• Title/Summary/Keyword: Metabolic Enzymes

검색결과 373건 처리시간 0.027초

Synthesis of (R,R)-2,3-butanediol from starch in a hybrid cell-free reaction system

  • Yi, Tong;Lim, Hye Jin;Lee, So Jeong;Lee, Kyung-Ho;Kim, Dong-Myung
    • Journal of Industrial and Engineering Chemistry
    • /
    • 제67권
    • /
    • pp.231-235
    • /
    • 2018
  • In this study, we demonstrate the conversion of starch to (R,R)-2,3-butanediol (2,3-BD) in a hybrid cell-free synthesis system containing a mixture of lysates derived from Escherichia coli (E. coli) and cyanobacteria. A sufficient pool of pyruvate required for the synthesis of 2,3-BD was generated by combining metabolic pathways of cyanobacteria and E. coli. Successful synthesis of 2,3-BD was achieved by additional modifications of the hybrid cell-free system with the enzymes required to convert pyruvate to 2,3-BD. The results demonstrate a new approach to harness biological pathways to expand the scope of cell-free metabolic engineering by cross-species combinations of cell lysates.

Modulation of L-Arginine-Arginase Metabolic Pathway Enzymes: Immunocytochemistry and mRNA Expression in Peripheral Blood and Tissue Levels in Head and Neck Squamous Cell Carcinomas in North East India

  • Srivastava, Shilpee;Ghosh, Sankar Kumar
    • Asian Pacific Journal of Cancer Prevention
    • /
    • 제16권16호
    • /
    • pp.7031-7038
    • /
    • 2015
  • Background: Arginine may play important roles in tumor progression by providing ornithine for polyamine biosynthesis, required for cell growth. The aim of this work was to determine the expression of arginine metabolic pathway enzymes in head and neck squamous cell carcinoma (HNSCC) in northeast India. Materials and Methods: The expressions of arginase isoforms (ARG1 and ARG2), ornithine aminotransferase (OAT) and ornithine decarboxylase (ODC) were examined in fifty paired HNSCC and adjacent non-tumor tissues by immunohistochemistry. Immunocytochemistry, semiquantitative reverse transcription sq-PCR and quantitative real-time qPCR were used to assess protein and mRNA expressions in peripheral blood of fifty HNSCC patients and hundred controls. Results: ARG1 and ODC protein and mRNA were strongly expressed in peripheral blood from HNSCC patients. No ARG2 expression was observed. In vivo, expression of ARG1, ARG2 and ODC was significantly higher in tumor than in non-tumor tissues. Most tumors expressed low levels of OAT, with no difference in tissues or blood, compared to controls. The absolute extent of maximal ARG1 upregulation with qPCR showed 6.23 fold increase in HNSCC. Conclusions: These findings strongly suggest that in HNSCCs, the ARG1 pathway is stimulated leading to the formation of polyamines as indicated by higher ODC expression, which promote tumor growth.

Effect of the Saponin Fraction of Korean Ginseng on the Ethanol Metabolism in the Animal Body

  • Joo, Chung-No;Kwak, Hahn-Shik
    • 고려인삼학회:학술대회논문집
    • /
    • 고려인삼학회 1987년도 Proceedings of Korea-Japan Panax Ginseng Symposium 1987 Seoul Korea
    • /
    • pp.47-58
    • /
    • 1987
  • Ethanol exerts different effects on hepatic cellular metabolism, depending mainly on the duration of its intake. In the presence of ethanol following an acute load, a number of hepatic functions are inhibited, including lipid oxidation and microsomal drug metabolism. In its early stages, chronic ethanol consumption produces adaptive metabolic changes in the endoplasmic reticulum which result in increased metabolism of ethanol and drugs and accelerated lipoprotein production. Prolongation of ethanol intake may result in injurious hepatic lesions such as alcoholic hepatitis and cirrhosis A number of such metabolic effects of ethanol are directly linked to the two major products of its oxidation; hydrogen and acetaldehyde. The excess hydrogen from ethanol unbalances the liver cell's chemistry. In the presence of excess hydrogen ions the process is turned in a different direction. In this study, it was attempted to observe the effect of ginseng saponins on alcohol Oehydrogenase(ADH), aldehyde dehydrogenase(ALDH) and microsomal ethanol oxidizing system(MEOS) in vivo as well as in vitro. Furthermore, the effect of ginseng saponin on the hydrogen balance in the liver and the hepatic cellular distribution of (1-14C) ethanol, its incorporation into acetaldehyde and lipids was also investigated. It seemed that ginseng saponin stimulated the above enzymes and other related enzymes in ethanol metabolism, resulting in a rapid removal of acetaldehyde and excess hydrogen from the animal body,

