scholarly journals Combining reverse genetics and nuclear magnetic resonance-based metabolomics unravels trypanosome-specific metabolic pathways

2015 ◽  
Vol 96 (5) ◽  
pp. 917-926 ◽  
Author(s):  
Frédéric Bringaud ◽  
Marc Biran ◽  
Yoann Millerioux ◽  
Marion Wargnies ◽  
Stefan Allmann ◽  
...  
Keyword(s):  
Nuclear Magnetic Resonance ◽  
Magnetic Resonance ◽  
Metabolic Pathways ◽  
Reverse Genetics ◽  
Nuclear Magnetic

Elucidation of metabolic pathways in glycogen-accumulating organisms with in vivo13C nuclear magnetic resonance

2007 ◽  
Vol 9 (11) ◽  
pp. 2694-2706 ◽  
Author(s):  
Paulo C. Lemos ◽  
Yu Dai ◽  
Zhiguo Yuan ◽  
Jurg Keller ◽  
Helena Santos ◽  
...  
Keyword(s):  
Nuclear Magnetic Resonance ◽  
Magnetic Resonance ◽  
Metabolic Pathways ◽  
Nuclear Magnetic

scholarly journals Catabolism of Glucose and Lactose in Bifidobacterium animalis subsp. lactis, Studied by13C Nuclear Magnetic Resonance

2013 ◽  
Vol 79 (24) ◽  
pp. 7628-7638 ◽  
Author(s):  
Irene González-Rodríguez ◽  
Paula Gaspar ◽  
Borja Sánchez ◽  
Miguel Gueimonde ◽  
Abelardo Margolles ◽  
...  
Keyword(s):  
Nuclear Magnetic Resonance ◽  
Magnetic Resonance ◽  
Metabolic Pathways ◽  
Transcriptional Analysis ◽  
Content Type ◽  
Bifidobacterium Animalis ◽  
Metabolic Flux Distribution ◽  
Nuclear Magnetic ◽  
In Cells

ABSTRACTBifidobacteria are widely used as probiotics in several commercial products; however, to date there is little knowledge about their carbohydrate metabolic pathways. In this work, we studied the metabolism of glucose and lactose in the widely used probiotic strainBifidobacterium animalissubsp.lactisBB-12 byin vivo13C nuclear magnetic resonance (NMR) spectroscopy. The metabolism of [1-13C]glucose was characterized in cells grown in glucose as the sole carbon source. Moreover, the metabolism of lactose specifically labeled with13C on carbon 1 of the glucose or the galactose moiety was determined in suspensions of cells grown in lactose. These experiments allowed the quantification of some intermediate and end products of the metabolic pathways, as well as determination of the consumption rate of carbon sources. Additionally, the labeling patterns in metabolites derived from the metabolism of glucose specifically labeled with13C on carbon 1, 2, or 3 in cells grown in glucose or lactose specifically labeled in carbon 1 of the glucose moiety ([1-13Cglucose]lactose), lactose specifically labeled in carbon 1 of the galactose moiety ([1-13Cgalactose]lactose), and [1-13C]glucose in lactose-grown cells were determined in cell extracts by13C NMR. The NMR analysis showed that the recovery of carbon was fully compatible with the fructose 6-phosphate, or bifid, shunt. The activity of lactate dehydrogenase, acetate kinase, fructose 6-phosphate phosphoketolase, and pyruvate formate lyase differed significantly between glucose and lactose cultures. The transcriptional analysis of several putative glucose and lactose transporters showed a significant induction of Balat_0475 in the presence of lactose, suggesting a role for this protein as a lactose permease. This report provides the firstin vivoexperimental evidence of the metabolic flux distribution in the catabolic pathway of glucose and lactose in bifidobacteria and shows that the bifid shunt is the only pathway involved in energy recruitment from these two sugars. On the basis of our experimental results, a model of sugar metabolism inB. animalissubsp.lactisis proposed.

19F Nuclear Magnetic Resonance as a Tool To Investigate Microbial Degradation of Fluorophenols to Fluorocatechols and Fluoromuconates

1998 ◽  
Vol 64 (4) ◽  
pp. 1256-1263 ◽  
Author(s):  
Marelle G. Boersma ◽  
Tatiana Y. Dinarieva ◽  
Wouter J. Middelhoven ◽  
Willem J. H. van Berkel ◽  
Joel Doran ◽  
...  
Keyword(s):  
Nuclear Magnetic Resonance ◽  
Magnetic Resonance ◽  
Metabolic Pathways ◽  
Whole Cells ◽  
Nmr Method ◽  
Cell Extracts ◽  
Nmr Characterization ◽  
Metabolite Profiles ◽  
Nuclear Magnetic

ABSTRACT A method was developed to study the biodegradation and oxidative biodehalogenation of fluorinated phenols by 19F nuclear magnetic resonance (NMR). Characterization of the 19F NMR spectra of metabolite profiles of a series of fluorophenols, converted by purified phenol hydroxylase, catechol 1,2-dioxygenase, and/or by the yeast-like fungus Exophiala jeanselmei, provided possibilities for identification of the 19F NMR chemical shift values of fluorinated catechol and muconate metabolites. As an example, the 19F NMR method thus defined was used to characterize the time-dependent metabolite profiles of various halophenols in either cell extracts or in incubations with whole cells of E. jeanselmei. The results obtained for these two systems are similar, except for the level of muconates observed. Altogether, the results of the present study describe a19F NMR method which provides an efficient tool for elucidating the metabolic pathways for conversion of fluorine-containing phenols by microorganisms, with special emphasis on possibilities for biodehalogenation and detection of the type of fluorocatechols and fluoromuconates involved. In addition, the method provides possibilities for studying metabolic pathways in vivo in whole cells.

