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Literature summary for 2.4.3.1 extracted from

  • Choi, Y.H.; Kim, J.H.; Park, J.H.; Lee, N.; Kim, D.H.; Jang, K.S.; Park, I.H.; Kim, B.G.
    Protein engineering of alpha2,3/2,6-sialyltransferase to improve the yield and productivity of in vitro sialyllactose synthesis (2014), Glycobiology, 24, 159-169.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BW25113 (DE3) cells Photobacterium damselae

Protein Variants

Protein Variants Comment Organism
I411A the mutant exhibits 169% of wild type activity Photobacterium damselae
I411T/L433T the mutant exhibits 5fold enhancement of the enzyme's specific activity compared with the wild type enzyme Photobacterium damselae
K135A the mutant exhibits 8% of wild type activity Photobacterium damselae
L112A the mutant exhibits 61% of wild type activity Photobacterium damselae
L433A the mutant exhibits 114% of wild type activity Photobacterium damselae
L433S the mutant exhibits 3fold enhancement of the enzyme's specific activity compared with the wild type enzyme Photobacterium damselae
L433T the mutant exhibits 3fold enhancement of the enzyme's specific activity compared with the wild type enzyme Photobacterium damselae
N157A the mutant exhibits 32% of wild type activity Photobacterium damselae
R136A the mutant exhibits 10% of wild type activity Photobacterium damselae
S142A the mutant exhibits 38% of wild type activity Photobacterium damselae
T344A the mutant exhibits 38% of wild type activity Photobacterium damselae
W347A the mutant exhibits 16% of wild type activity Photobacterium damselae

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.27
-
CMP-N-acetylneuraminate mutant enzyme L433T, at pH 8.0 and 22°C Photobacterium damselae
3.37
-
CMP-N-acetylneuraminate mutant enzyme I411T/L433T, at pH 8.0 and 22°C Photobacterium damselae
5.87
-
CMP-N-acetylneuraminate wild type enzyme, at pH 8.0 and 22°C Photobacterium damselae
6.06
-
CMP-N-acetylneuraminate mutant enzyme I411T, at pH 8.0 and 22°C Photobacterium damselae
7.62
-
lactose wild type enzyme, at pH 8.0 and 22°C Photobacterium damselae
8.15
-
CMP-N-acetylneuraminate mutant enzyme L433S, at pH 8.0 and 22°C Photobacterium damselae
10.7
-
CMP-N-acetylneuraminate mutant enzyme I411T/L433S, at pH 8.0 and 22°C Photobacterium damselae
27.3
-
lactose mutant enzyme I411T, at pH 8.0 and 22°C Photobacterium damselae
59.7
-
lactose mutant enzyme L433S, at pH 8.0 and 22°C Photobacterium damselae
61.1
-
lactose mutant enzyme L433T, at pH 8.0 and 22°C Photobacterium damselae
62
-
lactose mutant enzyme I411T/L433T, at pH 8.0 and 22°C Photobacterium damselae
75.7
-
lactose mutant enzyme I411T/L433S, at pH 8.0 and 22°C Photobacterium damselae

Organism

Organism UniProt Comment Textmining
Photobacterium damselae
-
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA column chromatography Photobacterium damselae

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
CMP-N-acetylneuraminate + lactose
-
Photobacterium damselae CMP + 6'-sialyllactose
-
?

Synonyms

Synonyms Comment Organism
alpha 2,6-sialyltransferase
-
Photobacterium damselae
alpha2,6ST
-
Photobacterium damselae

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.3
-
lactose wild type enzyme, at pH 8.0 and 22°C Photobacterium damselae
4.32
-
CMP-N-acetylneuraminate wild type enzyme, at pH 8.0 and 22°C Photobacterium damselae
10.7
-
CMP-N-acetylneuraminate mutant enzyme L433T, at pH 8.0 and 22°C Photobacterium damselae
10.9
-
CMP-N-acetylneuraminate mutant enzyme I411T, at pH 8.0 and 22°C Photobacterium damselae
16.9
-
CMP-N-acetylneuraminate mutant enzyme I411T/L433S, at pH 8.0 and 22°C Photobacterium damselae
18.8
-
CMP-N-acetylneuraminate mutant enzyme I411T/L433T, at pH 8.0 and 22°C Photobacterium damselae
20.3
-
CMP-N-acetylneuraminate mutant enzyme L433S, at pH 8.0 and 22°C Photobacterium damselae
25.4
-
lactose mutant enzyme I411T, at pH 8.0 and 22°C Photobacterium damselae
76.5
-
lactose mutant enzyme L433T, at pH 8.0 and 22°C Photobacterium damselae
85
-
lactose mutant enzyme L433S, at pH 8.0 and 22°C Photobacterium damselae
102
-
lactose mutant enzyme I411T/L433S, at pH 8.0 and 22°C Photobacterium damselae
110
-
lactose mutant enzyme I411T/L433T, at pH 8.0 and 22°C Photobacterium damselae

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.58
-
lactose wild type enzyme, at pH 8.0 and 22°C Photobacterium damselae
0.74
-
CMP-N-acetylneuraminate wild type enzyme, at pH 8.0 and 22°C Photobacterium damselae
0.93
-
lactose mutant enzyme I411T, at pH 8.0 and 22°C Photobacterium damselae
1.26
-
lactose mutant enzyme L433T, at pH 8.0 and 22°C Photobacterium damselae
1.35
-
lactose mutant enzyme I411T/L433S, at pH 8.0 and 22°C Photobacterium damselae
1.43
-
lactose mutant enzyme L433S, at pH 8.0 and 22°C Photobacterium damselae
1.58
-
CMP-N-acetylneuraminate mutant enzyme I411T/L433S, at pH 8.0 and 22°C Photobacterium damselae
1.78
-
lactose mutant enzyme I411T/L433T, at pH 8.0 and 22°C Photobacterium damselae
1.8
-
CMP-N-acetylneuraminate mutant enzyme I411T, at pH 8.0 and 22°C Photobacterium damselae
2.61
-
CMP-N-acetylneuraminate mutant enzyme L433S, at pH 8.0 and 22°C Photobacterium damselae
4.7
-
CMP-N-acetylneuraminate mutant enzyme L433T, at pH 8.0 and 22°C Photobacterium damselae
5.6
-
CMP-N-acetylneuraminate mutant enzyme I411T/L433T, at pH 8.0 and 22°C Photobacterium damselae