  • PDF

Effect of Chromium Stress on Antioxidative Enzymes and Malondialdehyde Content Activities in Leaves and Roots of Mangrove Seedlings Kandelia Candel (L.) Druce

  • Rahman, Mohammed Mahabubur;Rahman, Motiur M.;Islam, Kazi Shakila;Chongling, Yan
    • Journal of Forest and Environmental Science
    • /
    • 제26권3호
    • /
    • pp.171-179
    • /
    • 2010
  • Effect of chromium (Cr) stress on antioxidant enzyme activities and malondialdehyde (MDA) content were investigated in leaves and roots of mangrove (italic (L.) Druce) seedlings. Cr toxicity effects were also assessed on young seedlings. The seedlings were grown in green house condition for three months in nutrient solution with 0, 0.5, 1, 1.5, 2, 2.5, and 3 mg $L^{-1}$ $CrCl_3$. This study showed that Cr led to the change of antioxidant enzymes such as superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) and activities at different concentrations. The activity of antioxidant enzymes in leaves of K. candel seedlings indicates that enzymes engaged in antioxidant defense in certain level especially in low concentration of Cr treatments. The activities of SOD and POD were activated by Cr in the root level, while CAT activity was inhibited. CAT activity decreased in response to high concentrations of Cr. In the present study indicated that SOD in root was active in scavenging the superoxide produced by Cr. Both in roots and leaves, an increase in malondialdehyde (MDA) content was observed with increase in metal concentration and exposure periods. Our finding indicated that the high concentration of excessive Cr supply may interfere with several metabolic processes of seedlings, causing toxicity to plants as exhibited by chlorosis, necrosis, photosynthetic impairing and finally, plant death.

Computational identification of significantly regulated metabolic reactions by integration of data on enzyme activity and gene expression

  • Nam, Ho-Jung;Ryu, Tae-Woo;Lee, Ki-Young;Kim, Sang-Woo;Lee, Do-Heon
    • BMB Reports
    • /
    • 제41권8호
    • /
    • pp.609-614
    • /
    • 2008
  • The concentrations and catalytic activities of enzymes control metabolic rates. Previous studies have focused on enzyme concentrations because there are no genome-wide techniques used for the measurement of enzyme activity. We propose a method for evaluating the significance of enzyme activity by integrating metabolic network topologies and genome-wide microarray gene expression profiles. We quantified the enzymatic activity of reactions and report the 388 significant reactions in five perturbation datasets. For the 388 enzymatic reactions, we identified 70 that were significantly regulated (P-value < 0.001). Thirty-one of these reactions were part of anaerobic metabolism, 23 were part of low-pH aerobic metabolism, 8 were part of high-pH anaerobic metabolism, 3 were part of low-pH aerobic reactions, and 5 were part of high-pH anaerobic metabolism.

Tropane alkaloid의 생합성과 분자육종 (Metabolic Engineering of Medicinal Plants tov Tropane Alkaloid Production)

