Table 2.

CYPs involved in secondary metabolite biosynthesis in medicinal plants, and their application in metabolic engineering in heterologous system

CYP	Organism	Enzyme/reaction	Engineering method	Objective/modulations after engineering	References
CYP88D	Glycyrrhiza uralensis	β-amyrin 11-oxidase	Codon optimisation of CYP88D6 and CYP72A154 with co-expression of β-amyrin synthase encoding gene and AtNADPH + GuCYB5 expression in S. cerevisiae	Glycyrrhizin accumulation	Seki et al. (2008), Wang et al. (2019)
CYP71AV1	Artemisia annua	Amorpha-4,11-diene C-12 oxidase	Expressed in conjunction with CPR and N -terminus modification, high copy number plasmid and a strong inducible promoter in S. cerevisiae	Anti-malaria-drug precursor artemisinic acid, artemisinic alcohol, artemisinic aldehyde and dihydroartemisinic aldehyde production	Chen et al. (2017), Teoh et al. (2006)
CYP76B1	Helianthus tuberosus	7-ethoxycoumarin O-de-ethylase involved in double N-dealkylation of phenylurea	Ectopic constitutive expression in Nicotiana tabacum and Arabidopsis	Increase in tolerance for several phenylurea herbicides	Didierjean et al. (2002)
CYP87D20	Cucurbitaceae family	C11 carbonylase and C20 hydroxylase	Structural and data-driven approach using Rosetta software and GREMLIN	Creating a de novo pathway to produce mogrol	Li et al. (2019a)
CYP706B1	Gossypium arboreum	Cadinene-8-hydroxylase	N-terminal modification in E. coli	8-Hydroxycadinene production	Chang et al. (2007)
CYP75A	Camellia sinensis	Flavonoid 3,5-hydroxylase	Gene overexpression in conjunction with CPR in tobacco	Accumulation of catechin	Wang et al. (2014)
CYP73A1	Helianthus tuberosus	Trans-cinnamate 4-hydoxylase	N-terminus modification in S. cerevisiae	Trans-cinnamic acid hydroxylation product	Schoch, et al. (2003b)
P450 isoflavone synthase 1	Glycine max	Isoflavone synthase	Electron flux optimisation by fusion of CPR in E. coli	Genistein accumulation	Leonard & Koffas (2007)
CYP82D1.1	Scutellaria baicalensis	Flavone hydroxylases	N-terminal modification and truncation in E. coli	Improved yield of baicalein and scutellarein	Li et al. (2019b)
CYP716a47	Panax ginseng	Encodes protopanaxadiol synthase	Fusion with AtCPR in S. cerevisiae	Increase in protopanaxadiol production	Zhao et al. (2016)
CYP76AH15	Coleus forskohlii	hydroformylation (converts 13R‑manoyl oxide to 11‑oxo‑13R‑manoyl oxide)	SRSs (Substrate Recognition Sites) engineering in S. cerevisiae	Increased production of forskolin	Forman et al. (2018)
CYP76AH1	Salvia miltiorrhiza	Ferruginol synthase	In-vivo optimization of redox partners in S. cerevisiae	Ferruginol production	Guo et al. (2013)
CYP725A4	Taxus cuspidata	Taxadiene-5α-Hydroxylase	Optimizing expression of CYP, interaction with reduction partner and N-terminal modificaiton in E. coli	Increased oxygenated taxanes production	Biggs et al. (2016)
AmI2′H/CYP81E42	Astragalus membranaceus	Isoflavone 2′-hydroxylase	N-terminal and ORF modification in E. coli	Medicarpin malonyl glucoside accumulation	Chen et al. (2015)
CYP82Y2-like	Papaver bracteatum	1,2-dehydroreticuline synthase	Gene mining, protein mutagenesis, codon optimization, and heterologous expression in yeast	Bioconversion of (S)-reticuline to (R)-reticuline for morphine biosynthesis	Farrow et al. (2015), Galanie et al. (2015)
P450 SalSyn	Papaver somniferum	Salutaridine synthase	Co-expression of heterologous proteins and deletion of one host gene in yeast	Production of thebaine/ hydrocodone	Galanie et al. (2015)
CYP716a155	Rosmarinus officinalis	BA-specific lupeol C-28 oxidase	Cognate expression of CYP/CPR in S. cerevisiae	A potent anti-cancer agent, betulinic acid accumulation	An et al. (2020), Huang et al. (2019)