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Information on EC 4.3.1.1 - aspartate ammonia-lyase and Organism(s) Escherichia coli and UniProt Accession P0AC38
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4.3.1.1 aspartate ammonia-lyaseSpecify your search results
The taxonomic range for the selected organisms is: Escherichia coli
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea
Synonyms
aspartase, l-aspartase, aspartate ammonia-lyase, aspartate ammonia lyase, l-aspartate ammonia-lyase, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
ammonia-lyase, aspartate
-
-
-
-
aspartase
fumaric aminase
-
-
-
-
L-aspartase
-
-
-
-
maspase 1
-
insertion of a 15-peptide random peptide into the three domains of L-aspartase to enhance their mobility. After directed screening, the three isoforms of monomeric, dimeric and tetrameric enzyme (named maspase 1, maspase 2 and maspase 3) with the activity of L-aspartase are obtained
maspase 2
-
insertion of a 15-peptide random peptide into the three domains of L-aspartase to enhance their mobility. After directed screening, the three isoforms of monomeric, dimeric and tetrameric enzyme (named maspase 1, maspase 2 and maspase 3) with the activity of L-aspartase are obtained
maspase 3
-
insertion of a 15-peptide random peptide into the three domains of L-aspartase to enhance their mobility. After directed screening, the three isoforms of monomeric, dimeric and tetrameric enzyme (named maspase 1, maspase 2 and maspase 3) with the activity of L-aspartase are obtained
aspartase
-
-
-
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
PATHWAY SOURCE
PATHWAYS
-
-, -, -, -
SYSTEMATIC NAME
IUBMB Comments
L-aspartate ammonia-lyase (fumarate-forming)
-
CAS REGISTRY NUMBER
COMMENTARY
9027-30-9
-
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
L-aspartic acid alpha-amide
(2E)-4-amino-4-oxobut-2-enoate + NH3
catalyzed by mutant enzyme K327N, no activity with wild-type enzyme
-
-
?
fumarate + NH3
L-aspartate
important enzyme in production of L-aspartate
-
-
r
L-aspartate
fumarate + NH3
highly selective towards L-aspartate
-
-
r
additional information
?
-
-
no activity with D-aspartic acid or crotonic acid, overview substrate specificity
-
?
additional information
?
-
-
L-aspartate ammonia-lyase and fumarate hydratase share extensive sequence homology
-
-
?
additional information
?
-
-
L-aspartate ammonia-lyase and fumarate hydratase share extensive sequence homology
-
-
?
additional information
?
-
-
enzyme synthesis is subject to catabolite repression by glucose and is suppressed under aerobic conditions
-
-
?
additional information
?
-
-
enzyme is essential for the catabolism of the glutamate
-
-
?
additional information
?
-
-
aspartase, fumarase and argininosuccinase are structurally-related enzymes
-
-
?
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
fumarate + NH3
L-aspartate
important enzyme in production of L-aspartate
-
-
r
additional information
?
-
-
L-aspartate ammonia-lyase and fumarate hydratase share extensive sequence homology
-
-
?
additional information
?
-
-
L-aspartate ammonia-lyase and fumarate hydratase share extensive sequence homology
-
-
?
additional information
?
-
-
enzyme synthesis is subject to catabolite repression by glucose and is suppressed under aerobic conditions
-
-
?
additional information
?
-
-
enzyme is essential for the catabolism of the glutamate
-
-
?
additional information
?
