P450 in biotechnology: zinc driven ω-hydroxylation of p-nitrophenoxydodecanoic acid using P450 BM-3 F87A as a catalyst

Microbial P450 enzymes in biotechnology

“…In vivo, the electrons are supplied by the generation of NAD(P)H which is, however, far too expensive to be used in equimolar concentrations during technical applications. Direct electron supply from electrodes (Kazlauskaite et al 1996), use of electron mediators (Schwaneberg et al 2000), as well as enzymatic (Fernandez-Salguero et al 1993;Taylor et al 2000) and organometallic (Hollmann et al 2002) approaches have been suggested as sources of the required reduction equivalents.…”

Detection of 25-Hydroxyvitamin D3 with an Enzyme modified Electrode

“…Faulkner et al reported that Co(sep) 3+ can be used in a CYP4A1 system with rates comparable to those obtained with NADPH [26]. Another research group constructed a biosensor by immobilizing CYP3A4 and Co(sep) 3+ on a Nafion ® -modified electrode and reported relevant detection limits for the substrate of CYP3A4 [41]. Synthetic mediators like Co(sep) 3+ have multiple advantages such as facilitating quick, reversible electrochemistry, enhanced rates of reactions, and facile electrode design while maintaining flexibility in enzyme immobilization [43].…”

“…Cobalt sepulchrate trichloride (Co(sep) 3+ ), a non-native redox mediator, has been effectively utilized with other CYP450s [40][41][42]. Faulkner et al reported that Co(sep) 3+ can be used in a CYP4A1 system with rates comparable to those obtained with NADPH [26].…”

Unraveling Binding Effects of Cobalt(II) Sepulchrate with the Monooxygenase P450 BM-3 Heme Domain Using Molecular Dynamics Simulations

“…[20][21] Recently, fast advancements have been made toward cost effective catalysis in P450BM-3 by regeneration or substitution of expensive cofactor (NADPH/NADH) as a source of electrons. 13,17 In last decades, electrochemistry of P450BM-3 received considerable attention and various methods have investigated to drive catalytic cycle either by direct contact with electrodes [22][23] or using molecules as electron transfer (ET) mediators. In the latter case, small 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 4 soluble compounds, such as cobalt(III)sepulchrate (Co(III)Sep), shuttle electrons from electrodes or other inexpensive electron sources (e.g.…”

“…zinc dust) to enzyme redox site. 13,[24][25] However, little is known about the binding and ET mechanism of these mediators to the P450BM-3 at molecular level. To the best of our knowledge, only few experimental studies are devoted to identify the binding sites of ET mediators on the enzyme surface, which are relevant for ET mechanism in these systems.…”