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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 .
    View publication on PubMedView publication on EuropePMC

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

Engineering

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
-
-

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

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. Reac.
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 ?
-
?
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]
-
?

Subunits

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

Synonyms

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