-
-
aspartase, fumarase and argininosuccinase are structurally-related enzymes
-
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Ca2+
Cd2+
Mg2+
Mn2+
Zn2+
additional information
Mg2+
-
maximal activity: 1.0 mM for native enzyme, 2.2 mM for monomeric mutant enzyme maspase 1, 1.8 mM for dimeric mutant enzyme maspase 2, 1.6 mM for tetrameric mutant enzyme maspase 3
additional information
-
absolute requirement for divalent metal ion at high pH value
additional information
-
absolute requirement for divalent metal ion at higher pH
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
aspartate beta-semialdehyde
-
i.e. aspartic beta-semialdehyde.Inactivates as an active-site directed agent
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
alpha-methyl-DL-aspartate
-
activates wild-type enzyme, mutant C-terminal-truncated and acetylated enzyme
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
700
-
activity in Escherichia coli BLR (DE3) cells after cloning the enzyme in the pLATE31 plasmid, composition and highly efficient expression under the control of the T7 phage promoter, pH and temperature not specified in the publication
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
8.5
deamination of L-aspartate, wild-type enzyme and mutant enzyme K327N
7.5
8.2
8.5
8.8
additional information
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
7 - 9
pH 7.0: about 40% of maximal activity, pH 9.0: about 90% of maximal activity, substrate: L-aspartate, wild-type enzyme
7.5 - 9
pH 7.5: about 70% of maximal activity, pH 9.0: about 85% of maximal activity, substrate: L-aspartic acid, mutant enzyme K327N
additional information
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
30
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
52000
56000
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
tetramer
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
K327N
mutant enzyme catalyzes the deamination of L-aspartic acid alpha-amide, 13.5fold increase in Km-value for L-aspartate compared to wild-type value
K140I
-
Km value 10fold higher than wild type, comparable increase in Ki for competitive inhibitors
K55R
-
completeley inactive and insoluble protein, reactivation by an artificial chaperone system including beta-cyclodextrin and cetyltrimethylammonium bromide
additional information
additional information
-
design of enzyme variants by ligation of subdomains with a random hexapeptide loop, variant with highest activity is a monomer with high thermotolerance
additional information
-
mutant N217K/T233R/V367G through enzyme modification by direct evolution in order to enhance enzymic activity
additional information
-
construction of hybrid enzyme from alpha-aspartyl dipeptidase and L-aspartase. The hybrid enzyme can be used in synthesis of the precursor for aspartame
additional information
-
insertion of a 15-peptide random peptide into the three domains of L-aspartase to enhance their mobility. After directed screening, the three isoforms of monomeric, dimeric and tetrameric enzyme (named maspase 1, maspase 2 and maspase 3) with the activity of L-aspartase are obtained
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
50
55
additional information
50
-
83% loss of activity (wild-type), 70% loss of activity (mutant enzyme) after 45 min
50
-
30 min, native enzyme loses 83% of initial activity, monomeric mutant enzyme maspase 1 loses 23% of initial activity, dimeric mutant enzyme maspase 2 loses 48% of initial activity, tetrameric mutant enzyme maspase 3 loses 59% of initial activity
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
after incubation in the presence of guanidine hydrochloride at a concentration of 1.1 M complete inactivation
-
enzyme is fairly stable in the presence of high concentrations of ammonium sulfate, potassium phosphate, and KCl
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
2-mercaptoethanol and dithiothreitol protect enzyme against inactivation during storage
-
4C, in the presence of various salts thiol compounds, or glycerol stable for 2 weeks
-
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
although Corynebacterium glutamicum posses the aspartate ammonia-lyase gene (aspA), aspartate aminotransferase (AAT) is necessary to cell growth and L-lysine production, because the activity of Corynebacterium glutamicum aspartate ammonia-lyase (AAL) is too low to maintain the cell growth. Escherichia coli aspartate ammonia-lyase helps to restore the cell growth of aspB-deleted strain and also leads to the accumulation of L-lysine, L-glutamate, pyruvate and 2-oxoglutarate
