A two-component system gene SACE_0101 regulates copper homeostasis in Saccharopolyspora erythraea

Abstract

Background: Saccharopolyspora erythraea (S. erythraea) is a Gram-positive bacterium widely used for the production of erythromycin, a potent macrolide antibiotic. However, the mechanism behind erythromycin production is poorly understood. In the high erythromycin-producer strain S. erythraea HL3168 E3, the level of copper ions positively correlates with erythromycin production. To explain this correlation, we performed a genome-based comparison between the wild-type strain NRRL23338 and the mutant strain HL3168 E3, and further characterized the identified gene(s) by targeted genome editing, mRNA transcript analysis, and functional analysis. Results:The response regulator of the two-component system (TCS) encoded by the gene SACE_0101 in S. erythraea showed high similarity with CopR of TCS CopRS in Streptomyces coelicolor, which is involved in the regulation of copper metabolism. The deletion of SACE_0101 was beneficial for erythromycin synthesis most likely by causing changes in the intracellular copper homeostasis, leading to enhanced erythromycin production. In addition, Cu 2+ supplementation and gene expression analysis suggested that SACE_0101 may be involved in the regulation of copper homeostasis and erythromycin production. Conclusions:The mutation of SACE_0101 gene increased the yield of erythromycin, especially upon the addition of copper ions. Therefore, the two-component system gene SACE_0101 plays a crucial role in regulating copper homeostasis and erythromycin synthesis in S. erythraea.

Comparative genomic and transcriptomic analysis guides to further enhance the biosynthesis of erythromycin by an overproducer

“…The low transcription of SACE 2955-2956 implied that alternative operons (SACE 0142-0143) encoding the bd complex were operative or E3 preferred an alternative electron transport channel, such as through bc 1 -aa 3 complex. The low transcription of SACE 3011, which encodes a copper resistance gene, fitted the conclusion drawn in our recent research that appropriate exogenous addition of copper ions would increase the production of erythromycin in E3 (Qiao, Li, Ke, & Chu, 2020).…”

“…Regulator genes with significantly different transcriptional levels in Table S5 are candidates which can affect the transcription of erythromycin BGC. Besides, a two component system (TCS) regulating the copper homeostasis in S. erythraea was identified in our recent study based on the present comparative genomics (Qiao et al, 2020).…”

“…SACE 0101 enhanced the copper tolerance of E3 compared to NRRL23338. Gene deletion of SACE 0101 in NRRL23338 increased its erythromycin titer by 110% and the addition of copper enhanced the erythromycin titer of E3 by 17% (Qiao et al, 2020).…”

RclS Sensor Kinase Modulates Virulence of Pseudomonas capeferrum

“…In various antibiotic producers, predominantly members of the genus Streptomyces , diverse TCSs control the biosynthesis of secondary metabolites, including antibiotics [ 31 ]. For example, in actinomycetes Nonomuraea gerenzanensis and Saccharopolyspora erythraea (formerly known as Streptomyces erythraeus ) TCS negatively regulates the production of glycopeptide A40926 and erythromycin, respectively [ 32 , 33 ]. Besides mapping the RclS regulon genes by transcriptome analysis, our aim was the phenotypic determination of RclS sensor kinase influence on the pathogenic potential of P. capeferrum WCS358.…”

Comparative genomic and transcriptomic analysis guides to further enhance the biosynthesis of erythromycin by an overproducer

“…Regulator genes with significantly different transcriptional levels in Table S5 are candidates which can affect the transcription of erythromycin BGC. Besides, a two component system (TCS) regulating the copper homeostasis in S. erythraea was identified in our recent study based on the present comparative genomics (Qiao et al, 2020). Copper functioned as a cofactor of bc 1 -aa 3 terminal oxidase (Fujimoto et al, 2016).…”