Deregulation of Feedback Inhibition of Phosphoenolpyruvate Carboxylase for Improved Lysine Production in Corynebacterium glutamicum
| Autor: | Doinita Frank, Sugima Rappert, An-Ping Zeng, Rajesh Reddy Bommareddy, Zhen Chen |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2014 |
| Předmět: |
Mutant
Allosteric regulation Lysine Malates Biology Pentose phosphate pathway Applied Microbiology and Biotechnology Gene Expression Regulation Enzymologic Corynebacterium glutamicum Feedback Metabolic flux analysis Point Mutation Enzyme Inhibitors Aspartic Acid Carbon Isotopes Ecology Gene Expression Regulation Bacterial Enzyme assay Metabolic Flux Analysis Phosphoenolpyruvate Carboxylase Biochemistry Metabolic Engineering Isotope Labeling biology.protein bacteria Mutant Proteins Phosphoenolpyruvate carboxylase NADP Food Science Biotechnology |
| Popis: | Allosteric regulation of phosphoenolpyruvate carboxylase (PEPC) controls the metabolic flux distribution of anaplerotic pathways. In this study, the feedback inhibition of Corynebacterium glutamicum PEPC was rationally deregulated, and its effect on metabolic flux redistribution was evaluated. Based on rational protein design, six PEPC mutants were designed, and all of them showed significantly reduced sensitivity toward aspartate and malate inhibition. Introducing one of the point mutations (N917G) into the ppc gene, encoding PEPC of the lysine-producing strain C. glutamicum LC298, resulted in ∼37% improved lysine production. In vitro enzyme assays and 13 C-based metabolic flux analysis showed ca. 20 and 30% increases in the PEPC activity and corresponding flux, respectively, in the mutant strain. Higher demand for NADPH in the mutant strain increased the flux toward pentose phosphate pathway, which increased the supply of NADPH for enhanced lysine production. The present study highlights the importance of allosteric regulation on the flux control of central metabolism. The strategy described here can also be implemented to improve other oxaloacetate-derived products. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|