Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Metagenome Resource for D-Serine Utilization in a DsdA-Disrupted Escherichia coli

  • Lim, Mi-Young (Department of Biotechnology, Catholic University of Korea) ;
  • Lee, Hyo-Jeong (Department of Biotechnology, Catholic University of Korea) ;
  • Kim, Pil (Department of Biotechnology, Catholic University of Korea)
  • Received : 2010.12.23
  • Accepted : 2011.02.08
  • Published : 2011.04.28
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Abstract

To find alternative genetic resources for D-serine dehydratase (E.C. 4.3.1.18, dsdA) mediating the deamination of D-serine into pyruvate, metagenomic libraries were screened. The chromosomal dsdA gene of a wild-type Escherichia coli W3110 strain was disrupted by inserting the tetracycline resistance gene (tet), using double-crossover, for use as a screening host. The W3110 dsdA::tet strain was not able to grow in a medium containing D-serine as a sole carbon source, whereas wild-type W3110 and the complement W3110 dsdA::tet strain containing a dsdA-expression plasmid were able to grow. After introducing metagenome libraries into the screening host, a strain containing a 40-kb DNA fragment obtained from the metagenomic souce derived from a compost was selected based on its capability to grow on the agar plate containing D-serine as a sole carbon source. For identification of the genetic resource responsible for the D-serine degrading capability, transposon-${\mu}$ was randomly inserted into the 40-kb metagenome. Two strains that had lost their D-serine degrading ability were negatively selected, and the two 6-kb contigs responsible for the D-serine degrading capability were sequenced and deposited (GenBank code: HQ829474.1 and HQ829475.1). Therefore, new alternative genetic resources for D-serine dehydratase was found from the metagenomic resource, and the corresponding ORFs are discussed.

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References

  1. Chen, Z., W. Huang, S. R. Srinivas, C. R. Jones, V. Ganapathy, and P. D. Prasad. 2004. Serine racemase and D-serine transport in human placenta and evidence for a transplacental gradient for D-serine in humans. J. Soc. Gynecol. Investig. 11: 294-303. https://doi.org/10.1016/j.jsgi.2004.02.003
  2. Donnenberg, M. S. and J. B. Kaper. 1991. Construction of an eae deletion mutant of enteropathogenic Escherichia coli by using a positive-selection suicide vector. Infect. Immun. 59: 4310-4317.
  3. Grant, S. L., Y. Shulman, P. Tibbo, D. R. Hampson, and G. B. Baker. 2006. Determination of D-serine and related neuroactive amino acids in human plasma by high-performance liquid chromatography with fluorimetric detection. J. Chromatogr. B Analyt. Technol. Biomed. Life Sci. 844: 278-282. https://doi.org/10.1016/j.jchromb.2006.07.022
  4. Hagishita, T., T. Yoshida, Y. Izumi, and T. Mitsunaga. 1996. Efficient L-serine production from methanol and glycine by resting cells of Methylobacterium sp. strain MN43. Biosci. Biotechnol. Biochem. 60: 1604-1607. https://doi.org/10.1271/bbb.60.1604
  5. Hui, K. Y., E. M. Holleran, and J. Kovacs. 1989. Serine and Dserine position-1 substituted angiotensin II analogues. Synthesis using new 2,3,5,6-tetrafluorophenyl active esters and biological activities. Int. J. Pept. Protein Res. 34: 177-183.
  6. Izumi, Y., T. Yoshida, S. S. Miyazaki, T. Mitsunaga, T. Ohshiro, M. Shimao, A. Miyata, and T. Tanabe. 1993. L-Serine production by a methylotroph and its related enzymes. Appl. Microbiol. Biotechnol. 39: 427-432. https://doi.org/10.1007/BF00205027
  7. Jasin, M. and P. Schimmel. 1984. Deletion of an essential gene in Escherichia coli by site-specific recombination with linear DNA fragments. J. Bacteriol. 159: 783-786.
  8. Matsui, T., M. Sekiguchi, A. Hashimoto, U. Tomita, T. Nishikawa, and K. Wada. 1995. Functional comparison of D-serine and glycine in rodents: The effect on cloned NMDA receptors and the extracellular concentration. J. Neurochem. 65: 454-458.
  9. Miller, V. L. and J. J. Mekalanos. 1988. A novel suicide vector and its use in construction of insertion mutations: Osmoregulation of outer membrane proteins and virulence determinants in Vibrio cholerae requires toxR. J. Bacteriol. 170: 2575-2583.
  10. Nishikawa, T. 2005. Metabolism and functional roles of endogenous D-serine in mammalian brains. Biol. Pharm. Bull. 28: 1561-1565. https://doi.org/10.1248/bpb.28.1561
  11. Peters-Wendisch, P., M. Stolz, H. Etterich, N. Kennerknecht, H. Sahm, and L. Eggeling. 2005. Metabolic engineering of Corynebacterium glutamicum for L-serine production. Appl. Environ. Microbiol. 71: 7139-7144. https://doi.org/10.1128/AEM.71.11.7139-7144.2005
  12. Philippe, N., J. P. Alcaraz, E. Coursange, J. Geiselmann, and D. Schneider. 2004. Improvement of pCVD442, a suicide plasmid for gene allele exchange in bacteria. Plasmid 51: 246-255. https://doi.org/10.1016/j.plasmid.2004.02.003
  13. Sambrook, J. and D. W. Russell. 2000. Molecular Cloning: A Laboratory Manual, 3rd Ed. Cold Spring Harbor Laboratory Press, New York.
  14. Soberon, X., L. Covarrubias, and F. Bolivar. 1980. Construction and characterization of new cloning vehicles. IV. Deletion derivatives of pBR322 and pBR325. Gene 9: 287-305.
  15. Stolz, M., P. Peters-Wendisch, H. Etterich, T. Gerharz, R. Faurie, H. Sahm, H. Fersterra, and L. Eggeling. 2006. Reduced folate supply as a key to enhanced L-serine production with Corynebacterium glutamicum. Appl. Environ. Microbiol. 73: 750-755.
  16. Tsai, G. and J. Coyle. 2005. Methods for treating neuropsychiatric disorders. USA patent 6,974,821.
  17. Yasuda, E., N. Ma, and R. Semba. 2001. Immunohistochemical evidences for localization and production of D-serine in some neurons in the rat brain. Neurosci. Lett. 299: 162-164. https://doi.org/10.1016/S0304-3940(01)01502-6