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Literature summary for 6.2.1.16 extracted from

  • McQualter, R.B.; Petrasovits, L.A.; Gebbie, L.K.; Schweitzer, D.; Blackman, D.M.; Chrysanthopoulos, P.; Hodson, M.P.; Plan, M.R.; Riches, J.D.; Snell, K.D.; Brumbley, S.M.; Nielsen, L.K.
    The use of an acetoacetyl-CoA synthase in place of a beta-ketothiolase enhances poly-3-hydroxybutyrate production in sugarcane mesophyll cells (2015), Plant Biotechnol. J., 13, 700-707 .
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
recombinant enzyme expression Streptomyces lividans

Crystallization (Commentary)

Crystallization (Comment) Organism
purified recombinant enzyme SlAacS complexed with AMP and acetoacetate, hanging drop vapor diffusion method, mixing of 11 mg/ml protein and 1.5fold molar excess of buffered AMP and acetoacetate with a reservoir solution containing 0.7-1.2M K3citrate and 50 mM BTP-HCl, pH 7.0, 14°C, 1 week, X-ray diffraction structure determination and analysis, molecular replacement using full-length Salmonella enterica acetyl-CoA synthetase, PDB ID1PG4, stripped of waters and cofactors/substrates as search model Streptomyces lividans

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required Streptomyces lividans

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + acetoacetate + CoA Streptomyces lividans
-
AMP + diphosphate + acetoacetyl-CoA
-
?
ATP + acetoacetate + CoA Streptomyces lividans TK24
-
AMP + diphosphate + acetoacetyl-CoA
-
?

Organism

Organism UniProt Comment Textmining
Streptomyces lividans D6EQU8
-
-
Streptomyces lividans TK24 D6EQU8
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme Streptomyces lividans

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
4.91 6.8 purified recombinant enzyme, pH 7.5, 30°C Streptomyces lividans

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + acetoacetate + CoA
-
Streptomyces lividans AMP + diphosphate + acetoacetyl-CoA
-
?
ATP + acetoacetate + CoA
-
Streptomyces lividans TK24 AMP + diphosphate + acetoacetyl-CoA
-
?

Synonyms

Synonyms Comment Organism
AACS
-
Streptomyces lividans
Acetoacetyl-CoA synthetase
-
Streptomyces lividans
acsA1
-
Streptomyces lividans
SlAacS
-
Streptomyces lividans

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Streptomyces lividans

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
5.89
-
acetoacetate pH 7.5, 30°C, recombinant enzyme Streptomyces lividans
6.43
-
ATP pH 7.5, 30°C, recombinant enzyme Streptomyces lividans
8.16
-
CoA pH 7.5, 30°C, recombinant enzyme Streptomyces lividans

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
assay at Streptomyces lividans

Cofactor

Cofactor Comment Organism Structure
ATP
-
Streptomyces lividans

General Information

General Information Comment Organism
additional information in addition to the two catalytic states, additional conformations are observed crystallographically that likely play a role in allowing access and egress of substrates and products from the relatively buried active site. The enzyme shows conformational flexibility. Structure-function analysis, overview. The C-terminal domain undergoes a large conformational change in the catalytic mechanism of acyl-CoA synthetases, the C-terminal extension is important for catalytic activity, structure comparisons. One region from the N-terminal domain interacts is the so-called P-loop, a glycine-, serine-, and threonine-rich region that interacts with the phosphates of ATP. This P-loop adopts multiple conformations in the different crystal structures and may play an important role in the release of PPi and trigger the conformational change. Specifically, the main chain carbonyls of Ser272, Gly274, and Gly277 form direct or water-mediated hydrogen bonds with Asn637 and Ser640. Asn637 also interacts directly with Arg183 and Asp187, while the carbonyl of Gly639 and the carbonyl and side chain oxygens of Ser640 interact with Ser184, Asp187, and Arg188. Streptomyces lividans

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
23.6
-
CoA pH 7.5, 30°C, recombinant enzyme Streptomyces lividans
26.6
-
acetoacetate pH 7.5, 30°C, recombinant enzyme Streptomyces lividans
32.4
-
ATP pH 7.5, 30°C, recombinant enzyme Streptomyces lividans