Literature summary for 6.2.1.20 extracted from

Kaczmarzyk, D.; Hudson, E.P.; Fulda, M.
Arabidopsis acyl-acyl carrier protein synthetase AAE15 with medium chain fatty acid specificity is functional in cyanobacteria (2016), AMB Express, 6, doi: 10.1186/s13568-016-0178-z .

Cloned(Commentary)

Cloned (Comment)	Organism
gene AAE15 or At4g14070, recombinant expression of N-terminally His6-tagged enzyme, full-length or lacking the predicted plastidial targeting signal sequence, in Spodoptera frugiperda Sf9 cells via baculovirus transfection method, recombinant expression of Arabidopsis thaliana AAE15 in an AAS deletion mutant of Synechocystis sp. PCC 6803 resulting in changes in intracellular and extracellular free fatty acids pools in the cyanobacterial DELTAaas strain	Arabidopsis thaliana

Cloned (Comment)

Organism

gene AAE15 or At4g14070, recombinant expression of N-terminally His6-tagged enzyme, full-length or lacking the predicted plastidial targeting signal sequence, in Spodoptera frugiperda Sf9 cells via baculovirus transfection method, recombinant expression of Arabidopsis thaliana AAE15 in an AAS deletion mutant of Synechocystis sp. PCC 6803 resulting in changes in intracellular and extracellular free fatty acids pools in the cyanobacterial DELTAaas strain

Arabidopsis thaliana

Protein Variants

Protein Variants	Comment	Organism
additional information	generation of a N-terminally truncated enzyme mutant with deleted predicted plastidial targeting signal sequence by removing the sequence encoding a predicted plastidial targeting peptide increases the expression level significantly	Arabidopsis thaliana

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
chloroplast	the enzyme has a predicted plastidial targeting peptide	Arabidopsis thaliana	9507	-

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	required	Arabidopsis thaliana

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
ATP + a long-chain fatty acid + an [acyl-carrier protein]	Arabidopsis thaliana	-	AMP + diphosphate + a long-chain acyl-[acyl-carrier protein]	-	?

Organism

Organism	UniProt	Comment	Textmining
Arabidopsis thaliana	Q8W471	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant N-terminally His6-tagged enzyme, full-length or lacking the predicted plastidial targeting signal sequence, from Spodoptera frugiperda Sf9 cells by nickel affinity chromatography, gel filtration, and dialysis. Removing the sequence encoding a predicted plastidial targeting peptide increases the expression level significantly, like DELTAaas, DELTAaas:AAE15 secretes large amounts of fatty acids into the culture medium. Expression of AAE15 in the DELTAaas strain does not affect fatty acid profiles and does not have a significant complementation effect	Arabidopsis thaliana

Purification (Comment)

Organism

recombinant N-terminally His6-tagged enzyme, full-length or lacking the predicted plastidial targeting signal sequence, from Spodoptera frugiperda Sf9 cells by nickel affinity chromatography, gel filtration, and dialysis. Removing the sequence encoding a predicted plastidial targeting peptide increases the expression level significantly, like DELTAaas, DELTAaas:AAE15 secretes large amounts of fatty acids into the culture medium. Expression of AAE15 in the DELTAaas strain does not affect fatty acid profiles and does not have a significant complementation effect

Arabidopsis thaliana

Source Tissue

Source Tissue	Comment	Organism	Textmining
leaf	-	Arabidopsis thaliana	-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
0.0000667	-	purified recombinant enzyme, pH 8.0, 37°C, with substrate decanoate	Arabidopsis thaliana
0.0001027	-	purified recombinant enzyme, pH 8.0, 37°C, with substrate laurate	Arabidopsis thaliana
0.0001572	-	purified recombinant enzyme, pH 8.0, 37°C, with substrate myristate	Arabidopsis thaliana

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
ATP + a long-chain fatty acid + an [acyl-carrier protein]	-	Arabidopsis thaliana	AMP + diphosphate + a long-chain acyl-[acyl-carrier protein]	-	?
ATP + decanoate + an [acyl-carrier protein]	-	Arabidopsis thaliana	AMP + diphosphate + long-chain decanoyl-[acyl-carrier protein]	-	?
ATP + hexadecanoate + an [acyl-carrier protein]	low activity	Arabidopsis thaliana	AMP + diphosphate + hexadecanoyl-[acyl-carrier protein]	-	?
ATP + laurate + an [acyl-carrier protein]	-	Arabidopsis thaliana	AMP + diphosphate + lauroyl-[acyl-carrier protein]	-	?
ATP + myristate + an [acyl-carrier protein]	-	Arabidopsis thaliana	AMP + diphosphate + myristoyl-[acyl-carrier protein]	-	?
ATP + octanoate + an [acyl-carrier protein]	low activity	Arabidopsis thaliana	AMP + diphosphate + octanoyl-[acyl-carrier protein]	-	?
additional information	recombinant enzyme AAE15 has acyl-ACP synthetase activity in vitro with specificity for medium chain fatty acids, substrate specificity, overview. Low activity with fatty acids 18:0, 18:1, 18:2, and 18:3	Arabidopsis thaliana	?	-	?

Subunits

Subunits	Comment	Organism
?	x * 79000, recombinant truncated enzyme lacking the predicted plastidial targeting signal sequence, SDS-PAGE	Arabidopsis thaliana

Synonyms

Synonyms	Comment	Organism
AAE15	-	Arabidopsis thaliana
AAS	-	Arabidopsis thaliana
Acyl-ACP synthetase	-	Arabidopsis thaliana
Acyl-acyl carrier protein synthetase	-	Arabidopsis thaliana
At4g14070	-	Arabidopsis thaliana

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
37	-	assay at	Arabidopsis thaliana

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
8	-	assay at	Arabidopsis thaliana

Cofactor

Cofactor	Comment	Organism	Structure
ATP	-	Arabidopsis thaliana