Method development for the screening of yeast transporters using SSM-based electrophysiology

  • Steven Axel van der Hoek (Other)

Activity: Talks and presentationsConference presentations

Description

Industrial biotechnology uses cell factories to produce compounds of interest in a sustainable manner. During the metabolic engineering of microbes, issues such as feedback inhibition, product degradation, product toxicity and titer constraints due to biomass hinder the application of engineered organisms for compound production as a sustainable alternative to chemical synthesis (Figure 1A)1. Transporters could solve the problems encountered in metabolic engineering by secreting product and possibly improve performance by taking up precursors.
S. cerevisiae is an eukaryotic model organism and used as an industrial cell factory. Approximately 350 putative transporters have been identified, but for only a small fraction their role has been experimentally verified. The most studied transporters are related to the uptake of nutrients2,3. To make use of transporters in metabolic engineering, many more transporters related to compound efflux have to be functionally characterized.
Electrogenic transporters transport a net charge over the membrane (Figure 1B). Solid supported membrane (SSM)-based electrophysiology is a technique that makes use of this feature of electrogenic transporters and can thus determine the substrates of these transporters4. SSM-based electrophysiology has been scaled up to 96 well format and would therefore be a good platform for the development of a screening method.
In our lab, we are developing and optimizing a workflow to enable us to screen electrogenic transportersagainst compounds of interest using SSM-based electrophysiology. This methodology is also being developed with an eye on heterologous transporter screening. Here we describe our initial efforts and results.
Period18 Sept 201821 Sept 2020
Event titleITTS Inaugural Conference: Inaugural Conference of the International Transmembrane Transporter Society: The Transporter Transition
Event typeConference
LocationVienna, AustriaShow on map
Degree of RecognitionInternational