• 제목/요약/키워드: Metabolic pathway

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Cloning and Expression of the UDP-Galactose-4-Epimerase Gene (galE) Constituting the gal/lac Operon of Lactococcus lactis ssp. lactis ATCC7962

  • Lee, Jung-Min, Choi, Jae-Yeon;Lee, Jong-Hoon;Chang, Hae-Choon;Chung, Dae-Kyun;Kim, Jeong-Hwan;Lee, Hyong-Joo
    • Journal of Microbiology and Biotechnology
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    • 제9권4호
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    • pp.393-397
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    • 1999
  • The gene (galE) encoding UDP-galactose-4-epimerase, operative in the galactose metabolic pathway, was cloned together with the $\beta$-galactosidase gene (lacZ) from Lactococcus lactis ssp. lactis ATCC7962 (L. lactis 7962). galE was found to have a length of 981 bps and encoded a protein with a molecular mass of 36,209 Da. The deduced amino acid sequence showed a homology with GalE proteins from several other microorganisms. A Northern analysis demonstrated that galE was constitutively expressed by its own promoter. When galactose or lactose was added into medium, the galE transcription was induced by several upstream promoters. The structure of the gal/lac operon of L. lactis 7962 was partially characterized and the gene order around galE was galT-lacA-lacZ-galE-orfX.

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Biotransformation of Eugenol via Protocatechuic Acid by Thermophilic Geobacillus sp. AY 946034 Strain

  • Giedraityte, Grazina;Kalediene, Lilija
    • Journal of Microbiology and Biotechnology
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    • 제24권4호
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    • pp.475-482
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    • 2014
  • The metabolic pathway of eugenol degradation by thermophilic Geobacillus sp. AY 946034 strain was analyzed based on the lack of data about eugenol degradation by thermophiles. TLC, GC-MS, and biotransformation with resting cells showed that eugenol was oxidized through coniferyl alcohol, and ferulic and vanillic acids to protocatechuic acid before the aromatic ring was cleaved. The cell-free extract of Geobacillus sp. AY 946034 strain grown on eugenol showed a high activity of eugenol hydroxylase, feruloyl-CoA synthetase, vanillate-O-demethylase, and protocatechuate 3,4-dioxygenase. The key enzyme, protocatechuate 3,4-dioxygenase, which plays a crucial role in the degradation of various aromatic compounds, was purified 135-fold to homogeneity with a 34% overall recovery from Geobacillus sp. AY 946034. The relative molecular mass of the native enzyme was about $450{\pm}10$ kDa and was composed of the non-identical subunits. The pH and temperature optima for enzyme activity were 8 and $60^{\circ}C$, respectively. The half-life of protocatechuate 3,4-dioxygenase at the optimum temperature was 50 min.

카드뮴에 대한 녹조류 Selenastrum capricornutum (Chlorophyceae)의 세포활력도 및 peroxidase 활성도 변화 (Changes in Cellular Viability and Peroxidase Activities of Green Algae Selenastrum capricornutum (Chlorophyceae) to Cadmium)

  • 최은주;이상구;이승진;문성경;박용석;이기태
    • Environmental Analysis Health and Toxicology
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    • 제18권4호
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    • pp.295-303
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    • 2003
  • Physiological cellular activities responses to cadmium (Cd) exposure in green algae with several reductases activities and viability of the cell were examined. The cell division of green algae, Selenastrum capricornutum treated with 5ppm was significantly decreased than that of normal algae. The mean cell number of normal algal culture was as twice much as than that of algae at 6 days after Cd treatment. The cellular viability of algae was analysed by flow-cytometry with fluorescent dye after esterase reaction on cell membrane. The 85.35% of cellular viability of normal culture was decreased to 34.35% when algae was treated with 5 ppm of Cd at 6 days after treatment. It was considered that those method of flow-cytometry is useful tool for toxicity test on micro-organisms in the respect of identifying cellular viability. Also, the activities of both glutathione peroxidase (GPX) and ascorbate peroxidase (APX), which are indirectly react against oxidative stress through reduction of glutathione by Cd were significantly increased with 25%. It is considered that both GPX and APX are involved in the metabolic pathway of Cd -detoxification with similar portion in Selenasturm capricornutum.

