Metabolic Pathway Confirmation and Discovery Through ¹³C-labeling of Proteinogenic Amino Acids

JoVE 3583 1/26/2012

Le You¹, Lawrence Page², Xueyang Feng¹, Bert Berla¹, Himadri B. Pakrasi³, Yinjie J. Tang¹

¹Department of Energy, Environmental and Chemical Engineering, Washington University, ²Department of Biology, Washington University, ³Department of Energy, Environmental and Chemical Engineering and Department of Biology, Washington University

¹³C-isotope labeling is a useful technique for determining the cell central metabolism for various types of microorganisms. After cells have been cultured with a specific labeled substrate, GC-MS measurement can reveal functional metabolic pathways based on unique labeling patterns in proteinogenic amino acids.

Comprehensive Compositional Analysis of Plant Cell Walls (Lignocellulosic biomass) Part II: Carbohydrates

JoVE 1837 3/12/2010

Cliff E. Foster¹, Tina M. Martin¹, Markus Pauly²

¹Great Lakes Bioenergy Research Center, Michigan State University (MSU), ²Great Lakes Bioenergy Research Center and DOE-Plant Research Lab, Michigan State University (MSU)

Plant biomass is a major carbon-neutral renewable resource that could be used for the production of biofuels. Plant biomass consists mainly of cell walls, a structurally complex composite material termed lignocellulosics. Here we describe a protocol for a comprehensive analysis of the content and composition of wall derived carbohydrates.

Biochemical Measurement of Neonatal Hypoxia

JoVE 2948 8/24/2011

Megan S. Plank¹, Teleka C. Calderon¹, Yayesh Asmerom², Danilo S. Boskovic¹, Danilyn M. Angeles²

¹Division of Biochemistry, Department of Basic Sciences, Loma Linda University, ²Division of Physiology, Department of Basic Sciences, Loma Linda University

A method is described to measure biochemical markers of neonatal hypoxia-ischemia. The approach utilizes high pressure liquid chromatography (HPLC) and Gas Chromatography Mass Spectrometry (GC/MS).

Monitoring Plant Hormones During Stress Responses

JoVE 1127 6/15/2009

Marie J. Engelberth, Jurgen Engelberth

Department of Biology, University of Texas

A simple method is provided that allows for the rapid extraction and analysis of multiple plant hormones from small tissue samples. The procedure uses vapor phase extraction as the solemn purification step. Samples are analyzed by GC/MS with chemical ionization that produces mainly (M+1)+ ions.

Comprehensive Compositional Analysis of Plant Cell Walls (Lignocellulosic biomass) Part I: Lignin

JoVE 1745 3/11/2010

Cliff E. Foster¹, Tina M. Martin¹, Markus Pauly²

¹Great Lakes Bioenergy Research Center, Michigan State University (MSU), ²Great Lakes Bioenergy Research Center and DOE-Plant Research Lab, Michigan State University (MSU)

Plant biomass is a major carbon-neutral renewable resource that could be used for the production of biofuels. Plant biomass consists mainly of cell walls, a structurally complex composite material termed lignocellulosics. Here we describe a protocol for a comprehensive analysis of the content and composition of the polyphenolic lignin.

Stable Isotopic Profiling of Intermediary Metabolic Flux in Developing and Adult Stage Caenorhabditis elegans

JoVE 2288 2/27/2011

Marni J. Falk^1,2, Meera Rao*¹, Julian Ostrovsky*¹, Evgueni Daikhin¹, Ilana Nissim¹, Marc Yudkoff^1,2

¹Department of Pediatrics, The Children's Hospital of Philadelphia, ²Department of Pediatrics, University of Pennsylvania

Stable isotopic profiling by gas chromatography mass spectrometric analysis of intermediary metabolic flux is described in the nematode, Caenorhabditis elegans. Methods are detailed for assessing isotopic enrichment in carbon dioxide, organic acids, and amino acids following isotope exposure either during development on agar plates or during adulthood in liquid culture.

One-step Metabolomics: Carbohydrates, Organic and Amino Acids Quantified in a Single Procedure

JoVE 2014 6/25/2010

James D. Shoemaker

Edward A. Doisy Department of Biochemistry and Molecular Biology, Saint Louis University School of Medicine

The urease method of sample preparation for GC/MS analysis of intermediary metabolites is presented by its inventor. The method allows one-step follow-up of newborn screening for inborn errors by tandem mass spectrometry by quantifying carbohydrates, organic and amino acids all in a single process.

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JoVE 2268 6/15/2010

Robert S. Plumb

Pharmaceutical Business Operations, Waters Corporation

Assessing Hepatic Metabolic Changes During Progressive Colonization of Germ-free Mouse by ¹H NMR Spectroscopy

JoVE 3642 12/15/2011

Peter Heath¹, Sandrine Paule Claus²

¹School of Chemistry, Food and Pharmacy, The University of Reading, ²Department of Nutritional Sciences, The University of Reading

A progressive colonization procedure is described to further assess its impact on the host hepatic metabolism. Colonization is monitored non invasively by evaluating the urinary excretion of microbial co-metabolites by NMR-based metabolic profiling while hepatic metabolism is assessed by High Resolution Magic Angle Spinning (HR MAS) NMR profiling of intact biopsy.

Electroantennographic Bioassay as a Screening Tool for Host Plant Volatiles

JoVE 3931 5/06/2012

John J. Beck, Douglas M. Light, Wai S. Gee

Plant Mycotoxin Research, U.S. Department of Agriculture, Agricultural Research Service

A method to rapidly screen host plant volatiles by measurement of the electrophysiological response of adult navel orangeworm (Amyelois transitella) antennae to single components and blends via electroantennographic analysis is demonstrated.