This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Kobashi, N.
Right arrow Articles by Tanokura, M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Kobashi, N.
Right arrow Articles by Tanokura, M.

 Previous Article  |  Next Article 

Journal of Bacteriology, March 1999, p. 1713-1718, Vol. 181, No. 6
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Aspartate Kinase-Independent Lysine Synthesis in an Extremely Thermophilic Bacterium, Thermus thermophilus: Lysine Is Synthesized via alpha -Aminoadipic Acid Not via Diaminopimelic Acid

Nobuyuki Kobashi, Makoto Nishiyama,* and Masaru Tanokura*

Biotechnology Research Center, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan

Received 28 September 1998/Accepted 23 December 1998

An aspartate kinase-deficient mutant of Thermus thermophilus, AK001, was constructed. The mutant strain did not grow in a minimal medium, suggesting that T. thermophilus contains a single aspartate kinase. Growth of the mutant strain was restored by addition of both threonine and methionine, while addition of lysine had no detectable effect on growth. To further elucidate the lysine biosynthetic pathway in T. thermophilus, lysine auxotrophic mutants of T. thermophilus were obtained by chemical mutagenesis. For all lysine auxotrophic mutants, growth in a minimal medium was not restored by addition of diaminopimelic acid, whereas growth of two mutants was restored by addition of alpha -aminoadipic acid, a precursor of lysine in biosynthetic pathways of yeast and fungi. A BamHI fragment of 4.34 kb which complemented the lysine auxotrophy of a mutant was cloned. Determination of the nucleotide sequence suggested the presence of homoaconitate hydratase genes, termed hacA and hacB, which could encode large and small subunits of homoaconitate hydratase, in the cloned fragment. Disruption of the chromosomal copy of hacA yielded mutants showing lysine auxotrophy which was restored by addition of alpha -aminoadipic acid or alpha -ketoadipic acid. All of these results indicated that in T. thermophilus, lysine was not synthesized via the diaminopimelic acid pathway, believed to be common to all bacteria, but via a pathway using alpha -aminoadipic acid as a biosynthetic intermediate.


* Corresponding author. Mailing address for Makoto Nishiyama: Biotechnology Research Center, University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan. Phone: 81-3(3812)2111, ext. 3075. Fax: 81-3(5802)3326. E-mail: umanis{at}hongo.ecc.u-tokyo.ac.jp.


Journal of Bacteriology, March 1999, p. 1713-1718, Vol. 181, No. 6
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.



This article has been cited by other articles:

  • Fujiwara, K., Tsubouchi, T., Kuzuyama, T., Nishiyama, M. (2006). Involvement of the arginine repressor in lysine biosynthesis of Thermus thermophilus. Microbiology 152: 3585-3594 [Abstract] [Full Text]  
  • Miyazaki, J., Asada, K., Fushinobu, S., Kuzuyama, T., Nishiyama, M. (2005). Crystal Structure of Tetrameric Homoisocitrate Dehydrogenase from an Extreme Thermophile, Thermus thermophilus: Involvement of Hydrophobic Dimer-Dimer Interaction in Extremely High Thermotolerance. J. Bacteriol. 187: 6779-6788 [Abstract] [Full Text]  
  • Tsubouchi, T., Mineki, R., Taka, H., Kaga, N., Murayama, K., Nishiyama, C., Yamane, H., Kuzuyama, T., Nishiyama, M. (2005). Leader Peptide-mediated Transcriptional Attenuation of Lysine Biosynthetic Gene Cluster in Thermus thermophilus. J. Biol. Chem. 280: 18511-18516 [Abstract] [Full Text]  
  • Fukui, T., Atomi, H., Kanai, T., Matsumi, R., Fujiwara, S., Imanaka, T. (2005). Complete genome sequence of the hyperthermophilic archaeon Thermococcus kodakaraensis KOD1 and comparison with Pyrococcus genomes. Genome Res 15: 352-363 [Abstract] [Full Text]  
  • Miyazaki, T., Miyazaki, J., Yamane, H., Nishiyama, M. (2004). {alpha}-Aminoadipate aminotransferase from an extremely thermophilic bacterium, Thermus thermophilus. Microbiology 150: 2327-2334 [Abstract] [Full Text]  
  • Miyazaki, J., Kobashi, N., Nishiyama, M., Yamane, H. (2003). Characterization of Homoisocitrate Dehydrogenase Involved in Lysine Biosynthesis of an Extremely Thermophilic Bacterium, Thermus thermophilus HB27, and Evolutionary Implication of beta -Decarboxylating Dehydrogenase. J. Biol. Chem. 278: 1864-1871 [Abstract] [Full Text]  
  • Nishida, H., Narumi, I. (2002). Disruption analysis of DR1420 and/or DR1758 in the extremely radioresistant bacterium Deinococcus radiodurans. Microbiology 148: 2911-2914 [Abstract] [Full Text]  
  • Brinkman, A. B., Bell, S. D., Lebbink, R. J., de Vos, W. M., van der Oost, J. (2002). The Sulfolobus solfataricus Lrp-like Protein LysM Regulates Lysine Biosynthesis in Response to Lysine Availability. J. Biol. Chem. 277: 29537-29549 [Abstract] [Full Text]  
  • Miyazaki, J., Kobashi, N., Nishiyama, M., Yamane, H. (2001). Functional and Evolutionary Relationship between Arginine Biosynthesis and Prokaryotic Lysine Biosynthesis through {alpha}-Aminoadipate. J. Bacteriol. 183: 5067-5073 [Abstract] [Full Text]  
  • Nishida, H., Nishiyama, M., Kobashi, N., Kosuge, T., Hoshino, T., Yamane, H. (1999). A Prokaryotic Gene Cluster Involved in Synthesis of Lysine through the Amino Adipate Pathway: A Key to the Evolution of Amino Acid Biosynthesis. Genome Res 9: 1175-1183 [Abstract] [Full Text]