10.6084/m9.figshare.5325010.v1 Zhao C. Zhao C. Huang Y. Huang Y. Guo C. Guo C. Yang B. Yang B. Zhang Y. Zhang Y. Lan Z. Lan Z. Guan X. Guan X. Song Y. Song Y. Zhang X. Zhang X. Supplementary Material for: Heterologous Expression of Spinosyn Biosynthetic Gene Cluster in Streptomyces Species Is Dependent on the Expression of Rhamnose Biosynthesis Genes Karger Publishers 2017 Saccharopolyspora spinosa Spinosyn biosynthetic gene cluster Heterologous expression Streptomyces Rhamnose biosynthetic genes 2017-08-18 09:58:05 Dataset https://karger.figshare.com/articles/dataset/Supplementary_Material_for_Heterologous_Expression_of_Spinosyn_Biosynthetic_Gene_Cluster_in_Streptomyces_Species_Is_Dependent_on_the_Expression_of_Rhamnose_Biosynthesis_Genes/5325010 <p>Spinosyns are a group of macrolide insecticides produced by <i>Saccharopolyspora spinosa</i>. Although <i>S. spinosa</i> can be used for industrial-scale production of spinosyns, this might suffer from several limitations, mainly related to its long growth cycle, low fermentation biomass, and inefficient utilization of starch. It is crucial to generate a robust strain for further spinosyn production and the development of spinosyn derivatives. A BAC vector, containing the whole biosynthetic gene cluster for spinosyn (74 kb) and the elements required for conjugal transfer and site-specific integration, was introduced into different <i>Streptomyces</i> hosts in order to obtain heterologous spinosyn-producing strains. The exconjugants of different <i>Streptomyces</i> strains did not show spinosyn production unless the rhamnose biosynthesis genes from <i>S. spinosa</i> genomic DNA were present and expressed under the control of a strong constitutive <i>ermE</i>*<i>p</i> promoter. Using this heterologous expression system resulted in yields of 1 μg/mL and 1.5 μg/mL spinosyns in <i>Streptomyces coelicolor</i> and <i>Streptomyces lividans</i>, respectively. This report demonstrates spinosyn production in 2 <i>Streptomyces</i> strains and stresses the essential role of rhamnose in this process. This work also provides a potential alternative route for producing spinosyn analogs by means of genetic manipulation in the heterologous hosts.</p>