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Butenyl-spinosyns, a natural example of genetic engineering of antibiotic biosynthetic genes

  • Original Paper - JMBM
  • Published:
Journal of Industrial Microbiology and Biotechnology

Abstract

Spinosyns, a novel class of insect active macrolides produced by Saccharopolyspora spinosa, are used for insect control in a number of commercial crops. Recently, a new class of spinosyns was discovered from S. pogona NRRL 30141. The butenyl-spinosyns, also called pogonins, are very similar to spinosyns, differing in the length of the side chain at C-21 and in the variety of novel minor factors. The butenyl-spinosyn biosynthetic genes (bus) were cloned on four cosmids covering a contiguous 110-kb region of the NRRL 30141 chromosome. Their function in butenyl-spinosyn biosynthesis was confirmed by a loss-of-function deletion, and subsequent complementation by cloned genes. The coding sequences of the butenyl-spinosyn biosynthetic genes and the spinosyn biosynthetic genes from S. spinosa were highly conserved. In particular, the PKS-coding genes from S. spinosa and S. pogona have 91–94% nucleic acid identity, with one notable exception. The butenyl-spinosyn gene sequence codes for one additional PKS module, which is responsible for the additional two carbons in the C-21 tail. The DNA sequence of spinosyn genes in this region suggested that the S. spinosa spnA gene could have been the result of an in-frame deletion of the S. pogona busA gene. Therefore, the butenyl-spinosyn genes represent the putative parental gene structure that was naturally engineered by deletion to create the spinosyn genes.

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Acknowledgements

We would like to acknowledge the assistance of Dennis Duebelbeis and Paul Lewer who provided LC and LC/MS analysis of fermentations. We also acknowledge Dow AgroSciences Discovery management for enthusiastic support of this work.

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Authors and Affiliations

  1. Discovery Research, LLC, Dow AgroSciences, 9330 Zionsville Road., Indianapolis, IN, 46268-1054, USA

    Donald R. Hahn, Gary Gustafson, Clive Waldron & Jon Mitchell

  2. Department of Medical Microbiology and Immunology, Medical University of Ohio, 3055 Arlington Avenue, Toledo, OH, 43614-5806, USA

    Brian Bullard

  3. Pfizer Inc., 7000 Portage Road, Kalamazoo, MI, 49001, USA

    James D. Jackson

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  1. Donald R. Hahn
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  2. Gary Gustafson
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  3. Clive Waldron
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  5. James D. Jackson
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  6. Jon Mitchell
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Correspondence to Donald R. Hahn.

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Hahn, D.R., Gustafson, G., Waldron, C. et al. Butenyl-spinosyns, a natural example of genetic engineering of antibiotic biosynthetic genes. J IND MICROBIOL BIOTECHNOL 33, 94–104 (2006). https://doi.org/10.1007/s10295-005-0016-9

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  • DOI: https://doi.org/10.1007/s10295-005-0016-9

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