LX-2 human hepatic stellate cells were treated with GPS 1 h prior to transforming growth factor-β (TGF-β) stimulation, and murine hepatocyte AML12 cells were pretreated by GPS 1 h prior to ethanol treatment. GPS inhibited the expression of type I collagen (collagen I), α-smooth muscle actin (α-SMA) and tissue inhibitor of metal protease 1 in ethanol-fed mouse livers with mild fibrosis. In addition, the imbalanced lipid metabolism induced by chronic ethanol-feeding was ameliorated by GPS pretreatment, characterized by the modulation of lipid accumulation. Consistently, GPS inhibited the expression of collagen I and α-SMA in LX-2 cells stimulated by TGF-β. Inhibition of lipid synthesis and promotion of oxidation by GPS were also confirmed in ethanol-treated AML12 cells. GPS could prevent hepatic steatosis advancing to the inception of a mild fibrosis caused by chronic alcohol exposure, suggesting GPS might be a promising therapy for targeting the early stage of alcoholic liver disease.

The Use of Nafamostat Mesilate as an Anticoagulant during Continuous Renal Replacement Therapy for Children with a High Risk of Bleeding (출혈성 경향이 높은 소아환자의 지속성 신대체 요법시 사용되는 항응고제로서 Nafamostat mesilate의 사용)

  • Lee, Sang Taek;Cho, Heeyeon
    • Childhood Kidney Diseases
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    • v.18 no.2
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    • pp.98-105
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    • 2014
  • Purpose: Nafamostat mesilate (NM), a synthetic serine protease inhibitor, has been investigated as an anticoagulant for adult patients with a high risk of bleeding, who need chronic renal replacement therapy (CRRT). However, little is known about the use of NM as an anticoagulant in pediatric CRRT. The aim of this study was to evaluate the ideal dosage, efficacy, and safety of NM in pediatric CRRT. Methods: We conducted a retrospective study of 40 pediatric patients who had undergone at least 24 h of venovenous CRRTs between January 2011 and October 2013. We divided the patients according to risk of bleeding. Those at high risk received no anticoagulation (group 1) or NM as an anticoagulant (group 2), while those at low risk received heparin (group 3). Results: Forty patients (25 male and 15 female; mean age, $8.2{\pm}6.6$ years) were enrolled. The mean duration of CRRT was 13.0 days, and the survival rate was 57.5%. The mean hemofilter lifespan was 39.3 h in group 1 and 11.3 h in group 3. In group 2, hemofilter lifespan was extended from 7.5 h to 27.4 h after the use of NM (P =0.001). The mean hemofilter lifespan with NM was greater than with heparin (P =0.018). No patient experienced a major bleeding event during treatment with NM. Conclusion: NM may be a good alternative anticoagulant in pediatric patients with a high risk of bleeding requiring CRRT, and is not associated with bleeding complications.

Studies on the Preparation of Weanling Food from Soybean (Part 1) -Conditions for the digestion of soybean protein by Eezyme from Aspergillus- (대두를 이용한 이유식 제조에 관한 연구(제 1보) -효소를 이용한 대두단백질 분해 적정 조건결정 및 조제에 관하여-)

  • Kim, Z.U.;Cho, M.J.
    • Applied Biological Chemistry
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    • v.13 no.1
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    • pp.29-34
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    • 1970
  • In order to prepare digested Protein source for the Weanling Food from soybean, an attempt was made to decompose steamed soybean protein to amino acids and peptides by protease and cellulase produced from Aspergillus niger and Aspergillus sojae. In this paper, the optimum condition for digestion of soybean protein were studied and also investigated the effects of decolorization of it. As the results, followings were obtained; 1. As steaming conditions, a treatment under 15 lb of pressure and 10 minutes of heating shows most effective. 2. The optimum pH of Asp, sojae enzyme for the digestion of soybean protein is 6.0, while that of Asp. niger enzyme is 4.4. In successievly-decomposing with Asp. sojae and Asp. niger, it shows the most effective on ratio of water-soluble-nitrogen to total nitrogen and amino-nitrogen to total nitrogen than any other separate treatments. 3. The suitable amount of the enzyme solution to that of the soybean substrate paste, in volume, is 1 : 2. 4. Digestion ratio of soybean protein indicates the gradual and steady effects of increasing time of digestion, but 8 hour-digestion regarding to putrefaction was suitable. 5. The most effective decolorization was successively passed on culumns of active carbon and anion exchanger (Dowex 2-x-8) at room temperature. In separate treatments, the effective order of decolorization was as follows; (Dowex 2-x-8)>Active carborn>Amberite IR-120 6. The powder type of the soy protein source obtained by concentration below $60^{\circ}C$ contains 12.51% of moisture, 66.31% of protein, 4.25% of fat, 12.75% of carbohydrate, 4.18% of ash.