A Study on Transcriptome Analysis Using de novo RNA-sequencing to Compare Ginseng Roots Cultivated in Different Environments

  • Yang, Byung Wook
    • 한국자원식물학회:학술대회논문집
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    • 한국자원식물학회 2018년도 춘계학술발표회
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    • pp.5-5
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    • 2018
  • Ginseng (Panax ginseng C.A. Meyer), one of the most widely used medicinal plants in traditional oriental medicine, is used for the treatment of various diseases. It has been classified according to its cultivation environment, such as field cultivated ginseng (FCG) and mountain cultivated ginseng (MCG). However, little is known about differences in gene expression in ginseng roots between field cultivated and mountain cultivated ginseng. In order to investigate the whole transcriptome landscape of ginseng, we employed High-Throughput sequencing technologies using the Illumina HiSeqTM2500 system, and generated a large amount of sequenced transcriptome from ginseng roots. Approximately 77 million and 87 million high-quality reads were produced in the FCG and MCG roots transcriptome analyses, respectively, and we obtained 256,032 assembled unigenes with an average length of 1,171 bp by de novo assembly methods. Functional annotations of the unigenes were performed using sequence similarity comparisons against the following databases: the non-redundant nucleotide database, the InterPro domains database, the Gene Ontology Consortium database, and the Kyoto Encyclopedia of Genes and Genomes pathway database. A total of 4,207 unigenes were assigned to specific metabolic pathways, and all of the known enzymes involved in starch and sucrose metabolism pathways were also identified in the KEGG library. This study indicated that alpha-glucan phosphorylase 1, putative pectinesterase/pectinesterase inhibitor 17, beta-amylase, and alpha-glucan phosphorylase isozyme H might be important factors involved in starch and sucrose metabolism between FCG and MCG in different environments.

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Identification of Propentofylline Metabolites in Rats by Gas Chromatography/Mass Spectrometry

  • Kwon, Oh-Seung;Ryu, Jae-Chun
    • Archives of Pharmacal Research
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    • 제23권4호
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    • pp.374-380
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    • 2000
  • Propentofylline (PPF, 3-methyl-1-(5-oxohexyl)-7-propylxanthine) has been reported to be a compound for treatment of both vascular dementia and dementia of the Alzheimer type. The short half-life (about 15 min) of PPF at the terminal elimination phase and poor bioavailability after oral administration of PPF to rabbits (Kim et al., 1992) suggest in part that this drug takes the extensive first-pass metabolism in the liver. In addition, the metabolic pathway for PPF remains unclear. The objective of this experiment is to identify urinary metabolites of PPF in rats. For the identification of the metabolites, rat urine was collected after oral administration of 100${m}g/kg$ PPF. PPF metabolite, 3-methyl-1-(5-hydroxyhexyl)-7-propylxanthine, was synthesized and confirmed by gas chromatography/mass spectroscopy (GC/MS) and $^1H$ nuclear magnetic resonance spectroscopy. The urinary metabolites of PPF were extracted with diethyl ether and identified by electron impact and chemical ionization GC/MS. One urinary metabolite was confirmed to be 3-methyl-1-(5-hydroxyhexyl)-7-propylxanthine by synthesized authentic compound. Several metabolites of monohydroxy- and dihydroxy-PPF were identified based on mass fragmentation of both intact and trimethylsilylated derivatives of PPF metabolites and the novel structure of these metabolites is suggested based on mass spectra.

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아황산가스에 노출된 백서조직 lactic dehydrogenase의 isozyme변화에 관한 연구 (A study on the isozyme alterations of lactic dehydrogenase in the tissues of albino rat by the exposure in sulfur dioxide)

  • 권숙표
    • 약학회지
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    • 제13권4호
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    • pp.101-110
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    • 1969
  • The isozyme alteration of lactic dehydrogenase in the tissues of albino rat inhaled SO$_{2}$ were studied in vivo and in vitro, with the following results: (1) The H-type of LDH activity relatively dominated in the normal brain, heart and kidney tissues of rat, M-type in the normal lung, liver, and muscle tissues of the animal. (2) When rats inhale SO$_{2}$ in the concentration of 250 ppm, it appears that the M-type tends to predominate in the anaerobic tissues such as liver, kidney and muscle tissues and the H-type in the aerobic tissues such as brain and heart tissues. (3) When 5% SO$_{2}$ is introduced into tissue homogenates, LDH activities in the heart, lung, liver and muscle tissues are increased more than that of introducing room-air only. With sam treatment, LDH activity is decreased in the kidney tissue and no alteration is observed in the brain tissue. (4) Although, after the aeration of SO$_{2}$, the oxygen tension seems to bring decreases in the level of LDH activity in the anerobic tissues such as liver and muscle tissues, while, on the other hand, increases in the level of the activity in the aerobic tissues, such as the brain, heart and lung tissues. (5) Accordinglly, SO$_{2}$ affects LDH activities, its isozyme pattern of each organs, and their metabolic pathway by its absorption of the gas.

