Flavonoid hydroxylase from Catharanthus roseus: cDNA, heterologous expression, enzyme properties and cell-type specific expression in plants - PubMed (original) (raw)

Flavonoid hydroxylase from Catharanthus roseus: cDNA, heterologous expression, enzyme properties and cell-type specific expression in plants

M Kaltenbach et al. Plant J. 1999 Jul.

Free article

Abstract

We investigated the P450 dependent flavonoid hydroxylase from the ornamental plant Catharanthus roseus. cDNAs were obtained by heterologous screening with the CYP75 Hf1 cDNA from Petunia hybrida. The C. roseus protein shared 68-78% identity with other CYP75s, and genomic blots suggested one or two genes. The protein was expressed in Escherichia coli as translational fusion with the P450 reductase from C. roseus. Enzyme assays showed that it was a flavonoid 3', 5'-hydroxylase, but 3'-hydroxylated products were also detected. The substrate specificity was investigated with the C. roseus enzyme and a fusion protein of the Petunia hybrida CYP75 with the C. roseus P450 reductase. Both enzymes accepted flavanones as well as flavones, dihydroflavonols and flavonols, and both performed 3'- as well as 3'5'-hydroxylation. Kinetics with C. roseus cultures on the level of enzyme activity, protein and RNA showed that the F3'5'H was present in dark-grown cells and was induced by irradiation. The same results were obtained for cinnamic acid 4-hydroxylase and flavanone 3beta-hydroxylase. In contrast, CHS expression was strictly dependent on light, although CHS is necessary in the synthesis of the F3'5'H substrates. Immunohistochemical localization of F3'5'H had not been performed before. A comparison of CHS and F3'5'H in cotyledons and flower buds from C. roseus identified CHS expression preferentially in the epidermis, while F3'5'H was only detected in the phloem. The cell-type specific expression suggests that intercellular transport may play an important role in the compartmentation of the pathways to the different flavonoids.

PubMed Disclaimer

Cited by

Temperature desynchronizes sugar and organic acid metabolism in ripening grapevine fruits and remodels their transcriptome.
Rienth M, Torregrosa L, Sarah G, Ardisson M, Brillouet JM, Romieu C. Rienth M, et al. BMC Plant Biol. 2016 Jul 20;16(1):164. doi: 10.1186/s12870-016-0850-0. BMC Plant Biol. 2016. PMID: 27439426 Free PMC article.
The soybean F3'H protein is localized to the tonoplast in the seed coat hilum.
Toda K, Kuroiwa H, Senthil K, Shimada N, Aoki T, Ayabe S, Shimada S, Sakuta M, Miyazaki Y, Takahashi R. Toda K, et al. Planta. 2012 Jul;236(1):79-89. doi: 10.1007/s00425-012-1590-5. Epub 2012 Jan 19. Planta. 2012. PMID: 22258749
Transporters in plant sulfur metabolism.
Gigolashvili T, Kopriva S. Gigolashvili T, et al. Front Plant Sci. 2014 Sep 9;5:442. doi: 10.3389/fpls.2014.00442. eCollection 2014. Front Plant Sci. 2014. PMID: 25250037 Free PMC article. Review.
Hydroxylation of diverse flavonoids by CYP450 BM3 variants: biosynthesis of eriodictyol from naringenin in whole cells and its biological activities.
Chu LL, Pandey RP, Jung N, Jung HJ, Kim EH, Sohng JK. Chu LL, et al. Microb Cell Fact. 2016 Aug 5;15(1):135. doi: 10.1186/s12934-016-0533-4. Microb Cell Fact. 2016. PMID: 27495155 Free PMC article.
Heterologous expression of the flavonoid 3',5'-hydroxylase gene of Vinca major alters flower color in transgenic Petunia hybrida.
Mori S, Kobayashi H, Hoshi Y, Kondo M, Nakano M. Mori S, et al. Plant Cell Rep. 2004 Jan;22(6):415-21. doi: 10.1007/s00299-003-0709-3. Epub 2003 Sep 18. Plant Cell Rep. 2004. PMID: 14504908

Flavonoid hydroxylase from Catharanthus roseus: cDNA, heterologous expression, enzyme properties and cell-type specific expression in plants - PubMed (original) (raw)