• Title/Summary/Keyword: carbon metabolism

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Cloning and Expression of Serratia marcescens Coenzyme A(CoA) Transferase Gene in E. coli

  • Choi, Yong-Lark;Kim, Hae-Sun;Yoo, Ju-Soon;Kim, Yong-Gyun;Chung, Chung-Han
    • Journal of Life Science
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    • v.9 no.1
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    • pp.54-57
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    • 1999
  • We have got several clones from Serratia marcescens which stimulated the cells to use maltose as a carbon source in E. coli TP2139 (${\Delta}$lac, ${\Delta}$crp). One of the cloned genes, pCKB13, was further analyzed. In order to find whether the increased expression of the gene under the direction of maltose metabolism, we constructed several recombinant subclones. We have confirmed that the clone, pCKB13 codes Coenzyme A transferase gene by partial nucleotide sequencing in the terminal region. The enzyme activity of Coenzyme A transferase increased after introduction of the multicopy of the cloned gene in E. coli. The recombinant proteins expressed by multicopy and induction with IPTG, two polypeptide of 26-and 28-kDa, were confirmed by SDS-PAGE. Southern hybridization analysis confirmed that the cloned DNA fragment was originated from S. marcescens chromosomal DNA.

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Electrochemical Activation of Nitrate Reduction to Nitrogen by Ochrobactrum sp. G3-1 Using a Noncompartmented Electrochemical Bioreactor

  • Lee, Woo-Jin;Park, Doo-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.19 no.8
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    • pp.836-844
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    • 2009
  • A denitrification bacterium was isolated from riverbed soil and identified as Ochrobactrum sp., whose specific enzymes for denitrification metabolism were biochemically assayed or confirmed with specific coding genes. The denitrification activity of strain G3-1 was proportional to glucose/nitrate balance, which was consistent with the theoretical balance (0.5). The modified graphite felt cathode with neutral red, which functions as a solid electron mediator, enhanced the electron transfer from electrode to bacterial cell. The porous carbon anode was coated with a ceramic membrane and cellulose acetate film in order to permit the penetration of water molecules from the catholyte to the outside through anode, which functions as an air anode. A non-compartmented electrochemical bioreactor (NCEB) comprised of a solid electron mediator and an air anode was employed for cultivation of G3-1 cells. The intact G3-1 cells were immobilized in the solid electron mediator, by which denitrification activity was greatly increased at the lower glucose/nitrate balance than the theoretical balance (0.5). Metabolic stability of the intact G3-1 cells immobilized in the solid electron mediator was extended to 20 days, even at a glucose/nitrate balance of 0.1.

Adhesive Properties, Extracellular Protein Production, and Metabolism in the Lactobacillus rhamnosus GG Strain when Grown in the Presence of Mucin

  • Sanchez, Borja;Saad, Naima;Schmitter, Jean-Marie;Bressollier, Philippe;Urdaci, Maria C.
    • Journal of Microbiology and Biotechnology
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    • v.20 no.6
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    • pp.978-984
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    • 2010
  • This paper examines the probiotic bacterium Lactobacillus rhamnosus GG, and how it reacts to the presence of mucin in its extracellular milieu. Parameters studied included cell clustering, adhesion to mucin, extracellular protein production, and formation of final metabolites. L. rhamnosus GG was found to grow efficiently in the presence of glucose, N-acetylglucosamine, or mucin (partially purified or purified) as sole carbon sources. However, it was unable to grow using other mucin constituents, such as fucose or glucuronic acid. Mucin induced noticeable changes in all the parameters studied when compared with growth using glucose, including in the formation of cell clusters, which were easily disorganized with trypsin. Mucin increased adhesion of the bacterium, and modulated the production of extracellular proteins. SDS-PAGE revealed that mucin was not degraded during L. rhamnosus GG growth, suggesting that this bacterium is able to partially use the glucidic moiety of glycoprotein. This study goes some way towards developing an understanding of the metabolic and physiological changes that L. rhamnosus GG undergoes within the human gastrointestinal tract.