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Succinic Acid Production by Continuous Fermentation Process Using Mannheimia succiniciproducens LPK7

  • Oh, In-Jae;Lee, Hye-Won;Park, Chul-Hwan;Lee, Sang-Yup;Lee, Jin-Won
    • Journal of Microbiology and Biotechnology
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    • 제18권5호
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    • pp.908-912
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    • 2008
  • To achieve a higher succinic acid productivity and evaluate the industrial applicability, this study used Mannheimia succiniciproducens LPK7 (knock-out: ldhA, pflB, pta-ackA), which was recently designed to enhance the productivity of succinic acid and reduce by-product secretion. Anaerobic continuous fermentation of Mannheimia succiniciproducens LPK7 was carried out at different glucose feed concentrations and dilution rates. After extensive fermentation experiments, a succinic acid yield and productivity of 0.38 mol/mol and 1.77 g/l/h, respectively, were achieved with a glucose feed concentration of 18.0 g/l and $0.2\;h^{-1}$ dilution rate. A similar amount of succinic acid production was also produced in batch culture experiments. Therefore, these optimal conditions can be industrially applied for the continuous production of succinic acid. To examine the quantitative balance of the metabolism, a flux distribution analysis was also performed using the metabolic network model of glycolysis and the pentose phosphate pathway.

Rhizopus oryzae의 카드뮴 해독기작과 이에 관련된 동위효소의 변화 양상 (Detoxification Mechanism and Isoenzyme Pattern Changes against Cadmium in Rhizopus oryzae)

  • 이기성;김영호;박영식;박용근
    • 한국균학회지
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    • 제23권1호통권72호
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    • pp.86-91
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    • 1995
  • Rhizopus oryzae의 카드뮴 적응 및 해독기작과 이에 관련된 세포내 생리 생화학적 변화를 조사하였다. R. oryzae는 카드뮴을 첨가 배양하였을 때 카드뮴 영향하에서는 carbohydrate metabolic pathway에 관련된 효소 활성(MDH, GPI)이 촉진되고 과산화물 제거에 관여하는 효소가 새롭게 유도(CAT2)된 반면, lactate를 이용하는 효소(LDH, ADH)의 활성이 감소된 사실은 중금속 영향하에서 세포의 성장과 에너지 공급을 위해 에너지 수율이 낮은 lactate를 이용하는 경로보다는 에너지 수율이 높은 TCA cycle 경로에 작용하는 효소들과 독성과산화물 제거에 관여하는 효소의 더 많은 derepression이 필요하다는 것을 알 수 있었다.

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Liposoluble portion of the red alga Pyropia yezoensis protects alcohol induced liver injury in mice

  • Lee, Ji-Hyeok;Ahn, Ginne;Ko, Ju-Young;Kang, Nalae;Jung, Kyungsook;Han, Eui-Jeong;Kim, Gwang-Hoon;Kim, Hee Jeong;Choi, Cheol Soo;Jeon, You-Jin
    • ALGAE
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    • 제36권3호
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    • pp.219-229
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    • 2021
  • The hepatoprotective effect of liposoluble portion of Pyropia yezoensis (PYLP) was investigated against alcohol-induced liver injury in mice. Fatty acids were predominant in PYLP obtained from hexane fraction of 70% EtOH extract after ultrasonication. In particular, polyunsaturated fatty acids such as eicosapentaenoic acid and linoleic acid accounted for 56.91% of the total lipids. PYLP significantly reduced liver damage induced by the alcohol treatment in mice. PYLP treatment increased the activity of antioxidant enzymes including superoxide dismutase, catalase, and glutathion peroxidase by reducing thiobarbituric acid reactive substances. Histological observations showed that PYLP minimizes damage to living tissue induced by alcohol treatment by modulating the expression level of proteins involved in the anti-apoptotic signaling pathway. Our results suggest that PYLP, rich in polyunsaturated fatty acids extracted from the red alga P. yezoensis, will be useful as a potential liver protectant in the hangover industry.

Identification of HYIpro-3-1 Metabolites, a Novel Anti-Inflammatory Compound, in Human Liver Microsomes by Quadrupole-Orbitrap High-Resolution Mass Spectrometry

  • Bai, Honghao;Kim, Younah;Paudel, Sanjita;Lee, Eung-Seok;Lee, Sangkyu
    • Mass Spectrometry Letters
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    • 제12권4호
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    • pp.172-178
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    • 2021
  • HYIpro-3-1 is an adjuvant for preventing or treating inflammatory growth diseases. In this study, we identified the metabolic pathway of HYIpro-3-1 in human liver microsomes (HLMs) by quadrupole-orbitrap high-resolution mass spectrometry (HR-MS) and characterized the major human cytochrome P450 (CYP). Ten metabolites were identified, including one O-demethylation (M1), two O-demethylation and monohydroxylation (M2 and M3), and seven monohydroxylation metabolites (M4-M10). Based on the HR-MS2 spectra, the metabolites are divided into two groups of monohydroxylated metabolites according to the hydroxylation position. We verified that HYIpro-3-1 is metabolized by CYP in HLMs, CYP2B6 is mainly involved in O-demethylation, and various CYPs are involved in the monohydroxylation of HYIpro-3-1.