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Literature summary extracted from

  • North, R.A.; Kessans, S.A.; Atkinson, S.C.; Suzuki, H.; Watson, A.J.; Burgess, B.R.; Angley, L.M.; Hudson, A.O.; Varsani, A.; Griffin, M.D.; Fairbanks, A.J.; Dobson, R.C.
    Cloning, expression, purification, crystallization and preliminary X-ray diffraction studies of N-acetylneuraminate lyase from methicillin-resistant Staphylococcus aureus (2013), Acta Crystallogr. Sect. F, 69, 306-312.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.1.3.3 gene nanA, DNA and amino acid sequence determination and analysis, sequence comparison, expression in Escherichia coli strain BL21(DE3) Staphylococcus aureus

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
4.1.3.3 purified recombinant enzyme, hanging drop vapour diffusion method, 0.002-0.0025 ml of 10 mg/ml protein in 20 mM Tris-HCl, pH 8.0, is mixed with 0.0017-0.0022 ml of reservoir solution containing 25% w/v PEG 3350, 200 mM ammonium sulfate, 100 mM Bis-Tris pH 5.5, equilibration against 1 ml of reservoir solution, 8-20°C, method optimization, X-ray diffraction structure determination and analysis at 1.7 A resolution, molecular replacement Staphylococcus aureus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
4.1.3.3 N-acetyl-D-mannosamine + pyruvate Staphylococcus aureus the enzyme catalyzes the retro-aldol cleavage of N-acetylneuraminic acid to form N-acetyl-D-mannosamine and pyruvate N-acetylneuraminate
-
 r
4.1.3.3 N-acetyl-D-mannosamine + pyruvate Staphylococcus aureus MRSA252 the enzyme catalyzes the retro-aldol cleavage of N-acetylneuraminic acid to form N-acetyl-D-mannosamine and pyruvate N-acetylneuraminate
-
 r

Organism

EC Number Organism UniProt Comment Textmining
4.1.3.3 Staphylococcus aureus Q6GK01 subspecies aureus, methicillin-resistant, gene nanA
-
4.1.3.3 Staphylococcus aureus MRSA252 Q6GK01 subspecies aureus, methicillin-resistant, gene nanA
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.1.3.3 recombinant enzyme from Escherichia coli strain BL21(DE3) by anion exchange and hydrophobic interaction chromatography, and gel filtration to over 95% purity Staphylococcus aureus

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
4.1.3.3 11
-
 purified recombinant enzyme, pH 8.0, temperature not specified in the publication Staphylococcus aureus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.1.3.3 N-acetyl-D-mannosamine + pyruvate the enzyme catalyzes the retro-aldol cleavage of N-acetylneuraminic acid to form N-acetyl-D-mannosamine and pyruvate Staphylococcus aureus N-acetylneuraminate
-
 r
4.1.3.3 N-acetyl-D-mannosamine + pyruvate the enzyme catalyzes the retro-aldol cleavage of N-acetylneuraminic acid to form N-acetyl-D-mannosamine and pyruvate Staphylococcus aureus MRSA252 N-acetylneuraminate
-
 r
4.1.3.3 N-acetylneuraminate
-
 Staphylococcus aureus N-acetyl-D-mannosamine + pyruvate
-
 r
4.1.3.3 N-acetylneuraminate
-
 Staphylococcus aureus MRSA252 N-acetyl-D-mannosamine + pyruvate
-
 r

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
4.1.3.3 8
-
 assay at Staphylococcus aureus