A comparative study at bioprocess and metabolite levels of superhost strain Streptomyces coelicolor M1152 and its derivative M1581 heterologously expressing chloramphenicol biosynthetic gene cluster

Abstract

Microbial superhost strains should provide an ideal platform for the efficient homologous or heterologous phenotypic expression of biosynthetic gene clusters (BGCs) of new and novel bioactive molecules. Our aim in the current study was to perform a comparative study at the bioprocess and metabolite levels of the previously designed superhost strain Streptomyces coelicolor M1152 and its derivative strain S. coelicolor M1581 heterologously expressing chloramphenicol BGC. Parent strain M1152 was characterized by a higher specific growth rate, specific CO 2 evolution rate, and a higher specific L-glutamate consumption rate as compared with M1581. Intracellular primary central metabolites (nucleoside/sugar phosphates, amino acids, organic acids, and CoAs) were quantified using four targeted LC-MS/ MS-based methods. The metabolite pathways in the nonantibiotic producing S. coelicolor host strain were flooded with carbon from both carbon sources, whereas in antibiotic-producing strain, the carbon of L-glutamate seems to be draining out through excreting synthesized antibiotic. The 13 C-isotope-labeling experiments revealed the bidirectionality in the glycolytic pathway and reversibility in the nonoxidative part of PPP even with continuous uptake of D-glucose. The change in the primary metabolites due to the insertion of BGC disclosed a clear linkage between the primary and secondary metabolites.

Differences at Species Level and in Repertoires of Secondary Metabolite Biosynthetic Gene Clusters among Streptomyces coelicolor A3(2) and Type Strains of S. coelicolor and Its Taxonomic Neighbors

“…Its wild strain and derived strains have contributed to various biotechnological fields. It is notable that these strains are used as the hosts for heterogeneous secondary metabolite production [40][41][42][43][44]. In contrast to these applications, the classification of S. coelicolor A3(2) has been paid less attention.…”

Nutrient-depended metabolic switching during batch cultivation of Streptomyces coelicolor explored with absolute quantitative mass spectrometry-based metabolite profiling

“…Two strains of Streptomyces coelicolor were used in this study: S. coelicolor M145 and S. coelicolor M1146. The protocols for spore preparation, pre-germination, media preparation, and setting up the experiments were adopted from earlier reports (Wentzel et al 2012a ; Kumar and Bruheim 2021b ). The cultivations were conducted in 3 L bioreactors (Eppendorf Bioflo 320) with an initial working volume of 1.8 L and the inoculum was pre-germinated spores having ~ 1 × 10 9 CFU L −1 .…”

“…The central carbon metabolites (CCMs) carry most of the metabolic flux and provide precursors, reductive power, and energy used for biomass generation and secondary metabolite synthesis and their analysis can give real-time status of phenotype (Coze et al 2013 ; Kumar et al 2021 ; Rost et al 2020a ; Kumar and Bruheim 2021a , b ). Therefore, information about the intracellular metabolite concentrations provided by metabolome analysis will further enhance our understanding of the genotype–phenotype interactions both by direct single interpretation of metabolomics data but also when integrated with multi omics-data such as genomics (Xu et al 2017 ; Nguyen et al 2012 ), transcriptomics (Kim and Kim 2016 ; Yague et al 2014 ; Nieselt et al 2010 ) and proteomics (Thomas et al 2012 ).…”

“…We have developed a set of LC–MS/MS methods for the absolute quantitative metabolite profiling including the 13 C-isotope dilution approach for enhanced analytical accuracy (Supplementary Fig. S1) (Bartosova et al 2021 ; Kumar et al 2021 ; Rost et al 2020a ; Kumar and Bruheim 2021b ). The current study aimed to apply these methods for the quantification of CCMs of S. coelicolor M145 during the growth and antibiotic production phases in two nutrient-limited conditions (L-glutamate and phosphate limited).…”

“…S coelicolor M145 is a plasmid cured derivative strain of the wild type strain A3(2) (Kieser et al 2000 ) and a reference strain of a series of new derivative strains (Gomez-Escribano and Bibb 2011 ). Recently, we have published a comprehensive coverage of primary metabolome data by comparing super host strain S. coelicolor M1152 and its derivative S. coelicolor M1581 heterologously expressing a chloramphenicol BGC (Kumar and Bruheim 2021b ). The current study was designed to further decipher the distribution and participation of carbon from two substrates (D-glucose and L-glutamate) in primary metabolic pathways and their role in the growth and support of antibiotic biosynthesis.…”