Enhanced Heterologous Spinosad Production from a 79-kb Synthetic Multioperon Assembly
Research output: Contribution to journal › Research article › Contributed › peer-review
Contributors
Abstract
Refactoring biosynthetic pathways for enhanced secondary metabolite production is a central challenge for synthetic biology. Here we applied advanced DNA assembly methods and a uniform overexpression logic using constitutive promoters to achieve efficient heterologous production of the complex insecticidal macrolide spinosad. We constructed a 79-kb artificial gene cluster in which 23 biosynthetic genes were grouped into 7 operons, each with a strong constitutive promoter. Compared with the original gene cluster, the artificial gene cluster resulted in a 328-fold enhanced spinosad production in Streptomyces albus J1074. To achieve this goal, we applied the ExoCET DNA assembly method to build a plasmid from 13 GC-rich fragments with high efficiency in one step. Together with our previous direct cloning and recombineering tools, we present new synthetic biology options for refactoring large gene clusters for diverse applications.
Details
Original language | English |
---|---|
Pages (from-to) | 137-147 |
Number of pages | 11 |
Journal | ACS synthetic biology |
Volume | 8 |
Issue number | 1 |
Publication status | Published - 18 Jan 2019 |
Peer-reviewed | Yes |
External IDs
PubMed | 30590919 |
---|---|
ORCID | /0000-0002-4754-1707/work/142248102 |
Keywords
ASJC Scopus subject areas
Keywords
- biosynthesis, DNA engineering, gene clusters, heterologous expression, secondary metabolites, synthetic operons