Light-Controlled Fermentations for Microbial Chemical and Protein Production

Journal Article · Tue Mar 22 00:00:00 EDT 2022 · Journal of Visualized Experiments

DOI:https://doi.org/10.3791/63269· OSTI ID:1900140

Hoffman, Shannon M. ^[1]; Lalwani, Makoto A. ^[2]; Avalos, José L. ^[2]

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1. Princeton Univ., NJ (United States); Princeton University
2. Princeton Univ., NJ (United States)

Microbial cell factories offer a sustainable alternative for producing chemicals and recombinant proteins from renewable feedstocks. However, overburdening a microorganism with genetic modifications can reduce host fitness and productivity. This problem can be overcome by using dynamic control: inducible expression of enzymes and pathways, typically using chemical- or nutrient-based additives, to balance cellular growth and production. Optogenetics offers a non-invasive, highly tunable, and reversible method of dynamically regulating gene expression. Here, we describe how to set up light-controlled fermentations of engineered Escherichia coli and Saccharomyces cerevisiae for the production of chemicals or recombinant proteins. We discuss how to apply light at selected times and dosages to decouple microbial growth and production for improved fermentation control and productivity, as well as the key optimization considerations for best results. Additionally, we describe how to implement light controls for lab-scale bioreactor experiments. As a result, these protocols facilitate the adoption of optogenetic controls in engineered microorganisms for improved fermentation performance.

• Research Organization: Princeton Univ., NJ (United States)
• Sponsoring Organization: USDOE Office of Science (SC), Biological and Environmental Research (BER)
• Grant/Contract Number: SC0019363
• OSTI ID: 1900140
• Journal Information: Journal of Visualized Experiments, Journal Name: Journal of Visualized Experiments Journal Issue: 181; ISSN 1940-087X
• Publisher: MyJoVE Corp.
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
60 APPLIED LIFE SCIENCES
light controlled fermentations
methods
optogentics