Industrial enzymes protein engineering

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. 

Decolorization of azo dyes by WRF technology improvements will require integration of all major areas of industrial biotechnology novel enzymes and microorganisms, functional genomics, protein engineering, biomaterial development, bioprocess design and applications. 

MW 27,500) with no cofactors or metal ions reqnirement for its function, it displays Michaelis-Menten kinetics and it is secreted in large amounts by a wide variety of Bacillus species. Subtilisin is also among the most important industrial enzymes due to its use in laundry detergents. Protein engineering strategies for subtilisin have focused on a number of aspects, namely catalysis, substrate specificity, thermal and oxidative stability and pH profile. We will describe briefly each of these aspects. 

E. coli - [GASTROINTESTINAL AGENTS] (Vol 12) - [GENETICENGINEERING - PROCEDURES] (Vol 12) -industrial enzymes from [ENZYTffi APPLICATIONS - INDUSTRIAL] (Vol 9) -for protein engineering [PROTEIN ENGINEERING] (Vol 20)... 

PROTEIN ENGINEERING] (Vol 20) for new enzymes [ENZYME APPLICATIONS - INDUSTRIAL] (Vol 9)... 

Further advantages of biocatalysis over chemical catalysis include shorter synthesis routes and milder reaction conditions. Enzymatic reactions are not confined to in vivo systems - many enzymes are also available as isolated compounds which catalyze reactions in water and even in organic solvents [28]. Despite these advantages, the activity and stability of most wild-type enzymes do not meet the demands of industrial processes. Fortunately, modern protein engineering methods can be used to change enzyme properties and optimize desired characteristics. In Chapter 5 we will outline these optimization methods, including site-directed mutagenesis and directed evolution. 

In spite of their catalytic versatility and their capacity to transform a variety of pollutant compounds, peroxidases are not applied at large scale yet. The challenges that should be solved to use peroxidases for environmental purposes have been recently reviewed [146], Three main protein engineering challenges have been identified (a) the enhancement of operational stability, specifically hydrogen peroxide stability (see Chap. 11) (b) the increase of the enzyme redox potential in order to widen the substrate range (see Chap. 4) (c) the development of heterologous expression and industrial production (see Chap. 12). 

The time is ripe for the widespread application of biocatalysis in industrial organic synthesis and according to a recent estimate [113] more than 130 processes have been commercialised. Advances in recombinant DNA techniques have made it, in principle, possible to produce virtually any enzyme for a commercially acceptable price. Advances in protein engineering have made it possible, using techniques such as site directed mutagenesis and in vitro evolution, to manipulate enzymes such that they exhibit the desired substrate specificity, activity, stability, pH profile, etc. [114]. Furthermore, the development of effective immobilisation techniques has paved the way for optimising the performance and recovery and recycling of enzymes. 

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