• Title/Summary/Keyword: Acid Resistance

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Presence of Carbonic Anhydrase III-like Protein in Shaggy Sea Raven, Hemitripterus villosus (삼세기(Shaggy sea raven, Hemitripterus villosus)의 carbonic anhydrase III에 관한 연구)

  • Kweon, Rok Eun;Kho, Kang Hee
    • Journal of Life Science
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    • v.24 no.2
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    • pp.186-190
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    • 2014
  • Carbonic anhydrase isozymes (CAs) are widespread zinc-containing metalloenzyme family. The enzyme catalyzes the reversible interconversion of $CO_2$ and $HCO_3$. This reaction is the main role of CA enzymes in physiological conditions. CA III, one of the CA isozymes, has been identified in many tissues. It is distinguished from the other isozymes by several characteristics, particularly by a lower specific activity and by its resistance to acetazolamide. However, the physiological function of CA III in fish is unknown. In this study, we examined the detection of CAs in the Shaggy sea raven Hemitripterus villosus, using SDS-PAGE, isoelectric focusing (IEF), and western blot analysis. We detected a significant protein band with molecular weight about 30 kDa from the tissues of H. villosus by SDS-PAGE and western blotting. A specific band of CA III with pI 7.0 was detected by IEF and western blotting in gill and muscle. The immunoreaction of anti-CA III expressed in the gill of H. villosus was much stronger than other tissues. One explanation for this result is that the fish gill is the only organ that is exposed to the external environment and that plays an important role in acid-base relevant ion transfer, the transfer of $H^+$ and/or $HCO{_3}^-$, for the maintenance of systemic pH. This is the first report on the identification of a carbonic anhydrase III-like protein from H. villosus.

Probiotic Properties of Lactobacillus strains Isolated from Kimchi (김치로부터 분리된 Lactobacillus strains의 probiotic 특성)

  • Choi, Hye Jung;Lim, Bo Ram;Kim, Dong Wan;Kwon, Gi-Seok;Joo, Woo Hong
    • Journal of Life Science
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    • v.24 no.11
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    • pp.1231-1237
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    • 2014
  • The objective of this study was to evaluate the safety and functional properties of four potential probiotic strains isolated from Kimchi, traditional Korean fermented vegetables. Based on being higher tolerance to bile salts and showing higher acid resistance or hydrophobic properties, one Lactobacillus arizonensis strain (BCNU 9032) and three L. brevis strains (BCNU 9037, BCNU 9098 and BCNU 9101) were selected in the screening experiment. All strains can survived up to 99% after 3h culture in pH 2.5 and resistant to 1% bile salts. These strains also showed good antimicrobial activities against a number of food borne pathogens, especially against Escherichia coli and Shigella sonnei. The ability to lower cholesterol levels of L. arizonensis BCNU 9032 and L. brevis 9037 were demonstrated by bile salt hydrolytic activity and cholesterol assimilation tests. Moreover, L. brevis BCNU 9098 and BCNU 9101 showed higher adherence to Caco-2 cells (12.76 and 11.86%, respectively) than Lactobacillus rhamnosus GG, a commercial probiotic strain used worldwide. The results suggest that these strains could be used as probiotics.

Antifungal Activity of Bacterial Strains isolated from Tidal Mudflat and Salted Seafood (traditional Jeotgal) Against Six Major Plant Pathogens (갯벌 및 젓갈에서 분리한 세균의 작물 주요 병원균에 대한 항균활성 효과 검정)

  • Kim, Tack-Soo;Lee, Ga-Hyung;Kim, Gyun-Jang;Lee, Se-Won;Park, Kyung-Seok;Park, Jin-Woo
    • The Korean Journal of Pesticide Science
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    • v.14 no.4
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    • pp.421-426
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    • 2010
  • Many bacterial strains inhabit strong saline condition, such as tidal mudflat and salted seafoods, were identified and reported for the proposed protease activities and salt resistance; however antifungal activities against plant fungal pathogen have not well been studied until now. In this study, primary screening was performed for the isolation of promising strains against major plant pathogens like Sclerotinia sclerotiorum, Fusarium oxysporum, Phytophthora capsici, Botrytis cineria, Collectotrichum acutatum and Pythium ultimum. Totally 423 bacterial strain were isolated from laboratory media which was based on different morphological characteristics and all the strains were dual cultured against major fungal pathogens on PDA, finally 40 strains were selected as antifungal bacterial strain and identified by fatty acid phylogenic difference analysis from MIDI shorlock gas chromatography system. As a result, antifungal strains from tidal mudflat were 10 species of 6 genus. Paenibacillus macerans was dominant species; 5 strains among the 17 isolates from tidal mudflat. Antifungal strains from salted seafoods were 7 species of 3 genus and Bacillus atrophaeus was dominant species; 12 strains among the 23 isolates from salted fishes.

