Literature summary for 1.14.11.9 extracted from

Tu, Y.; Liu, F.; Guo, D.; Fan, L.; Zhu, Z.; Xue, Y.; Gao, Y.; Guo, M.
Molecular characterization of flavanone 3-hydroxylase gene and flavonoid accumulation in two chemotyped safflower lines in response to methyl jasmonate stimulation (2016), BMC Plant Biol., 16, 132 .

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Cloned(Commentary)

Cloned (Comment)	Organism
gene CtF3H, DNA and amino acid sequence determination and analysis, phylogenic analysis, quantitative real-time PCR enzyme expression analysis, functional recombinant expression of MBP-tagged enzyme in Escherichia coli BL21(DE3)pLysS from vector pMAL-C5x leading to production of dihydrokaempferol when naringenin is available the substrate. Recombinant expression of GFP-tagged enzyme in onion cells via transfromation of Agrobacterium tumefaciens strain GV3101, where it is localized both in the nucleus and in the cytosol	Carthamus tinctorius

Cloned (Comment)

Organism

gene CtF3H, DNA and amino acid sequence determination and analysis, phylogenic analysis, quantitative real-time PCR enzyme expression analysis, functional recombinant expression of MBP-tagged enzyme in Escherichia coli BL21(DE3)pLysS from vector pMAL-C5x leading to production of dihydrokaempferol when naringenin is available the substrate. Recombinant expression of GFP-tagged enzyme in onion cells via transfromation of Agrobacterium tumefaciens strain GV3101, where it is localized both in the nucleus and in the cytosol

Carthamus tinctorius

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
cytosol	-	Carthamus tinctorius	5829	-
additional information	subcellular localization study	Carthamus tinctorius	-	-
nucleus	-	Carthamus tinctorius	5634	-

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Fe2+	dependent on, required for catalysis, the ferrous iron binding site HxDxnH is formed by His78, His121, His211, Asp213, His221, Asp 223, His265, His267, and His279	Carthamus tinctorius

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
a (2S)-flavan-4-one + 2-oxoglutarate + O2	Carthamus tinctorius	-	a (2R,3R)-dihydroflavonol + succinate + CO2	-	?

Organism

Organism	UniProt	Comment	Textmining
Carthamus tinctorius	M9MTF3	a quinochalcone-type safflower line with orange-yellow flowers and a flavonol-type safflower line with white flowers, i.e. ZHH0119 line and XHH007 line, respectively	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant MBP-tagged enzyme from Escherichia coli BL21(DE3)pLysS by amylose affinity chromatography	Carthamus tinctorius

Source Tissue

Source Tissue	Comment	Organism	Textmining
flower	the transcriptome expression of CtF3H shows a diametrically opposed expression pattern in a quinochalcone-type safflower line (with orange-yellow flowers) and a flavonol-type safflower line (with white flowers) under external stimulation by methyl jasmonate (MeJA), which has been identified as an elicitor of flavonoid metabolites. High expression of CtF3H in quinochalcone-type safflower line	Carthamus tinctorius	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
a (2S)-flavan-4-one + 2-oxoglutarate + O2	-	Carthamus tinctorius	a (2R,3R)-dihydroflavonol + succinate + CO2	-	?
a (2S)-flavan-4-one + 2-oxoglutarate + O2	the 2-oxoglutarate binding site RxS is formed by Arg289 and Ser291	Carthamus tinctorius	a (2R,3R)-dihydroflavonol + succinate + CO2	-	?
additional information	no activity with kaempferol, dihydrokaempferol, quercetin, and dihydroquercetin	Carthamus tinctorius	?	-	?
naringenin + 2-oxoglutarate + O2	-	Carthamus tinctorius	dihydrokaempferol + succinate + CO2	-	?

Subunits

Subunits	Comment	Organism
?	x * 40720, sequence calculation, x * 40000, recombinant detagged enzyme, SDS-PAGE	Carthamus tinctorius

Synonyms

Synonyms	Comment	Organism
CtF3H	-	Carthamus tinctorius
F3H	-	Carthamus tinctorius
flavanone 3-hydroxylase	-	Carthamus tinctorius

pI Value

Organism	Comment	pI Value Maximum	pI Value
Carthamus tinctorius	sequence calculation	-	5.57

Expression

Organism	Comment	Expression
Carthamus tinctorius	the enzyme flavanone 3-hydroxylase is induced by methyl jasmonate. Further metabolite analysis shows the increasing tendency of quinochalcones and flavonols, such as hydroxysafflor yellow A, kaempferol-3-O-beta-D-glucoside, kaempferol-3-O-beta-rutinoside, rutin, carthamin, and luteolin, in the quinochalcone-type safflower line. Also, the accumulation of kaempferol-3-O-beta-rutinoside and kaempferol-3-O-beta-D-glucoside in flavonols-typed safflower line shows enhanced accumulation pattern after methyl jasmonate treatment. Other flavonols, such as kaempferol, dihydrokaempferol and quercetin-3-O-beta-D-glucoside, in flavonols-typed safflower line present downregulation in respons to the methyl jasmonate stimulus	up

General Information

General Information	Comment	Organism
evolution	the enzyme belongs to the Fe2+/2oxoglutarate-dependent dioxygenase superfamily	Carthamus tinctorius
physiological function	flavanone 3-hydroxylase (F3H) is one of the nuclear enzymes acting at the bifurcation of the flavonoid biosynthetic pathway, initiating catalysis of the 3-hydroxylation of (2S)-flavanones, such as naringenin to dihydroflavonols	Carthamus tinctorius