PhD-position | Artificial organelles for advanced understanding of protein folding

Background

Organelles are important subcellular chassis for biochemical conversions, because the enrichment of specialized enzymes allows for higher conversion rates. Their chemical environment can be uniquely tailored compared to the rest of the cell. By re-engineering organelles, a favorable redox or cofactor environment can be created to enhance biochemical pathways.

Oxidative protein folding presents such a compartmentalized pathway, located within the ER of eukaryotic cells. Strikingly, even in single cell eukaryotes such as yeast, several functionally redundant isoforms exist for each step. So far, their distinct functions and substrate specificity has been studied in deletion mutant cells, however, these studies are limited as during the lack of one isoform the cells compensate by the other isoforms.

Artificial organelles, such as synthetic ER-mimetic environments, have the potential to address current challenges in protein folding, enzyme production, and the sustainable synthesis of key proteins. Design and development of artificial organelles in yeast cells will enable novel solutions for production of recombinant proteins of high societal impact and high demand.

Research Objectives

This research aims to advance the understanding of protein folding mechanisms using synthetic mini-compartments and to develop novel approaches for engineering and production of proteins and enzymes. The project focuses on:

1) Creation of a synthetic yet contextualized ER mimetic environment in a cell-free setup to study the individual roles of redundant eukaryotic oxidative protein folding catalysts and their requirements during substrate folding kinetics and transport.

2) Design of tailored synthetic ER folding modules in vivo to meet the requirements for the identified best suited folding catalysts.

3) Using the gained knowledge and synthetic organelles for compartmentalized protein synthesis to accelerate the sustainable, cost-efficient production of recombinant proteins and enzymes.

Requirements

- Completed Master degree (or equivalent) in biotechnology, molecular biology, biochemistry, or similar study.

- Strong background and practical experience in molecular biology, synthetic biology and/or (microbial) cell engineering, and protein analysis requested

- Strong background and practical experience in in vitro protein synthesis (CFPS) and/or biochemical protein characterization desired

- Scientific interest in protein folding and yeast biotechnology desired.

Where to apply:

www.circularbioengineering.at