Insecticidal antibiotics, due to their high efficiency and environmental protection, meet the technical requirements of controlling crop diseases and insect during storage. In order to obtain new insecticidal antibiotics, numerous actionmycetes were separated from 162 soil samples collected from different districts by different methods. The high activity strain ASAGF58 was screened out by high throughput fermentation platform and mosquito larvae bioassay. Basing on the 16S rDNA sequencing and the construction of phylogenetic tree, ASAGF58 is preliminarily identified as a kind of Saccharopolyspora. Using HPLC and LC-MS, two bioactive components were extracted from the fermentation broth, whose mass-to-charge ratios were 758.483 8 and 772.471 2, respectively, which was consistent with butenyl- spinosyns, the metabolite of Saccharopolyspora pogona. A strain producing butenyl-spinosyns was obtained, which provided technical reserves for the research and development of efficient and green insecticidal compounds in China.