Metabolic engineering, the generation of organisms with genetically engineered alterations in their metabolism is one of the most rapidly growing fields of biotechnology and one with huge prospective benefits to industry, agriculture and the environment. This proposal is part of a project (coordinated by Dr Andrew Hanson of U. Florida and involving researchers at five universities) on engineering and analyzing non-photosynthetic one-carbon metabolism in plants. This cooperative agreement will bring together the resources and experience in plant biochemistry and in vivo NMR at NMSU with the facilities and expertise in advanced NMR methods of analysing biomolecules of the Bioprocess Engineering Group at NIST. The project involves analysis of changes in metabolism and gene expression in transgenically altered and naturally occuring mutant plant varieties of tobacco and maize and aims at an understanding of the regulation of an area of metabolism that is central to many current and future engineering projects. The analysis proposed here will use nuclear magnetic resonance (NMR) spectroscopy to establish and apply metabolic profiling methods, to analyze one-carbon (1-C) metabolic fluxes, and to critically test models of 1-C metabolism.