• Title/Summary/Keyword: PhaR

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Characterizations of Denitrifying Polyphosphate-accumulating Bacterium Paracoccus sp. Strain YKP-9

  • Lee, Han-Woong;Park, Yong-Keun
    • Journal of Microbiology and Biotechnology
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    • v.18 no.12
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    • pp.1958-1965
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    • 2008
  • A denitrifying polyphosphate-accumulating bacterium (YKP-9) was isolated from activated sludge of a 5-stage biological nutrient removal process with step feed system. This organism was a Gram-negative, coccus-shaped, facultative aerobic chemoorganotroph. It had a respiratory type of metabolism with oxygen, nitrate, and nitrite as terminal electron acceptors. The 16S rRNA gene sequence of strain YKP-9 was most similar to the 16S rRNA gene sequence of Paracoccus sp. OL18 (AY312056) (similarity level, 97%). Denitrifying polyphosphate accumulation by strain YKP-9 was examined under anaerobic-anoxic and anaerobic-oxic batch conditions. It was able to use external carbon sources for polyhydroxyalkanoates(PHA) synthesis and to release phosphate under anaerobic condition. It accumulated polyphosphate and grew a little on energy provided by external carbon sources under anoxic condition, but did neither accumulate polyphosphate nor grow in the absence of external carbon sources under anoxic condition. Cells with intracellular PHA cannot accumulate polyphosphate in the absence of external carbon sources under anoxic condition. Under oxic condition, it grew but could not accumulate polyphosphate with external carbon sources. Based on the results from this study, strain YKP-9 is a new-type denitrifying polyphosphate-accumulating bacterium that accumulates polyphosphate only under anoxic condition, with nitrate and nitrite as the electron acceptors in the presence of external carbon sources.

Antigenicity of Recombinant Human G-CSF (CJ-50001) (CJ-50001(rG-CSF)에 대한 항원성시험)

  • Baek, Nam-Jin;Kang, Jae-Ku;Kim, Dal-Hyun;Mok, K.-Hun;Kim, Je-Hak;Kim, Hyun-Su
    • Toxicological Research
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    • v.13 no.3
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    • pp.303-306
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    • 1997
  • Antigenic potential of genetically engineered human granulocyte colony-stimulating factor (CJ-50001) was assessed in guinea pigs and mice. In active systemic anaphylaxis (ASA) test, although CJ-50001 at 50 $\mu\textrm{g}$ /head induced anaphylactic responses, CJ-50001 5 $\mu\textrm{g}$ /head alone or 50 $\mu\textrm{g}$ / head with adjuvant did not induce anaphylactic responses. In passive systemic anaphylaxis test (PCA) or passive hemagglutination test (PHA), CJ-50001 did not induce positive responses. It is concluded that, in light of the fact that CJ-50001 was antigenic only in ASA but not in PCA or PHA and also that CJ-50001 is a foreign human recombinant protein to guinea pigs, CJ-50001 may not induce systemic allergic react-ion in its clinical use in human.

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Synthesis of Copolymeric PHA by Hydrogenophaga pseudoflava and Ralstonia eutropha H16 from Vari-ous Lactones and Their Microstructural Studies (락톤류로부터 Hydrogenophaga pseudoflava와 Ralstonia eutropha H16 두 세균에 의한 공중합 PHA의 합성 및 미세구조적 특성 연구)

  • Jang, Young-Ok;Nam, Won;Choi, Mun-Hwan;Song, Jae-Jun;Yoon, Sung-Chul
    • Microbiology and Biotechnology Letters
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    • v.28 no.2
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    • pp.71-79
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    • 2000
  • Two typess of copolyesters, poly(3-hydroxybutyric acid-co-4-hydroxy-butyric acid)[P(3HB-co-4HB] and poly(3-hydroxybutyric acid-co-3-hydroxyvaleric acid)[P(3HB-co-3HV)], with various monomer ratios and different degree of microstructural heterogeneity were synthesized from Ralstonia eutropha H16 and Hydrogenophaga pseudoflava by using ${\gamma}$-butyrolactone and ${\gamma}$-valerolactone, respectively. The two bacteria showed a large difference in the utilization of ${\gamma}$-butyrolactone for cell growth and PHA synthesis. H. pseudoflava synthesized P(3HB-co-4HB) copolyesters with a wide range of 4HB content from 13 to 96 mol% depending on culture conditions, whiel R. eutropha H16 was able to synthesize the copolyesters containing less than 20 mol% of 4HB. An increase in the 4HB content in the P(3HB-co-4HB) copolyesters synthesized by H. pseud-oflava induced an lowering of their melting temperatures as well as their enthalpies of fusion. The increase in the 4HB content, however, increased the rate of degradation by an extracellular P(3HB) depolymerase. NMR spectros-copy and differential scanning calorimetry showed that the P(3HB-co-4HB) copolyesters from H. pseudoflava were generally microstructurally heterogeneous. The P(3HB-co-4HB) copolyesters) synthesized by R. eutropha H16 were rather random copolymers showing less microstructural heterogeneity than those synthesized by H. pseudoflava. The NMR D value analysis suggested that the monomer distribution of the P(3HB-co-3HV) copolymers from the two bacteria were relatively random.