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Production of Vitamin $B_{12}$ by Using Protoplast Fusion between Bacillus natto and Bacillus megaterium (Bacillus natto 및 Bacillus megaterium의 원형질체 융합에 의한 Vitamin $B_{12}$의 생산)

  • Jin, Sung-Hyun;Park, Bub-Gyu;Roh, Myung-Hoon;Kim, Dong-Gyu;Ryu, Beung-Ho
    • Korean Journal of Food Science and Technology
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    • v.22 no.6
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    • pp.611-617
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    • 1990
  • This study was conducted to breed a high vitamin $B_{12}$ producer by the fusion of protoplasts between Bacillus natto and Bacillus megaterium. Auxotrophic mutants of Bacillus natto SH-34 ($thr^-try^-rif^r$) and Bacillus megaterium BK-13 ($arg^-ade^-lys^-str^r$) which showed high protease activity and production of vitamin $B_{12}$, respectively, were isolated for the fusion experiment. Protoplasts were induced by incubating the cells with lysis solution containing $500{\mu}/ml$ lysozyme, and the ratio of protoplast and regeneration formation were ranged from 99% and 67%, respectively. Fusion frequencies of fusants between Bacillus natto SH-34 and Bacillus megaterium BK-13 were appeared in the ranges of $1.0{\times}10^{-5}$ under the treatment of 30% PEG 6000 containing 3% PVP. The fusant, MNF-72 showed the highest product yield of $7.85{\mu}g/g-cell\;vitamin\;B_{12}$ in production medium. For the improvement of productivity, the immobilization of fusants with sodium alginate was carried out. In batch and continuous fermentation systems, the productivity were determined to be $0.58{\mu}g/ml.hr\;and\;0.80{\mu}g/ml.hr\;vitamin\;B_{12}$ under optimum condition, respectivity.

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Optimization of enzymatic hydrolysis of viscera waste proteins of black body fowl(Yeonsan Ogae) to produce peptides using a commercial protease and it's characters analysis (단백질 분해효소를 이용한 연산오계 내장 펩타이드 생산 최적화 및 특성분석)

  • Choi, So-Young;Kim, A-Yeon;Song, Yu-Rim;Ji, Joong-Gu;Yoo, Sun-Kyun
    • Journal of Digital Convergence
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    • v.14 no.1
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    • pp.253-262
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    • 2016
  • Yeonsan Ogae has been known as supporting health and high efficacy of treatment. In recent days, as the efficacy of functional peptides has known, the optimization of oligo peptides production and its characteristics from Ogae viscera has been performed. Response surface method was used to perform the optimizaion of enzyme hydrolysis. The range of processes was temperature (40, 50 and $60^{\circ}C$), pH(6.0, 7.0 and 8.0), and enzyme(1, 2 and 3%). The degree of hydrolysis, amono acids, molecular weight of products were analyzed. The optimum process of enzyme hydrolysis were determined as temperature $58^{\circ}C$, pH 7.5, and enzyme concetration 3%. At optimum conditions, the degree of hydrolysis after 2 h reaction was 75-80%. The total amino acids of amino acid and were 386.15 mg/100 g and 155.26 mg/100 g, respectively. The molecular weight of products by using Maldi-TOF was ranged from 300 to 1,000 Da.

