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Information on EC 4.1.1.4 - acetoacetate decarboxylase and Organism(s) Clostridium acetobutylicum and UniProt Accession P23670
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4.1.1.4 acetoacetate decarboxylaseSpecify your search results
Word Map
- 4.1.1.4
- clostridium
- acetobutylicum
- beijerinckii
-
solventogenesis
-
solventogenic
-
thiolase
- isopropanol
- acetoacetyl-coa
-
acetone-butanol-ethanol
-
primary-secondary
- phosphotransbutyrylase
-
solvent-producing
-
acidogenic
- synthesis
- analysis
The taxonomic range for the selected organisms is: Clostridium acetobutylicum
The expected taxonomic range for this enzyme is: Bacteria, Archaea, Eukaryota
The expected taxonomic range for this enzyme is: Bacteria, Archaea, Eukaryota
Synonyms
acetoacetate decarboxylase, aadase, acetoacetic acid decarboxylase, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
AAD
-
-
-
-
Acetoacetic decarboxylase
-
-
-
-
ADC
CP 28/CP 29
-
-
-
-
Decarboxylase, acetoacetate
-
-
-
-
Polymyxin MI
-
-
-
-
ADC
-
-
-
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
acetoacetate + H+ = acetone + CO2
the enzyme catalyzes the reaction with net racemization
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
decarboxylation
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
acetoacetate carboxy-lyase (acetone-forming)
-
CAS REGISTRY NUMBER
COMMENTARY
9025-03-0
-
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
2-Oxo-3-phenylpropionic acid
Acetophenone + CO2
-
i.e. phenylacetoacetate
-
-
?
Acetoacetate
?
-
induction by linear acids from C1 to C4, whereas branched acids and linear acids from C5 to C7 are not inducers
-
-
?
acetoacetate + H+
acetone + CO2
-
-
-
?
acetoacetate + H+
acetone + CO2
the AADC activity relies on measurement of CO2 produced as a result of AADC-catalyzed decarboxylation of acetoacetate
-
-
?
additional information
?
-
acetoacetate decarboxylase from Clostridium acetobutylicum can act as a biocatalyst for decarboxylation of levulinic acid
-
-
?
additional information
?
-
-
acetoacetate decarboxylase from Clostridium acetobutylicum can act as a biocatalyst for decarboxylation of levulinic acid
-
-
?
additional information
?
-
-
catalyzes the exchange of deuterium or tritium into the alpha position of cyclohexanone or 2-butanone
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
Acetoacetate
?
-
induction by linear acids from C1 to C4, whereas branched acids and linear acids from C5 to C7 are not inducers
-
-
?
additional information
?
-
acetoacetate decarboxylase from Clostridium acetobutylicum can act as a biocatalyst for decarboxylation of levulinic acid
-
-
?
additional information
?
-
-
acetoacetate decarboxylase from Clostridium acetobutylicum can act as a biocatalyst for decarboxylation of levulinic acid
-
-
?
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
levulinic acid
substrate inhibition is observed with levulinic acid concentration higher than 5 mM
2,4-Dinitrophenyl acetate
-
acetylation and complete inactivation of the enzyme
Acetic anhydride
-
acetylation and complete inactivation of the enzyme
Acetopyruvate
-
the enzyme inhibitor compound is an enamine, obtained by the tautomerization of the Schiff base initially formed from inhibitor and enzyme
Borohydride
-
borohydride plus acetoacetate or 2-oxopropane sulfonate irreversibly inhibits, without affecting the latent decarboxylase. Monovalent anions protect
HCN-
-
inhibitory synergism between cyanide and carbonyl compounds. The effectiveness in descending order: acetaldehyde, acetone, cyclohexanone, methyl ethyl ketone, 3-hexanone, diethyl ketone
Monovalent anions
-
at pH 7.0 and above, monovalent anions are entirely noncompetitive inhibitors, as the pH is lowered the inhibition becomes increasingly uncompetitive
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
2,2-azinobis(3-ethylbenzothiazoline-6-sulfonic acid)
activates about 2fold at 5-10 mM
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
4.1
acetoacetate
at 25°C, in 50 mM phosphate buffer, pH 5.95
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
165
acetoacetate
at 25°C, in 50 mM phosphate buffer, pH 5.95
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0007
2,4-Pentanedione
at 25°C, in 50 mM phosphate buffer, pH 5.95
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
29000
-
x * 29000, meniscus depletion method in presence of 6 M guanidium chloride or 4 M urea
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
?
