Protein engineering stability improvement

The most recent stages in the history of detergent enzymes has been the fine-tuning of the molecular structure by protein engineering to improve the enzyme performance and stability and the use of gene technology to enhance the production yield. The production of detergent lipases and cellulases would, indeed, not have been possible with the classical methods. 

The search for and the development of a useful biocatalyst with a suitable yield, specific activity and stability is, in the beginning, the task of microbiologists, biochemists, molecular biologists, protein engineers, etc. However, especially with respect to stability, the process engineer also has means available, among others immobilization, to improve the stability of biocatalysts. 

H. Bisgard-Frantzen, S. Carlsen, and J. M. Mikkelsen, Protein engineering of subtilisins to improve stability in detergent formulations, J. Biotechnol. 1993, 28, 55-68. 

Under these conditions, it is possible to evaluate protein stability from denaturation experiments. This is not a subject of simply academic interest the increasing use of proteins in everyday use, for example as additives in washing powders, has made protein stability a matter of great practical importance, the more so since protein engineering has been used to improve stability properties. 

The use of an LeuDH as an amino acid dehydrogenase showed a high L-enan-tiospecificity [24]. In this connection, an L-leucine dehydrogenase from Bacillus sphaericus has been applied very efficiently. The FDH from Candida boidinii is the preferred formate dehydrogenase for this process. The stability of this enzyme, which is available in technical quantities, has been remarkably improved by protein engineering and directed evolution [25], In particular the replacement of cys-... 

Formulation strategies, such as addition of specific functional excipients, freezedrying, protein engineering, and delivery strategies, have been used to improve the stability and circulation half-life and reduce immunogenicity, which in turn improve... 

---