Engineering cytochrome P450 biocatalysts for biotechnology, medicine and bioremediation

Abstract

Importance of the field: Cytochrome P450 enzymes comprise a superfamily of heme monooxygenases that are of considerable interest for the: 1) synthesis of novel drugs and drug metabolites, 2) targeted cancer gene therapy, 3) biosensor design, and 4) bioremediation. However, their applications are limited because cytochrome P450, especially mammalian P450 enzymes, show a low turnover rate and stability, and require a complex source of electrons through cytochrome P450 reductase and NADPH. Areas covered in this review: In this review, we discuss the recent progress towards the use of P450 enzymes in a variety of above-mentioned applications. We also present alternate and cost-effective ways to perform P450-mediated reaction, especially using peroxides. Furthermore, we expand upon the current progress in P450 engineering approaches describing several recent examples that are utilized to enhance heterologous expression, stability, catalytic efficiency, and utilization of alternate oxidants. What the reader will gain: The review will provide a comprehensive knowledge in the design of P450 biocatalysts for potentially practical purposes. Finally, we provide a prospective on the future aspects of P450 engineering and its applications in biotechnology, medicine, and bioremediation. Take home message: Because of its wide applications, academic and pharmaceutical researchers, environmental scientists, and health care providers are expected to gain current knowledge and future prospects of the practical use of P450 biocatalysts.

Use of Chemical Auxiliaries to Control P450 Enzymes for Predictable Oxidations at Unactivated C-H Bonds of Substrates

“…As a result, increased uncoupling rates are often observed due to uncontrolled water access to the active site [21]. One strategy to overcome these issues involves protein engineering, which has been used to improve enzymatic activity, change substrate specificity and alter product distributions [22,23]. As reviewed below, other research groups have attempted to control P450 selectivity and improve product predictability by using small molecules.…”

Protein Engineering Methods and Applications

“…Food industry applications (James & Simpson, 1996), (Kirk et al, 2002), (Akoh et al, 2008) Detergent industry applications (proteases) (Gupta et al, 2002) Environmental applications (Wiseman, 1993), (Cirino & Arnold, 2002), (Le Borgne & Quintero, 2003), (Ayala et al, 2008), (Cao et al, 2009) Medical applications (Buckel, 1996), (Filpula & McGuire, 1999), (Paques & Duchateau, 2007), (Nuttall & Walsh, 2008), (Liu et al, 2009), (Lam et al, 2003), (Zafir-Lavie et al, 2007), (Vazquez et al, 2009), (Olafsen & Wu, 2010) Biopolymer production applications (Chow et al, 2008), (Rehm, 2010), (Banta et al, 2010) Nanobiotechnology applications (Hamada et al, 2004), (Sarikaya et al, 2003) Applications with redox proteins and enzymes (Saab-Rincon & Valderrama, 2009), (Kumar, 2010) Applications with various industrially important enzymes (Martinkova & Kren, 2010), (Clapes et al, 2010), (Jordan & Wagschal, 2010), (Rao et al, 2009), (Marcaida et al, 2010).…”

Detection of 25-Hydroxyvitamin D3 with an Enzyme modified Electrode

“…These enzymes are capable of metabolizing a diverse range of chemicals using different biotransformation reactions and are associated with biosynthesis of steroids, vitamins and lipids, xenobiotic and drug metabolism [1][2][3][4][5][6][7][8]. Thus, there has been immense biotechnological interest in these enzymes, and they have been utilized for applications in bioelectronic devices, biochips, bioreactors, and biosensor technologies [9][10][11][12].…”