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Short-Chain-Length Polyhydroxyalkanoates: Synthesis in Metabolically Engineered Escherichia coli and Medical Applications

  • PARK, SI-JAE;CHOI, JONG-IL;LEE, SANG-YUP
    • Journal of Microbiology and Biotechnology
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    • v.15 no.1
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    • pp.206-215
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    • 2005
  • Polyhydroxyalkanoates (PHAs) are homo or hetero polyesters of (R)-hydroxyalkanoates accumulated in various microorganisms under growth-limiting condition in the presence of excess carbon source. They have been suggested as biodegradable substitutes for chemically synthesized polymers. Recombinant Escherichia coli is one of the promising host strains for the economical production of PHAs, and has been extensively investigated for the process development. The heterologous PHA biosynthetic pathways have been established through the metabolic engineering and inherent metabolic pathways of E. coli have been redirected to supply PHA precursors. Fermentation strategies for cultivating these recombinant E. coli strains have also been developed for the efficient production of PHAs. Nowadays, short-chain-length (SCL) PHAs are being re-invited due to its improved mechanical properties and possible applications in the biomedical area. In this article, recent advances in the development of metabolically engineered E. coli strains for the enhanced production of SCL-PHAs are reviewed. Also, medical applications of SCL-PHAs are discussed.

Validating a Xylose Regulator to Increase Polyhydroxybutyrate Production for Utilizing Mixed Sugars from Lignocellulosic Biomass Using Escherichia coli

  • Suk-Jin Oh;Hong-Ju Lee;Jeong Hyeon Hwang;Hyun Jin Kim;Nara-Shin;Sang-Ho Lee;Seung-Oh Seo;Shashi Kant Bhatia;Yung-Hun Yang
    • Journal of Microbiology and Biotechnology
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    • v.34 no.3
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    • pp.700-709
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    • 2024
  • Polyhydroxybutyrate (PHB) production from lignocellulosic biomass is economically beneficial. Because lignocellulosic biomass is a mixture rich in glucose and xylose, Escherichia coli, which prefers glucose, needs to overcome glucose repression for efficient biosugar use. To avoid glucose repression, here, we overexpressed a xylose regulator (xylR) in an E. coli strain expressing bktB, phaB, and phaC from Cupriavidus necator and evaluated the effect of xylR on PHB production. XylR overexpression increased xylose consumption from 0% to 46.53% and produced 4.45-fold more PHB than the control strain without xylR in a 1% sugar mixture of glucose and xylose (1:1). When the xylR-overexpressed strain was applied to sugars from lignocellulosic biomass, cell growth and PHB production of the strain showed a 4.7-fold increase from the control strain, yielding 2.58 ± 0.02 g/l PHB and 4.43 ± 0.28 g/l dry cell weight in a 1% hydrolysate mixture. XylR overexpression increased the expression of xylose operon genes by up to 1.7-fold. Moreover, the effect of xylR was substantially different in various E. coli strains. Overall, the results showed the effect of xylR overexpression on PHB production in a non-native PHB producer and the possible application of xylR for xylose utilization in E. coli.

The Effect of Vitamin B-Complex on Stress-induced Immune Alteration (비타민 B 복합제가 스트레스에 의해 유도된 면역변화에 미치는 영향)