Advanced Water Treatment of High Turbidity Source by Hybrid Process of Ceramic Ultrafiltration and Photocatalyst: 2. Effect of Photo-oxidation and Adsorption (세라믹 한외여과 및 광촉매 혼성공정에 의한 고탁도 원수의 고도정수처리: 2. 광산화와 흡착의 영향)

  • Cong, Gao-Si;Park, Jin-Yong
    • Membrane Journal
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    • v.21 no.2
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    • pp.201-211
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    • 2011
  • The effects of humic acid (HA), photo-oxidation and adsorption were investigated in hybrid process of ceramic ultrafiltration and photocatalyst for drinking water treatment. UF, photocatalyst, and UV radiation processes were investigated in viewpoints of membrane fouling resistance $(R_f)$, permeate flux (J), and total penneate volume $(V_{\Upsilon})$ at 2 and 4 mg/L of HA respectively. As decreasing HA, $R_f$ decreased dramatically and J increased, and finally $V_{\Upsilon}$ was the highest at 2 mg/L HA. Average treatment efficiencies of turbidity decreased as increasing HA, but treatment efficiency of HA was the highest at 4 mg/L HA. It was because most of HA was removed by membrane and some HA passing through the membrane was adsorbed or photo-oxidized by photocatalyst at low HA, and therefore treated water quality was almost same at 2 and 4 mg/L HA, but feed water quality was higher at 4 mg/L. At effect experiment of photo-oxidation and adsorption, J of UF + $TiO_2$ + UV process was maintained at the highest, and ultimately $(V_{\Upsilon})$ after 180 minutes' operation was the highest. As results of comparing the treatment efficiencies of turbidity and HA, photocatalyst adsorption had more important role than photo-oxidation when HA increased from 2 to 4 mg/L.

Anti-obesity Effect of Rhizoma Atractylodis Herbal Acupuncture in High Fat Diet-induced Obese ICR Mouse (고지방 식이로 유도된 비만 생쥐에서 창출약침의 항비만 효과)

  • Youh, Eun-Joo;Seo, Byung-Kwan;Nam, Sang-Soo;Kang, Sung-Keel
    • Journal of Acupuncture Research
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    • v.27 no.6
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    • pp.31-42
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    • 2010
  • Objectives : The aim of this study was to investigate the anti-obesity potential and mechanisms of action of Rhizoma Atractylodis(RA) herbal acupuncture in high fat diet- induced obese ICR mice. Methods : Sample solutions for herbal acupuncture were prepared from the Rhizoma Atractylodis water extract powder at concentration of 150mg/kg and 300mg/kg with distilled water. Five week-old ICR mice acclimatized to the laboratory environment for 1 week were allocated into four groups: regular diet group (RD), high fat diet group(HFD), groups fed HFD with 150mg/kg RA herbal acupuncture treatment (RAE 150) and with 300mg/kg RA herbal acupuncture treatment(RAE 300). Herbal acupuncture groups were injected with either 150mg/kg or 300mg/kg of Rhizoma Atractylodis(RA) subcutaneously onto both Sinsu($BL_{23}$) alternately on the same time everyday for 30days. Body weight, gross appearance of epididymal fat area, blood glucose, insulin, insulin resistance(HOMA-IR), non-esterified fatty acid, cholesterol, triglyceride, AST, ALT, histological analysis of white adipose tissue, gene expression responsible for adipocyte differentiation and AMPK activation were analyzed. Results : RA herbal acupuncture inhibited the development of weight gain, hyperglycemia, hyperinsulinemia, hyperlipidemia, increases of AST and ALT, and the enlargement of fat cell size induced by HFD. Also, RA herbal acupuncture inhibited the expression of PPAR-${\gamma}$, C/$EBP{\alpha}$, aP2, LPL, FAS, SCD-1 and enhanced the activation of AMP-activated protein kinase. Conclusions : The results of this study demonstrate that RA herbal acupuncture can exert the anti-obesity effect and it is partially mediated by activation of AMPK and inhibition of the gene expressions responsible for adipocyte differentiation. Further studies will be required to ascertain the nti-obesity effect and mechanisms of action of RA herbal acupuncture in animal models and human for aclinical application.

