Protein engineering synthetic applications

Since most synthetic applications require enzymes catalyzing nonnatural substrates, their properties often have to be improved. One way to achieve this is to optimize reaction conditions such as pH, temperature, solvents, additives, etc. [6-9]. Another way is to modulate the substrates without compromising the synthetic efficiency of the overall reaction [10]. In most cases for commercial manufacturing, however, the protein sequences have to be altered to enhance reactivity, stereoselectivity and stability. It was estimated that over 30 commercial enzymes worldwide have been engineered for industrial applications [11]. Precise prediction of which amino acids to mutate is difficult to achieve. Since the mid 1990s, directed evolution... 

While CPO is highly attractive for synthetic applications [57], protein engineering has been hampered by the inability to express the fungal enzyme in bacteria or yeast. Hager and coworkers, however, have used gene replacement technology to allow functional expression and production of mutants in the enzyme s natural host, Caldario-mycesjumago [110]. In an effort to understand the importance of the proximal thiolate... 

Protein engineering technology has many potential of applications. Synthetic enzymes will have a significant impact on chemistry. Bioactive membranes and biodegradable polymers, which should be materials harmonizing with the Earth, are also expected. In the area of... 

Although the formate/FDH system is, in principle, the most attractive for large-scale applications, the co-product carbon dioxide being very easily removed from the reaction mixture, the catalytic and operational features of native FDH enzymes are far from being optimal. In fact, they are usually characterized by a very low specific activity, limited chemical and thermal stability, and strict preference for the NADH cofactor. As these facts hamper the wide application of FDHs in the development of novel industrial synthetic processes, a huge effort has been carried out in the last years to improve the performance of this biocatalyst, mainly by protein engineering [4]. 

The synthetic applications of the biocatalytic reduction of C=0 double bonds are also described in detail in Chapter 4, while Chapter 6 describes the use of protein engineering to develop novel enzymes for the improved reduction of C=0 double bonds. 

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