  • Koh, Kyung-Bong;Lew, Soon-Hyung
    • Korean Journal of Psychosomatic Medicine
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    • v.7 no.2
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    • pp.196-202
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    • 1999
  • The purpose of the study was to determine the effect of vitamin B complex on stress-induced immune alteration. 21 medical students participated in the study 4 weeks before an academic examination period(baseline), 2 weeks before the exam period and during the exam period. Among them, 10 subjects were given vitamin B complex for 4 weeks, and 11 were not given vitamin B during the whole period. Cell-mediated immune function was measured by lymphocyte proliferative response to phytohemagglutinin(PHA) and interleukin-2(IL-2) production. Global assessment of recent stress(GARS) scale and symptom checklist-90-revised(SCL-90-R) were used to measure the level of subjective stress and psychopathology. Vitamin group had significantly lower scores of anxiety scale on SCL-90-R than non-vitamin group. No significant differences were found in lymphocyte proliferative response to PHA and IL-2 production between vitamin and non-vitamin groups during each period. There were no significant differences in change of of each of the two immune parameters over time as well as between vitamin and non-vitamin groups. However, lymphocyte proliferative response to PHA was significantly increased over time. In conclusion, it was suggested that vitamin B complex is likely to decrease anxiety level, and that exam stress might enhance lymphocyte proliferation regardless of vitamin B.

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Production of Medium-chain-length Poly (3-hydroxyalkanoates) by Pseudomonas sp. EML8 from Waste Frying Oil (Pseudomonas sp. EML8 균주를 이용한 폐식용류로부터 medium-chain-length poly(3-hydroxyalkanoates) 생합성)

  • Kim, Tae-Gyeong;Kim, Jong-Sik;Chung, Chung-Wook
    • Journal of Life Science
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    • v.31 no.1
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    • pp.90-99
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    • 2021
  • In this study, to reduce the production cost of poly(3-hydroxyalkanoates) (PHA), optimal cell growth and PHA biosynthesis conditions of the isolated strain Pseudomonas sp. EML8 were established using waste frying oil (WFO) as the cheap carbon source. Gas chromatography (GC) and GC mass spectrometry analysis of the medium-chain-length PHA (mcl-PHAWFO) obtained by Pseudomonas sp. EML8 of WFO indicated that it was composed of 7.28 mol% 3-hydrxoyhexanoate, 39.04 mol% 3-hydroxyoctanoate, 37.11 mol% 3-hydroxydecanoate, and 16.58 mol% 3-hydroxvdodecanoate monomers. When Pseudomonas sp. EML8 were culture in flask, the maximum dry cell weight (DCW) and the mcl-PHAWFO yield (g/l) were showed under WFO (20 g/l), (NH4)2SO4 (0.5 g/l), pH 7, and 25℃ culture conditions. Based on this, the highest DCW, mcl-PHAWFO content, and mcl-PHAWFO yield from 3-l-jar fermentation was obtained after 48 hr. Similar results were obtained using 20 g/l of fresh frying oil (FFO) as a control carbon source. In this case, the DCW, the mcl-PHAFFO content, and the mcl-PHAFFO yields were 2.7 g/l, 62 wt%, and 1.6 g/l, respectively. Gel permeation chromatography analysis confirmed the average molecular weight of the mcl-PHAWFO and mcl-PHAFFO to be between 165-175 kDa. Thermogravimetric analysis showed decomposition temperature values of 260℃ and 274.7℃ for mcl-PHAWFO and mcl-PHAFFO, respectively. In conclusion, Pseudomonas sp. EML8 and WFO could be suggested as a new candidate and substrate for the industrial production of PHA.

Response of Rabbit Appendix Cells to Specific Antigen and Mitogen (가토충양돌기세포(家兎蟲樣突起細胞)의 특이항원(特異抗原) 및 Mitogen에 대(對)한 반응(反應))

  • Ha, Tai-You
    • The Journal of the Korean Society for Microbiology
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    • v.10 no.1
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    • pp.1-8
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    • 1975
  • Despite a number of recent studies on appendix its function appears to remain unknown. The present studies were undertaken in order to extend and confirm the previous studies concerning the role of appendix in immune response. An early hemagglutinin response of mercaptoethanol sensitive antibody(IgM antibody) in rabbit injected intravenously(i.v.) with 200mcg of bovine gamma globulin(BGG) was abolished by lethal whole body irradiation(900 r), but preserved in animals whose appendix and bone marrow were shielded during irradiation. Late formation of mercaptoethanol resistant antibody(IgG antibody) and the development of memory in bone marrow shielded animals were not affected by irradiation of the appendix. Formation of either IgM or IgG antibody to sheep red blood cells(SRBC) injected i.v. as determined by direct plaque forming cell(DPFC) technique in spleen were effectively abolished by appendectomy, thymectomy, or both followed by irradiation. When bone marrow was shielded in combination with autologous appendix reconstitution, DPFC response was about 5 times greater than the sum of two. Lysed appendix cells failed to restore the response. Lethally irradiated rabbits restored with combination of autologous appendix and thymus cells showed DPFC responses which were essentially normal. Three pools of appendix were obtained by manual separation technique and were stimulated with soluble concanavalin A(Con A), phytohemagglutinin-P(PHA) and pokeweed mitogen(PWM). Rabbit appendix cells responded to Con A, PHA and PWM. Cells of thymus dependent area(TDA) of the appendix were relatively enriched in their response to T cell mitogens compared to dome and follicle cells. The PHA/Con A responsive ratio of appenix TDA subpopulation was high, indicating that Con A responsive cells have a wider distribution among appendix. This finding showed that interfollicular area of the appendix is thymus-dependent. The present studies confirmed other evidence that the rabbit appendix cells itself are unable to form antibody and T lymphocytes in appendix TDA may be heterogenous, and that the appendix cells are synergistic with either bone marrow or thymus cells in the early hemagglutinin on splenic antibody response to BGG or SRBC.

