• Title/Summary/Keyword: EH-1

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Production of Poly-3-Hydroxyalkanhoate by Haloarcular sp. EH-1 (Haloarcular sp. EH-1으로부터 생분해성 Poly-3-Hydroxyalkanoate의 생산)

  • 정명주;박형숙
    • Journal of Life Science
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    • v.9 no.6
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    • pp.737-742
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    • 1999
  • The extremely halophilic archaebacterium Haloarcular sp. EH-1 was isolated from solar salts. Haloarcular sp. EH-1 accumulated poly(3-hydroxyalkanoate) (PHA) as intracellular granules. PHA production in batch culture have been studied. The PHA was identified as poly(3-hydroxybutyric acid-co-3-hydroxyvaleric acid) (PHB/HV) of 3-hydroxybutyric acid and 3-hydroxyvaleric acid by the analysis of GC, IR and NMR. The melting temperature of PHB/HV was 152.46$^{\circ}C$, viscosity was 1.25 ㎗/g, and molecular weight was $1.44 X 10^5.$

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Cloning and Characterization of Zebrafish Microsomal Epoxide Hydrolase Based on Bioinformatics (생물정보학을 이용한 Zebrafish Microsomal Epoxide Hydrolase 클로닝 및 특성연구)

  • Lee Eun-Yeol;Kim Hee-Sook
    • Microbiology and Biotechnology Letters
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    • v.34 no.2
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    • pp.129-135
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    • 2006
  • A gene encoding for a putative microsomal epoxide hydrolase (mEH) of a zebrafish, Danio rerio, was cloned and characterized. The putative mEH protein of D. rerio exhibited sequence similarity with mammalian mEH and some other bacterial EHs. A structural model for the putative mEH was constructed using homology modeling based on the crystallographic templates, 1 qo7 and 1 ehy. The catalytic triad consisting of $Asp^{233}$, $Glu^{413}$, and $His^{440}$ was identified, and the characteristic features such as two tyrosine residues and oxyanion hole were found to be highly conserved. Based on bioinformatic analysis together with EH activity assay, the putative protein was annotated as mEH of D. rerio. Enantiopure styrene oxide with enantiopurity of 99%ee and yield of 33.5% was obtained from racemic styrene oxide by the enantioselective hydrolysis activity of recombinant mEH of D. rerio for 45 min.

A Study of Assessment for Fatigue Strength of EH Steels at Fillet Welded Joints using 1mm Stress Method (1mm 응력 기법을 적용한 EH 강재 필릿 용접 이음부 피로 강도 평가)

  • Xin, Wen-Jie;Oh, Dong-Jin;Kim, Young-Nam;Kim, Myung-Hyun
    • Journal of the Society of Naval Architects of Korea
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    • v.51 no.1
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    • pp.26-33
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    • 2014
  • In this study, Non-load-carrying EH Grade steels in fillet welded joints were evaluated with both the hot spot stress method and the 1mm stress method. The thickness effect criterion for fatigue strength evaluation of welded of welded steel structures recommendations of the IIW was used to evaluate the fatigue strength of EH40 and EH36 and Both EH40 and EH36 have been compared with FAT 125 curve recommended in the IIW. Furthermore, fatigue strength of the welded tow and the ground conditions for Non-load-carrying EH36 based on the 1mm stress method has been discussed.

Study on Antioxidant and Anti-inflammatory Effects of Components of Mahwangbujaseshin-tang (마황부자세신탕(麻黃附子細辛湯)의 각 구성약물별 항산화 및 항염 효능에 관한 연구)

