In the first part of this thesis, a system for incorporation of non-canonical amino acids into proteins in S. cerevisiae by suppression of in-frame amber stop codons was reconstructed and analyzed critically. The used E. coli tyrosyl-tRNA synthetase (TyrRS) mutants were analyzed comprehensively with respect to their activation of Tyr and the analogs using the ATP-PPi exchange assay. In the second part, the industrial relevant lipase from T. thermohydrosulfuricus was expressed in E. coli and used as a model protein for studies on genetic code engineering. Thereby, all phenylalanine, proline and tyrosine residues were substituted globally by corresponding non-canonical amino acid analogs using the supplementation based incorporation method. Some of the resulting lipase congeners show new and in parts significantly improved features in their enzymatic activity.      «

In the first part of this thesis, a system for incorporation of non-canonical amino acids into proteins in S. cerevisiae by suppression of in-frame amber stop codons was reconstructed and analyzed critically. The used E. coli tyrosyl-tRNA synthetase (TyrRS) mutants were analyzed comprehensively with respect to their activation of Tyr and the analogs using the ATP-PPi exchange assay. In the second part, the industrial relevant lipase from T. thermohydrosulfuricus was expressed in E. coli and use...     »