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Stimulatory Effect of Saponin from Panax ginseng on Function of Lymnphocytes in the Elderly

  • Liu, Jun-Da;Wang, Shu;Liu, Hong-Tao
    • Proceedings of the Ginseng society Conference
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    • 1998.06a
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    • pp.312-321
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    • 1998
  • We used the saponin Rgi extracted from Panax ginseng to study its effects on lymphocytes of 10 young and 19 elderly persons. The proliferate response of Iymphocytes cocultured for 72h with PHA and saponin was measured by using MTT method and the'H-TdR incorporation procedure. PHA and Rgl had stimulative effects on the phenotype of Iymphocytes (p<0.001). Rgl also increased the fluidity of lymphocyte membrane of the aged (p<0.001). The CD2s and CDfsRA positive cells of Iymphocytes in the elderly were lower than those of the young people,8.6clo $\pm$ 2.7olo vs 10.43% : 3.5%, 20.95% $\pm$ 15.5clQ vs 50.86% :4.3olo, respectively. More CDfsRO positive cell lymphocy populations were seen in the aged. The CEfsRO positive cells of the young people were 39.63% $\pm$ 3.2%. We discussed the cause of declined immune function of Lymphocytes of aged person and the mechanism of the effect of P. ginseng on Lymphocytes. Key words: Saponin, Lymphocytes, Aged person, Stimulatory effect, and Panax ginseng

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Removal Behavior of Biological Nitrogen and Phosphorus and Prediction of Microbial Community Composition with Its Function, in an Anaerobic-Anoxic System form Weak Sewage

  • LEE, JIN WOO;EUI SO CHOI;KYUNG IK GIL;HAN WOONG LEE;SANG HYON LEE;SOO YOOUN LEE;YONG KEUN PARK
    • Journal of Microbiology and Biotechnology
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    • v.11 no.6
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    • pp.994-1001
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    • 2001
  • An easier way of understanding the BNR system was proposed from the study on substrate, nutrient removal tendency, microbial community and its metabolic function by applying the municipal settled sewage. During the anaerobic period, the phosphorus release rate per VFACOD we varied depending on the phosphorus content in the sludge. When the phosphorus content in the sludge was $6\%$ VSS, according to influent VFACOD, the phosphorus release rate and PHA production were $0.35 gPO_4P/gVFACOD$ and 1.0 gPHA/gVFACOD, respectively. The $NO_3N$ requirement for the phosphorus uptake as an electron acceptor was about $0.5 gNO_3N/gPO_4P_{uptake}$ based on the proposed equation with PHA, biomass, production, and the concentration of phosphorus release/uptake. Bacterial-community analysis of the sludge, as determined by FISH and 16SrDNA characterization FISH, revealed that the beta-subclass proteobacteria were the most abundant group ($27.9\%$ of the proteobacteria-specific probe EUB338), and it was likely that representative of the beta-subclass played key roles in activated sludge. The next dominant group found was the gamma-protebacteria ($15.4\%$ of probe EUB338). 16S rDNA clone library analysis showed that the members of${\beta}$- and ${\gamma}$-proteobacteria were also the most abundant groups, and $21.5\%$ (PN2 and PN4) and $15.4\%$ (PN1 and PN5) of total clones were the genera of denitrifying bacteria and PAO, respectively. Prediction of the microbial community composition was made with phosphorus content (Pv, $\%$ P/VSS) in wasted sludge and profiles of COD, PHA, $PO_4P,\;and\;NO_3N$ in an anaerobic-anoxic SBR unit. Generally, the predicted microbial composition based upon metabolic function, i.e., as measured by stoichiometry, is fairly similar to that measure by the unculturable dependent method. In this study, a proposal was made on he microbial community composition that was more easily approached to analyze the reactor behavior.

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