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Literature summary for 1.2.1.B25 extracted from

  • Lin, F.; Das, D.; Lin, X.N.; Marsh, E.N.
    Aldehyde-forming fatty acyl-CoA reductase from cyanobacteria expression, purification and characterization of the recombinant enzyme (2013), FEBS J., 280, 4773-4781 .
    View publication on PubMed
Search for more literature for 1.2.1.B25

Activating Compound

Activating Compound Comment Organism Structure
bovine serum albumin activates 3fold at 1 mg/ml Synechococcus elongatus
CHAPS activates about by 75% at 0.2%, inhibitory at above 0.5% Synechococcus elongatus
Digitonin activates about at 0.1%, inhibitory at above 0.5% Synechococcus elongatus
octyl glucoside activates slightly about at 0.2%, inhibitory at above 0.5% Synechococcus elongatus
tris(hydroxypropyl)phosphine activates 2fold at 20 mM Synechococcus elongatus

Application

Application Comment Organism
biofuel production long-chain acyl-CoA reductases (ACRs) catalyze a key step in the biosynthesis of hydrocarbon waxes. As such they are attractive as components in engineered metabolic pathways for drop in biofuels. The slow turnover number measured for Synechococcus elongatus ACR poses a challenge for its use in biofuel applications where highly efficient enzymes are needed Synechococcus elongatus

Cloned(Commentary)

Cloned (Comment) Organism
recombinant overexpression of His-tagged enzyme in Escherichia coli Synechococcus elongatus

Inhibitors

Inhibitors Comment Organism Structure
CHAPS activates about by 75% at 0.2%, inhibitory at above 0.5% Synechococcus elongatus
Digitonin activates about at 0.1%, inhibitory at above 0.5% Synechococcus elongatus
dithiothreitol inhibits the enzyme at 5 mM Synechococcus elongatus
glutathione inhibits the enzyme at 5 mM Synechococcus elongatus
iodoacetamide inactivation at 1 mM, stearoyl-CoA protects, probably due to competition between the two reagents for the active site cysteine Synechococcus elongatus
additional information once acylated by stearoyl-CoA the enzyme is protected from inactivation by iodoacetamide and the cACR activity remains unchanged Synechococcus elongatus
octyl glucoside activates slightly about at 0.2%, inhibitory at above 0.5% Synechococcus elongatus
Triton inhibitory at above 0.5% Synechococcus elongatus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information steady-state kinetic analysis Synechococcus elongatus
0.0319
-
 stearoyl-CoA pH 7.5, 37°C, recombinant enzyme Synechococcus elongatus
0.0356
-
 NADPH pH 7.5, 37°C, recombinant enzyme Synechococcus elongatus

Localization

Localization Comment Organism GeneOntology No. Textmining
cytosol
-
 Synechococcus elongatus 5829
-

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+ activates, inhibitory as it causes precipitation of the acyl-CoA substrate at 5 mM Synechococcus elongatus
K+ activates about 10fold Synechococcus elongatus
Mg2+ best activating divalent cation Synechococcus elongatus
Mn2+ activates, inhibitory as it causes precipitation of the acyl-CoA substrate at 2 mM Synechococcus elongatus
additional information ACR requires divalent metal ions for activity. 2-mercaptoethanol and Na+ have no effect on activity Synechococcus elongatus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
stearoyl-CoA + NADPH Synechococcus elongatus
-
 octadecanal + CoA + NADP+
-
 ?
stearoyl-CoA + NADPH Synechococcus elongatus PCC7942
-
 octadecanal + CoA + NADP+
-
 ?

Organism

Organism UniProt Comment Textmining
Synechococcus elongatus
-
-
-
Synechococcus elongatus PCC7942
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged enzyme from Escherichia coli by nickel affinity chromatography Synechococcus elongatus

Reaction

Reaction Comment Organism Reaction ID
hexadecanal + CoA + NADP+ = hexadecanoyl-CoA + NADPH the enzyme operates by a Ping-Pong mechanism Synechococcus elongatus
a