  • Choi, Chul-Woo;Oh, Min-Seok
    • Journal of Korean Medicine Rehabilitation
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    • v.24 no.4
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    • pp.15-28
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    • 2014
  • Objectives This study was carried out to find out the Antioxidant and Anti-inflammatory Effects of Components of Mahwangbujaseshin-tang in LPS-Stimulated RAW264.7 Macrophages. Methods There are 5 experimental groups. ; normal, control, EH (Ephedrae Herba), ALRP (Aconiti Lateralis Radix Preparata) and AR (Asiasari Radix). The extract of EH, ALRP and AR ($100{\mu}g/ml$) was added to each group. We examined cytotoxicity, total phenolic contents, DPPH and ABTS free radical scavenging activity, Intracellular ROS (reactive oxygen species) production, NO (Total Nitric oxide), iNOS (inducible nitric oxide synthase), PGE2 (prostaglandin E2), COX-2 (cyclooxygenase-2), $IL-1{\beta}$ ($interleukin-1{\beta}$), IL-6 (interleukin-6), $TNF-{\alpha}$ (tumor necrosis factor-${\alpha}$), MMP-9 (matrix metalloproteinase-9), TIMP-1 (tissue inhibitor of metalloproteinase-1) and HO-1 (heme oxygenase-1) expression level. Results 1. Total phenolic contents of EH were in the highest level. 2. DPPH and ABTS free radical scavenging activity of EH was in the highest level. 3. ROS production was significantly decreased in AR. 4. NO production was significantly decreased in EH, ALRP, AR and iNOS expression was decreased in EH, AR. 5. PGE2 and COX-2 expression was decreased in EH, AR. 6. $IL-1{\beta}$ production was significantly decreased in EH, AR and IL-6 production was significantly decreased in AR. $TNF-{\alpha}$ production was significantly decreased in ALRP, AR. 7. MMP-9 and TIMP-1 production were significantly decreased in EH. 8. HO-1 expression was significantly increased in EH. 9. With simultaneous usage of SnPP which is expression inhibitor of HO-1, NO, $IL-1{\beta}$, IL-6 and $TNF-{\alpha}$ production were partially increased in EH, ALRP, AR. Conclusions According to this study, Components of Mahwangbujaseshin-tang have anti-oxidants and anti-inflammation effects in LPS-Stimulated RAW264.7 Macrophages.

Molecular Characterization of Epoxide Hydrolase from Aspergillus niger LK using Phylogenetic Analysis (진화적 유연관계 분석을 통한 Aspergillus niger LK의 Epoxide Hydrolase의 특성분석)

  • 김희숙;이은열;이수정;이지원
    • KSBB Journal
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    • v.19 no.1
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    • pp.42-49
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    • 2004
  • A gene coding for epoxide hydrolase (EH) of Aspergillus niger LK, a fungus possessing the enantioselective hydrolysis activity for racemic epoxides, was characterized by phylogenetic analysis. The deduced protein of A. niger LK epoxide hydrolase shares significant sequence similarity with several bacterial EHs and mammalian microsomal EHs (mEH) and belongs to the a/${\beta}$ hydrolase fold family. EH from A. niger LK had 90.6% identity with 3D crystal structure of lqo7 in Protein Data Bank. Sequence comparison with other source EHs suggested that Asp$\^$l92/, Asp$\^$374/ and His$\^$374/ constituted the catalytic triad. Based on the multiple sequence comparison of the functional and structural domain sequence, the phylogenetic tree between relevant epoxide hydrolases from various species were reconstructed by using Neighbor-Joining method. Genetic distances were so far as 1.841-2.682 but characteristic oxyanion hole and catalytic triad were highly conserved, which means they have diverged from a common ancestor.

The Effect of EH-HA(Ephedrae Herba Herbal Acupuncture) at ST36 on OVA-induced Asthma in Mice (마황(麻黃) 약침(藥鍼)의 천식억제(喘息抑制) 및 면역조절효과(免疫調節效果)에 대(對)한 실험적(實驗的) 연구(硏究))

