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Droplet-based microfluidic platform for high-throughput screening of Streptomyces.

Ran TuYue ZhangErbing HuaLikuan BaiHuamei HuangKaiyue YunMeng Wang
Published in: Communications biology (2021)
Streptomyces are one of the most important industrial microorganisms for the production of proteins and small-molecule drugs. Previously reported flow cytometry-based screening methods can only screen spores or protoplasts released from mycelium, which do not represent the filamentous stationary phase Streptomyces used in industrial cultivation. Here we show a droplet-based microfluidic platform to facilitate more relevant, reliable and rapid screening of Streptomyces mycelium, and achieved an enrichment ratio of up to 334.2. Using this platform, we rapidly characterized a series of native and heterologous constitutive promoters in Streptomyces lividans 66 in droplets, and efficiently screened out a set of engineered promoter variants with desired strengths from two synthetic promoter libraries. We also successfully screened out several hyperproducers of cellulases from a random S. lividans 66 mutant library, which had 69.2-111.4% greater cellulase production than the wild type. Our method provides a fast, simple, and powerful solution for the industrial engineering and screening of Streptomyces in more industry-relevant conditions.
Keyphrases
  • high throughput
  • wild type
  • small molecule
  • single cell
  • flow cytometry
  • heavy metals
  • wastewater treatment
  • dna methylation
  • transcription factor
  • gene expression
  • mass spectrometry
  • sensitive detection