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PMID: 18245290 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S.

Elucidation of an alternate isoleucine biosynthesis pathway in Geobacter sulfurreducens.

Journal of bacteriology ·Vol. 190 ·No. 7 ·2008-04-00 ·Pages 2266-74

Risso C, Van Dien SJ, Orloff A, Lovley DR, Coppi MV

Abstract

The central metabolic model for Geobacter sulfurreducens included a single pathway for the biosynthesis of isoleucine that was analogous to that of Escherichia coli, in which the isoleucine precursor 2-oxobutanoate is generated from threonine. 13C labeling studies performed in G. sulfurreducens indicated that this pathway accounted for a minor fraction of isoleucine biosynthesis and that the majority of isoleucine was instead derived from acetyl-coenzyme A and pyruvate, possibly via the citramalate pathway. Genes encoding citramalate synthase (GSU1798), which catalyzes the first dedicated step in the citramalate pathway, and threonine ammonia-lyase (GSU0486), which catalyzes the conversion of threonine to 2-oxobutanoate, were identified and knocked out. Mutants lacking both of these enzymes were auxotrophs for isoleucine, whereas single mutants were capable of growth in the absence of isoleucine. Biochemical characterization of the single mutants revealed deficiencies in citramalate synthase and threonine ammonia-lyase activity. Thus, in G. sulfurreducens, 2-oxobutanoate can be synthesized either from citramalate or threonine, with the former being the main pathway for isoleucine biosynthesis. The citramalate synthase of G. sulfurreducens constitutes the first characterized member of a phylogenetically distinct clade of citramalate synthases, which contains representatives from a wide variety of microorganisms.

MeSH Terms
Acetyl Coenzyme A/metabolism Bacterial Proteins/genetics,metabolism Biosynthetic Pathways Butyrates/metabolism Carbon Isotopes/metabolism Geobacter/genetics,metabolism Isoleucine/biosynthesis Malates/metabolism Pyruvic Acid/metabolism Threonine/metabolism Threonine Dehydratase/genetics,metabolism
Chemicals
Bacterial Proteins Butyrates Carbon Isotopes Malates Isoleucine Threonine alpha-ketobutyric acid Acetyl Coenzyme A Pyruvic Acid Threonine Dehydratase citramalate
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Risso Carla
Department of Microbiology, 203N Morrill Science Center IVN, University of Massachusetts Amherst, Amherst, MA 01003, USA. [email protected]
Van Dien Stephen J
Orloff Amber
Lovley Derek R
Coppi Maddalena V
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
1098-5530
DOI
10.1128/JB.01841-07
Email
[email protected]
Published
2008-04-00
Epub
2008-00-01
Pages
2266-74
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC2293191
Subset
IM
External Links
PubMed source DOI: 10.1128/JB.01841-07 PMC Full Text View journal details
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