expression of His-tagged wild-type enzyme and His-tagged mutant enzyme K327N in Escherichia coli
cloning in the pLATE31 plasmid composition and highly efficient expression under the control of the T7 phage promoter in Escherichia coli BLR (DE3) cell. The use of the expression system allows an increase in aspartase activity in cells by 100times as compared to the initial strain. The maximal specific activity of cells reached 0.700 mM/mg/min
-
RENATURED/Commentary
ORGANISM
UNIPROT
LITERATURE
denaturation by addition of high concentrations of guanidine-HCl leads to reversible dissociation and reassembly of the tetrameric structure
-
renaturation by dialysis against 50 mM Tris-HCl buffer ,pH 8.0, at 4C for 2 days
-
reversible denaturation is influenced by various environmental factors
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
biotechnology
synthesis
biotechnology
-
enhancement of recombinant protein production in Escherichia coli by coproduction of aspartase. The excretion of acetate by the aerobic growth of Escherichia coli on glucose is a manifestation of imbalanced flux between glycolysis and the tricarboxylic acid (TCA) cycle. This may restrict the production of recombinant proteins in E. coli, due to the limited amounts of precursor metabolites produced in TCA cycle. To approach this issue, an extra supply of intermediate metabolites in TCA cycle is made by conversion of aspartate to fumarate, a reaction mediated by the activity of L-aspartate ammonia-lyase
synthesis
-
industrial production of L-aspartic acid using polyurethane-immobilized cells containing aspartase
synthesis
-
enzyme or whole cells are used for the industrial production of L-aspartate, which together with L-phenylalanine is the basis of the sweetener aspartame
synthesis
-
production of L-aspartic acid, which is used as an intermediate for aspartame, an artificial sweetener. It is also used as acidulant in pharmaceuticals and foods
synthesis
-
construction of hybrid enzyme from alpha-aspartyl dipeptidase and L-aspartase. The hybrid enzyme can be used in synthesis of the precursor for aspartame. Synthesizing aspartate from fumarate and NH4+, and then taking advantage of the catalytic action of alpha-aspartyl dipeptidase, the precursor of aspartame is produced
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Hanson, K.R.; Havir, E.A.
The enzymic elimination of ammonia
The Enzymes, 3rd Ed. (Boyer, P. D. , ed. )
7
75-166
1972
Anas platyrhynchos, Bacterium cadaveris, Escherichia coli, Frog, Gallus gallus, shark, Yersinia pestis
-
Suzuki, S.; Yamaguchi, J.; Tokushige, M.
Purification and molecular properties of aspartate from Escherichia coli
Biochim. Biophys. Acta
321
369-381
1973
Escherichia coli
Chibata, I.; Tosa, T.; Sato, T.;Sano, R.; Yamamoto, K.; Matuo, Y.
Immobilized L-aspartic acid
Methods Enzymol.
34
405-411
1974
Escherichia coli
Mizuta, K.; Tokushige, M.
Role of sulfhydryl groups in aspartase from Escherichia coli
Biochim. Biophys. Acta
403
221-231
1975
Escherichia coli
Tokushige, M.; Mizuta, K.
Activation of aspartase by glycerol
Biochem. Biophys. Res. Commun.
68
1082-1087
1976
Escherichia coli
Mizuta, K.; Tokushige, M.
Alteration of enzymatic properties upon trypsin-mediated activation
Biochim. Biophys. Acta
452
253-261
1976
Escherichia coli
Tokushige, M.; Eguchi, G.; Hirata, F.
Reversible denaturation of Escherichia coli aspartase
Biochim. Biophys. Acta
480
479-488
1977
Escherichia coli
Tokushige, M.; Eguchi, G.
Denaturant-mediated reactivation of aspartase, which has been otherwise irreversibly inactivated by various causes
Biochim. Biophys. Acta
522
243-250
1978
Escherichia coli
Yumoto, N.; Tokushige, M.; Hayashi, R.
Trypsin-mediated activation releasing carboxy-terminal peptides
Biochim. Biophys. Acta
616
319-328
1980
Escherichia coli
Watanabe, Y.; Iwakura, M.; Togushige, M.; Eguchi, G.
Subunit arrangement of Escherichia coli aspartase
Biochim. Biophys. Acta
661
261-266
1981
Escherichia coli
Takagi, J.S.; Ida, N.; Tokushige, M.; Sakamoto, H.; Shimura, Y.
Cloning and nucleotide sequence of the aspartase gene of Escherichia coli W
Nucleic Acids Res.
