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Literature summary for 3.4.21.26 extracted from

  • Shetty, R.; Vestergaard, M.; Jessen, F.; Haegglund, P.; Knorr, V.; Koehler, P.; Prakash, H.S.; Hobley, T.J.
    Discovery, cloning and characterisation of proline specific prolyl endopeptidase, a gluten degrading thermo-stable enzyme from Sphaerobacter thermophiles (2017), Enzyme Microb. Technol., 107, 57-63 .
    View publication on PubMed
Search for more literature for 3.4.21.26

Application

Application Comment Organism
food industry the enzyme can be used during mashing to produce gluten free beer Sphaerobacter thermophilus

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli DE3 cells Sphaerobacter thermophilus

General Stability

General Stability Organism
the purified enzyme is active both before and after cleaving with thrombin Sphaerobacter thermophilus

Inhibitors

Inhibitors Comment Organism Structure
Al3+
-
 Sphaerobacter thermophilus
Ca2+ 83.9% residual activity at 1 mM Sphaerobacter thermophilus
Cu2+ 91.2% residual activity at 1 mM Sphaerobacter thermophilus
EDTA 91.2% residual activity at 30 mM Sphaerobacter thermophilus
iodoacetate 98.3% residual activity at 1 mM Sphaerobacter thermophilus
K+ 88.8% residual activity at 1 mM Sphaerobacter thermophilus
Mg2+ 90.1 % residual activity at 1 mM Sphaerobacter thermophilus
Mn2+ 90.4% residual activity at 1 mM Sphaerobacter thermophilus
Na+ 92.3% residual activity at 1 mM Sphaerobacter thermophilus
phenylmethylsulfonyl fluoride 96% residual activity at 1 mM Sphaerobacter thermophilus
Zn2+ 91.3% residual activity at 1 mM Sphaerobacter thermophilus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.5
-
 benzyloxycarbonyl-Gly-L-Pro-4-nitroanilide at pH 6.6 and 60°C Sphaerobacter thermophilus

Metals/Ions

Metals/Ions Comment Organism Structure
Co2+ the enzyme is activated and stabilized by Co2+ (1 mM) Sphaerobacter thermophilus

Organism

Organism UniProt Comment Textmining
Sphaerobacter thermophilus D1C7Y4
-
-
Sphaerobacter thermophilus DSM 20745 D1C7Y4
-
-
Sphaerobacter thermophilus S 6022 D1C7Y4
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA column chromatography Sphaerobacter thermophilus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
alpha-gliadin + H2O i.e. PQPQLPYPQPQLPY Sphaerobacter thermophilus ?
-
 ?
alpha-gliadin + H2O i.e. PQPQLPYPQPQLPY Sphaerobacter thermophilus DSM 20745 ?
-
 ?
alpha-gliadin + H2O i.e. PQPQLPYPQPQLPY Sphaerobacter thermophilus S 6022 ?
-
 ?
barley malt + H2O
-
 Sphaerobacter thermophilus ?
-
 ?
barley malt + H2O
-
 Sphaerobacter thermophilus DSM 20745 ?
-
 ?
barley malt + H2O
-
 Sphaerobacter thermophilus S 6022 ?
-
 ?
benzyloxycarbonyl-Gly-L-Pro-4-nitroanilide + H2O
-
 Sphaerobacter thermophilus benzyloxycarbonyl-Gly-L-Pro + 4-nitroaniline
-
 ?
benzyloxycarbonyl-Gly-L-Pro-4-nitroanilide + H2O
-
 Sphaerobacter thermophilus DSM 20745 benzyloxycarbonyl-Gly-L-Pro + 4-nitroaniline
-
 ?
benzyloxycarbonyl-Gly-L-Pro-4-nitroanilide + H2O
-
 Sphaerobacter thermophilus S 6022 benzyloxycarbonyl-Gly-L-Pro + 4-nitroaniline
-
 ?
gamma-hordein + H2O i.e. SQQQFPQPQQPFPQQP Sphaerobacter thermophilus ?
-
 ?
gamma-hordein + H2O i.e. SQQQFPQPQQPFPQQP Sphaerobacter thermophilus DSM 20745 ?
-
 ?
gamma-hordein + H2O i.e. SQQQFPQPQQPFPQQP Sphaerobacter thermophilus S 6022 ?
-
 ?

Subunits

Subunits Comment Organism
? x * 77000, SDS-PAGE Sphaerobacter thermophilus

Synonyms

Synonyms Comment Organism
POP
-
 Sphaerobacter thermophilus
proline specific prolyl endopeptidase
-
 Sphaerobacter thermophilus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
63
-
-
 Sphaerobacter thermophilus

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
50 75 the enzyme shows high activity in the temperature range 50-75°C Sphaerobacter thermophilus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
50 63 the residual activity after 1 h incubation at 63°C is higher than 75%. The enzyme activity is very stable between the temperatures ranging from 50 to 63°C after incubation for 30 min. The activity decreases rapidly at temperature above 63°C. About 65% activity of the enzyme is lost when enzyme is exposed to 70-80°C for 30 min at pH 6.6. No activity is lost after 2 h incubation up to 30°C. However between 40 and 63°C, 30% of the enzyme activity is lost upon incubation for 1 h and a further 50% lost after 2 h of incubation. A higher temperature (70 and 80°C), the enzyme activity is completely lost after 2 h of incubation Sphaerobacter thermophilus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
60
-
 benzyloxycarbonyl-Gly-L-Pro-4-nitroanilide at pH 6.6 and 60°C Sphaerobacter thermophilus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.6
-
-
 Sphaerobacter thermophilus

pH Range

pH Minimum pH Maximum Comment Organism
5 8 the enzyme is most active from pH 5.0-8.0 Sphaerobacter thermophilus