  • Song, Hyong-Gun;Lee, Hyeon
    • Journal of Acupuncture Research
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    • v.23 no.1
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    • pp.1-14
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    • 2006
  • Background : Recently, so many people are suffered from the allergic or auto-immune disease, and the representative disease is just Allergic Asthma. It is because human immune function has been decreased. Many treatments were done to treat this disease, and many methods were studied to increase immune function and to suppress the asthma. But, the effect of asthma-suppression and improvement of immune response of EH-HA(Ephedrae Herba Herbal Acupuncture) has not been studied in detail. To study the effects of EH-HA, we injected EH-HA at Joksamni(ST36) of C57BL/6 mice. Objectives : The purpose of this study is to investigate the effect of asthma-suppression and improvement of immune response of EH-HA(Ephedrae Herba Herbal Acupuncture). EH-HA was done at Joksamni(ST36) of the mice with ovalbumin-induced asthma. Methods : C57BL/6 mice were sensitized and challenged with OVA(ovalbumin) for 12 weeks. Two experimental groups were treated with different concentrations(1%, 0.1%) of EH-HA at Joksamni(ST36) for the later 8 weeks(3times/week). Results : 1. The lung weight of the group treated with EH-HA decreased significantly compared with that of control group. 2. The total cells in lung, total leukocytes and eosinophils in BALF of the group treated with EH-HA decreased significantly compared with those of control group. 3. Eosinophils in BALF of the group treated with EH-HA in photomicrographs decreased significantly compared with those of control group. 4. The concentrations of IL-13, IgE, IL-4 in serum and IL-4 in BALF of the group treated with EH-HA decreased significantly compared with those of control group. 5. The numbers of $Gr-1^+/CD11b^+\;cells,\;CD3e^-/CCR3^+\;cells,\;CD4^+\;cells,\;CD8^+\;cells,\;CD3e^+/CD69^+\;cells\;and\;IgE^+/B220^+\;cells$ in lung of the group treated with EH-HA decreased significantly compared with those of control group. 6. In RT-PCR, the mRNA expression of IL-4, IL-5 and IL-13 in the group treated with EH-HA decreased compared with those of control group. Conclusion : These results suggested that EH-HA at Joksamni(ST36) in C57BL/6mice may be effective to OVA-induced asthma of C57BL/6 mice.

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Progesterone Effects on Microsomal Epoxide Hydrolase and Glutathione S-transferease mRNA Levels in Rats (랫드 간 Epoxide Hydrolase와 Glutathione S-Transferase 유전자 발현에 미치는 Progesterone의 효과)

  • Cho, Joo-Youn;Kim, Sang-Geon
    • The Korean Journal of Pharmacology
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    • v.32 no.2
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    • pp.233-241
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    • 1996
  • Previous studies have shown that glucocorticoid suppresses microsomal epoxide hydrolase(EH) gene expression and that EH expression is altered during pregnancy. The effects of progesterone on the expression of rat EH and certain glutathione S-transferase(GST) genes were examined in this study. Northern RNA blot analysis revealed that progesterone was effective in increasing hepatic EH mRNA levels at 12 h to 48 h after treatment with a maximal 9-fold increase being noted at 12 h time point. Nonetheless, multiple daily treatment with progesterone rather caused minimal relative increases in EH mRNA levels. GST Ya and Yb1/2 mRNA levels were also transiently elevated at 12 h after progesterone treatment, followed by gradual decreases from the maximal Increases at day 1, 2 and 5 post-treatment. These changes in EH and GST mRNA levels were noted only at a relatively high dose of progesterone. Furthermore, immunoblot analyses showed that rats treated with progesterone for 5 days failed to show EH or GST induction, indicating that progesterone-induced alterations in EH and GST mRNA levels do not reflect bona fide induction of the detoxifying enzymes. Concomitant progesterone treatment of rats with the known EH inducers including ketoconazole and clotrimazole failed to additively nor antagonistically alter EH mRNA levels. In contrast, dexamethasone substantially reduced ketoconazole- or clotrimazole-inducible EH expression. These results showed that progesterone stimulates the EH, GST Ya and Yb1/2 gene expression at early times followed by marked reduction in the RNA levels from the maximum after multiple treatment and that the changes in mRNA do not necessarily reflect induction of the proteins.

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Characterization of Hexane Biodegradation by Rhodococcus sp. EH741 (Rhodococcus sp. EH741에 의한 Hexane 생분해 특성)

  • Lee, Eun-Hee;Cho, Kyung-Suk
    • Journal of Korean Society of Environmental Engineers
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    • v.28 no.2
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    • pp.144-149
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    • 2006
  • As a strain EH741, having an excellent hexane degradability, was isolated from bacterial consortium using hexane as a sole carbon and energy source. EH741 was identified as a Rhodococcus sp. and the addition of a surfactant Pluronic F68(PF68), for increasing hexane solubility couldn't enhance the specific growth rate of the isolate EH741 n the mineral salt medium supplemented with hexane as a sole carbon source(hexane-BH medium). In the hexane-BH medium, the maximum specific growth rate(${\mu}_{max}$) of this strain was $0.04h^{-1}$, and the maximum hexane degradation rate($V_{max}$) and saturation constant($K_s$) were$161{\mu}mol{\cdot}g-DCW^{-1}{\cdot}h^{-1}$ and 10.5 mM, respectively. Rhodococcus sp. EH741 was one of excellent microorgamisms for hexane biodegradation processes.

