Oxidoreductases represent the foundation of white biotechnology industry, which is forecasted to be the major driver for the global growth of chemical industry. However, development of new biotechnology based products and processes requires a substantial expansion of oxidoreductase-mediated transformations through discovery or generation of novel activities as well as expansion of substrate scope and optimization of selectivities for existing biocatalysts. The interdisciplinary project aims to identify novel oxidoreductases for industrial relevant transformations. We will use a dual focus strategy to identify new reactivities and selectivities: i) novel oxidoreductases of the Red Sea Brine Pool proteome database and from electroactive microbes will be expressed and tested and ii) unnatural amino acid incorporation methods, which were recently established at KAUST will be used to create de novo activities. Following successful expression, enzymes will be analyzed in parallel systems developed at TUM. Candidate enzymes and mutants showing novel features of interest will be structurally characterized. Based on these protein X-Ray structures or homology models, promising mutation positions will be identified. A combination of iterative saturation mutagenesis (ISM), selectivity/activity screening and mutant characterization will be used to generate new generations of optimized mutant enzymes. Such a computationally guided evolution strategy will significantly reduce the numbers of test required to identify highly selective mutants. The project team combines the complimentary expertise of four research groups at KAUST (biocatalysis and biocomputation) and TUM (structural biology and biochemical engineering).
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