Literature summary for 5.5.1.12 extracted from

Zhou, K.; Peters, R.J.
Investigating the conservation pattern of a putative second terpene synthase divalent metal binding motif in plants (2009), Phytochemistry, 70, 366-369.

Cloned(Commentary)

Cloned (Comment)	Organism
Site-directed mutagenesis is carried out via PCR amplification with overlapping mutagenic primers, and the mutant genes verify by complete sequencing. The resulting wild type and mutant genes are then transferred via directional recombination to the T7-promoter and N-terminal 6xhis fusion expression vector pDEST17. Use of the pDEST17 vector results in a 25 amino acid residue linker between the 6 His-tag and the cloned protein.	Abies grandis

Cloned (Comment)

Organism

Site-directed mutagenesis is carried out via PCR amplification with overlapping mutagenic primers, and the mutant genes verify by complete sequencing. The resulting wild type and mutant genes are then transferred via directional recombination to the T7-promoter and N-terminal 6xhis fusion expression vector pDEST17. Use of the pDEST17 vector results in a 25 amino acid residue linker between the 6 His-tag and the cloned protein.

Abies grandis

Protein Variants

Protein Variants	Comment	Organism
D621A	eliminates class I activity	Abies grandis
T769A	enzyme activity	Abies grandis
T769G	enzyme activity	Abies grandis
T769S	enzyme activity	Abies grandis

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
additional information	-	geranylgeranyl diphosphate	mutant T769A, value not determined	Abies grandis
0.0012	-	geranylgeranyl diphosphate	wild-type	Abies grandis
0.0015	-	geranylgeranyl diphosphate	mutant T769G	Abies grandis
0.0015	-	geranylgeranyl diphosphate	mutant T769S	Abies grandis

Metals/Ions

Metals/Ions	Comment	Organism	Structure
K+	100 mM included in assay medium	Abies grandis
Mg2+	terpene synthases (TPS) require divalent metal ion co-factors, typically magnesium	Abies grandis

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
geranylgeranyl diphosphate	Abies grandis	class II terpene synthase reaction	copalyl diphosphate	-	r

Organism

Organism	UniProt	Comment	Textmining
Abies grandis	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
AgAS is purified using Ni-NTA His-bind resin in batch mode	Abies grandis

Source Tissue

Source Tissue	Comment	Organism	Textmining
cell culture	-	Abies grandis	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
geranylgeranyl diphosphate	class II terpene synthase reaction	Abies grandis	copalyl diphosphate	-	r

Synonyms

Synonyms	Comment	Organism
abietadiene synthase	class II synthase	Abies grandis
AGAS	-	Abies grandis

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
30	-	assay at	Abies grandis

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
additional information	-	geranylgeranyl diphosphate	mutant T769A, value below 10000	Abies grandis
0.003	-	geranylgeranyl diphosphate	mutant T769G	Abies grandis
0.03	-	geranylgeranyl diphosphate	mutant T769S	Abies grandis
0.4	-	geranylgeranyl diphosphate	wild-type	Abies grandis

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.2	-	assay at	Abies grandis