The catalytic activity of cytochrome P450cam towards styrene oxidation is increased by site-specific mutagenesis. by Nickerson, D, Harford-Cross, C, Fulcher, SR, Wong, L

Sesquiterpene oxidation by evolved CYP102A1 (P450BM3) monooxygenase by Cao, Y

“…Site-directed, site-saturation and random mutagenesis have been applied to create CYP102A1 variants to expand the substrate range and explore new reaction types. …”

P450(BM3) (CYP102A1): connecting the dots. by Whitehouse, C, Bell, S, Wong, L

“…Crystal structures have provided a basis for several of the catalytic effects brought about by mutagenesis, while changes to reduction potentials, inter-domain electron transfer rates and catalytic parameters have yielded functional insights. …”

The dimerization of Pseudomonas putida cytochrome P450cam: practical consequences and engineering of a monomeric enzyme. by Nickerson, D, Wong, L

“…To eliminate any ambiguity introduced by dimer formation and to simplify handling and storage of the enzyme, site-directed mutagenesis was used to identify C334 as the single cysteine residue responsible for the formation of the disulfide linkage and to engineer a monomeric enzyme by substituting an alanine in its place. …”

The crystal structures of 4-methoxybenzoate bound CYP199A2 and CYP199A4: structural changes on substrate binding and the identification of an anion binding site. by Bell, S, Yang, W, Tan, AB, Zhou, R, Johnson, E, Zhang, A, Zhou, W, Rao, Z, Wong, L

“…The methoxy group is located over the heme iron, accounting for the high activity and selectivity of these enzymes for oxidative demethylation of the substrate. Mutagenesis studies on CYP199A4 highlight the involvement of hydrophobic (Phe185) and hydrophilic (Arg92, Ser95 and Arg243) amino acid residues in the binding of para-substituted benzoates by these enzymes.…”

Mutations of glutamate-84 at the putative potassium-binding site affect camphor binding and oxidation by cytochrome p450cam. by Westlake, A, Harford-Cross, C, Donovan, J, Wong, L

“…This behaviour has been investigated by mutagenesis of Glu84, a surface residue which forms part of the cation-binding site. …”

Catalytic reductive dehalogenation of hexachloroethane by molecular variants of cytochrome P450cam (CYP101). by Walsh, M, Kyritsis, P, Eady, N, Hill, H, Wong, L

“…The hexachloroethane dehalogenation activity of CYP101 has been investigated by mutagenesis. The effects of active-site polarity and volume were probed by combinations of active-site mutations. …”

Enantioselective remote functionalisation of cyclic amines by engineered P450BM3 variants by Zhang, Y

“…In Chapter 3, the iterative docking-guided mutagenesis (IDGM) approach is explored. Comparative analysis of the correct binding poses and unwanted poses leads to targeted mutation design to disfavour unwanted poses and increase both the activity and product selectivity of amine oxidation. …”

Cyclic amine functionalisation by engineered cytochrome P450BM3 by Li, Y

“…A combination of substrate engineering via introducing substituents (methoxy and methyl), N-protecting groups (phenyl, benzyl, acyl, Boc, mesyl, tosyl, and isopropylsulfonyl) and auxiliary carbocycles (cyclopentane and cycloheptane), and protein engineering by site-directed and site-saturation mutagenesis at active site residues identified by screening with a 48 P450_BM3 variant library of diverse substrate pocket topology and polarity, have enabled the oxidative diversification of these core motifs in numerous FDA-approved drugs. …”

Engineering terpenoid oxidation by cytochrome P450BM3 for fine chemical synthesis by Chen, W

“…To design altered steroid binding orientations to broaden the regioselectivity, the structures of wild type P450BM3 and the steroid aromatase CYP19A1 are overlaid to identify key regions for scanning glycine mutagenesis which greatly expanded the product range of AD, DHEA and TST. …”

Protein-protein recognition in biological systems exhibiting highly-conserved tertiary structure: cytochrome P450 by Johnson, E, Eachan Oliver Daniel Johnson

“…</p> <p>The putative iron-sulfur biosynthesis ferredoxin PuxB had been engineered by rational mutagenesis to support catalysis by CYP199A2. It was confirmed this arose from improved protein-protein recognition. …”