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Literature summary extracted from

  • Leal, A.R.; Cruz, R.; Bur, D.; Huesgen, P.F.; Faro, R.; Manadas, B.; Wlodawer, A.; Faro, C.; Simoes, I.
    Enzymatic properties, evidence for in vivo expression, and intracellular localization of shewasin D, the pepsin homolog from Shewanella denitrificans (2016), Sci. Rep., 6, 23869 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.4.23.1 expressed in Escherichia coli C41(DE3) cells Shewanella denitrificans

Protein Variants

EC Number Protein Variants Comment Organism
3.4.23.1 D37A inactive Shewanella denitrificans

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.4.23.1 additional information not inhibited by Pefabloc, EDTA, E-64, amstatin, and dithiohtreitol Shewanella denitrificans
3.4.23.1 pepstatin complete inhibition at 0.001 mM Shewanella denitrificans

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.4.23.1 0.00867
-
 ((7-methoxycoumarin-4-yl)acetyl)-Lys-Lys-Pro-Ala-Glu-Phe-Phe-Ala-Leu-Lys-(2,4-dinitrophenyl) at pH 4.0 and 37°C Shewanella denitrificans

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
3.4.23.1 cytoplasm
-
 Shewanella denitrificans 5737
-

Organism

EC Number Organism UniProt Comment Textmining
3.4.23.1 Shewanella denitrificans Q12R32
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.4.23.1 Co2+-carged His-Trap HP column chromatography, pepstatin-agarose affinity chromatography, and Mono Q column chromatography Shewanella denitrificans

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.4.23.1 ((7-methoxycoumarin-4-yl)acetyl)-Lys-Lys-Pro-Ala-Glu-Phe-Phe-Ala-Leu-Lys-(2,4-dinitrophenyl) + H2O
-
 Shewanella denitrificans ((7-methoxycoumarin-4-yl)acetyl)-Lys-Lys-Pro-Ala-Glu-Phe + Phe-Ala-Leu-Lys-(2,4-dinitrophenyl)
-
 ?
3.4.23.1 additional information no activity with ((7-methoxycoumarin-4-yl)acetyl)-Lys-Leu-His-Pro-Glu-Val-Leu-Phe-Val-Leu-Glu-Lys-(2,4-dinitrophenyl), Arg-Glu-(EDANS)-Ile-His-Pro-Phe-His-Leu-Val-Ile-His-Thr-Lys-(DABCYL)-Arg, ((7-methoxycoumarin-4-yl)acetyl)-Lys-Ser-Glu-Val-Asn-Leu-Asp-Ala-Glu-Phe-Lys-(2,4-dinitrophenyl), or ((7-methoxycoumarin-4-yl)acetyl)-Lys-Ala-Leu-Ile-Pro-Ser-Tyr-Lys-Trp-Ser-Lys-(2,4-dinitrophenyl) Shewanella denitrificans ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.4.23.1 ? x * 50000, SDS-PAGE Shewanella denitrificans
3.4.23.1 ? x * 49025, calculated from amino acid sequence Shewanella denitrificans

Synonyms

EC Number Synonyms Comment Organism
3.4.23.1 shewasin D
-
 Shewanella denitrificans

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.4.23.1 30 37
-
 Shewanella denitrificans

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
3.4.23.1 10 40 maximum enzyme activity is detected between 30°C and 37°C. Above this temperature the activity abruptly decreases, with no detectable activity remaining at 50°C. At lower temperatures the activity decreases gradually, with the enzyme retaining 30% activity at 10°C Shewanella denitrificans

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.4.23.1 3.51
-
 ((7-methoxycoumarin-4-yl)acetyl)-Lys-Lys-Pro-Ala-Glu-Phe-Phe-Ala-Leu-Lys-(2,4-dinitrophenyl) at pH 4.0 and 37°C Shewanella denitrificans

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.4.23.1 3.5 4
-
 Shewanella denitrificans

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.4.23.1 3 5 maximum activity towards this substrate at pH values between pH 3.5 and 4.0, with a drastic decrease in activity of 50% and 70% at pH values 3.0 and 5.0, respectively Shewanella denitrificans

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.4.23.1 400
-
 ((7-methoxycoumarin-4-yl)acetyl)-Lys-Lys-Pro-Ala-Glu-Phe-Phe-Ala-Leu-Lys-(2,4-dinitrophenyl) at pH 4.0 and 37°C Shewanella denitrificans