Institute of Biomolecule Reconstruction (iBR), Department of Life Science and Biochemical Engineering, Sun Moon University, Asan-si, Chungnam, Korea.
Department of BT-Convergent Pharmaceutical Engineering, Sun Moon University, Asan-si, Chungnam, Korea.
Biotechnol Appl Biochem. 2023 Jun;70(3):1035-1043. doi: 10.1002/bab.2418. Epub 2022 Dec 13.
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.
螺旋霉素是由土壤放线菌（Saccharopolyspora spinosa）产生的一种高效杀虫剂，由 A 和 D 两种螺旋霉素混合而成。由于野生型土壤放线菌产率低，因此人们对提高螺旋霉素的产量产生了浓厚的兴趣。在本研究中，我们设计并构建了 pIBR-SPN 载体。pIBR-SPN 是一种整合载体，可用于将外源基因导入土壤放线菌的染色体。我们合成了不同组合的编码法呢醇胺和鼠李糖的基因，并用于构建不同的重组质粒。开发了以下重组菌株：携带 pIBR-SPN 载体的 S. spinosa（仅携带载体）、携带法呢醇胺基因的 S. spinosa pIBR-SPN F、携带鼠李糖基因的 S. spinosa pIBR-SPN R、携带法呢醇胺和鼠李糖基因的 S. spinosa pIBR-SPN FR、携带法呢醇胺、鼠李糖和 SAM（S-腺苷甲硫氨酸合成酶）基因的 S. spinosa pIBR-SPN FRS 以及携带 pIBR-SPN FR 的 S. spinosa MUV。在这些重组菌株中，S. spinosa pIBR-SPN FR 产生了 1394±163 mg/L 的螺旋霉素，比野生型提高了 13 倍。S. spinosa MUV pIBR-SPN FR 产生了 1897（±129）mg/L 的螺旋霉素，比 S. spinosa MUV 提高了 7 倍，比野生型提高了 17 倍。