  • 윤대진
    • Journal of Plant Biotechnology
    • /
    • 제29권3호
    • /
    • pp.199-207
    • /
    • 2002
  • The tropane alkaloids hyoscyamine (its racemic form being atropine) and scopolamine are used medicinally as anticholinergic agents that act on the parasympathetic nerve system. Because they differ in their actions on the central nervous system, currently there is a 10-fold higher commercial demand for scopolamine, in the N-butylbromide form, than there is for hyoscyamine and atropine combined. Several solanaceous species have been used as the commercial sources of these alkaloids, but the scopolamine contents in these plants often are much lower than those of hyoscyamine. For this reason there has been long-standing interest in increasing the scopolamine contents of cultivated medicinal plants. Naturally occurring and artificial interspecific hybrids of Duboisia have high scopolamine contents and are cultivated as a commercial source of scopolamine in Australia and other countries. Anther culture combined with conventional interspecific hybridization also has been used to breed high scopolamine-containing plants in the genera Datura and Hyoscyamus, but without much success. The use of recombinant DNA technology for the manipulation of metabolic processes in cells promises to provide important contributions to basic science, agriculture, and medicine. In this review, I introduce on the enzymes and genes involved in tropane alkaloid biosynthesis and current progress in metabolic engineering approaches for tropane alkaloid, especially scopolamine, production.

Isolation, Characterization, and Metabolic Profiling of Ceratorhiza hydrophila from the Aquatic Plant Myriophyllum spicatum

  • Elsaba, Yasmin M.;Boroujerdi, Arezue;Abdelsalam, Asmaa
    • Mycobiology
    • /
    • 제50권2호
    • /
    • pp.110-120
    • /
    • 2022
  • The goal of the present study was to investigate the antibacterial properties, enzyme production, and metabolic profiling of a new Ceratorhiza hydrophila strain isolated from the submerged aquatic plant Myriophyllum spicatum. Furthermore, the fungus' morphological characterization and DNA sequencing have been described. The fungus has been identified and submitted to the GenBank as Ceratorhiza hydrophila isolate EG19 and the fungus ID is MK387081. The enzyme analyses showed its ability to produce protease and cellulase enzymes. According to the CSLI standard, the ethyl acetate extract of C. hydrophila showed intermediate antibacterial activity against Streptococcus pneumonia, Micrococcus luteus, and Staphylococcus aureus. Metabolic profiling has been carried out using 700 MHz NMR spectroscopy. Based on the 1H and 1H-13C heteronuclear single quantum coherence (HSQC) NMR data and NMR databases, 23 compounds have been identified. The identified metabolites include 31% amino acids, 9% sugars, 9% amines, 4% sugar alcohols, and 4% alkaloids. This is the first report for the metabolic characterization of C. hydrophila, which gave preliminary information about the fungus. It is expected that our findings not only will pave the way to other perspectives in enormous applications using C. hydrophila as a new promising source of antimicrobial agents and essential metabolites, but also it will be valuable in the classification and chemotaxonomy of the species.

Cloning, Expression, and Characterization of a Cold-Adapted Shikimate Kinase from the Psychrophilic Bacterium Colwellia psychrerythraea 34H

  • Nugroho, Wahyu Sri Kunto;Kim, Dong-Woo;Han, Jong-Cheol;Hur, Young Baek;Nam, Soo-Wan;Kim, Hak Jun
    • Journal of Microbiology and Biotechnology
    • /
    • 제26권12호
    • /
    • pp.2087-2097
    • /
    • 2016
  • Most cold-adapted enzymes possess higher $K_m$ and $k_{cat}$ values than those of their mesophilic counterparts to maximize the reaction rate. This characteristic is often ascribed to a high structural flexibility and improved dynamics in the active site. However, this may be less convincing to cold-adapted metabolic enzymes, which work at substrate concentrations near $K_m$. In this respect, cold adaptation of a shikimate kinase (SK) in the shikimate pathway from psychrophilic Colwellia psychrerythraea (CpSK) was characterized by comparing it with a mesophilic Escherichia coli homolog (EcSK). The optimum temperatures for CpSK and EcSK activity were approximately $30^{\circ}C$ and $40^{\circ}C$, respectively. The melting points were $33^{\circ}C$ and $45^{\circ}C$ for CpSK and EcSK, respectively. The ${\Delta}G_{H_2O}$ (denaturation in the absence of denaturing agent) values were 3.94 and 5.74 kcal/mol for CpSK and EcSK, respectively. These results indicated that CpSK was a cold-adapted enzyme. However, contrary to typical kinetic data, CpSK had a lower $K_m$ for its substrate shikimate than most mesophilic SKs, and the $k_{cat}$ was not increased. This observation suggested that CpSK may have evolved to exhibit increased substrate affinity at low intracellular concentrations of shikimate in the cold environment. Sequence analysis and homology modeling also showed that some important salt bridges were lost in CpSK, and higher Arg residues around critical Arg 140 seemed to increase flexibility for catalysis. Taken together, these data demonstrate that CpSK exhibits characteristics of cold adaptation with unusual kinetic parameters, which may provide important insights into the cold adaptation of metabolic enzymes.

