Table 2.

Protein engineering of P450s.

P450	Gene ID	Source	Substrates	Products	Key Residues and Mutants	Mechanism of action	Results	References
CYP72A63	11410378	Medicago truncatula	11-oxo-β-amyrin	glycyrrhetol	L509I	Increases the hydrophobicity of the active pocket ends	Yield is 90.6%	[9]
CYP72A63	11410378	Medicago truncatula	glycyrrhetol	glycyrrhetinic acid (GA)	T338S	Improve the hydrophilicity of the binding pocket	GA increased by 7.1-fold	[9]
CYP72A63	11410378	Medicago truncatula	11-oxo-β-amyrin	29-OH-11-oxo-β-amyrin	L398I	Rotate the substrate 180° to reverse the relative positions of C-30 and C-29	100% regional selectivity	[9]
CYP72A63	11410378	Medicago truncatula	11-oxo-β-amyrin	GA	T338S/W205A	Eliminate limitations on proton transfer	36.4 ± 3.0 mg/L	[9]
CYP72A154	371940465	Glycyrrhiza uralensis	β-amyrin	30-OH-β-amyrin	L149/L398	L149 and L398 determined the substrate orientation, which controlled its product regioselectivity	Further enhance the production of high-value triterpenoid compounds	[4]
CYP72A62v2	11409246	Medicago truncatula	β-amyrin	30-OH-β-amyrin	V398L	L398 determined the substrate orientation, which controlled its product regioselectivity	Changed from being regioselective at C-29 to producing only C-30 oxidation products	[4]
CYP87D20	101213710	cucumber	cucurbitadienol	11-H- cucurbitadienol	E286A	E286A plays a role in shuttling protons into the active site	Decreases the production of 11C-Cuol and 11C-20H-Cuol	[45]
TwCYP712K1	2081630515	Tripterygium wilfordii	friedelin	polpunonic acid	S110F	Maked the substrate closer to the reaction centre	S110F increased the production of polpunonic acid	[46]