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 Kuzuyama, T.
Right arrow Articles by Seto, H.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Kuzuyama, T.
Right arrow Articles by Seto, H.

 Previous Article  |  Next Article 

Journal of Bacteriology, February 2000, p. 891-897, Vol. 182, No. 4
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Cloning and Characterization of 1-Deoxy-D-Xylulose 5-Phosphate Synthase from Streptomyces sp. Strain CL190, Which Uses both the Mevalonate and Nonmevalonate Pathways for Isopentenyl Diphosphate Biosynthesis

Tomohisa Kuzuyama, Motoki Takagi, Shunji Takahashi,dagger and Haruo Seto*

Institute of Molecular and Cellular Biosciences, University of Tokyo, Bunkyo-ku, Tokyo 113-0032, Japan

Received 31 August 1999/Accepted 18 November 1999

In addition to the ubiquitous mevalonate pathway, Streptomyces sp. strain CL190 utilizes the nonmevalonate pathway for isopentenyl diphosphate biosynthesis. The initial step of this nonmevalonate pathway is the formation of 1-deoxy-D-xylulose 5-phosphate (DXP) by condensation of pyruvate and glyceraldehyde 3-phosphate catalyzed by DXP synthase. The corresponding gene, dxs, was cloned from CL190 by using PCR with two oligonucleotide primers synthesized on the basis of two highly conserved regions among dxs homologs from six genera. The dxs gene of CL190 encodes 631 amino acid residues with a predicted molecular mass of 68 kDa. The recombinant enzyme overexpressed in Escherichia coli was purified as a soluble protein and characterized. The molecular mass of the enzyme was estimated to be 70 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and 130 kDa by gel filtration chromatography, suggesting that the enzyme is most likely to be a dimer. The enzyme showed a pH optimum of 9.0, with a Vmax of 370 U per mg of protein and Kms of 65 µM for pyruvate and 120 µM for D-glyceraldehyde 3-phosphate. The purified enzyme catalyzed the formation of 1-deoxyxylulose by condensation of pyruvate and glyceraldehyde as well, with a Km value of 35 mM for D-glyceraldehyde. To compare the enzymatic properties of CL190 and E. coli DXP synthases, the latter enzyme was also overexpressed and purified. Although these two enzymes had different origins, they showed the same enzymatic properties.

* Corresponding author. Mailing address: Institute of Molecular and Cellular Biosciences, University of Tokyo, Bunkyo-ku, Tokyo 113-0032, Japan. Phone: 81-3-5841-7839. Fax: 81-3-5841-8485. E-mail: haseto{at}imcbns.iam.u-tokyo.ac.jp.

dagger Present address: Department of Biochemistry, Chiba University, School of Medicine, Inohana, Chiba 260-8670, Japan.

