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Literature summary for 3.5.1.1 extracted from

  • Maggi, M.; Chiarelli, L.R.; Valentini, G.; Scotti, C.
    Engineering of Helicobacter pylori L-asparaginase: characterization of two functionally distinct groups of mutants (2015), PLoS ONE, 10, e0117025.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) pLysS cells Helicobacter pylori

Protein Variants

Protein Variants Comment Organism
M121C mutants has nearly 2.5fold reduced catalytic activity compared to the wild type enzyme Helicobacter pylori
M121C/T169M the mutant has a preserved efficiency versus L-asparagine but is completely unable to carry out L-glutamine hydrolysis Helicobacter pylori
Q63E the mutant endows with a similar catalytic efficiency versus L-asparagine and halved glutaminase efficiency with respect to the wild type enzyme Helicobacter pylori
T169M mutants has nearly 4fold reduced catalytic activity compared to the wild type enzyme Helicobacter pylori
T16D inactive Helicobacter pylori
T95E inactive Helicobacter pylori

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
37000
-
x * 37000, SDS-PAGE Helicobacter pylori

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-asparagine + H2O Helicobacter pylori strongly preferred substrate L-aspartate + NH3
-
?
L-asparagine + H2O Helicobacter pylori CCUG 17874 strongly preferred substrate L-aspartate + NH3
-
?
L-glutamine + H2O Helicobacter pylori
-
L-glutamic acid + NH3
-
?
L-glutamine + H2O Helicobacter pylori CCUG 17874
-
L-glutamic acid + NH3
-
?

Organism

Organism UniProt Comment Textmining
Helicobacter pylori I0ZIE0
-
-
Helicobacter pylori CCUG 17874 I0ZIE0
-
-

Purification (Commentary)

Purification (Comment) Organism
HisTrap column chromatography and Superdex S200 gel filtration Helicobacter pylori

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-asparagine + H2O strongly preferred substrate Helicobacter pylori L-aspartate + NH3
-
?
L-asparagine + H2O strongly preferred substrate Helicobacter pylori CCUG 17874 L-aspartate + NH3
-
?
L-glutamine + H2O
-
Helicobacter pylori L-glutamic acid + NH3
-
?
L-glutamine + H2O
-
Helicobacter pylori CCUG 17874 L-glutamic acid + NH3
-
?

Subunits

Subunits Comment Organism
? x * 37000, SDS-PAGE Helicobacter pylori

Synonyms

Synonyms Comment Organism
ASNase
-
Helicobacter pylori
HPA
-
Helicobacter pylori
L-ASNase
-
Helicobacter pylori
type II L-asparaginase
-
Helicobacter pylori

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
53
-
the wild type enzyme shows 50% activity after 10 min incubation at 53°C Helicobacter pylori

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.83
-
L-asparagine mutant enzyme M121C, at pH 7.5 and 37°C Helicobacter pylori
6.53
-
L-asparagine mutant enzyme Q63E, at pH 7.5 and 37°C Helicobacter pylori
7.4
-
L-asparagine mutant enzyme T169M, at pH 7.5 and 37°C Helicobacter pylori
13.95
-
L-asparagine mutant enzyme M121C/T169M, at pH 7.5 and 37°C Helicobacter pylori
14.5
-
L-glutamine mutant enzyme Q63E, at pH 7.5 and 37°C Helicobacter pylori
19.26
-
L-asparagine wild type enzyme, at pH 7.5 and 37°C Helicobacter pylori
22.1
-
L-glutamine wild type enzyme, at pH 7.5 and 37°C Helicobacter pylori