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Literature summary for 3.2.1.83 extracted from

  • Matard-Mann, M.; Bernard, T.; Leroux, C.; Barbeyron, T.; Larocque, R.; Prechoux, A.; Jeudy, A.; Jam, M.; Nyvall Collen, P.; Michel, G.; Czjzek, M.
    Structural insights into marine carbohydrate degradation by family GH16 kappa-carrageenases (2017), J. Biol. Chem., 292, 19919-19934 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Zobellia galactanivorans
expressed in Escherichia coli C43(DE3) cells Pseudoalteromonas carrageenovora

Crystallization (Commentary)

Crystallization (Comment) Organism
hanging drop vapor diffusion method, 28-29% PEG 3350, 100 mM MES, pH 6.5, and 0.3 M NaNO3 Zobellia galactanivorans
mutant enzyme E168D in complex with kappa-neocarratetraose, hanging drop vapor diffusion method, using 1.0 M sodium citrate and 100 mM cacodylate at pH 6.5 Pseudoalteromonas carrageenovora

Protein Variants

Protein Variants Comment Organism
E168D inactive Pseudoalteromonas carrageenovora
Q171A the mutant shows 157% of wild type activity Pseudoalteromonas carrageenovora
R151A the mutant shows 199% of wild type activity Pseudoalteromonas carrageenovora
R196A the mutant shows 3% of wild type activity Pseudoalteromonas carrageenovora
R260A the mutant shows less than 1% of wild type activity Pseudoalteromonas carrageenovora
R92A the mutant shows 102% of wild type activity Pseudoalteromonas carrageenovora
W266A the mutant shows 5% of wild type activity Pseudoalteromonas carrageenovora

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.000385
-
kappa-carrageen mutant enzyme Q171A, at pH 7.0 and 40°C Pseudoalteromonas carrageenovora
0.000683
-
kappa-carrageen wild type enzyme, at pH 7.0 and 40°C Pseudoalteromonas carrageenovora
0.000822
-
kappa-carrageen wild type enzyme, at pH 7.0 and 40°C Zobellia galactanivorans
0.000899
-
kappa-carrageen mutant enzyme R92A, at pH 7.0 and 40°C Pseudoalteromonas carrageenovora
0.00115
-
kappa-carrageen mutant enzyme R196A, at pH 7.0 and 40°C Pseudoalteromonas carrageenovora
0.00121
-
kappa-carrageen mutant enzyme R151A, at pH 7.0 and 40°C Pseudoalteromonas carrageenovora
0.00263
-
kappa-carrageen mutant enzyme W266A, at pH 7.0 and 40°C Pseudoalteromonas carrageenovora

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
kappa-carrageen + H2O Zobellia galactanivorans
-
kappa-neocarrahexaose + kappa-neocarratetraose
-
?
kappa-carrageen + H2O Pseudoalteromonas carrageenovora
-
kappa-neocarratetraose + kappa-neocarrabiose
-
?
kappa-carrageen + H2O Pseudoalteromonas carrageenovora ATCC 43555T
-
kappa-neocarratetraose + kappa-neocarrabiose
-
?
kappa-carrageen + H2O Zobellia galactanivorans DsijT
-
kappa-neocarrahexaose + kappa-neocarratetraose
-
?

Organism

Organism UniProt Comment Textmining
Pseudoalteromonas carrageenovora P43478
-
-
Pseudoalteromonas carrageenovora ATCC 43555T P43478
-
-
Zobellia galactanivorans G0L921
-
-
Zobellia galactanivorans DsijT G0L921
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni affinity column chromatography and Sephadex gel filtration Pseudoalteromonas carrageenovora
Ni affinity column chromatography and Sephadex gel filtration Zobellia galactanivorans

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
kappa-carrageen + H2O
-
Zobellia galactanivorans kappa-neocarrahexaose + kappa-neocarratetraose
-
?
kappa-carrageen + H2O
-
Zobellia galactanivorans DsijT kappa-neocarrahexaose + kappa-neocarratetraose
-
?
kappa-carrageen + H2O
-
Pseudoalteromonas carrageenovora kappa-neocarratetraose + kappa-neocarrabiose
-
?
kappa-carrageen + H2O
-
Pseudoalteromonas carrageenovora ATCC 43555T kappa-neocarratetraose + kappa-neocarrabiose
-
?

Synonyms

Synonyms Comment Organism
CgkA
-
Pseudoalteromonas carrageenovora
CgkA
-
Zobellia galactanivorans
CgkAGH16
-
Pseudoalteromonas carrageenovora
CgkAGH16
-
Zobellia galactanivorans

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
6.13
-
kappa-carrageen mutant enzyme R196A, at pH 7.0 and 40°C Pseudoalteromonas carrageenovora
27.36
-
kappa-carrageen mutant enzyme W266A, at pH 7.0 and 40°C Pseudoalteromonas carrageenovora
120.17
-
kappa-carrageen mutant enzyme Q171A, at pH 7.0 and 40°C Pseudoalteromonas carrageenovora
135.84
-
kappa-carrageen wild type enzyme, at pH 7.0 and 40°C Pseudoalteromonas carrageenovora
183.2
-
kappa-carrageen mutant enzyme R92A, at pH 7.0 and 40°C Pseudoalteromonas carrageenovora
478.79
-
kappa-carrageen mutant enzyme R151A, at pH 7.0 and 40°C Pseudoalteromonas carrageenovora
914.65
-
kappa-carrageen wild type enzyme, at pH 7.0 and 40°C Zobellia galactanivorans

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
5310
-
kappa-carrageen mutant enzyme R196A, at pH 7.0 and 40°C Pseudoalteromonas carrageenovora
10400
-
kappa-carrageen mutant enzyme W266A, at pH 7.0 and 40°C Pseudoalteromonas carrageenovora
199000
-
kappa-carrageen wild type enzyme, at pH 7.0 and 40°C Pseudoalteromonas carrageenovora
204000
-
kappa-carrageen mutant enzyme R92A, at pH 7.0 and 40°C Pseudoalteromonas carrageenovora
312000
-
kappa-carrageen mutant enzyme Q171A, at pH 7.0 and 40°C Pseudoalteromonas carrageenovora
397000
-
kappa-carrageen mutant enzyme R151A, at pH 7.0 and 40°C Pseudoalteromonas carrageenovora
1110000
-
kappa-carrageen wild type enzyme, at pH 7.0 and 40°C Zobellia galactanivorans