Anti-diabetic effect of purple corn extract on C57BL/KsJ db/db mice

  • Huang, Bo;Wang, Zhiqiang;Park, Jong Hyuk;Ryu, Ok Hyun;Choi, Moon Ki;Lee, Jae-Yong;Kang, Young-Hee;Lim, Soon Sung
    • Nutrition Research and Practice
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    • v.9 no.1
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    • pp.22-29
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    • 2015
  • BACKGROUND/OBJECTIVES: Recently, anthocyanins have been reported to have various biological activities. Furthermore, anthocyanin-rich purple corn extract (PCE) ameliorated insulin resistance and reduced diabetes-associated mesanginal fibrosis and inflammation, suggesting that it may have benefits for the prevention of diabetes and diabetes complications. In this study, we determined the anthocyanins and non-anthocyanin component of PCE by HPLC-ESI-MS and investigated its anti-diabetic activity and mechanisms using C57BL/KsJ db/db mice. MATERIALS/METHODS: The db/db mice were divided into four groups: diabetic control group (DC), 10 or 50 mg/kg PCE (PCE 10 or PCE 50), or 10 mg/kg pinitol (pinitol 10) and treated with drugs once per day for 8 weeks. During the experiment, body weight and blood glucose levels were measured every week. At the end of treatment, we measured several diabetic parameters. RESULTS: Compared to the DC group, Fasting blood glucose levels were 68% lower in PCE 50 group and 51% lower in the pinitol 10 group. Furthermore, the PCE 50 group showed 2-fold increased C-peptide and adiponectin levels and 20% decreased HbA1c levels, than in the DC group. In pancreatic islets morphology, the PCE- or pinitol-treated mice showed significant prevention of pancreatic ${\beta}$-cell damage and higher insulin content. Microarray analyses results indicating that gene and protein expressions associated with glycolysis and fatty acid metabolism in liver and fat tissues. In addition, purple corn extract increased the phosphorylation of AMP-activated protein kinase (AMPK) and decreased phosphoenolpyruvate carboxykinase (PEPCK), glucose 6-phosphatase (G6pase) genes in liver, and also increased glucose transporter 4 (GLUT4) expressions in skeletal muscle. CONCLUSIONS: Our results suggested that PCE exerted anti-diabetic effects through protection of pancreatic ${\beta}$-cells, increase of insulin secretion and AMPK activation in the liver of C57BL/KsJ db/db mice.

A Study on the Galactooligosaccharides Produced from Transgalactosylation with ${\beta}-Galactosidase$ of Aspergillus and Kluyveromyces (A. oryzae와 K. fragilis 유래의 ${\beta}-Galactosidase$ 혼합처리에 의한 전이 Galactooligo당의 생성에 관한 연구)

  • Moon, Ji-Woong;Huh, Kyeong-Tak;Lee, Su-Won;Lee, Jai-Young
    • Korean Journal of Food Science and Technology
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    • v.27 no.3
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    • pp.271-280
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    • 1995
  • These studies examined the production of oligosaccharides by ${\beta}-galactosidase$originated from Aspergillus oryzae and Kluyveromyces fragilis mixed. In addition, heat resistance and acid stability of transgalactosylated oligosaccharides were measured. When ${\beta}-galactosidase$ from A. oryzae, K. fragilis and mixed ${\beta}-galactosidase$ were added to 30%(w/v) lactose solution, maximum production of transgalactosylated oligosaccharides were 26.9%, 37.05 and 27.2%, respectively. The ratios of disacchride, trisacchride and tetrasaccharide in transgalactosylated oligosaccharides were 20.5 : 5.4 : 0.6, 20.4 : 10.5 : 4.2 and 21.0 : 4.1 : 1.9, respectively. Nine different oligosaccharides were recovered with 30% and 40% ethanol fractions. When the 30% ethanol fraction was treated at $150^{\circ}C$ for 10 min more than 90% of oligosaccharides remained stable. More than 90% of the oligosaccharides were stable at $130^{\circ}C$ for 3 min with pH 3.0, whereas there of Kluyveromyces was more than 90% with pH 3.5.

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Cloning and Overexpression of a Paenibacillus ${\beta}-Glucanase$ in Pichia pastoris: Purification and Characterization of the Recombinant Enzyme

  • Yang, Peilong;Shi, Pengjun;Wang, Yaru;Bai, Yingguo;Meng, Kun;Luo, Huiying;Yuan, Tiezheng;Yao, Bin
    • Journal of Microbiology and Biotechnology
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    • v.17 no.1
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    • pp.58-66
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    • 2007
  • Isolation, expression, and characterization of a novel $endo-{\beta}-1,3(4)-D-glucanase$ with high specific activity and homology to Bacillus lichenases is described. One clone was screened from a genomic library of Paenibacillus sp. F-40, using lichenan-containing plates. The nucleotide sequence of the clone contains an ORF consisting of 717 nucleotides, encoding a ${\beta}-glucanase$ protein of 238 amino acids and 26 residues of a putative signal peptide at its N-terminus. The amino acid sequence showed the highest similarity of 87% to other ${\beta}-1,3-1,4-glucanases$ of Bacillus. The gene fragment Bg1 containing the mature glucanase protein was expressed in Pichia pastoris at high expression level in a 3-1 high-cell-density fermenter. The purified recombinant enzyme Bg1 showed activity against barley ${\beta}-glucan$, lichenan, and laminarin. The gene encodes an $endo-{\beta}-1,3(4)-D-glucanase$ (E. C. 3.2.1.6). When lichenan was used as substrate, the optimal pH was 6.5, and the optimal temperature was $60^{\circ}C$. The $K_m,\;V_{max},\;and\;k_{cat}$ values for lichenan are 2.96mg/ml, $6,951{\mu}mol/min{\cdot}mg,\;and\;3,131s^{-1}$, respectively. For barley ${\beta}-glucan$ the values are 3.73mg/ml, $8,939{\mu}mol/min{\cdot}mg,\;and\;4,026s^{-1}$, respectively. The recombinant Bg1 had resistance to pepsin and trypsin. Other features of recombinant Bg1 including temperature and pH stability, and sensitivity to some metal ions and chemical reagents were also characterized.

