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Core-Shell Droplet-Based Microfluidic Screening System for Filamentous Fungi.
Changtai Zhang1,2, Xiaohui Wu1,2, Fuqiang Song1,2
1Science Center for Future Foods, Jiangnan University, 1800 Lihu Road, Wuxi, Jiangsu 214122, China.
ACS Sensors
|August 21, 2023
Summary
A new high-throughput screening system using droplet microfluidics optimizes protein production in filamentous fungi. This method enhances the discovery of superior strains for biotechnology applications.
Area of Science:
- Biotechnology
- Microbiology
- Bioprocess Engineering
Background:
- Filamentous fungi are valuable hosts for producing drugs, proteins, and chemicals.
- Current screening methods for optimizing production in these fungi are limited by low throughput.
Purpose of the Study:
- To develop a universal high-throughput system for optimizing protein production in filamentous fungi.
- To overcome limitations of existing screening methods for enhanced bioprocess development.
Main Methods:
- Utilized droplet microfluidics to encapsulate large mutant strain pools in biocompatible core-shell microdroplets for prolonged culture.
- Employed self-assembled split green fluorescent protein (GFP) for characterizing secretory capacity and isolating high-producing strains based on fluorescence signals.
Main Results:
- Successfully applied the platform to optimize alpha-amylase secretion in *Aspergillus niger*, isolating a strain with a 2.02-fold higher secretion capacity.
- The system enables analysis of over 10^5 single cells per hour, facilitating ultrahigh-throughput screening.
Conclusions:
- The developed droplet microfluidics system offers a broadly applicable solution for ultrahigh-throughput screening of filamentous fungi.
- This method can significantly accelerate the identification of improved fungal hosts for large-scale production of biotechnology-relevant proteins.