Carbon-13 Nuclear Magnetic Resonance for Analysis of Metabolic Pathways

2013 ◽  
pp. 415-445
Author(s):  
Craig R. Malloy ◽  
Elizabeth Maher ◽  
Isaac Marin-Valencia ◽  
Bruce Mickey ◽  
Ralph J. DeBerardinis ◽  
...  
Keyword(s):  
Nuclear Magnetic Resonance ◽  
Magnetic Resonance ◽  
Metabolic Pathways ◽  
Nuclear Magnetic

NMR-based Metabolomic Techniques Identify the Anticancer Effects of Three Polyphyllins in HepG2 Cells

2021 ◽  
Vol 17 ◽  
Author(s):  
Feng Su ◽  
Haibo Wang ◽  
Yifan Wang ◽  
Lv Ye ◽  
Peixi Zhu ◽  
...  
Keyword(s):  
Nuclear Magnetic Resonance ◽  
Magnetic Resonance ◽  
Metabolic Pathways ◽  
Hepg2 Cells ◽  
Chinese Herb ◽  
Principal Component ◽  
Metabolic Phenotypes ◽  
H Nmr ◽  
Anticancer Effects ◽  
Nuclear Magnetic

Background: Rhizoma Paridis (RP) is a traditional Chinese herb used for the treatment of tumors, detoxification and hemostasia. Studies show the main components of RP are Polyphyllin I (PPI), polyphyllin VI (PPVI), and polyphyllin VII (PPVII). However, the pharmaco-mechanisms of these compounds are not clear. Objective: By used 1 H nuclear magnetic resonance (1 H-NMR) based metabolomics approach to identify the Anticancer effects of PPI, PPVI and PPVII in HepG2 cells. Methods 1 H nuclear magnetic resonance (1 H-NMR) based metabolomics approach was applied to investigate the toxicological effect of PPI, PPVI, PPVII on HepG2 cells. Multivariate statistical analysis was employed to examine the metabolic changes and abnormal metabolic pathways, including Principal Component Analysis (PCA), Partial Least Squares Discriminant Analysis (PLS-DA), and orthogonal PLS-DA (OPLS-DA). Results: The results showed that the effects of metabolic phenotypes were affected separately by PPI, PPVI, and PPVII. The metabolic phenotypes were also changed over time. The characteristic metabolites were varied by affecting different polyphylins, which were identified by the reconstructed OPLSDA loading plots. According to the characteristic metabolites, the mainly disturbed metabolic pathways were found, such as alanine, aspartate and glutamate metabolism, pyruvate metabolism, glycine, serine, and threonine metabolism. Conclusion: The current work could allow us to understand the therapeutic effect of RP in metabolism. It also indicated that RP would be a promising candidate for liver cancer treatment.

scholarly journals Nuclear magnetic resonance spectroscopy of biofluids for osteoarthritis

2020 ◽  
Author(s):  
Emily J Clarke ◽  
James R Anderson ◽  
Mandy J Peffers
Keyword(s):  
Nuclear Magnetic Resonance ◽  
Nmr Spectroscopy ◽  
Magnetic Resonance ◽  
Synovial Fluid ◽  
Metabolic Pathways ◽  
Research Area ◽  
Resonance Spectroscopy ◽  
World Population ◽  
Chondrocyte Phenotype ◽  
Nuclear Magnetic

Abstract Background Osteoarthritis is a common degenerative musculoskeletal disease of synovial joints. It is characterized by a metabolic imbalance resulting in articular cartilage degradation, reduced elastoviscosity of synovial fluid and an altered chondrocyte phenotype. This is often associated with reduced mobility, pain and poor quality of life. Subsequently, with an ageing world population, osteoarthritis is of increasing concern to public health. Nuclear magnetic resonance (NMR) spectroscopy can be applied to characterize the metabolomes of biofluids, determining changes associated with osteoarthritis pathology, identifying potential biomarkers of disease and alterations to metabolic pathways. Sources of data A comprehensive search of PubMed and Web of Science databases using combinations of the following keywords: ‘NMR Spectroscopy’, ‘Blood’, ‘Plasma’, ‘Serum’, ‘Urine’, ‘Synovial Fluid’ and ‘Osteoarthritis’ for articles published from 2000 to 2020. Areas of agreement The number of urine metabolomics studies using NMR spectroscopy to investigate osteoarthritis is low, whereas the use of synovial fluid is significantly higher. Several differential metabolites have previously been identified and mapped to metabolic pathways involved in osteoarthritis pathophysiology. Areas of controversy Conclusions are sometimes conservative or overinflated, which may reflect the variation in reporting standards. NMR metabolic experimental design may require further consideration, as do the animal models used for such studies. Growing points There are various aspects which require improvement within the field. These include stricter adherence to the Metabolomics Standards Initiative, inclusive of the standardization of metabolite identifications; increased utilization of integrating NMR metabolomics with other ‘omic’ disciplines; and increased deposition of raw experimental files into open access online repositories, allowing greater transparency and enabling additional future analyses. Areas timely for developing research Overall, this research area could be improved by the inclusion of more heterogeneous cohorts, reflecting varying osteoarthritis phenotypes, and larger group sizes ensuring studies are not underpowered. To correlate local and systemic environments, the use of blood for diagnostic purposes, over the collection of synovial fluid, requires increased attention. This will ultimately enable biomarkers of disease to be determined that may provide an earlier diagnosis, or provide potential therapeutic targets for osteoarthritis, ultimately improving patient prognosis.

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