Physiological Characteristics and Anti-diabetic Effect of Lactobacillus plantarum KI69 (Lactobacillus plantarum KI69의 생리적 특성 및 항당뇨 효과)

  • Kim, Seulki;Lim, Sang-Dong
    • Journal of Dairy Science and Biotechnology
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    • v.37 no.4
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    • pp.223-236
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    • 2019
  • This study aimed to investigate the physiological characteristics and anti-diabetic effects of Lactobacillus plantarum KI69. The α-amylase and α-glucosidase inhibitory activity of L. plantarum KI69 was 91.17±2.23% and 98.71±4.23%, respectively. The propionic, acetic, and butyric acid contents of the MRS broth inoculated with L. plantarum KI69 were 8.78±1.12 ppm, 1.34±0.07% (w/v), and 0.876±0.003 g/kg, respectively. L. plantarum KI69 showed higher sensitivity to penicillin-G, oxacillin, and chloramphenicol among 16 different antibiotics and showed the highest resistance to ampicillin and vancomycin. The strain showed higher β-galactosidase, β-glucosidase, and N-acetyl-β-glucosaminidase activities than other enzymes. Additionally, it did not produce carcinogenic enzymes, such as β-glucuronidase. The survival rate of L. plantarum KI69 in 0.3% bile was 96.42%. Moreover, the strain showed a 91.45% survival rate at pH 2.0. It was resistant to Escherichia coli, Salmonella Typhimurium, Listeria monocytogenes, and Staphylococcus aureus with the rates of 15.44%, 50.79%, 58.62%, and 37.85%, respectively. L. plantarum (25.85%) showed higher adhesion ability than the positive control L. rhamnosus GG (20.87%). These results demonstrate that L. plantarum KI69 has a probiotic potential with anti-diabetic effects.

Induction of Heat Shock Protein 70 Inhibits Tumor Necrosis $Factor{\alpha}-induced$ Lipid Peroxidation in Rat Mesangial Cells (Heat Shock Protein 70이 흰쥐 배양 혈관간 세포에서 관찰되는 $TNF{\alpha}$에 의한 지질과산화에 미치는 보호 효과)

  • Ha, Hun-Joo;Park, Young-Mee;Ahn, Young-Soo;Kim, Kyung-Hwan
    • The Korean Journal of Pharmacology
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    • v.31 no.3
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    • pp.323-331
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    • 1995
  • Monocyte/macrophage infiltration is the well known initial features associated with the development of glomerular disease including non-immune mediated nephropathy. Tumor necrosis factor ${\alpha}(TNF{\alpha})$, a cytokine produced primarily by monocyte/macrophage, exhibits similar effects as observed at the initial stages and during the progression of glomerular injury. Because the mesangial cells are target cells for glomerular injury, the present study examined the effect of $TNF{\alpha}$ on glomerular mesangial cell membrane lipid peroxidation as an index of cytotoxicity attributing to $TNF{\alpha}$. Primary culture of rat mesangial cell was established by incubation of glomeruli isolated from male Sprague-Dawley rat kidneys utilizing a standard sieving method. The levels of lipid peroxides in the mesangial cells were quantitated by malondialdehyde- thiobarbituric acid adduct formation. During an 8 hour incubation at $37^{\circ}C$, $TNF{\alpha}$ at 10 to 10,000 units/ml increased the levels of lipid peroxides dose dependently. Western blot analysis demonstrated that a short thermal stress induced heat shock response and the synthesis of heat shock protein 70(hsp70) in this mesangial cells. Further, this induction of hsp 70 prevented increase of lipid peroxides in the mesangial cells exposed to $TNF{\alpha}$. These data suggest that $TNF{\alpha}-induced$ lipid peroxidation in the mesangial cells may have pathophysiological relevance to glomerular injury and prior induction of heat shock response may play a role in the cellular resistance against $TNF{\alpha}-induced$ glomerular injury.

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