Journal of Bacteriology, February 2000, p. 891-897, Vol. 182, No. 4
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Cordoba, E., Salmi, M., Leon, P. (2009). Unravelling the regulatory mechanisms that modulate the MEP pathway in higher plants. J Exp Bot 60: 2933-2943 [Abstract] [Full Text]  
  • Xiang, S., Usunow, G., Lange, G., Busch, M., Tong, L. (2007). Crystal Structure of 1-Deoxy-D-xylulose 5-Phosphate Synthase, a Crucial Enzyme for Isoprenoids Biosynthesis. J. Biol. Chem. 282: 2676-2682 [Abstract] [Full Text]  
  • Guevara-Garcia, A., San Roman, C., Arroyo, A., Cortes, M. E., de la Luz Gutierrez-Nava, M., Leon, P. (2005). Characterization of the Arabidopsis clb6 Mutant Illustrates the Importance of Posttranscriptional Regulation of the Methyl-D-Erythritol 4-Phosphate Pathway. Plant Cell 17: 628-643 [Abstract] [Full Text]  
  • Cassera, M. B., Gozzo, F. C., D'Alexandri, F. L., Merino, E. F., del Portillo, H. A., Peres, V. J., Almeida, I. C., Eberlin, M. N., Wunderlich, G., Wiesner, J., Jomaa, H., Kimura, E. A., Katzin, A. M. (2004). The Methylerythritol Phosphate Pathway Is Functionally Active in All Intraerythrocytic Stages of Plasmodium falciparum. J. Biol. Chem. 279: 51749-51759 [Abstract] [Full Text]  
  • Tabernero, L., Rodwell, V. W., Stauffacher, C. V. (2003). Crystal Structure of a Statin Bound to a Class II Hydroxymethylglutaryl-CoA Reductase. J. Biol. Chem. 278: 19933-19938 [Abstract] [Full Text]  
  • Bailey, A. M., Mahapatra, S., Brennan, P. J., Crick, D. C. (2002). Identification, cloning, purification, and enzymatic characterization of Mycobacterium tuberculosis 1-deoxy-D-xylulose 5-phosphate synthase. Glycobiology 12: 813-820 [Abstract] [Full Text]  
  • Rodriguez-Concepcion, M., Boronat, A. (2002). Elucidation of the Methylerythritol Phosphate Pathway for Isoprenoid Biosynthesis in Bacteria and Plastids. A Metabolic Milestone Achieved through Genomics. Plant Physiol. 130: 1079-1089 [Full Text]  
  • Hedl, M., Sutherlin, A., Wilding, E. I., Mazzulla, M., McDevitt, D., Lane, P., Burgner, J. W. II, Lehnbeuter, K. R., Stauffacher, C. V., Gwynn, M. N., Rodwell, V. W. (2002). Enterococcus faecalis Acetoacetyl-Coenzyme A Thiolase/3-Hydroxy-3-Methylglutaryl-Coenzyme A Reductase, a Dual-Function Protein of Isopentenyl Diphosphate Biosynthesis. J. Bacteriol. 184: 2116-2122 [Abstract] [Full Text]  
  • Altincicek, B., Kollas, A.-K., Sanderbrand, S., Wiesner, J., Hintz, M., Beck, E., Jomaa, H. (2001). GcpE Is Involved in the 2-C-Methyl-D-Erythritol 4-Phosphate Pathway of Isoprenoid Biosynthesis in Escherichia coli. J. Bacteriol. 183: 2411-2416 [Abstract] [Full Text]  
  • Kaneda, K., Kuzuyama, T., Takagi, M., Hayakawa, Y., Seto, H. (2001). An unusual isopentenyl diphosphate isomerase found in the mevalonate pathway gene cluster from Streptomyces sp. strain CL190. Proc. Natl. Acad. Sci. USA 10.1073/pnas.020472198v1 [Abstract] [Full Text]  
  • Hahn, F. M., Eubanks, L. M., Testa, C. A., Blagg, B. S. J., Baker, J. A., Poulter, C. D. (2001). 1-Deoxy-D-Xylulose 5-Phosphate Synthase, the Gene Product of Open Reading Frame (ORF) 2816 and ORF 2895 in Rhodobacter capsulatus. J. Bacteriol. 183: 1-11 [Abstract] [Full Text]  
  • Estévez, J. M., Cantero, A., Romero, C., Kawaide, H., Jiménez, L. F., Kuzuyama, T., Seto, H., Kamiya, Y., León, P. (2000). Analysis of the Expression of CLA1, a Gene That Encodes the 1-Deoxyxylulose 5-Phosphate Synthase of the 2-C-Methyl-D-Erythritol-4-Phosphate Pathway in Arabidopsis. Plant Physiol. 124: 95-104 [Abstract] [Full Text]  
  • Takagi, M., Kuzuyama, T., Takahashi, S., Seto, H. (2000). A Gene Cluster for the Mevalonate Pathway from Streptomyces sp. Strain CL190. J. Bacteriol. 182: 4153-4157 [Abstract] [Full Text]  
  • Rohdich, F., Wungsintaweekul, J., Lüttgen, H., Fischer, M., Eisenreich, W., Schuhr, C. A., Fellermeier, M., Schramek, N., Zenk, M. H., Bacher, A. (2000). Biosynthesis of terpenoids: 4-Diphosphocytidyl-2-C-methyl-D-erythritol kinase from tomato. Proc. Natl. Acad. Sci. USA 10.1073/pnas.140209197v1 [Abstract] [Full Text]  
  • Kuzuyama, T., Takahashi, S., Takagi, M., Seto, H. (2000). Characterization of 1-Deoxy-D-xylulose 5-Phosphate Reductoisomerase, an Enzyme Involved in Isopentenyl Diphosphate Biosynthesis, and Identification of Its Catalytic Amino Acid Residues. J. Biol. Chem. 275: 19928-19932 [Abstract] [Full Text]  
  • Estevez, J. M., Cantero, A., Reindl, A., Reichler, S., Leon, P. (2001). 1-Deoxy-D-xylulose-5-phosphate Synthase, a Limiting Enzyme for Plastidic Isoprenoid Biosynthesis in Plants. J. Biol. Chem. 276: 22901-22909 [Abstract] [Full Text]  
  • Kaneda, K., Kuzuyama, T., Takagi, M., Hayakawa, Y., Seto, H. (2001). An unusual isopentenyl diphosphate isomerase found in the mevalonate pathway gene cluster from Streptomyces sp. strain CL190. Proc. Natl. Acad. Sci. USA 98: 932-937 [Abstract] [Full Text]  
  • Rohdich, F., Wungsintaweekul, J., Luttgen, H., Fischer, M., Eisenreich, W., Schuhr, C. A., Fellermeier, M., Schramek, N., Zenk, M. H., Bacher, A. (2000). Biosynthesis of terpenoids: 4-Diphosphocytidyl-2-C-methyl-D-erythritol kinase from tomato. Proc. Natl. Acad. Sci. USA 97: 8251-8256 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»