Isoflavone Daidzein: Chemistry and Bacterial Metabolism

  • Kim, Mi-Hyang;Han, Jae-Hong;Kim, Soo-Un
    • Journal of Applied Biological Chemistry
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    • v.51 no.6
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    • pp.253-261
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    • 2008
  • Isoflavone daidzein is a phytoestrogen widely distributed in Leguminosae and is especially rich in the soybean. The C6-C3 (rings B and C) unit of isoflavones is derived from the phenylpropanoid pathway and the remaining C6 (ring A) unit is from the polyketide pathway. This unique carbon skeleton is the result of isomerization of the flavone catalyzed by the isoflavone synthase, a cytochrome P450 enzyme. The isoflavones daidzein and genistein are present in the plant mostly in the glucosylated forms. However, in the human intestine, the glycosidic linkage is broken, and the free form is uptaked into blood stream. The free form is further metabolized into various reduction products to end up at the equol, which is known to have the most potent estrogenic effect among the metabolites. Several human intestinal bacteria that can convert daidzein into equol have been described, and the study into the chemistry and biochemistry of the daizein reduction would be rewarding to the improvement of the human health.

Crystal Structure of Acyl-CoA Oxidase 3 from Yarrowia lipolytica with Specificity for Short-Chain Acyl-CoA

  • Kim, Sangwoo;Kim, Kyung-Jin
    • Journal of Microbiology and Biotechnology
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    • v.28 no.4
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    • pp.597-605
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    • 2018
  • Acyl-CoA oxidases (ACOXs) play important roles in lipid metabolism, including peroxisomal fatty acid ${\beta}$-oxidation by the conversion of acyl-CoAs to 2-trans-enoyl-CoAs. The yeast Yarrowia lipolytica can utilize fatty acids as a carbon source and thus has extensive biotechnological applications. The crystal structure of ACOX3 from Y. lipolytica (YlACOX3) was determined at a resolution of $2.5{\AA}$. It contained two molecules per asymmetric unit, and the monomeric structure was folded into four domains; $N{\alpha}$, $N{\beta}$, $C{\alpha}1$, and $C{\alpha}2$ domains. The cofactor flavin adenine dinucleotide was bound in the dimer interface. The substrate-binding pocket was located near the cofactor, and formed at the interface between the $N{\alpha}$, $N{\beta}$, and $C{\alpha}1$ domains. Comparisons with other ACOX structures provided structural insights into how YlACOX has a substrate preference for short-chain acyl-CoA. In addition, the structure of YlACOX3 was compared with those of medium- and long-chain ACOXs, and the structural basis for their differences in substrate specificity was discussed.

Electrochemical and Biochemical Analysis of Ethanol Fermentation of Zymomonas mobilis KCCM11336

  • Jeon, Bo-Young;Hwang, Tae-Sik;Park, Doo-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.19 no.7
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    • pp.666-674
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    • 2009
  • An electrochemical bioreactor (ECB) composed of a cathode compartment and an air anode was used in this study to characterize the ethanol fermentation of Zymomonas mobilis. The cathode and air anode were constructed of modified graphite felt with neutral red (NR) and a modified porous carbon plate with cellulose acetate and porous ceramic membrane, respectively. The air anode operates as a catalyst to generate protons and electrons from water. The growth and ethanol production of Z. mobilis were 50% higher in the ECB than were observed under anoxic nitrogen conditions. Ethanol production by growing cells and the crude enzyme of Z. mobilis were significantly lower under aerobic conditions than under other conditions. The growing cells and crude enzyme of Z. mobilis did not catalyze ethanol production from pyruvate and acetaldehyde. The membrane fraction of crude enzyme catalyzed ethanol production from glucose, but the soluble fraction did not. NADH was oxidized to $NAD^+$in association with $H_2O_2$reduction, via the catalysis of crude enzyme. Our results suggested that NADH/$NAD^+$balance may be a critical factor for ethanol production from glucose in the metabolism of Z. mobilis, and that the metabolic activity of both growing cells and crude enzyme for ethanol fermentation may be induced in the presence of glucose.

Computer Models on Oxygenation Process in the Pulmonary Circulation by Gas Diffusion

  • Chang, Keun-Shik;Bae, Hwang
    • International Journal of Vascular Biomedical Engineering
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    • v.4 no.1
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    • pp.9-16
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    • 2006
  • In this article we introduce computer models that have been developed in the past to determine the concentration of metabolic gases, the oxygen and carbon dioxide, along the pulmonary circulation. The terminal concentration of these gases in the arterial blood is related with the total change of the partial pressure of the same gases in the alveoli for the time beginning with inspiration and ending with expiration. It is affected not only by the ventilation-perfusion ratio and the gas diffusion capacity of the lung membrane but also by the pulmonary defect such as shunt, dead space, diffusion impairment and ventilation-perfusion mismatch. Some pathological pulmonary symptoms such as ARDS and CDPD can be understood through the mathematical models of these pulmonary dysfunctions. Quantitative study on the blood oxygenation process using various computer models is therefore of foremost importance in order to monitor not only the pulmonary health but also the cardiac output and cell metabolism. Reviewed in this paper include the basic and advanced methods that enable numerical study on the gas exchange and on the arterial oxygenation process, which might depend on the various heart and lung physiological conditions listed above.

