Amanda van Tilburg
Lipid rafts in Bacillus subtilis as production platforms for lantibiotics
Recently, it has become clear that bacteria, similar as eukaryotic cells, also contain specific microdomains in their cytoplasmic membranes which are known as lipid rafts. Lipid rafts are thought to increase the efficiency of reaction pathways by the spatial concentration of protein partners into lipid raft domains and by limiting the occurrence of cross-reactions.
Lipid rafts for improved production yields of industrial relevant compounds.
The major goal of my PhD project is to improve bacteria as production hosts for various industrial relevant compounds by using the beneficial effects of lipid rafts. We will focus on the production of antimicrobial peptides (lantibiotics) by Bacillus subtilis. The enzymes involved in the production of lantibiotics will be fused to specific tags for recognition of the lipid raft domains. By shielding the reaction pathway from the rest of the intracellular metabolism, we expect to obtain higher production yields.
My research includes the following techniques:
- general molecular cloning techniques;
- heterologous expression of lantibiotics by B. subtilis strains, antimicrobial activity assays to analyze the presence of antimicrobial peptides, and verification of the lantibiotic peptides by SDS-PAGE and mass spectrometry;
- lipid raft domain extraction and analysis for the presence of the enzymes involved in lantibiotic production.
Lopez, D. Molecular composition of functional microdomains in bacterial membranes. Chemistry and Physics of Lipids 192 (2015): 3–11.