13
2063-2074
1985
Escherichia coli
Tokushige, M.
Aspartate ammonia-lyase
Methods Enzymol.
113
618-627
1985
Escherichia coli, Hafnia alvei, Pseudomonas fluorescens
Takagi, J.S.; Tokushige, M.; Shimura, Y.; Kanehisa, M.
L-Aspartate ammonia-lyase and fumarate hydratase share extensive sequence homology
Biochem. Biophys. Res. Commun.
138
568-572
1986
Escherichia coli, Pseudomonas fluorescens
Woods, S.A.; Miles, J.S.; Roberts, R.E.; Guest, J.R.
Structural and functional relationships between fumarase and aspartase
Biochem. J.
237
547-557
1986
Escherichia coli, Serratia marcescens
Fusee, M.
Industrial production of L-aspartic acid using polyurethane-immobilized cells containing aspartase
Methods Enzymol.
136
463-471
1987
Escherichia coli
-
Falzone, C.J.; Karsten, W.E.; Conley, J.D.; Viola, R.E.
L-Aspartase from Escherichia coli: substrate specificity and role of divalent metal ions
Biochemistry
27
9089-9093
1988
Escherichia coli
Woods, S.A.; Miles, J.S.; Guest, J.R.
Sequence homologies between argininosuccinase, aspartase and fumarase: A family of structurally-related enzymes
FEMS Microbiol. Lett.
51
181-186
1988
Escherichia coli, Pseudomonas fluorescens
-
Lu, J.; Zhang, J.; Zhang, H.; Wang, X.
Studies on properties of mutants of aspartase from Escherichia coli W
Ann. N. Y. Acad. Sci.
864
631-635
1988
Escherichia coli
Karsten, W.E.; Viola, R.E.
Kinetic studies of L-aspartase from Escherichia coli: pH-dependent activity changes
Arch. Biochem. Biophys.
287
60-67
1991
Escherichia coli, Hafnia alvei
Ma, L.; Cao, S.G.; Yan, B.X.; Zuo, H.T.; Ding, Z.T.; Meng, Q.F.; Cheng, Y.H.
Induced rebuilding of aspartase conformation
Ann. N. Y. Acad. Sci.
672
60-65
1992
Escherichia coli
Murase, S.; Yumoto, N.
Characterization of three types of aspartase activated by site-directed mutagenesis, limited proteolysis, and acetylation
J. Biochem.
114
735-739
1993
Escherichia coli
Shi, W.; Kidd, R.; Giorgianni, F.; Schindler, J.F.; Viola, R.E.; Farber, G.K.
Crystallization and preliminary X-ray studies of L-aspartase from Escherichia coli
J. Mol. Biol.
234
1248-1249
1993
Escherichia coli
Zhang, H.Y.; Zhang, J.; Lin, L.; Du, W.Y.; Lu, J.
Enhancement of stability and activity of aspartase by random and site-directed mutagenesis
Biochem. Biophys. Res. Commun.
192
15-21
1993
Escherichia coli
Paulsen, J.; Hustedt, H.
Extractive purification of aspartase from Escherichia coli K12
Methods Enzymol.
228
590-599
1994
Escherichia coli, Pseudomonas fluorescens
Zhang, H.; Wang, X.; Zhang, J.; Lu, J.; Du, W.
Purification of L-aspartase by gene fusion
Ann. N. Y. Acad. Sci.
799
429-433
1996
Escherichia coli
Sjstrm, I.; Grndahl, H.; Falk, G.; Kronvall, G.; Ullberg, M.
Purification and characterization of a plasminogen-binding protein from Haemophilus influenzae. Sequence determination reveals identity with aspartase
Biochim. Biophys. Acta
1324
182-190
1997
Bacillus subtilis, Escherichia coli, Haemophilus influenzae, Haemophilus influenzae HI-23459, Serratia marcescens
Shi, W.; Dunbar, J.; Jayasekera, M.M.K.; Viola, R.E.; Farber, G.K.