Chronic Toxicity of the Triazole Fungicide Tebuconazole on a Heterocystous, Nitrogen-Fixing Rice Paddy Field Cyanobacterium, Westiellopsis prolifica Janet

  • Nirmal Kumar, J.I.;Bora, Anubhuti;Amb, Manmeet Kaur
    • Journal of Microbiology and Biotechnology
    • /
    • 제20권7호
    • /
    • pp.1134-1139
    • /
    • 2010
  • This study explored the chronic effects of different doses of the triazole fungicide tebuconazole on the growth, and metabolic and enzymatic functions of the filamentous paddy field cyanobacterium, Westiellopsis prolifica Janet. The growth of the cyanobacterium was determined by an estimation of the change in pigment contents. Chlorophyll-a, carotenoids, and accessory pigments such as phycocyanin, allophycocyanin, and phycoerythrin were shown to decline over a 16-day period by a factor of 92%, 93%, 83%, 95%, and 100%, respectively, with increasing doses of the fungicide. Metabolic and enzymatic activities were also adversely affected. Over the 16 days, a gradual rise in total phenol content was recorded when Westiellopsis prolifica Janet was treated with 60 ppm of the fungicide, despite the reduction in carbohydrates, proteins, and amino acids by 96%, 92%, and 90%, respectively. Moreover, the enzymes nitrate reductase (NR), glutamine synthetase (GS), and succinate dehydrogenase (SDH) also registered reductions of 93%, 90%, and 98%, respectively. This study indicates that tebuconazole, although an important fungicide used extensively in rice fields, exhibits an inhibitory effect on the growth and metabolic activities of Westiellopsis prolifica Janet and hence possibly on other varieties as well.

Vitellogenin Induction and Histo-metabolic Changes Following Exposure of Cyprinus carpio to Methyl Paraben

  • Barse, A.V.;Chakrabarti, T.;Ghosh, T.K.;Pal, A.K.;Kumar, Neeraj;Raman, R.P.;Jadhao, S.B.
    • Asian-Australasian Journal of Animal Sciences
    • /
    • 제23권12호
    • /
    • pp.1557-1565
    • /
    • 2010
  • Methyl paraben (MP), which is used as a preservative in pharmaceutical and cosmetic (shampoo) products, foods and beverages, enters into the aquatic environment and can pose a potential fish health hazard. In this experiment, effects of MP were evaluated in adult male common carp (Cyprinus carpio) by exposing them to fractions (1/$143^{rd}$ to 1/29th) of the $LC_{50}$ dose with every change of water for 28 days. Vitellogenin induction, metabolic enzymes, somatic indices and bioaccumulation were studied at weekly intervals. The $96^{th}$ h $LC_{50}$ of MP in fingerlings was 120 mg/L. Compared to the control, except for increases (p<0.01) in alkaline phosphatase (EC 3.1.3.1), alanine aminotransferase (EC 2.6.1.2) and liver size, there were decreases (p<0.01) in activity of acid phosphatase (EC 3.1.3.2), aspartate aminotransferase (EC 2.6.1.1), and testiculosomatic index following exposure to any dose of MP. Vitellogenin induction was significantly higher (p<0.01) in exposed than unexposed (control) fish. The bioaccumulation of MP in testis, liver, brain, gills and muscle tissues of fish increased significantly (p<0.01) with increase of dose from 0.84 ppm to 1.68 ppm. Dose and duration of exposure (p<0.01) indicated that an exposure period of 1 to 2 weeks was sufficient to cause changes in the quantifiable parameters studied. Fish exposed to 4.2 ppm MP became lethargic after the $26^{th}$ d. Histologically, degeneration, vacuolization and focal necrotic changes in liver and fibrosis-like changes in testicular tissue were noted.