Molecules and Cells

Korean Society for Molecular and Cellular Biology (한국분자세포생물학회)
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Metabolic Engineering of Rational Screened Saccharopolyspora spinosa for the Enhancement of Spinosyns A and D Production

  • Jha, Amit Kumar (Institute of Biomolecule Reconstruction (iBR), Department of Pharmaceutical Engineering, Sun Moon University) ;
  • Pokhrel, Anaya Raj (Institute of Biomolecule Reconstruction (iBR), Department of Pharmaceutical Engineering, Sun Moon University) ;
  • Chaudhary, Amit Kumar (Institute of Biomolecule Reconstruction (iBR), Department of Pharmaceutical Engineering, Sun Moon University) ;
  • Park, Seong-Whan (Life Research Institute Dongbu Farm Hannong Co., Ltd.) ;
  • Cho, Wan Je (Life Research Institute Dongbu Farm Hannong Co., Ltd.) ;
  • Sohng, Jae Kyung (Institute of Biomolecule Reconstruction (iBR), Department of Pharmaceutical Engineering, Sun Moon University)
  • Received : 2014.06.16
  • Accepted : 2014.08.19
  • Published : 2014.10.31
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Abstract

Spinosyns A and D are potent ingredient for insect control with exceptional safety to non-target organisms. It consists of a 21-carbon tetracyclic lactone with forosamine and tri-Omethylated rhamnose which are derived from S-adenosyl-methionine. Although previous studies have revealed the involvement of metK1 (S-adenosylmethionine synthetase), rmbA (glucose-1-phosphate thymidylyltransferase), and rmbB (TDP-D-glucose-4, 6-dehydratase) in the biosynthesis of spinosad, expression of these genes into rational screened Saccharopolyspora spinosa (S. spinosa MUV) has not been elucidated till date. In the present study, S. spinosa MUV was developed to utilize for metabolic engineering. The yield of spinosyns A and D in S. spinosa MUV was $244mgL^{-1}$ and $129mgL^{-1}$, which was 4.88-fold and 4.77-fold higher than that in the wild-type ($50mgL^{-1}$ and $27mgL^{-1}$), respectively. To achieve the better production; positive regulator metK1-sp, rmbA and rmbB genes from Streptomyces peucetius, were expressed and co-expressed in S. spinosa MUV under the control of strong $ermE^*$ promoter, using an integration vector pSET152 and expression vector pIBR25, respectively. Here-with, the genetically engineered strain of S. spinosa MUV, produce spinosyns A and D up to $372/217mgL^{-1}$ that is 7.44/8.03-fold greater than that of wild type. This result demonstrates the use of metabolic engineering on rationally developed high producing natural variants for the production.

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References

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