Storage Stability

Storage Stability Organism
-80°C, purified cACR is quite unstable and tends to precipitate after 3-4 days when stored at 4°C, although it can be stored for prolonged periods at -80°C without loss of activity Synechococcus elongatus
4°C, purified cACR is quite unstable and tends to precipitate after 3-4 days when stored at 4°C, although it can be stored for prolonged periods at -80°C without loss of activity Synechococcus elongatus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
dodecanoyl-CoA + NADPH low activity Synechococcus elongatus dodecanal + CoA + NADP+
-
 ?
dodecanoyl-CoA + NADPH low activity Synechococcus elongatus PCC7942 dodecanal + CoA + NADP+
-
 ?
eicosanoyl-CoA + NADPH
-
 Synechococcus elongatus eicosanal + CoA + NADP+
-
 ?
hexadecanoyl-CoA + NADPH + H+ low activity Synechococcus elongatus hexadecanal + CoA + NADP+
-
 ?
hexadecanoyl-CoA + NADPH + H+ low activity Synechococcus elongatus PCC7942 hexadecanal + CoA + NADP+
-
 ?
additional information the enzyme cACR is specific for NADPH and specifically catalyzes the reduction of fatty acyl-CoA esters to the corresponding aldehydes, rather than alcohols. Stearoyl-CoA is the most effective substrate, being reduced more rapidly than either longer or shorter chain acyl-CoAs. The enzyme is acylated on incubation with stearoyl-CoA, suggesting that the reduction occurs through an enzyme-thioester intermediate, cf. EC 1.2.1.50, formation of an acyl thioester on an active site cysteinyl residue as an intermediate in the mechanism of reduction. No activity with octanoyl-CoA Synechococcus elongatus ?
-
 ?
additional information the enzyme cACR is specific for NADPH and specifically catalyzes the reduction of fatty acyl-CoA esters to the corresponding aldehydes, rather than alcohols. Stearoyl-CoA is the most effective substrate, being reduced more rapidly than either longer or shorter chain acyl-CoAs. The enzyme is acylated on incubation with stearoyl-CoA, suggesting that the reduction occurs through an enzyme-thioester intermediate, cf. EC 1.2.1.50, formation of an acyl thioester on an active site cysteinyl residue as an intermediate in the mechanism of reduction. No activity with octanoyl-CoA Synechococcus elongatus PCC7942 ?
-
 ?
oleoyl-CoA + NADPH low activity Synechococcus elongatus (9Z)-octadecenal + CoA + NADP+
-
 ?
stearoyl-CoA + NADPH
-
 Synechococcus elongatus octadecanal + CoA + NADP+
-
 ?
stearoyl-CoA + NADPH best substrate. When the enzyme is incubated with both stearoyl-CoA and NADPH, the acyl-enzyme accumulates to comprise about 50% of the enzyme. This observation suggests that the rates of acylation and reduction are approximately the same Synechococcus elongatus octadecanal + CoA + NADP+
-
 ?
stearoyl-CoA + NADPH
-
 Synechococcus elongatus PCC7942 octadecanal + CoA + NADP+
-
 ?
stearoyl-CoA + NADPH best substrate. When the enzyme is incubated with both stearoyl-CoA and NADPH, the acyl-enzyme accumulates to comprise about 50% of the enzyme. This observation suggests that the rates of acylation and reduction are approximately the same Synechococcus elongatus PCC7942 octadecanal + CoA + NADP+
-
 ?

Subunits

Subunits Comment Organism
? x * 42626, mass spectrometry and sequence calculation Synechococcus elongatus

Synonyms

Synonyms Comment Organism
Acr
-
 Synechococcus elongatus
aldehyde-forming fatty acyl-CoA reductase
-
 Synechococcus elongatus
cACR
-
 Synechococcus elongatus
cyanobacterial acyl-CoA reductase
-
 Synechococcus elongatus
long-chain ACR
-
 Synechococcus elongatus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Synechococcus elongatus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.006
-
 stearoyl-CoA pH 7.5, 37°C, recombinant enzyme Synechococcus elongatus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
-
 Synechococcus elongatus

Cofactor

Cofactor Comment Organism Structure
additional information no activity with NADH Synechococcus elongatus
NADPH specific for Synechococcus elongatus

General Information

General Information Comment Organism
physiological function long-chain acyl-CoA reductases (ACRs) catalyze a key step in the biosynthesis of hydrocarbon waxes Synechococcus elongatus