Protein Engineering Challenges

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). 

Moore GL, Maranas CD. Computational challenges in combinatorial library design for protein engineering. AlCHE J. 2004 50 262-272. 

With the abundant number (> 16 000) of three-dimensional structures in the Brookhaven Protein Data Bank, a challenging but promising task in protein engineering is the synthesis of novel biocatalysts by assembling individual functional modules (substrate binding sites, catalytic centers etc.), or by introducing a... 

Leal MCMR, Cammarota MC, Freire MG et al. (2002) Hydrolytic enzymes as coadjuvants in the anaerobic treatment of dairy wastewaters. Braz J Chem Eng 19(2) 175-180 Leisola M, Turunen O (2007) Protein engineering opportunities and challenges. Appl Microbiol Biotechnol 75(6) 1225-1232... 

Apart from the usually low activity and sometimes insufficient selectivity of P450s towards steroids (which can be improved by means of protein engineering), the low solubility of steroid compounds in water (1-100 pM [334]) represents a challenging problem for the establishment of whole-cell biocatalysis. Consequently, several promising reaction-engineering techruques that were applied for biotransformations of other hydrophobic compoimds have also been tested with steroid substrates. Among these are (1) biphasic reaction setups with an organic phase, which serves as substrate reservoir, (2) surfactant-... 

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