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Literature summary for 3.2.1.157 extracted from

  • Rebuffet, E.; Barbeyron, T.; Jeudy, A.; Jam, M.; Czjzek, M.; Michel, G.
    Identification of catalytic residues and mechanistic analysis of family GH82 iota-carrageenases (2010), Biochemistry, 49, 7590-7599.
    View publication on PubMed
Search for more literature for 3.2.1.157

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli Origami (DE3) pLysS, BL21 (DE3), C41 (DE3), and C43 (DE3) cells Alteromonas fortis

Protein Variants

Protein Variants Comment Organism
D247A the enzymatic activity is completely abolished Alteromonas fortis
E245Q the enzymatic activity is completely abolished Alteromonas fortis
E310Q the Km value is reduced by a factor of about 4, the reduction in kcat by 50fold results in a substantial decrease in the overall enzyme efficiency (about 10fold) Alteromonas fortis
H281A the mutant shows 7fold decreased Km and 180fold decreased kcat value compared to the wild type enzyme with 4% relative efficiency Alteromonas fortis
Q222E the enzymatic activity is abolished Alteromonas fortis
Q222K the enzymatic activity is abolished Alteromonas fortis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.2
-
 iota-carrageenan mutant enzyme H281A, at 40°C in 10 mM Tris, pH 8.0, and 100 mM NaCl Alteromonas fortis
2.4
-
 iota-carrageenan mutant enzyme E310Q, at 40°C in 10 mM Tris, pH 8.0, and 100 mM NaCl Alteromonas fortis
8.5
-
 iota-carrageenan wild type enzyme, at 40°C in 10 mM Tris, pH 8.0, and 100 mM NaCl Alteromonas fortis

Organism

Organism UniProt Comment Textmining
Alteromonas fortis
-
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-Sepharose column chromatography, Source 15S column chromatography, gel filtration Alteromonas fortis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
iota-carrageenan + H2O
-
 Alteromonas fortis iota-neocarratetraose sulfate + iota-neocarrahexaose sulfate
-
 ?

Synonyms

Synonyms Comment Organism
Cgi
-
 Alteromonas fortis
iotase
-
 Alteromonas fortis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.1
-
 iota-carrageenan mutant enzyme H281A, at 40°C in 10 mM Tris, pH 8.0, and 100 mM NaCl Alteromonas fortis
13.8
-
 iota-carrageenan mutant enzyme E310Q, at 40°C in 10 mM Tris, pH 8.0, and 100 mM NaCl Alteromonas fortis
726.6
-
 iota-carrageenan wild type enzyme, at 40°C in 10 mM Tris, pH 8.0, and 100 mM NaCl Alteromonas fortis

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.42
-
 iota-carrageenan mutant enzyme H281A, at 40°C in 10 mM Tris, pH 8.0, and 100 mM NaCl Alteromonas fortis
5.75
-
 iota-carrageenan mutant enzyme E310Q, at 40°C in 10 mM Tris, pH 8.0, and 100 mM NaCl Alteromonas fortis
85.18
-
 iota-carrageenan wild type enzyme, at 40°C in 10 mM Tris, pH 8.0, and 100 mM NaCl Alteromonas fortis