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Open Access Research

Metabolic engineering of the purine biosynthetic pathway in Corynebacterium glutamicum results in increased intracellular pool sizes of IMP and hypoxanthine

Susanne Peifer1, Tobias Barduhn1, Sarah Zimmet1, Dietrich A Volmer2, Elmar Heinzle1 and Konstantin Schneider1*

Author Affiliations

1 Biochemical Engineering Institute, Saarland University, Campus A1.5, 66123, Saarbrücken, Germany

2 Institute of Bioanalytical Chemistry, Saarland University, Campus B2.2, 66123, Saarbrücken, Germany

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Microbial Cell Factories 2012, 11:138  doi:10.1186/1475-2859-11-138

Published: 24 October 2012

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 gCDW-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 gCDW-1). Targeted metabolite profiling of the purine biosynthetic pathway further revealed a metabolite shift towards the formation of the corresponding nucleobase hypoxanthine (102 μmol gCDW-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.

Keywords:
Purine accumulation; Metabolic engineering; Corynebacterium glutamicum; Targeted metabolomics; Metabolic flux analysis