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Time-course transcriptome analysis-guided identification of a four-component system, SbrH1-R that regulates milbemycin biosynthesis by influencing gene cluster expression, precursor supply, and antibiotic efflux

ABSTRACT: Streptomyces bingchenggensis is a soil bacterium that produces milbemycins, which have been applied widely as insecticides and anthelmintics in agricultural and veterinary field. To gain further insights into the regulatory mechanisms in Streptomyces bingchenggensis and mine useful targets for future metabolic engineering to improve milbemycin production, we have compared two strains（the parental strain and high-yielding strain）by comparative transcriptome analysis.This work provides beneficial targets and corresponding engineering methods to facilitate high-production of milbemycins and other polyketide pesticides.

ORGANISM(S): Streptomyces bingchenggensis

PROVIDER: GSE186852 | GEO | 2022/03/02

REPOSITORIES: GEO

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Project description:Biofilms are ubiquitous in natural, medical, and engineering environments. While most antibiotics that primarily aim to inhibit cell growth may result in bacterial drug resistance, biofilm inhibitors do not affect cell growth and there is less chance of developing resistance. This work sought to identify novel, non-toxic and potent biofilm inhibitors from Streptomyces bacteria for reducing the biofilm formation of Pseudomonas aeruginosa PAO1. Out of 4300 Streptomyces strains, one species produced and secreted peptide(s) to inhibit P. aeruginosa biofilm formation by 93% without affecting the growth of planktonic cells. Global transcriptome analyses (DNA microarray) revealed that the supernatant of the Streptomyces 230 strain induced phenazine, pyoverdine, and pyochelin synthesis genes. Electron microscopy showed that the supernatant of Streptomyces 230 strain reduced the production of polymeric matrix in P. aeruginosa biofilm cells, while the Streptomyces species enhanced swarming motility of P. aeruginosa. Therefore, current study suggests that Streptomyces bacteria are an important resource of biofilm inhibitors as well as antibiotics. For the microarray experiments, P. aeruginosa were inoculated in 25 0ml of LB medium in 1000 ml shake flasks with overnight cultures that were diluted 1:100. Streptomyces 230 strain culture media was added in at 1% . Cells were cultured with 10g of glass wool in LB at 37M-BM-0C with 100 rpm shaking for 7 hrs. Cells were immediately chilled with dry ice and 95% ethanol (to prevent RNA degradation) for 30 sec before centrifugation in 50 ml centrifuge tubes at 13,000 g for 2 min; cell pellets were frozen immediately with dry ice and stored -80M-BM-0C. RNA was isolated using Qiagen RNeasy mini Kit (Valencia, CA, USA). RNA quality was assessed by Agilent 2100 bioanalyser using the RNA 6000 Nano Chip (Agilent Technologies, Amstelveen, The Netherlands), and quantity was determined by ND-1000 Spectrophotometer (NanoDrop Technologies, Inc., DE, USA).