Reverse engineering of industrially relevant phenotypes in yeast: An integrated approach

Reverse engineering is the study of discovering the structure, function and operation of a device or system with the express aim to reconstruct its key functionalities. This principle is applied to many disciplines, from military, through computer engineering, to health, but also in metabolic engineering. In this context, reverse metabolic engineering examines a particular functionality or phenotype of a cell or culture and subsequently aims to reconstruct it, with the aid of targeted genetic modification, in another cell or culture. Even with increasing knowledge on targeted metabolic engineering, microbial production platforms for fuels and chemicals are often obtained by non-targeted approaches, such as mutagenesis or evolutionary engineering. Reverse engineering of the interesting traits of these microbial platforms not only provides the potential to implement and combine them in other hosts, but also allows for the protection of the resulting intellectual property. The major challenge in reverse metabolic engineering is the elucidation of the molecular mechanisms underlying the phenotype of the strains of interest. In this thesis, various techniques were evaluated for their application in reverse metabolic engineering of a diverse range of industrially relevant phenotypes in the yeast Saccharomyces cerevisiae. Simultaneously, the different analytical methods that were used in these studies were evaluated for their individual and combined contributions