-
x * 29000, meniscus depletion method in presence of 6 M guanidium chloride or 4 M urea
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
AAD complexed with 2,4-pentanedione, hanging drop vapor diffusion method, using 18-20% (v/v) glycerol, 40 mM phosphate buffer, pH 5.95, 100 mM sarcosine and 14-15% (w/v) PEG 3350
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
E61Q
the catalytic activity of the mutant shows a decrease in kcat (about 20fold with no change in Km)
E76Q
R29Q
the catalytic activity of Arg29Gln does not increase at pH values above the wild type optimum for AAD of about 5.4
K115C
-
mutant enzymes K115C and K115Q are catalytically inactive at pH 5.95. Mutant enzymes K116C, K116N and K116R have reduced but significant activities
K115Q
-
mutant enzymes K115C and K115Q are catalytically inactive at pH 5.95. Mutant enzymes K116C, K116N and K116R have reduced but significant activities
additional information
-
the butanol ratio increases from 70% to 80.05%, with acetone production reduced to approximately 0.21 g/l in the adc-disrupted mutant 2018adc
E76Q
the catalytic activity of the mutant shows a decrease in kcat, the mutant exhibits a slight downward shift of the pH optimum
E76Q
the optimum pH value for the enzymatic activity remains essentially unchanged in the E76Q mutation
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
80
-
30 min: 50% loss of activity, without addition of acetylacetone, stable in presence of 1 mM acetylacetone
85
-
30 min: 86% loss of activity, without addition of acetylacetone, stable in presence of 10 mM acetylacetone
additional information
additional information
-
biphasic thermal inactivation. Acetylacetone protects more effectively in preventing the rapid phase than in preventing the slow phase of activity loss
additional information
-
some enzyme is expressed and an equal amount is latent and may be measured only after its exposure to heat
additional information
-
acetylacetone protects against thermal inactivation
additional information
-
acetylacetone protects against thermal inactivation
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
rather easily inactivated by urea, guanidinium chloride and SDS
-
the enzyme dissociates into subunit dimers at pH 8 in 4 M urea solution at low temperature. The subunit dimers can be reassociated to form native, active enzyme by diluting the urea solution with phosphate buffer at pH 6.0 in the presence of dithiothreitol
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
ammonium sulfate precipitation, DEAE-Sepharose column chromatography, and S-200 gel filtration
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
gene aadc, cloning in Escherichia coli strain DH5alpha, expression as GFP-tagged enzyme
recombinant expression of His6-tagged enzyme in Escherichia coli strain BL21(DE3), best induction over 24 h at 20°C with 1 mM IPTG
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
synthesis
acetoacetate decarboxylase from Clostridium acetobutylicum can act as a biocatalyst for decarboxylation of levulinic acid in an enzymatic system for synthesis of 2-butanone from levulinic acid
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Rozzel, J.D.; Benner, S.A.
Stereochemical imperative in enzymic decarboxylation. Stereochemical course of the decarboxylation catalyzed by acetoacetate decarboxylase
J. Am. Chem. Soc.
106
4937-4941
1984
Clostridium acetobutylicum
-
Benner, S.A.; Rozzel, J.D.
Stereospecificity and stereochemical infidelity of acetoacetate decarboxylase (AAD)
J. Am. Chem. Soc.
103
993-994
1981
Clostridium acetobutylicum
-
Kluger, R.; Nakaoka, K.
Inhibition of acetoacetate decarboxylase by ketophosphonates. Structural and dynamic probes of the active site
Biochemistry
13
910-914
1974
Clostridium acetobutylicum
Fridovich, I.
Acetoacetate decarboxylase
The Enzymes, 3rd Ed. (Boyer, P. D. , ed. )
6
255-270
1972
Clostridium acetobutylicum
-
Autor, A.P.; Fridovich, I.
The thermal inactivation of acetoacetate decarboxylase
J. Biol. Chem.
245
5223-5227
1970
Clostridium acetobutylicum
Autor, A.P.; Fridovich, I.
The interactions of acetoacetate decarboxylase with carbonyl compounds, hydrogen cyanide, and an organic mercurial
J. Biol. Chem.
245
5214-5222
1970
Clostridium acetobutylicum
Westheimer, F.H.
Acetoacetate decarboxylase from Clostridium acetobutylicum
Methods Enzymol.
14
231-241
1969
Clostridium acetobutylicum
-
Tagaki, W.; Guthrie, J.P.; Westheimer, F.H.
Acetoacetate decarboxylase. Reaction with acetopyruvate
Biochemistry
7
905-913
1968
Clostridium acetobutylicum
Tagaki, W.; Westheimer, F.H.
Acetoacetate decarboxylase. Catalysis of hydrogen-deuterium exchange in acetone
Biochemistry
7
901-905
1968
Clostridium acetobutylicum
Tagaki, W.; Westheimer, F.H.
Acetoacetate decarboxylase. The molecular weight of the enzyme and subunits
Biochemistry
7
895-900
1968
Clostridium acetobutylicum
Tagaki, W.; Westheimer, F.H.
Acetoacetate decarboxylase. Reassociation of subunits
Biochemistry
7
891-894
1968
Clostridium acetobutylicum
Fridovich, I.
