A Streptomyces coelicolor host for the heterologous expression of Type III polyketide synthase genes

作者：Thanapipatsiri Anyarat; Claesen Jan; Gomez Escribano Juan Pablo; Bibb Mervyn; Thamchaipenet Arinthip^*

来源：Microbial Cell Factories, 2015, 14(1): 145.

DOI：10.1186/s12934-015-0335-0

• Streptomyces
• Expression host
• Type III polyketide synthases

Background: Recent advances in genome sequencing, combined with bioinformatic analysis, has led to the identification of numerous novel natural product gene clusters, particularly in actinomycetes of terrestrial and marine origin. Many of these gene clusters encode uncharacterised Type III polyketide synthases. To facilitate the study of these genes and their potentially novel products, we set out to construct an actinomycete expression host specifically designed for the heterologous expression of Type III PKS genes and their gene clusters.
Results: A derivative of Streptomyces coelicolor A3(2) designed for the expression of Type III polyketide synthase (PKS) genes was constructed from the previously engineered expression strain S. coelicolor M1152 [Delta act Delta red Delta cpk Delta cda rpoB(C1298T)] by removal of all three of the endogenous Type III PKS genes (gcs, srsA, rppA) by PCR targeting. The resulting septuple deletion mutant, M1317, proved to be an effective surrogate host for the expression of actinobacterial Type III PKS genes: expression of the reintroduced gcs gene from S. coelicolor and of the heterologous rppA gene from Streptomyces venezuelae under the control of the constitutive ermE* promoter resulted in copious production of germicidin and flaviolin, respectively.
Conclusions: The newly constructed expression host S. coelicolor M1317 should be particularly useful for the discovery and analysis of new Type III polyketide metabolites.

• 出版日期2015-9-16