Literature summary for 5.4.99.39 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 .

Search for more literature for 5.4.99.39

Cloned(Commentary)

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

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

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/ Products (Substrates)

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

Organism	UniProt	Comment	Textmining
Gentiana straminea	A0A023U990	-	-
Gentiana straminea	C1K2M2	-	-

Reaction

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	a

Source Tissue

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 and Products (Substrate)

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

Synonyms

Synonyms	Comment	Organism
betaAS	-	Gentiana straminea
GsAS1	-	Gentiana straminea
GsAS2	-	Gentiana straminea

Expression

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

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