Protein engineering proteases

Many enzymes have been the subject of protein engineering studies, including several that are important in medicine and industry, eg, lysozyme, trypsin, and cytochrome P450. SubtiHsin, a bacterial serine protease used in detergents, foods, and the manufacture of leather goods, has been particularly well studied (68). This emphasis is in part owing to the wealth of stmctural and mechanistic information that is available for this enzyme. 

In 1989, two enzymes based on genetic engineering techniques were introduced, ie, a cloned alkaline protease (IBIS) and a protein engineered Subtihsin Novo (Genencor, California). Lipase and ceUulase types of detergent enzymes have also begun to appear. 

Pig. 7. Storage stabHity of proteases in European powder detergent with activated bleach system. ( ) Traditional protease (--) protein-engineered... 

Proteases are used in many industrial areas as well as basic research. They are classified by their mechanism of catalysis. Proteases are used in the pharmacological, food and other consumer industries to convert raw materials into a final product or to alter properties of the raw material. In biomedical research, proteases are used to study the structure of other proteins and for nthesis of peptides. The choice of a protease for an application depends in part on its specificity for peptide bonds, activity and stability. Technical advances in protein engineering have enabled alteration of these properties and allowed proteases to be used more effectively. Some easily obtained proteases can be modified so that they can substitute for proteases whose supply is limited. 

With subtihsin nature has provided us with a model system for protein engineering studies (Wells and Estell, 1988). Being a small single domain serine protease... 

Siezen, R.J., de Vos, W.M., Lennissen, J.A.M. and Dijkstra, B.W. (1991) Homology modelling and Protein Engineering strategy of subtilases, the family of subtilisinlike serine proteases. Protein Engineering, 4, 719-737. 

Aehle, W., Sobek, H., Amory, A., Vetter, R., Wilke, D. Schomburg,D. (1993). Rational protein engineering and industrial application structure prediction by homology and rational design of protein-variants with improved washing performance the alkaline protease from Bacillus alcalophilus. Journal of Biotechnology, 28, 31-40. 

Teplyakov, A. V., van der Laan, J. M., Lammers, A. A., Kelders, H., Kalk, K. H., Misset, O., Mulleners, L.J. Dijkstra, B. W. (1992). Protein engineering of the high-alkaline serine protease PB92 from Bacillus alcalophilus functional and structural consequences of mutation at the S4 substrate binding pocket. Protein Engineering, 5, 413-20. 

Protein engineering (continued) differential and uniform binding energy changes 438,439 dissection of catalytic triad of serine proteases 450-452 domain structure analysis... 

Subtilisin (see also serine proteases) 27, 28, 30, 450-454, 476 prosequence 540 protein engineering specificity of 452-454... 

Serine proteases - [EGGS] (Vol 8) -in coagulation [BLOOD, COAGULANTS AND ANTICOAGULANTS] (Vol 4) -protein engineering of [PROTEIN ENGINEERING] (Vol 20)... 

E. Liberation of Biologically Functional Peptides IV Genetic Engineering of Milk Proteins and Proteases... 

Protein engineering has allowed for the development of many new protease inhibitors with increased potency and specificity and for diverse mechanisms of action. Because of the relatively shallow active sites, homology, and broad specificity of many proteases, larger molecules are attractive inhibitors in that they can bnry more surface area during binding and hopefully gain more potency and specificity. 

Walker B, Bayley H (1994) A pore-forming protein with a protease-activated trigger. In Protein Engineering 7 91-97. 

---