P450 biotechnology

Abstract

Cytochrome P450s accept a broad spectrum of substrates and catalyze a vast variety of chemical transformations under mild reaction conditions. Therefore, P450s are considered as attractive candidates for the synthesis of valuable compounds. Moreover, these enzymes have a great potential for the development of biosensors, as well as in bioremediation. However, the use of P450s in industrial processes is a challenging task because of their multicomponent nature, relatively low activity, and the need for the reducing cofactors nicotinamide adenine dinucleotide phosphate (NAD(P)H). Over the last two decades, the fundamental understanding of P450 systems has greatly improved and tremendous progress has been made towards industrial application of these complex systems. In this chapter, we highlight several basic aspects and recent advances in P450 biotechnology, including the exploitation of the natural diversity of P450s, their engineering to achieve higher activity, enhanced stability, and altered selectivity. Considerable progress has also been made in terms of electron flow optimization between P450s and their redox partners and implementation of cofactor regeneration, thus overcoming the need for stoichiometric addition of NAD(P)H. Finally, several examples of established P450-based whole-cell biocatalysts are discussed, with special focus on microbial de novo synthesis of plant secondary metabolites and the generation of transgenic plants.