Increasing production of spinosad inSaccharopolyspora spinosaby metabolic engineering

Abstract Spinosad, a combination of spinosyn A and D produced by Saccharopolyspora spinosa , is a highly efficient pesticide. There has been a considerable interest in the improvement of spinosad production because of a low yield achieved by wild‐type S. spinosa . In this study, we designed and constructed a pIBR‐SPN vector. pIBR‐SPN is an integrative vector that can be used to introduce foreign genes into the chromosome of S. spinosa . Different combinations of genes encoding forasamine and rhamnose were synthesized and used for the construction of different recombinant plasmids. The following recombinant strains were developed: S. spinosa pIBR‐SPN (only the vector), S. spinosa pIBR‐SPN F (forosamine genes), S. spinosa pIBR‐SPN R (rhamnose genes), S. spinosa pIBR‐SPN FR (forosamine and rhamnose genes), S. spinosa pIBR‐SPN FRS (forosamine, rhamnose, and SAM [S‐adenosyl‐L‐methionine synthetase] genes), and S. spinosa MUV pIBR‐SPN FR. Among these recombinant strains, S. spinosa pIBR‐SPN FR produced 1394 ± 163 mg/L spinosad, which was 13‐fold higher than the wild‐type. S. spinosa MUV pIBR‐SPN FR produced 1897 (±129) mg/L spinosad, which was seven‐fold higher than S. spinosa MUV and 17‐fold higher than the wild‐type strain.