Cloned (Comment) | Organism |
---|---|
gene GsAS1, gene GsAS1 possesses introns, phylogenetic tree, recombinant expression in Pichia pastoris under the control of the methanol-inducible AOX1 promoter, heterologously expressed GsAS1 generates less beta-amyrin in yeast than does GsAS2, constitutive overexpression of GsAS2 results in a 3fold increase in oleanolic acid accumulation, quantitative RT-PCR expression analysis | Gentiana straminea |
gene GsAS2, DNA and amino acid sequence determination and analysis, the gene GsAS2 lacks introns, phylogenetic tree, recombinant expression in Pichia pastoris under the control of the methanol-inducible AOX1 promoter, heterologously expressed GsAS2 generates more beta-amyrin in yeast than does GsAS1, constitutive overexpression of GsAS2 results in a 5.7fold increase in oleanolic acid accumulation, quantitative RT-PCR expression analysis | Gentiana straminea |
Protein Variants | Comment | Organism |
---|---|---|
G257W | site-directed mutagenesis | Gentiana straminea |
H560Y | site-directed mutagenesis, the mutation of the key residue of GsAS1 to that of GsAS2 results in 38% increased catalytic efficiency compared to wild-type GsAS1 | Gentiana straminea |
additional information | RNAi-directed suppression of GsAS1 in Gentiana straminea decreasing oleonolic acid levels by 65.9% | Gentiana straminea |
additional information | RNAi-directed suppression of GsAS2 in Gentiana straminea decreasing oleonolic acid levels by 21.0% | Gentiana straminea |
Y560F | site-directed mutagenesis, the mutation of the key residue of GsAS2 results in 71.15% reduced catalytic efficiency compared to wild-type GsAS2 | Gentiana straminea |
Y560H | site-directed mutagenesis, the mutation of the key residue of GsAS2 results in 41.3% reduced catalytic efficiency compared to wild-type GsAS2 | Gentiana straminea |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
(3S)-2,3-epoxy-2,3-dihydrosqualene | Gentiana straminea | - |
beta-amyrin | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Gentiana straminea | A0A023U990 | - |
- |
Gentiana straminea | C1K2M2 | - |
- |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
(3S)-2,3-epoxy-2,3-dihydrosqualene = beta-amyrin | molecular substrate docking and catalytic reaction mechanism analysis, overview. Residue Y560 is responsible for the high catalytic efficiency of enzyme GsAS2 compared to GsAS1 | Gentiana straminea |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
leaf | - |
Gentiana straminea | - |
additional information | GsAS1 transcript levels in roots are 2.3fold higher than the levels in either the leaves or stems. GsAS2 expression is 13.3fold higher than GsAS1 expression in roots and 1.6fold higher in stems | Gentiana straminea | - |
additional information | GsAS2 transcript levels in roots are 27.5fold greater compared to leaves and 18.6fold higher than in stems. GsAS2 expression is 13.3fold higher than GsAS1 expression in roots and 1.6fold higher in stems | Gentiana straminea | - |
root | highest enzyme level | Gentiana straminea | - |
root | highest enzyme level, very high GsAS2 activity in roots | Gentiana straminea | - |
stem | - |
Gentiana straminea | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(3S)-2,3-epoxy-2,3-dihydrosqualene | - |
Gentiana straminea | beta-amyrin | - |
r |
Synonyms | Comment | Organism |
---|---|---|
betaAS | - |
Gentiana straminea |
GsAS1 | - |
Gentiana straminea |
GsAS2 | - |
Gentiana straminea |
Organism | Comment | Expression |
---|---|---|
Gentiana straminea | RNAi-directed suppression of GsAS1 in Gentiana straminea decreases oleonolic acid levels by 65.9% | down |
Gentiana straminea | RNAi-directed suppression of GsAS2 in Gentiana straminea decreases oleonolic acid levels by 21.0% | down |
Gentiana straminea | GsAS1 transcription is 9.8fold induced by methyl jasmonate (MeJA) treatment with increased oleanolic acid content, while salicylic acid has no effect | up |
Gentiana straminea | GsAS2 transcription is 12.2fold induced by methyl jasmonate (MeJA) treatment with increased oleanolic acid content, while salicylic acid has no effect | up |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme is a member of the oxidosqualene cyclase (OSC) family | Gentiana straminea |
malfunction | RNAi-directed suppression of GsAS1 in Gentiana straminea decreases oleonolic acid levels by 65.9% | Gentiana straminea |
malfunction | RNAi-directed suppression of GsAS2 in Gentiana straminea decreasing oleonolic acid levels by 21.0% | Gentiana straminea |
additional information | the intronless beta-amyrin synthase gene GSAS2 is more efficient in oleanolic acid accumulation than its paralogue GsAS1 in Gentiana straminea. Both enzymes harbor two (alpha/alpha) barrel domains connected by loops, as well as three smaller beta structures | Gentiana straminea |
physiological function | GsAS2 plays a more important role than GsAS1 in oleanolic acid biosynthesis in Gentiana straminea. Wild-type GsAS1 and mutant GsAS1 H560Y show 7-11% of wild-type GSAS2 activity | Gentiana straminea |