• Title/Summary/Keyword: lysate

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In vitro Antioxidative Properties of Lactobacilli

  • Kim, H.S.;Chae, H.S.;Jeong, S.G.;Ham, J.S.;Im, S.K.;Ahn, C.N.;Lee, J.M.
    • Asian-Australasian Journal of Animal Sciences
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    • 제19권2호
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    • pp.262-265
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    • 2006
  • The antioxidative properties of lactobacilli originating from humans (Lactobacillus acidophilus KCTC 3111, Lactobacillus jonsonii KCTC 3141, Lactobacillus acidophilus KCTC 3151, and Lactobacillus brevis KCTC 3498) were investigated using in vitro methods, including inhibition of lipid peroxidation, resistance to hydrogen peroxide and hydroxyl radical, hydroxyl radical scavenging activity, and glutathione peroxidase (GPX) activity. L. acidophilus KCTC 3111 showed the highest inhibition of lipid peroxidation in both intact cells (49.7%) and cell lysate (65.2%). This strain exhibited resistance to hydrogen peroxide and hydroxyl radical, which was viable for 7 h in the concentration of 1.0 mM hydrogen peroxide. In addition, this strain showed high hydroxyl radical scavenging activity. In the GPX activity assay, the highest activity was measured in L. brevis 3498. GPX activity of L acidophilus 3111 was lower than that of L. brevis 3498.

Overexpression of Arylsulfatase in E. coli and Its Application to Desulfatation of Agar

  • Lim, Jae-Myung;Jang, Yeon-Hwa;Kim, Hyeung-Rak;Kim, Young-Tae;Choi, Tae-Jin;Kim, Joong-Kyun;Nam, Soo-Wan
    • Journal of Microbiology and Biotechnology
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    • 제14권4호
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    • pp.777-782
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    • 2004
  • The arylsulfatase gene (astA, 984 bp ORF) from the P. carrageenovora genome was amplified by PCR and subcloned into the pET21a vector. When the constructed plasmid pAST-A1 (6.4 kb) was introduced into E. coli BL21(DE3), the transformant on the LB plate containing IPTG showed a hydrolyzing activity for 4-methylumbelliferyl sulfate and p-nitrophenyl sulfate. The highest arylsulfatase activity (2.1 unit/ml) was obtained at 10 mM IPTG. Most arylsulfatase activity was found in the cell lysate, whereas no significant activity was detected in the culture supernatant. The molecular weight of the recombinant enzyme was estimated to be 33.1 kDa by SDS-PAGE. After the reaction of agar with arylsulfatase for 12 h at $40^{\circ}C$, the gel strength of the agar increased by 2-fold, and 73% of the sulfate in the agar had been removed. This result suggests that arylsulfatase expressed in E. coli could be useful in the production of electrophoretic grade agarose.

Purification and Characterization of an Antiviral Ribosome-inactivating Protein from Chenopodium album L.

  • Cho, Kang-Jin;Lee, Si-Myung;Kim, Yeong-Tae;Hwang, Young-Soo
    • Journal of Applied Biological Chemistry
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    • 제43권3호
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    • pp.125-130
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    • 2000
  • An antiviral protein (CAP30) with ribosome-inactivating activity was purified from the leaves of Chenopodium album L. through ammonium sulfate precipitation and column chromatography using S-Sepharose, Blue-Sepharose, FPLC Suprose12 HR, and FPLC Mono-S. The molecular wight of CAP30 was estimated to be 30kD. CAP30 was thermostable, maintaing its activity even after incubation at $70^{\circ}C$ for 30 min, and was stable in the pH range of 6 to 9. In a cell-free in vitro translation system using rabbit reticulocyte lysate, protein synthesis was inhibited by the addition of CAP30 with an $IC_{50}$ of 2.26pM. The comparison of N-terminal amino acid sequences of this protein with known ribosome-inactivating proteins (RIPs) revealed that it had some sequence homology with PAP-S and PAP-R from pokeweed (Phytolacca americana)and dodecandrin from P. dodecandra, but had no sequence homology with RIPs from other plants belonging to different orders. The mosaic symptoms on tobacco leaves caused by cucumber mosaic virus infection was completely inhibited by 100 ng/ml of the pure CAP30 protein.

