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Literature summary extracted from

  • Liu, Y.; Zhao, Z.; Xue, Z.; Wang, L.; Cai, Y.; Wang, P.; Wei, T.; Gong, J.; Liu, Z.; Li, J.; Li, S.; Xiang, F.
    An intronless beta-amyrin synthase gene is more efficient in oleanolic acid accumulation than its paralog in Gentiana straminea (2016), Sci. Rep., 6, 33364 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
5.4.99.39 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
5.4.99.39 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

EC Number Protein Variants Comment Organism
5.4.99.39 G257W site-directed mutagenesis Gentiana straminea
5.4.99.39 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
5.4.99.39 additional information RNAi-directed suppression of GsAS1 in Gentiana straminea decreasing oleonolic acid levels by 65.9% Gentiana straminea
5.4.99.39 additional information RNAi-directed suppression of GsAS2 in Gentiana straminea decreasing oleonolic acid levels by 21.0% Gentiana straminea
5.4.99.39 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
5.4.99.39 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/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
5.4.99.39 (3S)-2,3-epoxy-2,3-dihydrosqualene Gentiana straminea
-
 beta-amyrin
-
 r

Organism

EC Number Organism UniProt Comment Textmining
5.4.99.39 Gentiana straminea A0A023U990
-
-
5.4.99.39 Gentiana straminea C1K2M2
-
-

Reaction

EC Number Reaction Comment Organism Reaction ID
5.4.99.39 (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
a

Source Tissue

EC Number Source Tissue Comment Organism Textmining
5.4.99.39 leaf
-
 Gentiana straminea
-
5.4.99.39 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
-
5.4.99.39 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
-
5.4.99.39 root highest enzyme level Gentiana straminea
-
5.4.99.39 root highest enzyme level, very high GsAS2 activity in roots Gentiana straminea
-
5.4.99.39 stem
-
 Gentiana straminea
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5.4.99.39 (3S)-2,3-epoxy-2,3-dihydrosqualene
-
 Gentiana straminea beta-amyrin
-
 r

Synonyms

EC Number Synonyms Comment Organism
5.4.99.39 betaAS
-
 Gentiana straminea
5.4.99.39 GsAS1
-
 Gentiana straminea
5.4.99.39 GsAS2
-
 Gentiana straminea

Expression

EC Number Organism Comment Expression
5.4.99.39 Gentiana straminea RNAi-directed suppression of GsAS1 in Gentiana straminea decreases oleonolic acid levels by 65.9% down
5.4.99.39 Gentiana straminea RNAi-directed suppression of GsAS2 in Gentiana straminea decreases oleonolic acid levels by 21.0% down
5.4.99.39 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
5.4.99.39 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

EC Number General Information Comment Organism
5.4.99.39 evolution the enzyme is a member of the oxidosqualene cyclase (OSC) family Gentiana straminea
5.4.99.39 malfunction RNAi-directed suppression of GsAS1 in Gentiana straminea decreases oleonolic acid levels by 65.9% Gentiana straminea
5.4.99.39 malfunction RNAi-directed suppression of GsAS2 in Gentiana straminea decreasing oleonolic acid levels by 21.0% Gentiana straminea
5.4.99.39 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
5.4.99.39 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