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Antimicrobial Agents and Chemotherapy, January 2005, p. 230-240, Vol. 49, No. 1
0066-4804/05/$08.00+0     doi:10.1128/AAC.49.1.230-240.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Molecular Cloning, Sequence Analysis, and Heterologous Expression of the Phosphinothricin Tripeptide Biosynthetic Gene Cluster from Streptomyces viridochromogenes DSM 40736

Joshua A. V. Blodgett, Jun Kai Zhang, and William W. Metcalf^*

Department of Microbiology, University of Illinois, Urbana, Illinois

Received 4 June 2004/ Returned for modification 16 August 2004/ Accepted 20 September 2004

A fosmid library from genomic DNA of Streptomyces viridochromogenes DSM 40736 was constructed and screened for the presence of genes known to be involved in the biosynthesis of phosphinothricin tripeptide (PTT). Eight positives were identified, one of which was able to confer PTT biosynthetic capability upon Streptomyces lividans after integration of the fosmid into the chromosome of this heterologous host. Sequence analysis of the 40,241-bp fosmid insert revealed 29 complete open reading frames (ORFs). Deletion analysis demonstrated that a minimum set of 24 ORFs were required for PTT production in the heterologous host. Sequence analysis revealed that most of these PTT genes have been previously identified in either S. viridochromogenes or S. hygroscopicus (or both), although only 11 out of 24 of these ORFs have experimentally defined functions. Three previously unknown genes within the cluster were identified and are likely to have roles in the stepwise production of phosphonoformate from phosphonoacetaldehyde. This is the first report detailing the entire PTT gene cluster from any producing streptomycete.

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* Corresponding author. Mailing address: Department of Microbiology, University of Illinois, B103 Chemical and Life Sciences Laboratory, 601 S. Goodwin Ave., Urbana, IL 61801. Phone: (217) 244-1943. Fax: (217) 244-6697. E-mail: metcalf{at}uiuc.edu.

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Antimicrobial Agents and Chemotherapy, January 2005, p. 230-240, Vol. 49, No. 1
0066-4804/05/$08.00+0     doi:10.1128/AAC.49.1.230-240.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

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