Light-Controlled Fermentations for Microbial Chemical and Protein Production.

TitleLight-Controlled Fermentations for Microbial Chemical and Protein Production.
Publication TypeJournal Article
Year of Publication2022
AuthorsHoffman, SM, Lalwani, MA, Avalos, JL
JournalJ Vis Exp
Issue181
Date Published2022 03 22
ISSN1940-087X
KeywordsEscherichia coli, Fermentation, Metabolic Engineering, Recombinant Proteins, Saccharomyces cerevisiae
Abstract

<p>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. These protocols facilitate the adoption of optogenetic controls in engineered microorganisms for improved fermentation performance.</p>

DOI10.3791/63269
Alternate JournalJ Vis Exp
PubMed ID35404352