Butenyl-spinosyns, a natural example of genetic engineering of antibiotic biosynthetic genes |
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Authors: | Donald R Hahn Gary Gustafson Clive Waldron Brian Bullard James D Jackson Jon Mitchell |
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Institution: | (1) Discovery Research, LLC, Dow AgroSciences, 9330 Zionsville Road., Indianapolis, IN 46268-1054, USA;(2) Department of Medical Microbiology and Immunology, Medical University of Ohio, 3055 Arlington Avenue, Toledo, OH 43614-5806, USA;(3) Pfizer Inc., 7000 Portage Road, Kalamazoo, MI 49001, USA |
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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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Keywords: | Spinosyn Macrolide Polyketide Saccharopolyspora pogona |
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