Expression of human cytochrome P450 enzymes in yeast and bacteria and relevance to studies on catalytic specificity

Engineering yeast for high-level production of stilbenoid antioxidants

“…As C4H used in resveratrol pathway is a membrane-associated plant-derived P450 enzyme, which requires an electron carrier for optimal activity, we hypothesized that C4H activity may be limiting resveratrol production. Plant-derived P450s have previously been reported to poorly accept electrons directly from yeast endogenous electron carriers24. Thus, we upgraded the strain ST4978 (basic resveratrol pathway) with overexpression of cytochrome P450 reductase ( AtATR2 ) from A. thaliana or cytochrome B5 ( CYB5 ) from S. cerevisiae or both (Fig.…”

Engineering yeast for high-level production of stilbenoid antioxidants

“…As C4H used in resveratrol pathway is a membrane-associated plant-derived P450 enzyme, which requires an electron carrier for optimal activity, we hypothesized that C4H activity may be limiting resveratrol production. Plant-derived P450s have previously been reported to poorly accept electrons directly from yeast endogenous electron carriers24. Thus, we upgraded the strain ST4978 (basic resveratrol pathway) with overexpression of cytochrome P450 reductase ( AtATR2 ) from A. thaliana or cytochrome B5 ( CYB5 ) from S. cerevisiae or both (Fig.…”

CYP79F1 and CYP79F2 have distinct functions in the biosynthesis of aliphatic glucosinolates in Arabidopsis

“…In the E. coli expression system, this is, however, easily overcome by supplementing with eukaryotic NADPH cytochrome P450 reductases. The additional ability of the yeast system to perform post‐translational modifications of eukaryotic proteins is not a concern here as cytochromes P450 are generally not post‐translationally modified (Guengerich et al ., 1993). As a consequence thereof, catalytic specificity of cytochromes P450 have never been reported to be affected by post‐translational modification.…”

Substrates, Inducers, Inhibitors and Structure-Activity Relationships of Human Cytochrome P450 2C9 and Implications in Drug Development

“…Tolbutamide is less potent than second-generation sulphonylureas such as glipizide and glyburide and it is not routinely used due to a higher incidence of adverse effects compared to newer second-generation sulfonylureas such as glyburide [1014]. Tolbutamide is mainly metabolized via methylhydroxylation by CYP2C9 to 4-OH-tolbutamide [968][969][970]1015] and thus is often used as a probe substrate for determining CYP2C9 activity (Fig. (123)).…”

Engineering yeast for high-level production of stilbenoid antioxidants

“…As C4H used in resveratrol pathway is a membrane-associated plant-derived P450 enzyme, which requires an electron carrier for optimal activity, we hypothesized that C4H activity may be limiting resveratrol production. Plant-derived P450s have previously been reported to poorly accept electrons directly from yeast endogenous electron carriers24. Thus, we upgraded the strain ST4978 (basic resveratrol pathway) with overexpression of cytochrome P450 reductase ( AtATR2 ) from A. thaliana or cytochrome B5 ( CYB5 ) from S. cerevisiae or both (Fig.…”

CYP79F1 and CYP79F2 have distinct functions in the biosynthesis of aliphatic glucosinolates in Arabidopsis

“…In the E. coli expression system, this is, however, easily overcome by supplementing with eukaryotic NADPH cytochrome P450 reductases. The additional ability of the yeast system to perform post‐translational modifications of eukaryotic proteins is not a concern here as cytochromes P450 are generally not post‐translationally modified (Guengerich et al ., 1993). As a consequence thereof, catalytic specificity of cytochromes P450 have never been reported to be affected by post‐translational modification.…”

Substrates, Inducers, Inhibitors and Structure-Activity Relationships of Human Cytochrome P450 2C9 and Implications in Drug Development

“…Tolbutamide is less potent than second-generation sulphonylureas such as glipizide and glyburide and it is not routinely used due to a higher incidence of adverse effects compared to newer second-generation sulfonylureas such as glyburide [1014]. Tolbutamide is mainly metabolized via methylhydroxylation by CYP2C9 to 4-OH-tolbutamide [968][969][970]1015] and thus is often used as a probe substrate for determining CYP2C9 activity (Fig. (123)).…”

Engineering yeast for high-level production of stilbenoid antioxidants

“…As C4H used in resveratrol pathway is a membrane-associated plant-derived P450 enzyme, which requires an electron carrier for optimal activity, we hypothesized that C4H activity may be limiting resveratrol production. Plant-derived P450s have previously been reported to poorly accept electrons directly from yeast endogenous electron carriers24. Thus, we upgraded the strain ST4978 (basic resveratrol pathway) with overexpression of cytochrome P450 reductase ( AtATR2 ) from A. thaliana or cytochrome B5 ( CYB5 ) from S. cerevisiae or both (Fig.…”

CYP79F1 and CYP79F2 have distinct functions in the biosynthesis of aliphatic glucosinolates in Arabidopsis

“…In the E. coli expression system, this is, however, easily overcome by supplementing with eukaryotic NADPH cytochrome P450 reductases. The additional ability of the yeast system to perform post‐translational modifications of eukaryotic proteins is not a concern here as cytochromes P450 are generally not post‐translationally modified (Guengerich et al ., 1993). As a consequence thereof, catalytic specificity of cytochromes P450 have never been reported to be affected by post‐translational modification.…”

Substrates, Inducers, Inhibitors and Structure-Activity Relationships of Human Cytochrome P450 2C9 and Implications in Drug Development

“…Tolbutamide is less potent than second-generation sulphonylureas such as glipizide and glyburide and it is not routinely used due to a higher incidence of adverse effects compared to newer second-generation sulfonylureas such as glyburide [1014]. Tolbutamide is mainly metabolized via methylhydroxylation by CYP2C9 to 4-OH-tolbutamide [968][969][970]1015] and thus is often used as a probe substrate for determining CYP2C9 activity (Fig. (123)).…”