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Arginase Inhibition by Ethylacetate Extract of Caesalpinia sappan Lignum Contributes to Activation of Endothelial Nitric Oxide Synthase

  • Shin, Woo-Sung;Cuong, To Dao;Lee, Jeong-Hyung;Min, Byung-Sun;Jeon, Byeong-Hwa;Lim, Hyun-Kyo;Ryoo, Sung-Woo
    • The Korean Journal of Physiology and Pharmacology
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    • 제15권3호
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    • pp.123-128
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    • 2011
  • Caesalpinia sappan (C. sappan) is a medicinal plant used for promoting blood circulation and removing stasis. During a screening procedure on medicinal plants, the ethylacetate extract of the lignum of C. sappan (CLE) showed inhibitory activity on arginase which has recently been reported as a novel therapeutic target for the treatment of cardiovascular diseases such as atherosclerosis. CLE inhibited arginase II activity prepared from kidney lysate in a dose-dependent manner. In HUVECs, inhibition of arginase activity by CLE reciprocally increased NOx production through enhancement of eNOS dimer stability without any significant changes in the protein levels of eNOS and arginase II expression. Furthermore, CLE-dependent arginase inhibition resulted in increase of NO generation and decrease of superoxide production on endothelium of isolated mice aorta. These results indicate that CLE augments NO production on endothelium through inhibition of arginase activity, and may imply their usefulness for the treatment of cardiovascular diseases associated with endothelial dysfunction.

Phage Conversion for β-Lactam Antibiotic Resistance of Staphylococcus aureus from Foods

  • Lee, Young-Duck;Park, Jong-Hyun
    • Journal of Microbiology and Biotechnology
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    • 제26권2호
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    • pp.263-269
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    • 2016
  • Temperate phages have been suggested to carry virulence factors and other lysogenic conversion genes that play important roles in pathogenicity. In this study, phage TEM123 in wild-type Staphylococcus aureus from food sources was analyzed with respect to its morphology, genome sequence, and antibiotic resistance conversion ability. Phage TEM123 from a mitomycin C-induced lysate of S. aureus was isolated from foods. Morphological analysis under a transmission electron microscope revealed that it belonged to the family Siphoviridae. The genome of phage TEM123 consisted of a double-stranded DNA of 43,786 bp with a G+C content of 34.06%. A bioinformatics analysis of the phage genome identified 43 putative open reading frames (ORFs). ORF1 encoded a protein that was nearly identical to the metallo-β-lactamase enzymes that degrade β-lactam antibiotics. After transduction to S. aureus with phage TEM123, the metallo-β-lactamase gene was confirmed in the transductant by PCR and sequencing analyses. In a β-lactam antibiotic susceptibility test, the transductant was more highly resistant to β-lactam antibiotics than S. aureus S133. Phage TEM123 might play a role in the transfer of β-lactam antibiotic resistance determinants in S. aureus. Therefore, we suggest that the prophage of S. aureus with its exotoxin is a risk factor for food safety in the food chain through lateral gene transfer.

Carbamoyl Phosphate Synthetase I이 요소회로 유전자를 발현하는 CHO 세포 주의 세포 성장과 재조합 Erythropoietin의 생산에 미치는 영향 (Effects of Carbamoyl Phosphate Synthetase I against Cell Growth and Production of Recombinant Erythropoietin in Urea Cycle Enzyme Expressing CHO Cell Line)

  • 조수미;김나영;김형진;김홍진
    • 약학회지
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    • 제51권3호
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    • pp.214-218
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    • 2007
  • In the previous reports, we developed the CO5 by introducing genes for the first and second urea cycle enzymes, carbamoyl phosphate synthetase I (CPS I) and ornithine transcarbamoylase (OTC) into the IBE cell lines producing erythropoietin (EPO). The CO5 have been found out to have 15-20% higher cell growth rate and produce 2-times more EPO than the parental cell line, IBE. To investigate the role of CPS I in CO5 cell line for the cell growth and amount of EPO, we knock-downed CPS I gene expression via siRNA treatment. Expression level of EPO in cell lysate of CO5 was 3-5 fold higher than that of IBE. After siRNA treatment, the cell growth of CO5 was decreased 8-21% and the EPO productivity in the cell Iysate was significantly decreased. However, these changes of the cell growth and EPO productivity were not observed in IBE. These results indicate that CPS I gene expression is important for the increased cell growth and EPO productivity of CO5 cell line.

Effect of Linker for Immobilization of Glutathione on BSA-Assembled Controlled Pore Glass Beads

  • Chen, Li-Hua;Choi, Young-Seo;Park, Jung-Won;Kwon, Joseph;Wang, Rong-Shun;Lee, Tae-Hoon;Ryu, Sung-Ho;Park, Joon-Won
    • Bulletin of the Korean Chemical Society
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    • 제25권9호
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    • pp.1366-1370
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    • 2004
  • Controlled pore glass bead was modified with bovine serum albumin (BSA), and glutathione (GSH) was immobilized through three kinds of linkers on top of BSA. Bis(3-sulfo-N-hydroxysuccinimide suberate) sodium salt $(BS^3)$, N-hydroxysuccinimide 3-(2-pyridyldithio)propionate (SPDP), or N-hydroxysuccinimide 4-maleimidobutyrate (GMBS) was introduced into the BSA-bound matrix. Subsequently, GSH was immobilized by addition of thiol side chain into the maleimido moiety, replacing a disulfide group, or formation of an amide group upon releasing 3-sulfo-N-hydroxysuccimide group. It was observed that conjugation methodology played a critical role for activity of the immobilized GSH. SDS-PAGE chromatogram showed that the matrix of glutathione immobilized on BSA through GMBS manifested high selectivity towards glutathione-S-transferase (GST) in cell lysate.