Optimization of the Reaction Conditions for (R)-Phenyl-1,2-ethanediol Preparation by Recombinant Epoxide Hydrolase from Caulobacter crescentus (재조합 Caulobacter crescentus 에폭사이드 가수분해효소를 이용한 광학활성 Diol 제조 조건의 최적화)

  • Lee, Ok-Kyung;Kim, Hee-Sook
    • Microbiology and Biotechnology Letters
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    • v.35 no.4
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    • pp.285-291
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    • 2007
  • Enantioconvergent hydrolysis process for the preparation of chiral diol from racemic epoxides by using the recombinant Caulobacter crescentus epoxide hydrolase (CcEH) in Escherichia coli BL21 (DE3) was optimized. For the optimization, the effects of detergent, temperature and product inhibition on the enantiopurity and the yield of diol were investigated. (R)-phenyl-1,2-ethanediol with 92% enantiomeric excess and 56% yield from 20 mM racemic styrene oxide was obtained by using the recombinant CcEH at the optimal condition of $10^{\circ}C$ and the addition of 2% (w/v) Tween 80. At 50 mM racemic styrene oxide was used as a substrate, (R)-phenyl-1,2-ethanediol was obtained with 87% enantiomeric excess and 77% yield. Racemic phenyl-1,2-ethanediol, (R)-phenyl-1,2-ethanediol and (S)-phenyl-1,2-ethanediol dramatically inhibited the hydrolytic activity of the recombinant CcEH. These results suggested that another EH with the regioselectivity on ${\beta}$-position of (R)-enantiomer and without feedback inhibition by products would be needed as the partner EH of C. crescentus EH.

Purification of Thiazole- and Pyrazine-inducible Microsomal Epoxide Hydrolase: Induction of Epoxide Hydrolase-related Novel 43 kDa Protein (Thiazole 또는 Pyrazine유도성 Microsomal Epoxide Hydrolase의 순수정제: Epoxide Hydrolase-관련성 43 kDa 단백질의 유도증가)

  • Kim, Sang-Geon
    • The Korean Journal of Pharmacology
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    • v.29 no.2
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    • pp.275-282
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    • 1993
  • Liver microsomal epoxide hydrolase (mEH) is active in the detoxification of epoxide-containing reactive intermediate. Previous studies in this laboratory have shown that thiazole and pyrazine are efficacious inducers of mEH in rats with large increases in mEH mRNA levels (Carcinogensis, Kim et al, 1993). mEH was purified to electrophoretic homogeneity from thiazole-induced rat hepatic microsomes using DEAE-cellulose column chromatography whereas another protein $({\sim}43\;kDa)$ was co-purified with mEH from pyrazine-induced rat hepatic micrsomes (200 mg/kg body weight/day, ip, 3d). The antibody raised from a rabbit against mEH protein purified from thiazole-induced rat hepatic microsomes appeared to specifically recognize mEH protein in rat hepatic microsomes, as assessed by immunoblotting analysis. Immunoblotting analyses revealed a 10- and 7-fold increase in mEH levels in the hepatic microsomes isolated from thiazole- and pyrazine-treated rats, respectively. Moreover, immunoblotting analysis showed cross-reactivity of the mEH antibody with a 43 kDa protein in pyrazine-induced rat hepatic microsomes and with co-purified 43 kDa protein in purified fractions. The ratio between the 43 kDa protein and mEH in pyrazine-induced rat microsomes or in purified fractions was ${\sim}1$ to 15. N-terminal amino acid sequence analysis of both purified rat mEH and 43 kDa protein revealed that 10 out of 12 amino acids in N-terminus of the 43 kDa protein were identical with the mEH sequence with two amino acid residues of the 43 kDa protein undetermined. Either thiazole or pyrazine treatment, however, failed to increase the levels of mEH protein in rabbits while pyrazine caused elevation of the 43 kDa protein in this species, as determined by irnrnunoblotting analysis. These results demonstrated that treatment of rats with either thiazole or pyrazine causes elevation in hepatic mEH expiession whereas pyrazine treatment results in induction of another mEH-related 43 kDa protein and that a distinct species difference exists between rats and rabbits in the induction of mEH by these xenobiotics.

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