A study of the interaction of acetoacetic decarboxylase with several inhibitors
J. Biol. Chem.
243
1043-1051
1968
Clostridium acetobutylicum
Neece, M.S.; Fridovich, I.
Acetoacetic decarboxylase. Activation by heat
J. Biol. Chem.
242
2939-2944
1967
Clostridium acetobutylicum
O'Leary, M.H.; Westheimer, F.H.
Acetoacetate decarboxylase. Selective acetylation of the enzyme
Biochemistry
7
913-919
1968
Clostridium acetobutylicum
Ballongue, J.; Amine, J.; Masion, E.; Petitdemange, H.; Gay, R.
Induction of acetoacetate decarboxylase in Clostridium acetobutylicum
FEMS Microbiol. Lett.
29
273-277
1985
Clostridium acetobutylicum
-
Gerischer, U.; Durre, P.
Cloning, sequencing, and molecular analysis of the acetoacetate decarboxylase gene region from Clostridium acetobutylicum
J. Bacteriol.
172
6907-6918
1990
Clostridium acetobutylicum
Highbarger, L.A.; Gerlt, J.A.; Kenyon, G.L.
Mechanism of the reaction catalyzed by acetoacetate decarboxylase. Importance of lysine 116 in determining the pKa of the active-site lysine 115
Biochemistry
35
41-46
1996
Clostridium acetobutylicum
Petersen, D.J.; Bennett, G.N.
Purification of acetoacetate decarboxylase from Clostridium acetobutylicum ATCC 824 and cloning of the acetoacetate decarboxylase gene in Escherichia coli
Appl. Environ. Microbiol.
56
3491-3498
1990
Clostridium acetobutylicum
Ravagnani, A.; Jennert, K.C.B.; Steiner, E.; Grunberg, R.; Jefferies, J.R.; Wilkinson, S.R.; Young, D.I.; Tidswell, E.C.; Brown, D.P.; Youngman, P.; Morris, J.G.; Young M.
Spo0A directly controls the switch from acid to solvent production in solvent-forming clostridia
Mol. Microbiol.
37
1172-1185
2000
Clostridium acetobutylicum, Clostridium beijerinckii, Clostridium beijerinckii AA243, Clostridium beijerinckii NCIMB 8052, Ruminiclostridium cellulolyticum
Hanai, T.; Atsumi, S.; Liao, J.C.
Engineered synthetic pathway for isopropanol production in Escherichia coli
Appl. Environ. Microbiol.
73
7814-7818
2007
Clostridium acetobutylicum
Jojima, T.; Inui, M.; Yukawa, H.
Production of isopropanol by metabolically engineered Escherichia coli
Appl. Microbiol. Biotechnol.
77
1219-1224
2008
Clostridium acetobutylicum (P23670)
Jiang, Y.; Xu, C.; Dong, F.; Yang, Y.; Jiang, W.; Yang, S.
Disruption of the acetoacetate decarboxylase gene in solvent-producing Clostridium acetobutylicum increases the butanol ratio
Metab. Eng.
11
284-291
2009
Clostridium acetobutylicum, Clostridium acetobutylicum EA 2018
Ho, M.C.; Menetret, J.F.; Tsuruta, H.; Allen, K.N.
The origin of the electrostatic perturbation in acetoacetate decarboxylase
Nature
459
393-397
2009
Clostridium acetobutylicum (P23670), Chromobacterium violaceum (Q7NSA6)
Ishikita, H.
Origin of the pKa shift of the catalytic lysine in acetoacetate decarboxylase
FEBS Lett.
584
3464-3468
2010
Clostridium acetobutylicum (P23670), Chromobacterium violaceum (Q7NSA6)
Han, B.; Gopalan, V.; Ezeji, T.C.
Acetone production in solventogenic Clostridium species: new insights from non-enzymatic decarboxylation of acetoacetate
Appl. Microbiol. Biotechnol.
91
565-576
2011
Clostridium acetobutylicum (P23670), Clostridium beijerinckii (A6M020), Clostridium beijerinckii NCIMB 8052 (A6M020), Clostridium beijerinckii NCIMB 8052
Min, K.; Kim, S.; Yum, T.; Kim, Y.; Sang, B.I.; Um, Y.
Conversion of levulinic acid to 2-butanone by acetoacetate decarboxylase from Clostridium acetobutylicum
Appl. Microbiol. Biotechnol.
97
5627-5634
2013
Clostridium acetobutylicum (P23670), Clostridium acetobutylicum
Hoenicke, D.; Luetke-Eversloh, T.; Liu, Z.; Lehmann, D.; Liebl, W.; Ehrenreich, A.
Chemostat cultivation and transcriptional analyses of Clostridium acetobutylicum mutants with defects in the acid and acetone biosynthetic pathways
Appl. Microbiol. Biotechnol.
98
9777-9794
2014
Clostridium acetobutylicum
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