Literature summary for 3.4.21.B55 extracted from

Zeng, J.; Gao, X.; Dai, Z.; Tang, B.; Tang, X.F.
Effects of metal ions on stability and activity of hyperthermophilic pyrolysin and further stabilization of this enzyme by modification of a Ca2+-binding site (2014), Appl. Environ. Microbiol., 80, 2763-2772.

Search for more literature for 3.4.21.B55

Cloned(Commentary)

Cloned (Comment)	Organism
expression in Escherichia coli. At high temperatures, pyrolysin proform (Pls) is converted to mature pyrolysin (mPls) via autoprocessing of both the N- and C-terminal propeptides	Pyrococcus furiosus

Protein Variants

Protein Variants	Comment	Organism
D55A	after teatment at 95°C for 12 h the mutant enzyme displays similar levels of residual activity to that of the wild-type enzyme. When incubated with 2 mM EDTA at 95°C for 10 min, the mutant enzyme is completely inactivated, whereas the wild-type enzyme displays a residual activity of 52.6%	Pyrococcus furiosus
D55A/D58A	mutant enzyme is capable of maturation at 95°C and also exhibits thermal stability similar to wild-type enzyme in the absence of chelating agents	Pyrococcus furiosus
D55A/D58A/E59A	the proform of the mutant enzyme is unable to mature at 95°C	Pyrococcus furiosus
D55A/E59A	less stable than wild-type enzyme under non-chelating conditions	Pyrococcus furiosus
D55N/D58N/E59Q	less stable than wild-type enzyme under non-chelating conditions	Pyrococcus furiosus
D58A	after teatment at 95°C for 12 h the mutant enzyme displays similar levels of residual activity to that of the wild-type enzyme. When incubated with 2 mM EDTA at 95°C for 10 min, the mutant enzyme retains 9.6% of its original activity, whereas the wild-type enzyme displays a residual activity of 52.6%	Pyrococcus furiosus
D58A/E59A	less stable than wild-type enzyme under non-chelating conditions	Pyrococcus furiosus
D818N/D820N	half-life of the mutant enzyme at 95°C is 18 h, compared to 12 h measured for the wild-type enzyme. Inactivation of the enzymes at 100-115°C results in retention of a higher level of its original activity by the mutant enzyme compared to wild-type enzyme. The mutant enzyme displays higher specific activity towards azocasein than wild-type enzyme in the temperature range of 50-110°C. The mutant enzyme also shows higher initial velocity of hydrolysis than wild-type enzyme with azocasein or succinyl-Ala-Ala-Pro-Lys-4-nitroanilide as substrate	Pyrococcus furiosus
E59A	after teatment at 95°C for 12 h the mutant enzyme exhibits a slightly lower residual activity than wild-type enzyme. When incubated with 2 mM EDTA at 95°C for 10 min, the mutant enzyme is completely inactivated, whereas the wild-type enzyme displays a residual activity of 52.6%	Pyrococcus furiosus
R276A	after teatment at 95°C for 12 h the mutant enzyme displays similar levels of residual activity to that of the wild-type enzyme. When incubated with 2 mM EDTA at 95°C for 10 min, the mutant enzyme retains 9.6% of its original activity, whereas the wild-type enzyme displays a residual activity of 52.6%	Pyrococcus furiosus

General Stability

General Stability	Organism
binding of Ca2+ increases the stability of the PPC domain and, in turn, stabilizes the global structure of the enzyme by modulating interdomain interactions	Pyrococcus furiosus
supplementation of Na+, Ca2+, or Mg2+ salts at concentrations similar to those in seawater destabilizes recombinant pyrolysin. Ca2+ and Mg2+ are stronger denaturants than Na+ for pyrolysin. Ca2+ also acts a stabilizer of pyrolysin by binding to the enzyme at specific sites. The destabilizing effect of metal ions on pyrolysin appears to be related to the disturbance of surface electrostatic interactions of the enzyme	Pyrococcus furiosus

Organism

binding of Ca2+ increases the stability of the PPC domain and, in turn, stabilizes the global structure of the enzyme by modulating interdomain interactions

Pyrococcus furiosus

supplementation of Na+, Ca2+, or Mg2+ salts at concentrations similar to those in seawater destabilizes recombinant pyrolysin. Ca2+ and Mg2+ are stronger denaturants than Na+ for pyrolysin. Ca2+ also acts a stabilizer of pyrolysin by binding to the enzyme at specific sites. The destabilizing effect of metal ions on pyrolysin appears to be related to the disturbance of surface electrostatic interactions of the enzyme

Pyrococcus furiosus

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
extracellular	-	Pyrococcus furiosus	-	-

Metals/Ions

Metals/Ions	Comment	Organism
Ca2+	supplementation of Na+, Ca2+, or Mg2+ salts at concentrations similar to those in seawater leads to an increase in enzyme activity. The activity level increases with the increase of the salt concentrations from 0.1 mM to 10 mM	Pyrococcus furiosus
Mg2+	supplementation of Na+, Ca2+, or Mg2+ salts at concentrations similar to those in seawater leads to an increase in enzyme activity. The activity level increases with the increase of the salt concentrations from 5 mM to 50 mM	Pyrococcus furiosus
Na+	supplementation of Na+, Ca2+, or Mg2+ salts at concentrations similar to those in seawater leads to an increase in enzyme activity. The activity level increases with the increase of the salt concentrations from 40 mM to 600 mM	Pyrococcus furiosus

Organism

Organism	UniProt	Comment	Textmining
Pyrococcus furiosus	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
-	Pyrococcus furiosus

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
azocasein + H2O	-	Pyrococcus furiosus	?	-	?
succinyl-Ala-Ala-Pro-Lys-4-nitroanilide + H2O	-	Pyrococcus furiosus	succinyl-Ala-Ala-Pro-Lys + 4-nitroaniline	-	?

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
95	-	assay at	Pyrococcus furiosus
110	-	wild-type enzyme and mutant enzyme D818N/D820N	Pyrococcus furiosus

Temperature Range [°C]

Temperature Minimum [°C]	Temperature Maximum [°C]	Comment	Organism
90	120	90°C: about 60% of maximal activity, 120°C: about 70% of maximal activity	Pyrococcus furiosus

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
95	-	half-life of wild-type enzyme: 12 h, half-life of mutant enzyme D818N/D820N is 18 h	Pyrococcus furiosus
105	-	20 min, wild-type enzyme loses about 45% of its activity, mutant enzyme D818N/D820N loses about 30% of its activity	Pyrococcus furiosus
110	-	5 min, wild-type enzyme loses about 30% of its activity, mutant enzyme D818N/D820N loses about 20% of its activity	Pyrococcus furiosus
115	-	5 min, wild-type enzyme loses about 75% of its activity, mutant enzyme D818N/D820N loses about 70% of its activity	Pyrococcus furiosus

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.8	-	assay at	Pyrococcus furiosus