Droplet-based Microfluidic Platform for High-throughput Screening of Streptomyces

Overview

Journal Commun Biol

Specialty Biology

Date 2021 Jun 1

PMID 34059751

Citations 18

Authors

Ran Tu

Yue Zhang

Erbing Hua

Likuan Bai

Huamei Huang

Kaiyue Yun

Meng Wang

Affiliations

Soon will be listed here.

Abstract

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.

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