The structure of L-aspartate ammonia-lyase from Escherichia coli
Biochemistry
36
9136-9144
1997
Escherichia coli (P0AC38), Escherichia coli
Giorgianni, F.; Beranova, S.; Wesdemiotis, C.; Viola, R.E.
Mapping the mechanism-based modification sites in L-aspartase from Escherichia coli
Arch. Biochem. Biophys.
341
329-336
1997
Escherichia coli
Kawata, Y.; Tamura, K.; Yano, S.; Mizobata, T.; Nagai, J.; Esaki, N.; Soda, K.; Tokushige, M.; Yumoto, N.
Purification and characterization of thermostable aspartase from Bacillus sp. YM55-1
Arch. Biochem. Biophys.
366
40-46
1999
Bacillus sp. (in: Bacteria), Bacillus sp. (in: Bacteria) YM55-1, Escherichia coli, Pseudomonas fluorescens
Viola, R.E.
L-aspartase: new tricks from an old enzyme
Adv. Enzymol. Relat. Areas Mol. Biol.
74
295-341
2000
Escherichia coli, Hafnia alvei, Haemophilus influenzae, Pseudomonas fluorescens
Jayasekera, M.M.; Viola, R.E.
Recovery of catalytic activity from an inactive aggregated mutant of l-aspartase
Biochem. Biophys. Res. Commun.
264
596-600
1999
Escherichia coli
Wang, L.J.; Kong, X.D.; Zhang, H.Y.; Wang, X.P.; Zhang, J.
Enhancement of the activity of l-aspartase from Escherichia coli W by directed evolution
Biochem. Biophys. Res. Commun.
276
346-349
2000
Escherichia coli
Kong, X.; Li, Z.; Gou, X.; Zhu, S.; Zhang, H.; Wang, X.; Zhang, J.
A monomeric L-aspartase obtained by in vitro selection
J. Biol. Chem.
277
24289-24293
2002
Escherichia coli
Tosa, T.; Shibatani, T.
Industrial applications of immobilized biocatalysts in Japan
Ann. N. Y. Acad. Sci.
750
364-375
1995
Escherichia coli
Sheng, Y.; Li, S.; Gou, X.; Kong, X.; Wang, X.; Sun, Y.; Zhang, J.
The hybrid enzymes from alpha-aspartyl dipeptidase and L-aspartase
Biochem. Biophys. Res. Commun.
331
107-112
2005
Escherichia coli
Asano, Y.; Kira, I.; Yokozeki, K.
Alteration of substrate specificity of aspartase by directed evolution
Biomol. Eng.
22
95-101
2005
Escherichia coli (P0AC38), Escherichia coli
Gou, X.j.; Li, S.; Kong, X.d.; Liu, W.; Sun, Y.h.; Zhang, J.
Directed evolution of L-aspartase by mobility of domains
Chem. Res. Chin. Univ.
20
50-54
2004
Escherichia coli, Escherichia coli J2
-
Wang, Z.W.; Chen, Y.; Chao, Y.P.
Enhancement of recombinant protein production in Escherichia coli by coproduction of aspartase
J. Biotechnol.
124
403-411
2006
Escherichia coli
Novikov, A.; Derbikov, D.; Shaposhnikova, O.; Gubanova, T.; Kameneva, S.; Yanenko, A.
The highly efficient expression of the aspartase gene (L-aspartate ammonia-lyase) in Escherichia coli cells
Appl. Biochem. Microbiol.
51
751-756
2015
Escherichia coli, Escherichia coli VKPM V-7188
-
Xu, J.; Zhang, J.; Guo, Y.; Zhang, W.
Genetically modifying aspartate aminotransferase and aspartate ammonia-lyase affects metabolite accumulation in L-lysine producing strain derived from Corynebacterium glutamicum ATCC13032
J. Mol. Catal. B
113
82-89
2015
Escherichia coli (P0AC38), Escherichia coli MG1655 (P0AC38)
-
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