title: Regulation of acetate metabolism in Corynebacterium glutamicum: transcriptional control of the isocitrate lyase and malate synthase genes.
abstract: In the amino-acid-producing microorganism Corynebacterium glutamicum, the specific activities of the acetate-activating enzymes acetate kinase and phosphotransacetylase and those of the glyoxylate cycle enzymes isocitrate lyase and malate synthase were found to be high when the cells were grown on acetate (0.8, 2.9, 2.1, and 1.8 U/mg protein, respectively). When the cells were grown on glucose or on other carbon sources such as lactate, succinate, or glutamate, the specific activities were two- to fourfold (acetate kinase and phosphotransacetylase) and 45- to 100-fold (isocitrate lyase and malate synthase) lower, indicating that the synthesis of the four enzymes is regulated by acetate in the growth medium. A comparative Northern (RNA) analysis of the C. glutamicum isocitrate lyase and malate synthase genes (aceA and aceB) and transcriptional cat fusion experiments revealed that aceA and aceB are transcribed as 1.6- and 2.7-kb monocistronic messages, respectively, and that the regulation of isocitrate lyase and malate synthase synthesis is exerted at the level of transcription from the respective promoters. Surprisingly, C. glutamicum mutants defective in either acetate kinase or phosphotransacetylase showed low specific activities of the other three enzymes (phosphotransacetylase, isocitrate lyase, and malate synthase or acetate kinase, isocitrate lyase, and malate synthase, respectively) irrespective of the presence or absence of acetate in the medium. This result and a correlation of a high intracellular acetyl coenzyme A concentration with high specific activities of isocitrate lyase, malate synthase, acetate kinase, and phosphotransacetylase suggest that acetyl coenzyme A or a derivative thereof may be a physiological trigger for the genetic regulation of enzymes involved in acetate metabolism of C. glutamicum.
authors.affiliation: Institut für Biotechnologie 1, Forschungszentrum Jülich, D-52425 Jülich, Germany.
authors.forename: V F
authors.initials: VF
authors.lastname: Wendisch
authors.forename: M
authors.initials: M
authors.lastname: Spies
authors.forename: D J
authors.initials: DJ
authors.lastname: Reinscheid
authors.forename: S
authors.initials: S
authors.lastname: Schnicke
authors.forename: H
authors.initials: H
authors.lastname: Sahm
authors.forename: B J
authors.initials: BJ
authors.lastname: Eikmanns
mesh_terms: D000084:Acetate Kinase / Q000235:genetics / Q000378:metabolism; D000085:Acetates / Q000378:metabolism*; D000105:Acetyl Coenzyme A / Q000378:metabolism; D019677:Artificial Gene Fusion; D015152:Blotting, Northern; D003001:Cloning, Molecular; D003352:Corynebacterium / Q000235:genetics* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D005947:Glucose / Q000378:metabolism; D018698:Glutamic Acid / Q000378:metabolism; D007522:Isocitrate Lyase / Q000235:genetics* / Q000378:metabolism; D007773:Lactates / Q000378:metabolism; D008292:Malate Synthase / Q000235:genetics* / Q000378:metabolism; D010709:Phosphate Acetyltransferase / Q000235:genetics / Q000378:metabolism; D010957:Plasmids; D015183:Restriction Mapping; D019802:Succinic Acid / Q000378:metabolism; D014158:Transcription, Genetic; D014170:Transformation, Genetic
chemical_list: D000085:Acetates; D007773:Lactates; D018698:Glutamic Acid; D000105:Acetyl Coenzyme A; D019802:Succinic Acid; D010709:Phosphate Acetyltransferase; D008292:Malate Synthase; D000084:Acetate Kinase; D007522:Isocitrate Lyase; D005947:Glucose