Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 3.5.1.46 extracted from

  • Negoro, S.; Ohki, T.; Shibata, N.; Mizuno, N.; Wakitani, Y.; Tsurukame, J.; Matsumoto, K.; Kawamoto, I.; Takeo, M.; Higuchi, Y.
    X-ray crystallographic analysis of 6-aminohexanoate-dimer hydrolase: molecular basis for the birth of a nylon oligomer-degrading enzyme (2005), J. Biol. Chem., 280, 39644-39652.
    View publication on PubMed
Search for more literature for 3.5.1.46

Cloned(Commentary)

Cloned (Comment) Organism
expression of the His-tagged EII and the EII-EII'-hybrid Hyb24 in Escherichia coli strains JM109 and KP3998, respectively Arthrobacter sp.

Crystallization (Commentary)

Crystallization (Comment) Organism
purified recombinant Hyb24 by sitting drop vapor diffusion from 0.1 M MES, pH 6.5, 2.0–2.2 M ammonium sulfate, 0.1–0.2 M lithium sulfate, at 10°C, 2 ml of sample mixed with 2 ml of reservoir solution, preparation of HgCl2 heavy atom derivatives, X-ray diffraction structure determination and analysis at 1.8 A resolution Arthrobacter sp.

Protein Variants

Protein Variants Comment Organism
D181E site-directed mutagenesis of EII, the mutant shows reduced activity compared to the wild-type enzyme Arthrobacter sp.
D181H site-directed mutagenesis of EII, the mutant shows highly reduced activity compared to the wild-type enzyme Arthrobacter sp.
D181K site-directed mutagenesis of EII, nearly inactive mutant Arthrobacter sp.
D181N site-directed mutagenesis of EII, the mutant shows reduced activity compared to the wild-type enzyme Arthrobacter sp.
T3A/P4R/T5S/S8Q/D15G construction of a hybrid of isozymes EII and EII', termed Hyb24, by five amino acid replacement in EII', the mutant shows the same activity as EII' Arthrobacter sp.

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
N-(6-aminohexanoyl)-6-aminohexanoate + H2O Arthrobacter sp. the enzyme is responsible for the degradation of nylon-6 industrial production by-products 6-aminohexanoate
-
 ?

Organism

Organism UniProt Comment Textmining
Arthrobacter sp.
-
 formerly Flavobacterium sp., strain K172, enzyme form EII and cryptic enzyme form EII'
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged EII from Escherichia coli strain by nickel affinity chromatography, recombinant EII-EII'-hybrid Hyb24 from Escherichia coli strain KP3998 by anion exchange chromatography, gel filtration, and again anion exchange chromatography, both enzymes to homogeneity Arthrobacter sp.

Reaction

Reaction Comment Organism Reaction ID
N-(6-aminohexanoyl)-6-aminohexanoate + H2O = 2 6-aminohexanoate Ser112 is a nucleophilic catalytic residue, Asp181 and Asn226 are also required Arthrobacter sp.
a

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
4.16
-
 purified recombinant wild-type EII Arthrobacter sp.

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-nitrophenyl acetate + H2O
-
 Arthrobacter sp. 4-nitrophenol + acetate
-
 ?
4-nitrophenylbutyrate + H2O
-
 Arthrobacter sp. 4-nitrophenol + butyrate
-
 ?
additional information substrate specificity of EII and mutant Hyb24, no activity with D-Ala-D-Ala Arthrobacter sp. ?
-
 ?
N-(6-aminohexanoyl)-6-aminohexanoate + H2O the enzyme is responsible for the degradation of nylon-6 industrial production by-products Arthrobacter sp. 6-aminohexanoate
-
 ?
N-(6-aminohexanoyl)-6-aminohexanoate + H2O isozyme EII' shows only about 0.5% of the activity of EII Arthrobacter sp. 6-aminohexanoate
-
 ?

Subunits

Subunits Comment Organism
More analysis of domain structures of EII, cryptic enzyme form EII', and EII-EII'-hybrid Hyb24 Arthrobacter sp.

Synonyms

Synonyms Comment Organism
nylon oligomer hydrolase
-
 Arthrobacter sp.

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
 assay at Arthrobacter sp.

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
 assay at Arthrobacter sp.