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Differently expressed genes of soybean by ambient heat stress

  • Jung, Inuk;Kim, Jin Hyeon;Jung, Woosuk
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.156-156
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    • 2017
  • Plants are grown under constitutive changing of environmental conditions and response to external conditions at both protein and transcription level. The effects of heat on plant growth are broad and influence the yield directly. Heat stresses could be classified depend on intensity and duration. Fundamental changes of growth condition by climate change maybe or maybe not classified as a stress on plant growth. The effects of a short and unanticipated impact of elevated heat on plant could be different with those of under longer extension of ambient heat. To examine differently expressed gene sets by ambient heat stress of soybean, we grow the soybean in normal condition for three weeks. After that, soybean plants move to growth chamber. The temperature of growth chamber increase up to $9^{\circ}C$ for four days. We have extracted mRNA and micro RNA every 24 hours and carried RNA sequence analysis. We found major metabolic pathways affected by ambient heat stress. Mainly carbon metabolism, translation machinery and amino acid synthesis are affected. We discussed the expression patterns of genes of heat sensing and hormone responses.

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Alteration of Gas Exchange in Rice Leaves Infected with Magnaporthe grisea

  • Yun, Sung-Chul;Kim, Pan-Gi;Park, Eun-Woo
    • The Plant Pathology Journal
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    • v.16 no.5
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    • pp.257-263
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    • 2000
  • Infection with rice blast fungus (Magnaporthe grisea) significantly reduced foliar net photosynthesis (A) of rice cultivars: Ilpoom, Hwasung, and Choochung in greenhouse experiments. By measuring the amount of diseased leaf area with a computer image analysis system, the relation between disease severity (DS) and net photosynthetic rate was curvilinearly correlated (r=0.679). Diseased leaves with 35% blast symptom can be predicted to have a 50% reduction of photosynthesis. The disease severity was linearly correlated (r=0.478) with total chlorophyll (chlorophyll a and chlorophyll b) per unit leaf area(TC). Light use efficiency was reduced by the fungal infection according to the light response curves. However, dark respiration (Rd) did not change after the fungal infection (p=0.526). Since the percent of reduction in photosynthesis greatly exceeded the percent of leaf area covered by blast lesions, loss of photosynthetic tissue on an area basis could not by itself account for the reduced photosynthesis. Quantitative photosynthetic reduction can be partially explained by decreasing TC, but cannot be explained by decreasing Rd. By photosynthesis (A)-internal CO$_2$ concentration (C$_i$ curve analysis, it was suggested that the fungal infection reduced ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) activity, ribulose-1,5-bisphosphate (RuBP) regeneration, and inorganic phosphate regeneration. Thus, the reduction of photosynthesis by blast infection was associated with decreased TC and biochemical capacity, which comprises all carbon metabolism after CO$_2$ enters through the stomata.

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The Pentose Phosphate Pathway as a Potential Target for Cancer Therapy

  • Cho, Eunae Sandra;Cha, Yong Hoon;Kim, Hyun Sil;Kim, Nam Hee;Yook, Jong In
    • Biomolecules & Therapeutics
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    • v.26 no.1
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    • pp.29-38
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    • 2018
  • During cancer progression, cancer cells are repeatedly exposed to metabolic stress conditions in a resource-limited environment which they must escape. Increasing evidence indicates the importance of nicotinamide adenine dinucleotide phosphate (NADPH) homeostasis in the survival of cancer cells under metabolic stress conditions, such as metabolic resource limitation and therapeutic intervention. NADPH is essential for scavenging of reactive oxygen species (ROS) mainly derived from oxidative phosphorylation required for ATP generation. Thus, metabolic reprogramming of NADPH homeostasis is an important step in cancer progression as well as in combinational therapeutic approaches. In mammalian, the pentose phosphate pathway (PPP) and one-carbon metabolism are major sources of NADPH production. In this review, we focus on the importance of glucose flux control towards PPP regulated by oncogenic pathways and the potential therein for metabolic targeting as a cancer therapy. We also summarize the role of Snail (Snai1), an important regulator of the epithelial mesenchymal transition (EMT), in controlling glucose flux towards PPP and thus potentiating cancer cell survival under oxidative and metabolic stress.