Petri, Kathrin; Walter, Frederik; Persicke, Marcus; Rückert, Christian ; Kalinowski, Jörn: A novel type of N-acetylglutamate synthase is involved in the first step of arginine biosynthesis in Corynebacterium glutamicum. In: BMC Genomics. Jg.14 H. 1. 2013
Inhalt
- Results
- Discovering a gene responsible for the first step of arginine biosynthesis
- Validating the gene function of cg3035 by heterologous complementation, gene deletion, metabolome analyses, and enzyme assays
- Cg3035 establishes a novel class of NAGS genes
- Discussion
- Metabolite profiling analyses provide new insight into arginine biosynthesis
- In vivo and invitro experiments confirm the ability of Cg3035 to acetylate l-glutamate
- Cg3035 – A novel form of N-acetylglutamate synthase
- Conclusions
- Methods
- Bacterial strains, growth conditions, plasmids, and oligonucleotides
- DNA isolation, manipulation, and analysis
- Construction of defined chromosomal deletions
- Construction of expression vectors
- Generation of a genomic library from C. glutamicum ATCC 13032 and complementation of E. coli JM109 ∆argA
- Cell harvesting, metabolite extraction, and sample preparation
- Sample preparation for qualitative LC-MS analysis
- Sample preparation for quantitative LC-MS analysis
- HPLC-ESI-QTOF conditions
- Processing of LC-MS(/MS) Data
- Preparation of crude protein extracts of bacterial cultures
- Measurement of NAGS enzyme activity
- Additional files
- Competing interests
- Authors' contributions
- Acknowledgements & funding
- References