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Literature summary for 2.5.1.18 extracted from

  • Kontur, W.S.; Olmsted, C.N.; Yusko, L.M.; Niles, A.V.; Walters, K.A.; Beebe, E.T.; Vander Meulen, K.A.; Karlen, S.D.; Gall, D.L.; Noguera, D.R.; Donohue, T.J.
    A heterodimeric glutathione S-transferase that stereospecifically breaks lignins beta(R)-aryl ether bond reveals the diversity of bacterial beta-etherases (2019), J. Biol. Chem., 294, 1877-1890 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expressed as His6-tagged protein in Escherichia coli BL21(DE3) cells Novosphingobium aromaticivorans

Protein Variants

Protein Variants Comment Organism
N14A the mutant of subunit A has a kcat value about 20fold lower and a Km value about 12.5fold higher than that of the wild type enzyme Novosphingobium aromaticivorans
S14A the mutant of subunit B has the same kcat and Km values as the wild type enzyme Novosphingobium aromaticivorans
S14A/S15A the double mutant has a kcat value similar to that of the wild type enzyme and a Km value that is 5-6fold higher than that of the wild type enzyme Novosphingobium aromaticivorans
S15A the mutant of subunit A has a kcat value similar to that of the wild type enzyme and a Km value that is 5-6fold higher than that of the wild type enzyme Novosphingobium aromaticivorans

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.02
-
beta-(2-methoxyphenoxy)-gamma-hydroxypropiovanillone wild type enzyme, at pH 8.0 and 30°C Novosphingobium aromaticivorans
0.022
-
beta-(2-methoxyphenoxy)-gamma-hydroxypropiovanillone mutant enzyme S14A, at pH 8.0 and 30°C Novosphingobium aromaticivorans
0.1
-
beta-(2-methoxyphenoxy)-gamma-hydroxypropiovanillone mutant enzyme S15A, at pH 8.0 and 30°C Novosphingobium aromaticivorans
0.12
-
beta-(2-methoxyphenoxy)-gamma-hydroxypropiovanillone mutant enzyme S14A/S15A, at pH 8.0 and 30°C Novosphingobium aromaticivorans
0.25
-
beta-(2-methoxyphenoxy)-gamma-hydroxypropiovanillone mutant enzyme N14A, at pH 8.0 and 30°C Novosphingobium aromaticivorans

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
59000
-
gel filtration Novosphingobium aromaticivorans

Organism

Organism UniProt Comment Textmining
Novosphingobium aromaticivorans Q2G4B4 and Q2G4B5 subunits BaeA and BaeB
-
Novosphingobium aromaticivorans DSM 12444 Q2G4B4 and Q2G4B5 subunits BaeA and BaeB
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA column chromatography Novosphingobium aromaticivorans

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
glutathione + beta-(2-methoxyphenoxy)-gamma-hydroxypropiovanillone the enzyme stereospecifically cleaves the beta(R)-aryl ether bond of the di-aromatic compound beta-(2-methoxyphenoxy)-gamma-hydroxypropiovanillone Novosphingobium aromaticivorans beta-(S)-hydroxypropiovanillonyl-glutathione + guaiacol
-
?
glutathione + beta-(2-methoxyphenoxy)-gamma-hydroxypropiovanillone the enzyme stereospecifically cleaves the beta(R)-aryl ether bond of the di-aromatic compound beta-(2-methoxyphenoxy)-gamma-hydroxypropiovanillone Novosphingobium aromaticivorans DSM 12444 beta-(S)-hydroxypropiovanillonyl-glutathione + guaiacol
-
?

Subunits

Subunits Comment Organism
heterodimer 1 * 29400 + 1 * 27000, SDS-PAGE Novosphingobium aromaticivorans

Synonyms

Synonyms Comment Organism
BaeAB
-
Novosphingobium aromaticivorans
beta-etherase
-
Novosphingobium aromaticivorans
glutathione S-transferase
-
Novosphingobium aromaticivorans
GST
-
Novosphingobium aromaticivorans
Saro_2873/Saro_2872
-
Novosphingobium aromaticivorans

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.14
-
beta-(2-methoxyphenoxy)-gamma-hydroxypropiovanillone mutant enzyme N14A, at pH 8.0 and 30°C Novosphingobium aromaticivorans
2.4
-
beta-(2-methoxyphenoxy)-gamma-hydroxypropiovanillone mutant enzyme S15A, at pH 8.0 and 30°C Novosphingobium aromaticivorans
2.8
-
beta-(2-methoxyphenoxy)-gamma-hydroxypropiovanillone mutant enzyme S14A, at pH 8.0 and 30°C Novosphingobium aromaticivorans
2.8
-
beta-(2-methoxyphenoxy)-gamma-hydroxypropiovanillone mutant enzyme S14A/S15A, at pH 8.0 and 30°C Novosphingobium aromaticivorans
2.9
-
beta-(2-methoxyphenoxy)-gamma-hydroxypropiovanillone wild type enzyme, at pH 8.0 and 30°C Novosphingobium aromaticivorans

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.5
-
beta-(2-methoxyphenoxy)-gamma-hydroxypropiovanillone mutant enzyme N14A, at pH 8.0 and 30°C Novosphingobium aromaticivorans
23
-
beta-(2-methoxyphenoxy)-gamma-hydroxypropiovanillone mutant enzyme S14A/S15A, at pH 8.0 and 30°C Novosphingobium aromaticivorans
23
-
beta-(2-methoxyphenoxy)-gamma-hydroxypropiovanillone mutant enzyme S15A, at pH 8.0 and 30°C Novosphingobium aromaticivorans
130
-
beta-(2-methoxyphenoxy)-gamma-hydroxypropiovanillone mutant enzyme S14A, at pH 8.0 and 30°C Novosphingobium aromaticivorans
150
-
beta-(2-methoxyphenoxy)-gamma-hydroxypropiovanillone wild type enzyme, at pH 8.0 and 30°C Novosphingobium aromaticivorans