Document Detail


Biosynthetic production of 13C-labeled amino acids with site-specific enrichment.
MedLine Citation:
PMID:  7035446     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
We have developed two Escherichia coli strains for the production of specifically labeled amino acids suitable for high resolution nuclear magnetic resonance experiments. The 13C atoms from the enriched carbon sources, [1-13C]lactate, [1,4-13C2]succinate, and [1-13C]-acetate, are incorporated into the amino acids producing multilabeled molecules with relatively few instances of adjacent enriched carbons. This greatly simplifies the resultant spectra as compared to the extensively spin-coupled spectra of uniformly enriched samples. No isotopic enrichment was found at carbon positions expected to be unenriched by consideration of the major biosynthetic pathways. Utilizing 90% enriched precursors, most positions were enriched to approximately 85% except for those positions derived from acetate and those affected by the carbon interchange of the pentose phosphate shunt. In both of these cases, the enrichment level fell to 70%. The only result enrichment at the delta-methyl carbon of isoleucine. For the bacterial strain in question, the main pathway of isoleucine biosynthesis appears not to be from threonine but from an alternate precursor, possibly glutamate.
Authors:
D M LeMaster; J E Cronan
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  257     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  1982 Feb 
Date Detail:
Created Date:  1982-04-20     Completed Date:  1982-04-20     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  1224-30     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Amino Acids / biosynthesis*
Carbon Isotopes
Escherichia coli / metabolism*
Isotope Labeling / methods
Mass Spectrometry
Mutation
Species Specificity
Grant Support
ID/Acronym/Agency:
AI-10186/AI/NIAID NIH HHS; GM-22778/GM/NIGMS NIH HHS; GM-22797/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
0/Amino Acids; 0/Carbon Isotopes

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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