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 Heinzelmann, E.
Right arrow Articles by Schwartz, D.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Heinzelmann, E.
Right arrow Articles by Schwartz, D.
Agricola
Right arrow Articles by Heinzelmann, E.
Right arrow Articles by Schwartz, D.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, August 2001, p. 3603-3609, Vol. 67, No. 8
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.8.3603-3609.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

The Phosphinomethylmalate Isomerase Gene pmi, Encoding an Aconitase-Like Enzyme, Is Involved in the Synthesis of Phosphinothricin Tripeptide in Streptomyces viridochromogenes

E. Heinzelmann, G. Kienzlen, S. Kaspar, J. Recktenwald, W. Wohlleben, and D. Schwartz*

Mikrobiologie/Biotechnologie, Eberhard-Karls-Universität Tübingen, D-72076 Tübingen, Germany

Received 31 January 2001/Accepted 7 May 2001

Streptomyces viridochromogenes Tü494 produces the antibiotic phosphinothricin tripeptide (PTT). In the postulated biosynthetic pathway, one reaction, the isomerization of phosphinomethylmalate, resembles the aconitase reaction of the tricarboxylic acid (TCA) cycle. It was speculated that this reaction is carried out by the corresponding enzyme of the primary metabolism (C. J. Thompson and H. Seto, p. 197-222, in L. C. Vining and C. Stuttard, ed., Genetics and Biochemistry of Antibiotic Production, 1995). However, in addition to the TCA cycle aconitase gene, a gene encoding an aconitase-like protein (the phosphinomethylmalate isomerase gene, pmi) was identified in the PTT biosynthetic gene cluster by Southern hybridization experiments, using oligonucleotides which were derived from conserved amino acid sequences of aconitases. The deduced protein revealed high similarity to aconitases from plants, bacteria, and fungi and to iron regulatory proteins from eucaryotes. Pmi and the S. viridochromogenes TCA cycle aconitase, AcnA, have 52% identity. By gene insertion mutagenesis, a pmi mutant (Mapra1) was generated. The mutant failed to produce PTT, indicating the inability of AcnA to carry out the secondary-metabolism reaction. A His-tagged protein (Hispmi*) was heterologously produced in Streptomyces lividans. The purified protein showed no standard aconitase activity with citrate as a substrate, and the corresponding gene was not able to complement an acnA mutant. This indicates that Pmi and AcnA are highly specific for their respective enzymatic reactions.

* Corresponding author. Mailing address: Mikrobiologie/Biotechnologie, Eberhard-Karls-Universität Tübingen, Auf der Morgenstelle 28, D-72076 Tübingen, Germany. Phone: 49 7071 29-74638. Fax: 49 7071 29-5979. E-mail: schwartz{at}molbio.biol.biologie.uni-tuebingen.de.

Applied and Environmental Microbiology, August 2001, p. 3603-3609, Vol. 67, No. 8
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.8.3603-3609.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Eys, S., Schwartz, D., Wohlleben, W., Schinko, E. (2008). Three Thioesterases Are Involved in the Biosynthesis of Phosphinothricin Tripeptide in Streptomyces viridochromogenes Tu494. Antimicrob. Agents Chemother. 52: 1686-1696 [Abstract] [Full Text]  
  • Muller, C., Nolden, S., Gebhardt, P., Heinzelmann, E., Lange, C., Puk, O., Welzel, K., Wohlleben, W., Schwartz, D. (2007). Sequencing and Analysis of the Biosynthetic Gene Cluster of the Lipopeptide Antibiotic Friulimicin in Actinoplanes friuliensis. Antimicrob. Agents Chemother. 51: 1028-1037 [Abstract] [Full Text]  
  • Schwartz, D., Grammel, N., Heinzelmann, E., Keller, U., Wohlleben, W. (2005). Phosphinothricin Tripeptide Synthetases in Streptomyces viridochromogenes Tu494. Antimicrob. Agents Chemother. 49: 4598-4607 [Abstract] [Full Text]  
  • Heinzelmann, E., Berger, S., Muller, C., Hartner, T., Poralla, K., Wohlleben, W., Schwartz, D. (2005). An acyl-CoA dehydrogenase is involved in the formation of the {Delta}cis3 double bond in the acyl residue of the lipopeptide antibiotic friulimicin in Actinoplanes friuliensis. Microbiology 151: 1963-1974 [Abstract] [Full Text]  
  • Blodgett, J. A. V., Zhang, J. K., Metcalf, W. W. (2005). Molecular Cloning, Sequence Analysis, and Heterologous Expression of the Phosphinothricin Tripeptide Biosynthetic Gene Cluster from Streptomyces viridochromogenes DSM 40736. Antimicrob. Agents Chemother. 49: 230-240 [Abstract] [Full Text]  
  • Schwartz, D., Berger, S., Heinzelmann, E., Muschko, K., Welzel, K., Wohlleben, W. (2004). Biosynthetic Gene Cluster of the Herbicide Phosphinothricin Tripeptide from Streptomyces viridochromogenes Tu494. Appl. Environ. Microbiol. 70: 7093-7102 [Abstract] [Full Text]  
  • Schmutz, E., Hennig, S., Li, S.-M., Heide, L. (2004). Identification of a topoisomerase IV in actinobacteria: purification and characterization of ParYR and GyrBR from the coumermycin A1 producer Streptomyces rishiriensis DSM 40489. Microbiology 150: 641-647 [Abstract] [Full Text]  
  • Heinzelmann, E., Berger, S., Puk, O., Reichenstein, B., Wohlleben, W., Schwartz, D. (2003). A Glutamate Mutase Is Involved in the Biosynthesis of the Lipopeptide Antibiotic Friulimicin in Actinoplanes friuliensis. Antimicrob. Agents Chemother. 47: 447-457 [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»