title: Metabolic engineering of the purine biosynthetic pathway in Corynebacterium glutamicum results in increased intracellular pool sizes of IMP and hypoxanthine.
abstract: BACKGROUND
  Purine nucleotides exhibit various functions in cellular metabolism. Besides serving as building blocks for nucleic acid synthesis, they participate in signaling pathways and energy metabolism. Further, IMP and GMP represent industrially relevant biotechnological products used as flavor enhancing additives in food industry. Therefore, this work aimed towards the accumulation of IMP applying targeted genetic engineering of Corynebacterium glutamicum.
  
  
  RESULTS
  Blocking of the degrading reactions towards AMP and GMP lead to a 45-fold increased intracellular IMP pool of 22 μmol g(CDW)⁻¹. Deletion of the pgi gene encoding glucose 6-phosphate isomerase in combination with the deactivated AMP and GMP generating reactions, however, resulted in significantly decreased IMP pools (13 μmol g(CDW)⁻¹). Targeted metabolite profiling of the purine biosynthetic pathway further revealed a metabolite shift towards the formation of the corresponding nucleobase hypoxanthine (102 μmol g(CDW)⁻¹) derived from IMP degradation.
  
  
  CONCLUSIONS
  The purine biosynthetic pathway is strongly interconnected with various parts of the central metabolism and therefore tightly controlled. However, deleting degrading reactions from IMP to AMP and GMP significantly increased intracellular IMP levels. Due to the complexity of this pathway further degradation from IMP to the corresponding nucleobase drastically increased suggesting additional targets for future strain optimization.
authors.affiliation: Biochemical Engineering Institute, Saarland University, Campus A1.5, 66123 Saarbrücken, Germany.
authors.forename: Susanne
authors.initials: S
authors.lastname: Peifer
authors.forename: Tobias
authors.initials: T
authors.lastname: Barduhn
authors.forename: Sarah
authors.initials: S
authors.lastname: Zimmet
authors.forename: Dietrich A
authors.initials: DA
authors.lastname: Volmer
authors.forename: Elmar
authors.initials: E
authors.lastname: Heinzle
authors.forename: Konstantin
authors.initials: K
authors.lastname: Schneider
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D057486:Carbon Cycle; D016000:Cluster Analysis; D048230:Corynebacterium glutamicum / Q000235:genetics / Q000378:metabolism*; D017353:Gene Deletion; D005838:Genotype; D005956:Glucose-6-Phosphate Isomerase / Q000235:genetics / Q000378:metabolism; D019271:Hypoxanthine / Q000378:metabolism*; D007291:Inosine Monophosphate / Q000096:biosynthesis*; D060847:Metabolic Engineering; D016297:Mutagenesis, Site-Directed; D025341:Principal Component Analysis; D011687:Purines / Q000096:biosynthesis*
chemical_list: D001426:Bacterial Proteins; D011687:Purines; D007291:Inosine Monophosphate; D019271:Hypoxanthine; D005956:Glucose-6-Phosphate Isomerase; C030985:purine