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http://dx.doi.org/10.4014/jmb.1909.09022

Heterologous Expression of Daptomycin Biosynthetic Gene Cluster Via Streptomyces Artificial Chromosome Vector System  

Choi, Seunghee (Department of Biological Engineering, Inha University)
Nah, Hee-Ju (Department of Biological Engineering, Inha University)
Choi, Sisun (Department of Biological Engineering, Inha University)
Kim, Eung-Soo (Department of Biological Engineering, Inha University)
Publication Information
Journal of Microbiology and Biotechnology / v.29, no.12, 2019 , pp. 1931-1937 More about this Journal
Abstract
The heterologous expression of the Streptomyces natural product (NP) biosynthetic gene cluster (BGC) has become an attractive strategy for the activation, titer improvement, and refactoring of valuable and cryptic NP BGCs. Previously, a Streptomyces artificial chromosomal vector system, pSBAC, was applied successfully to the precise cloning of large-sized polyketide BGCs, including immunosuppressant tautomycetin and antibiotic pikromycin, which led to stable and comparable production in several heterologous hosts. To further validate the pSBAC system as a generally applicable heterologous expression system, the daptomycin BGC of S. roseosporus was cloned and expressed heterologously in a model Streptomyces cell factory. A 65-kb daptomycin BGC, which belongs to a non-ribosomal polypeptide synthetase (NRPS) family, was cloned precisely into the pSBAC which resulted in 28.9 mg/l of daptomycin and its derivatives in S. coelicolor M511(a daptomycin non-producing heterologous host). These results suggest that a pSBAC-driven heterologous expression strategy is an ideal approach for producing low and inconsistent Streptomyces NRPS-family NPs, such as daptomycin, which are produced low and inconsistent in native host.
Keywords
Streptomyces artificial chromosome; daptomycin; biosynthetic gene cluster; heterologous expression;
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