Recombinant Glycyrrhiza echinata Licodione synthase (CYP93B1)
Description
Introduction
Recombinant Glycyrrhiza echinata Licodione synthase, also known as CYP93B1, is a cytochrome P450 monooxygenase enzyme that plays a crucial role in the biosynthesis of isoflavonoids and retrochalcones in Glycyrrhiza echinata (licorice) cells . Isoflavonoids possess various medicinal properties, including antioxidant, antiviral, and anti-inflammatory activities .
Discovery and Isolation
CYP93B1 was identified and isolated from a cDNA library of elicited G. echinata cells . Researchers obtained eight P450 fragments (Ge-1 to Ge-8) from the cDNA library using a PCR-based method . Full-length P450 cDNAs, CYP93B1 and CYP81E1, corresponding to the fragments Ge-5 and Ge-3, were then cloned .
Function and Mechanism
CYP93B1 functions as a (2S)-flavanone 2-hydroxylase (F2H) and licodione synthase . It catalyzes the hydroxylation of (2S)-flavanones, such as liquiritigenin and naringenin, at the C-2 position, leading to the formation of 2-hydroxyisoflavanones . These products can then be converted to isoflavones like daidzein and genistein through acid treatment . Licodione synthase activity increases in cells treated with yeast extract .
The enzyme can employ both 5-deoxyflavanone (liquiritigenin) and 5-hydroxyflavanone (naringenin) as substrates . In G. echinata microsomes, CYP93B1 competes with isoflavone synthase (IFS) for the same substrates .
Enzyme Assays and Products
G. inflata expresses an LMT gene, GiLMT1, that impacts the biosynthesis of Licochalcone A (LCA) . GiLMT1 can add a methyl group to licodione, an intermediate in LCA biosynthesis .
Table 1: Enzyme reaction products formed by recombinant GiLMT1 protein with licodione and its isomer as substrates
| Substrate | Product |
|---|---|
| Licodione | 2′-O-methyllicodione (P1) |
| Licodione Isomer | P2 |
Regulation
Histone deacetylases (HDACs) like GiSRT2 negatively regulate flavonoid biosynthesis . When GiSRT2 expression is suppressed, Licochalcone A (LCA) and total flavonoid levels increase .
Product Specs
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Target Background
This enzyme catalyzes the formation of licodione and 2-hydroxynaringenin from (2S)-liquiritigenin and (2S)-naringenin, respectively. It also converts eriodictyol to luteolin.
KEGG: ag:BAA22423
