Curriculum

•  Seit 12/21: Promotion am Lehrstuhl für Chemie Biogener Rohstoffe
• 2018-2021: Masterstudium Molekulare Biotechnologie, Technische Universität München
  • • Masterarbeit am Lehrstuhl für Chemie Biogener Rohstoffe: „Entwicklung von enzymatischenAssays für die Identifizierung von neuartigen Enzymen in Mikrofluidik‐basierten in vitroTranskriptions‐ Translationssystemen
• 2014- 2018: Bachelorstudium Molekulare Biotechnologie, Technische Universität München
  • • Bachelorarbeit am Lehrstuhl für Mikrobiologie: „Gerichtete Mutagenese einer GH10 Xylanase aus Clostridium stercorarium zur Thermostabilisierung“

Forschung

Acceleration of enzyme engineering through application of cell free protein expression platforms

Location: Garching lab

It is my goal to accelerate enzyme engineering through the application of cell free protein synthesis platforms (CFPS). By making use of CFPS, several challenges in enzyme engineering associated with heterologous gene expression can be overcome. First, CFPS does not require the transformation of cells with circular DNA molecules. Therefore, transformation efficiency is not the bottleneck that defines enzyme variant library size. Instead, the throughput of the screening procedure becomes limiting. In addition to this, CFPS allows for the direct expression of linear DNA templates and therefore greatly reduces the bias and effort of variant library construction. Further, the open nature of the system enables the addition of factors without the need to cross a cellular membrane, thereby improving control over expression conditions and cofactor loading. In order to facilitate ultra-high throughput screening, I employ a droplet microfluidic device capable of sorting water in oil emulsion droplets at a rate of 1,000 droplets per second. Single DNA molecules are encapsulated into picoliter-sized droplets alongside a CFPS reaction mix and assay reagents. In order to produce a measurable result, each DNA molecule is amplified within the droplet and expressed in concert with a biochemical assay reaction. Droplets that display high signal intensities are sorted and analyzed to inform further enzyme engineering efforts. This workflow allows for the exploration of vast sequence-function spaces only accessible through cell free protein synthesis.