The two-component system CepRS regulates the cephamycin C biosynthesis in Streptomyces clavuligerus F613-1

Abstract

During industrial fermentation, Streptomyces clavuligerus F613-1 simultaneously produces primary product clavulanic acid (CA) and cephamycin C. The cephamycin C biosynthetic gene cluster and pathway have been basically elucidated and the CcaR positive regulator was found to control the cephamycin genes expression. However, additional mechanisms of regulation cannot be excluded. The BB341_RS13780/13785 gene pair in S. clavuligerus F613-1 (annotated as SCLAV_2960/2959 in S. clavuligerus ATCC27064) encodes a bacterial two-component system (TCS) and were designated as CepRS (for cep hamycin r egulator/ s ensor). CepRS significantly affects cephamycin C production but only slightly affects CA production. To further understand the regulation of cephamycin C biosynthesis, the cepRS genes were deleted from S. clavuligerus F613-1. The deletion mutant resulted in decreased cephamycin C production but had no phenotypic effects. Real-time quantitative polymerase chain reaction analysis revealed that CepRS regulates the expression of most genes involved in cephamycin C biosynthesis, with electrophoretic mobility shift assays showing that CepR interacts with the cefD - cmcI intergenic region. These results demonstrate that the CepR response regulator serves as a transcriptional activator of cephamycin C biosynthesis, which may provide an approach for metabolic engineering methods for CA production by S. clavuligerus F613-1 in future. Electronic supplementary material The online version of this article (10.1186/s13568-019-0844-z) contains supplementary material, which is available to authorized users.

Comparative Genomics and Metabolomics Analyses of Clavulanic Acid-Producing Streptomyces Species Provides Insight Into Specialized Metabolism

“…Our results also show similarities and differences in the overall specialized metabolic capabilities of CA-producing Streptomyces species under different nutritional conditions, which, to the best of our knowledge, is the first report on the subject. Although the current study did not examine or address regulation, we would like to point out that many of the genes known to control Ceph-C and CA production in S. clavuligerus are also conserved in the two other producers (Liras et al, 2008; Ferguson et al, 2016; Fu et al, 2019b). It has been noted that deciphering the complete CA biosynthetic pathway in S. clavuligerus is challenging due to the presence of the 5 S clavam biosynthetic pathway.…”

Global Chromosome Topology and the Two-Component Systems in Concerted Manner Regulate Transcription in Streptomyces

“…Streptomyces two-component systems are known to control antibiotic production, central metabolism, morphology, and differentiation ( 36 ). Although numerous TCSs in Streptomyces affecting these processes have been well described (e.g., CepRS [ 44 ], MacRS [ 45 ], PhoRP [ 46 , 47 ], OsdKR [ 48 ], AfsQ1/Q2 [ 49 ], and DraRK [ 50 ]), the identification of signals to which TCSs respond has been successful for only a few TCSs (e.g., phosphate concentrations for PhoPR [ 46 ], nitrogen concentrations for AfsQ1/Q2 [ 49 ] and DraRK [ 50 ], iron availability for AbrA1/A2 [ 51 ], and redox stress for SenSR [ 52 ]). Here, we have established a role for SatKR in regulating growth rate, differentiation, and antibiotic production in S. coelicolor , but we were unable to predict the SatKR activation signal on the basis of our data.…”

Global chromosome topology and the two-component systems in concerted manner regulate transcription in Streptomyces

“…Although numerous TCSs in Streptomyces affecting these processes have been well-described ( e.g. CepRS (52), MacRS (53), PhoRP (54, 55), OsdKR (56), AfsQ1/Q2 (57), DraRK (58)), the identification of signals to which TCSs respond has only been successful for a few TCSs (e.g. phosphate concentrations for PhoPR (54), nitrogen concentrations for AfsQ1/Q2 (57) and DraRK (58), iron availability for AbrA1/A2 (59), and redox stress for SenSR (60)).…”