loading page

High-throughput GPCR-based autocrine screening for secondary metabolite production in yeast
  • +3
  • Tatyana Saleski,
  • Huadong Peng,
  • Bettina Lengger,
  • Jinglin Wang,
  • Michael Krogh Jensen,
  • Emil D. Jensen
Tatyana Saleski
Danmarks Tekniske Universitet The Novo Nordisk Foundation Center for Biosustainability
Author Profile
Huadong Peng
Danmarks Tekniske Universitet The Novo Nordisk Foundation Center for Biosustainability
Author Profile
Bettina Lengger
Danmarks Tekniske Universitet The Novo Nordisk Foundation Center for Biosustainability
Author Profile
Jinglin Wang
Danmarks Tekniske Universitet The Novo Nordisk Foundation Center for Biosustainability
Author Profile
Michael Krogh Jensen
Danmarks Tekniske Universitet The Novo Nordisk Foundation Center for Biosustainability
Author Profile
Emil D. Jensen
Danmarks Tekniske Universitet The Novo Nordisk Foundation Center for Biosustainability

Corresponding Author:emdaje@biosustain.dtu.dk

Author Profile

Abstract

Biosensors are valuable tools in accelerating the test phase of the design-build-test-learn cycle of cell factory development, as well as in bioprocess monitoring and control. G protein-coupled receptor (GPCR)-based biosensors enable cells to sense a wide array of molecules and environmental conditions in a specific manner. Due to the extracellular nature of their sensing, GPCR-based biosensors require compartmentalization of distinct genotypes when screening production levels of a strain library to ensure that detected levels originate exclusively from the strain under assessment. Here, we explore the integration of production and sensing modalities into a single Saccharomyces cerevisiae strain and compartmentalization using three different methods: (1) cultivation in microtiter plates, (2) spatial separation on agar plates, and (3) encapsulation in water-in-oil-in-water double emulsion droplets, combined with analysis and sorting via a fluorescence-activated cell sorting (FACS) machine. Employing tryptamine and serotonin as proof-of-concept target molecules, we optimize biosensing conditions and demonstrate the ability of the autocrine screening method to enrich for high producers, showing the enrichment of a serotonin-producing strain over a non-producing strain. These findings illustrate a workflow that can be adapted to screening for a wide range of complex chemistry at high throughput using commercially available microfluidic systems.
Submitted to Biotechnology and Bioengineering
16 Jun 2024Review(s) Completed, Editorial Evaluation Pending
16 Jun 2024Editorial Decision: Revise Minor
24 Jun 2024Submission Checks Completed
24 Jun 2024Assigned to Editor
24 Jun 2024Reviewer(s) Assigned
27 Jun 2024Editorial Decision: Accept