publication . Doctoral thesis . 2012

Advances in directed monooxygenase evolution : from diversity generation and flow cytometry screening to tailor-made monooxygenases

Ruff, Anna Joëlle;
Open Access English
  • Published: 01 Jan 2012
  • Publisher: Publikationsserver der RWTH Aachen University
  • Country: Germany
Directed Evolution became a powerful tool for proteins engineers to generate tailor-made biocatalyst. Directed protein evolution consist of the following three consecutive main steps, which are performed in iterative cycles; Step 1 the gene diversity generation, Step 2 the screening for improved variants and Step 3 the isolation of gene encoding for improved proteins. In this thesis, methodological advancements in the two key steps of the directed evolution, the diversity generation (SeSaM method) and high throughput screening (flow cytometer based screening technology) were performed and complemented by the evolution of P450 BM3 for alternative cofactor systems...
free text keywords: Ortspezifische Mutagenese, Gerichtete Evolution, Mutagenese, Durchflusscytometrie, Fluoreszenzaktivierter Zellsortierer, Proteindesign, Cytochrom P-450, Monooxygenasen, Biowissenschaften, Biologie, In-vitro-Evolution, gelenkte Evolution, P450 BM3, Vielfaltsgenerierung, Mutanten-Bibliotheken, directed evolution, random mutagenesis, flow cytometrie, ddc:570
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