Purification and Characterization of an Antimicrobial Substance from Bacillus subtilis HH28 Antagonistic to Bacillus cereus (Bacillus cereus를 억제하는 Bacillus subtilis HH28의 항균물질 정제와 특성규명)

  • Cha, Hyun A;Chung, Dawn;Hong, Sung Wook;Chung, Kun Sub
    • Microbiology and Biotechnology Letters
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    • v.42 no.4
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    • pp.393-401
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    • 2014
  • A bacterium producing antimicrobial substance was isolated from cheonggukjang. The bacterium was identified as a strain of Bacillus subtilis by 16S rDNA sequencing and designated as Bacillus subtilis HH28. The antimicrobial substance produced from Bacillus subtilis HH28 was purified by 0-80% ammonium sulfate precipitation, DEAE-sepharose FF column chromatography, and Sephacryl S-200 HR gel chromatography. The molecular weight of the purified antimicrobial substance was estimated to be approximately 3,500 Da using Tricine sodium dodecyl sulfate-polyacrylamide gel electrophoresis and direct detection analysis. Antimicrobial substance from B. subtilis HH28 not only inhibited B. cereus, but also Listeria monocytogenes and Vibrio parahaemolyticus. The purified antimicrobial substance was stable at $40-80^{\circ}C$, and between pH 2 and 8. Antimicrobial activity of the purified substance was completely destroyed by treatment of protease, proteinase K, and pronase E, indicating that it is proteinaceous.

Inhibitory Effects of Deer Antler Herbal-Acupuncture Solution on Cathepsin S and L Activity in RA Model Mice

  • Ahn, Hyung-Jun;Yoon, Jong-Hwa;Kim, Kyung-Ho;Lee, Seung-Deok;Kim, Kap-Sung
    • Journal of Acupuncture Research
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    • v.22 no.2
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    • pp.71-81
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    • 2005
  • Objective : Dear antler (Cervus korean TEMMINCK var. mantchuricus Swinhoe) used for traditional immunosuppressive and immuno-activating action. The effect of deer antler herbal-acupuncture(DAH) solution, prepared by water extract method, on cathepsin activities in bone tissues (cartilage and synovial) cells from mouse rheumatoid arthritis (RA) model was studied. The cysteine endoprotease cathepsin mediates degradation of the MHC class II invariant chain (Ii) in human and mouse antigen-presenting cells. The studies described here examine the functional significance of cathepsin inhibition on autoantigen presentation and organ-specific autoimmune diseases in a murine model for RA. Methods : An animal model for RA in BALB/c mice thymectomized 3 days after birth (3d-Tx) was constructed All 3d-Tx BALB/c mice developed autoimmune lesions in the bone tissue cells, starting at 3 weeks of age, and the disease mediated by CD4+ T cells was chronic and progressive. Significant inhibitory effects of DAH solution on cathepsin S and L were observed in each organ in a dose-dependent manner. Moreover, we confirmed that cathepsin S and L activity in each organ were clearly inhibited by DAB solution. When we examined the inhibitory effects of DAH solution against autoantigen-specific T cell responses in vitro, in regional lymph node cells, but not in spleens, from model mice, a significant inhibitory effect of DAB solution was observed in a dose-dependent manner. DAH solution do not block T cell proliferation to Con A, indicated that the dose of DAB solution 10 to $20\;{\mu}g/m{\ell}$ was sufficient to inactivate the autoantigen-specific T cell responses in vitro. In vivo therapeutic effects of DAB solution were examined in a murine model for RA, autoantigen-specific (C-II-specific) T cell response were significantly inhibited in LNCs from DAH solution-treated mice. Results : Iinhibition of cathepsin S and L in vivo alters autoantigen presentation and development of organ-specific autoimmunity in RA model. Conclusion : These data identify selective inhibition of cysteine protease cathepsin S and L as a potential therapeutic strategy for autoimmune disease process such RA. Thus, DAH solution will served as a potent anti-inflammatory and anti-arthritic agents for treatment of human RA.

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