한국에서 분리한 agrobacterium tumefaciens T7의 특성과 biovar.결정 (Characterization and biovar. cetermination of agrobacterium tumefaciens T7 isolated in Korea)

  • 이윤;김창진;김성훈;유익동;민태익
    • 미생물학회지
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    • 제25권1호
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    • pp.17-22
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    • 1987
  • For the purpose of securing of strains which can be usefully utilized to study symbiosis between Rhizobium and legume plant, A. tumefaciens T7 was isolated and characterized and then subgroup biovar was determined. A. tumefaciens T7 induced smooth tumor like nopaline type one and did not grow at $37^{\circ}C$ and in the presence of 2% NaCl on yeast extract mannitol medium. The strain was able to grow on the New and Kerr selective media and utilize erythritol but not phenylalanine, tryptophan, and tartarate as a sole carbon source. Negative results were obtained from 3-keto-lactose production and oxidase test. The strain produced alkalifrom malonate and citrate and showed acid litmus milk reaction At least two large plasmids were detected in the cell lysate. According to all of these results, it could be concluded that subdivision of isolated strain was biovar 2.

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총명탕(聰明湯)과 산사총명탕(山査聰明湯)이 CT105로 유도된 Alzheimer's Disease 병태 모델에 미치는 영향 (Effects of ChongMyung-Tang and SansaChongMyung-Tang Extract on the Alzheimer's Disease Model Induced dy CT105)

  • 이상룡;정인철
    • 동의생리병리학회지
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    • 제20권1호
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    • pp.138-148
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    • 2006
  • This research investigated the effect of the CMT and SCMT on Alzheimer's disease. The effects of the CMT and SCMT extract on expression of proinflammatory cytokine(IL-$1{\beta}$, IL-6, TNF-$\alpha$) in the THP-1 cell; amyloid precursor proteins(APP), acetylcholinesterase(AChE) mRNA of PC-12 cells treated with CT105; the AChE activity and the APP production of PC-12 cell lysate treated with CT105 were investigated. The CMT and SCMT extract suppressed overexpression of IL-$1{\beta}$, IL-6, TNF-$\alpha$ in the THP-1 cell treated dy LPS; the expression of APP, AChE mRNA in PC-12 cells treated with CT105; the AChE activity and the production of APP in PC-12 cell Iysate treated with CT105 significantly. This study suggest that CMT and SCMT may be effective for the prevention and treatment of Alzheimer's disease.

Vibrio vulnificus에 대한 경구용백신 CJ-50002의 일반약리작용 (General Pharmacology of CJ-50002, an Oral Vaccine against Vibrio vulnificus Infection)

  • 김영훈;정성목;신재규;최재묵;이나경;박완제;이윤하;이영수
    • Biomolecules & Therapeutics
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    • 제7권1호
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    • pp.89-96
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    • 1999
  • CJ-50002 is an oral vaccine against V.vulnificus infection composed of whole cell lysate of V. vulnificus. The general pharmacological properties of CJ-50002 were evaluated in various animals and in vitro system. CJ-50002 at oral doses of 0.2, 2 and 20 mg/kg had no effect on general behavior in mice, chromo- and electro-convulsions in mice, writhing syndrome induced by acetic acid in mice, body temperature in rats, charcoal meal propulsion in mice and urine and electrolytes excretion in rats. However, oral administration of CJ-50002 at dose of 20 mg/kg prolonged the hexobarbital-inuced sleeping inducing time in mice. In anesthetized dogs, CJ-50002 showed no effect on blood pressure, heart rate and ECG but decreased the respiratory rate and femoral blood flow at dose of 20 mg/kg. p.o. CJ-50002 had no effect on the contractile response of the isolated guinea pig ileum to various spasmogen at concentrations of 0.2, 2 and 20 $\mu\textrm{g}$/ml, respectively. Since these pharmacological effects of CJ-500o2 were observed at dose much greater than those in clinical use (approximately 0.16 mg/kg, p.o.), it is likely that this vaccine may be relatively free of undesirable effects in clinical practice.

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