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Literature summary extracted from

  • Yang, P.; Wang, J.; Pang, Q.; Zhang, F.; Wang, J.; Wang, Q.; Qi, Q.
    Pathway optimization and key enzyme evolution of N-acetylneuraminate biosynthesis using an in vivo aptazyme-based biosensor (2017), Metab. Eng., 43, 21-28 .
    View publication on PubMed

Application

EC Number Application Comment Organism
2.5.1.56 synthesis design of an in vivo NeuAc biosensor and application for genetic screening of a mutant library of NeuAc producers. A Ni2+-based selection system couples the cell growth with in vivo NeuAc concentration. The final strain produces up to 8.31 g/l NeuAc in minimal medium using glucose as sole carbon source Escherichia coli

Protein Variants

EC Number Protein Variants Comment Organism
2.5.1.56 A506G/G420A increase in specific activity Escherichia coli
2.5.1.56 C914T decrease in specific activity Escherichia coli
2.5.1.56 C914T/G1018A no significant change in specific activity Escherichia coli
2.5.1.56 G1018A no significant change in specific activity Escherichia coli
2.5.1.56 G283A no significant change in specific activity Escherichia coli
2.5.1.56 T74C increase in specific activity Escherichia coli
2.5.1.56 T74C/A506G increase in specific activity Escherichia coli

Organism

EC Number Organism UniProt Comment Textmining
2.5.1.56 Escherichia coli
-
-
-
2.5.1.56 Escherichia coli K1
-
-
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.5.1.56 17.93
-
mutant C914T, pH 7.0, 37°C Escherichia coli
2.5.1.56 26.5
-
mutant G1018A, pH 7.0, 37°C Escherichia coli
2.5.1.56 27.47
-
mutant C914T/G1018A, pH 7.0, 37°C Escherichia coli
2.5.1.56 28.11
-
wild-type, pH 7.0, 37°C Escherichia coli
2.5.1.56 28.96
-
mutant G283A, pH 7.0, 37°C Escherichia coli
2.5.1.56 30.76
-
mutant T74C, pH 7.0, 37°C Escherichia coli
2.5.1.56 32.48
-
mutant T74C/A506G, pH 7.0, 37°C Escherichia coli
2.5.1.56 35.71
-
mutant A506G/G420A, pH 7.0, 37°C Escherichia coli

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.5.1.56 phosphoenolpyruvate + N-acetyl-D-mannosamine + H2O
-
Escherichia coli phosphate + N-acetylneuraminate
-
?
2.5.1.56 phosphoenolpyruvate + N-acetyl-D-mannosamine + H2O
-
Escherichia coli K1 phosphate + N-acetylneuraminate
-
?