Abstract
L-threonine is an essential amino acid which is widely used in feed and pharmaceutical industries. We recently engineered Corynebacterium glutamicum R102 (AHVr) for improved production of L-threonine. Inactivation of genes metX and dapA encoding dihydrodipicolinate synthase and homoserine O-acetyltransferase, respectively, was firstly conducted by homologous recombination, which differed from the common random mutagenesis method. Then operon gene hom-thrB (O) and export gene thrE (E) from R102 were over-expressed alone or together to obtain a series of recombinant strains. qPCR was employed to evaluate the transcript quantification of the target genes. In flask fermentation, the newly constructed strain R102Δ metXΔdapA (pEC-Box) was able to accumulate 3.35 g threonine/L compared with 1.80 g threonine/L of strain R102 (AHVr).
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Wang, X. and P. J. Quinn (2010) Lipopolysaccharide: Biosynthetic pathway and structure modification. Prog. Lipid Res. 49: 97–107.
Leuchtenberger, W., K. Huthmacher, and K. Drauz (2005) Biotechnological production of amino acids and derivatives: Current status and prospects. Appl. Microbiol. Biotechnol. 69: 1–8.
Takors, R., B. Bathe, M. Rieping, S. Hans, R. Kelle, and K. Huthmacher (2007) Systems biology for industrial strains and fermentation processes—example: Amino acids. J. Biotechnol. 129: 181–190.
Willis, L. B., P. A. Lessard, and A. J. Sinskey (2005) Synthesis of L-Threonine and Branched-Chain Amino Acids. pp. 511–526. In: Eggeling, L. and M. Bott (eds.). Handbook of corynebacterium glutamicum. CRC Press Taylor & Francis Group, Boca Raton, USA.
Simic, P., H. Sahm, and L. Eggeling (2001) L-threonine export: Use of eptides to identify a new translocator from Corynebacterium glutamicum. J. Bacteriol. 183: 5317–5324.
Archer, J. A. C., D. E. Solow-Cordero, and A. J. Sinskey (1991) A C-erminal deletion in Corynebacterium glutamicum homoserine dehydrogenase abolishes allosteric inhibition by L-threonine. Gene. 107: 53–59.
Eikmanns, B. J., N. Thum-Schmitz, L. Eggeling, K. U. Lüdtke, and H. Sahm (1994) Nucleotide sequence, expression and transcriptional analysis f the Corynebacterium glutamicum gltA gene encoding citrate synthase. Microbiol. 140: 1817–1828.
van der Rest, M. E., C. Lange, and D. Molenaar (1999) A heat shock following electroporation induces highly efficient transformation ofCorynebacterium glutamicum with xenogeneic plasmid DNA. Appl.Microbiol. Biotechnol. 52: 541–545.
Schäfer, A., A. Tauch, W. Jäger, J. Kalinowski, G. Thierbach, and A. Pühler (1994) Small mobilizable multi-purpose cloning vectors derivedfrom the Escherichia coli plasmids pK18 and pK19: Selection of defined deletions in the chromosome of Corynebacterium glutamicum. Gene. 145: 69–73.
Kiyoshi, N. (1973) Process for producing L-threonine and L-lysine. US patent 3,732,144.
Hartmann, M., A. Tauch, L. Eggeling, B. Bathe, B. Mockel, A. Pühler, and J. Kalinowski (2003) Identification and characterization of the last two unknown genes, dapC and dapF, in the succinylase branch of the L-lysine biosynthesis of Corynebacterium glutamicum. J. Biotechnol. 104: 199–211.
Eikmanns, B. J., M. Metzger, D. Reinscheid, M. Kircher, and H. Sahm (1991) Amplification of three threonine biosynthesis genes in Corynebacterium glutamicum and its influence on carbon flux in different strains. Appl. Microbiol. Biotechnol. 34: 617–622.
Diesveld, R., N. Tietze, O. Fürst, A. Reth, B. Bathe, H. Sahm, and L. Eggeling (2009) Activity of exporters of Escherichia coli in Corynebacterium glutamicum, and their use to increase L-threonine production. J. Mol. Microbiol. Biotechnol. 16: 198–207.
Bellmann, A., M. Vrlji, M. Pátek, H. Sahm, R. Krämer, and L. Eggeling (2001) Expression control and specificity of the basic amino acid exporter LysE of Corynebacterium glutamicum. Microbiol. 147: 1765–1774.
Lee, K. H., J. H. Park, T. Y. Kim, H. U. Kim, and S. Y. Lee (2007) Systems metabolic engineering of Escherichia coli for L-threonine production. Mol. Syst. Biol. 3: 149.
Hüser, A. T., C. Chassagnole, N. D. Lindley, M. Merkamm, A. Guyonvarch, V. Eliáková, M. Pátek, J. Kalinowski, I. Brune, A. Pühler, and A. Tauch (2005) Rational design of a Corynebacterium glutamicum characterization by metabolic flux analysis and genome-wide transcriptional profiling. Appl. Environ. Microbiol. 71: 3255–3268.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Lv, Y., Wu, Z., Han, S. et al. Construction of recombinant Corynebacterium glutamicum for L-threonine production. Biotechnol Bioproc E 17, 16–21 (2012). https://doi.org/10.1007/s12257-011-0360-8
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12257-011-0360-8