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Literature summary for 3.5.1.1 extracted from

  • Kotzia, G.A.; Labrou, N.E.
    L-Asparaginase from Erwinia chrysanthemi 3937: cloning, expression and characterization (2006), J. Biotechnol., 127, 657-669.
    View publication on PubMed

Application

Application Comment Organism
medicine the enzyme can be efficiently immobilized on epoxy-activated Sepharose CL-6B. The immobilized enzyme retains most of its activity (60%) and shows high stability at 4°C. The approach offers the possibility of designing an L-asparaginase from Erwinia chrysanthemi bioreactor that can be operated over a long period of time with high efficiency, which can be used in leukaemia therapy Dickeya chrysanthemi

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli BL21(DE3)pLysS Dickeya chrysanthemi
expression in Escherichia coli strain BL21(DE3) Dickeya chrysanthemi

Protein Variants

Protein Variants Comment Organism
additional information immobilization of the purified recombinant enzyme on epoxy-activated resin, the immobilized enzyme retains 60% of maximal activity and is highly stable at 4°C, utilization as bioreactor Dickeya chrysanthemi

General Stability

General Stability Organism
the enzyme can be efficiently immobilized on epoxy-activated Sepharose CL-6B. The immobilized enzyme retains most of its activity (60%) and shows high stability at 4°C Dickeya chrysanthemi

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information kinetics Dickeya chrysanthemi
0.058
-
L-asparagine pH 8, 37°C Dickeya chrysanthemi
0.8
-
Nalpha-acetyl-Asn pH 8, 37°C Dickeya chrysanthemi
6.7
-
L-Gln pH 8.2, 37°C Dickeya chrysanthemi
18.8
-
succinamic acid pH 8, 37°C Dickeya chrysanthemi

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-asparagine + H2O Dickeya chrysanthemi
-
L-aspartate + NH3
-
?
L-asparagine + H2O Dickeya chrysanthemi 3937
-
L-aspartate + NH3
-
?

Organism

Organism UniProt Comment Textmining
Dickeya chrysanthemi
-
-
-
Dickeya chrysanthemi Q6Q4F3
-
-
Dickeya chrysanthemi 3937
-
-
-
Dickeya chrysanthemi 3937 Q6Q4F3
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Dickeya chrysanthemi
recombinant enzyme from Escherichia coli strain BL21(DE3) in a single step procedure to homogeneity by cation exchange Dickeya chrysanthemi

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-asparagine + H2O
-
Dickeya chrysanthemi L-aspartate + NH3
-
?
L-asparagine + H2O
-
Dickeya chrysanthemi 3937 L-aspartate + NH3
-
?
L-Gln + H2O
-
Dickeya chrysanthemi L-Glu + NH3
-
?
L-Gln + H2O
-
Dickeya chrysanthemi 3937 L-Glu + NH3
-
?
Nalpha-acetyl-L-Asn + H2O
-
Dickeya chrysanthemi Nalpha-acetyl-L-Asp + NH3
-
?
Nalpha-acetyl-L-Asn + H2O
-
Dickeya chrysanthemi 3937 Nalpha-acetyl-L-Asp + NH3
-
?
succinamic acid + H2O
-
Dickeya chrysanthemi succinate + NH3
-
?
succinamic acid + H2O
-
Dickeya chrysanthemi 3937 succinate + NH3
-
?

Synonyms

Synonyms Comment Organism
ASNase
-
Dickeya chrysanthemi
L-ASNase
-
Dickeya chrysanthemi
L-asparaginase
-
Dickeya chrysanthemi

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
37
-
particularly stable at Dickeya chrysanthemi
46
-
enzyme shows higher thermal stability at acidic to neutral pH values, lower stability is observed at alkaline pH. The inactivation profiles at pH 5.5 and 7 are biphasic and shows two clear transitions with inflection points corresponding to Tm values of 46.6°C and 62.5°C at pH 5.5, and 46.4 and 57.2°C at pH 7, respectively Dickeya chrysanthemi
47
-
enzyme shows higher thermal stability at acidic to neutral pH values, lower stability is observed at alkaline pH. The inactivation profiles at pH 5.5 and 7 are biphasic and shows two clear transitions with inflection points corresponding to Tm values of 46.6°C and 62.5°C at pH 5.5, and 46.4 and 57.2°C at pH 7, respectively Dickeya chrysanthemi
57
-
enzyme shows higher thermal stability at acidic to neutral pH values, lower stability is observed at alkaline pH. The inactivation profiles at pH 5.5 and 7 are biphasic and shows two clear transitions with inflection points corresponding to Tm values of 46.6°C and 62.5°C at pH 5.5, and 46.4 and 57.2°C at pH 7, respectively Dickeya chrysanthemi
63
-
enzyme shows higher thermal stability at acidic to neutral pH values, lower stability is observed at alkaline pH. The inactivation profiles at pH 5.5 and 7 are biphasic and shows two clear transitions with inflection points corresponding to Tm values of 46.6°C and 62.5°C at pH 5.5, and 46.4 and 57.2°C at pH 7, respectively Dickeya chrysanthemi

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.0043
-
L-Gln pH 8.2, 37°C Dickeya chrysanthemi
0.0108
-
Nalpha-acetyl-Asn pH 8, 37°C Dickeya chrysanthemi
0.0128
-
L-asparagine pH 8, 37°C Dickeya chrysanthemi
0.0239
-
succinamic acid pH 8, 37°C Dickeya chrysanthemi

pI Value

Organism Comment pI Value Maximum pI Value
Dickeya chrysanthemi pI calculated from sequence is 8.57
-
8.6