Differential expression and functional characterization of the NADPH cytochrome P450 reductase genes from Nothapodytes foetida

Three unique NADPH:cytochrome P450 reductase (CPR) cDNAs have been isolated from a Nothapodytes foetida cDNA library and characterized. Phylogenetic analysis showed that NfCPR1 is a class I isoform, whereas NfCPR2 and NfCPR3 are class II isoforms. Both NfCPR1 and NfCPR2 transcripts were detected in all examined organs of N. foetida, with the highest level for NfCPR1 being in the seeds whereas for NfCPR2 predominantly in leaves. In contrast, NfCPR3 transcripts were only detected in flower buds and seeds at almost equal expression levels. Moreover, NfCPR1 expression did not change during wounding treatment, whereas NfCPR2 and NfCPR3 were induced in response to wounding. Microsomes isolated from insect cells co-expressing NfCPR2 and cytochrome P450 enzyme geraniol 10-hydroxylase (G10H) enhanced the production of eriodictyol from naringenin approximately 11-fold relative to control G10H-only insect cells, indicating the supportive role of NfCPR2 for G10H activity in insect cells.

► We isolated three unique NADPH:cytochrome P450 reductase (CPR) cDNAs from Nothapodytes foetida.
► Phylogenetic analysis showed that NfCPR1 is a class I isoform, whereas NfCPR2 and NfCPR3 are class II isoforms.
► NfCPR2 and NfCPR3 were induced in response to wounding.
► NfCPR2 and geraniol 10-hydroxylase were co-expressed in insect cells to examine its ability to support cytochrome P450 hydroxylase activity in vitro.