Document Detail

Biosynthetic production of 13C-labeled amino acids with site-specific enrichment.
MedLine Citation:
PMID:  7035446     Owner:  NLM     Status:  MEDLINE    
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.
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
Amino Acids / biosynthesis*
Carbon Isotopes
Escherichia coli / metabolism*
Isotope Labeling / methods
Mass Spectrometry
Species Specificity
Grant Support
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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