BRENDA - Enzyme Database

Involvement of two distinct N-acetylglucosaminyltransferases and a dual-function deacetylase in neomycin biosynthesis

Yokoyama, K.; Yamamoto, Y.; Kudo, F.; Eguchi, T.; ChemBioChem 9, 865-869 (2008)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Cloned (Commentary)
Organism
2.4.1.283
co-expressed with molecular chaperone GroES and GroEL in Escherichia coli BL21(DE3) cells
Streptomyces sp.
2.4.1.283
expressed as a His-tagged fusion protein in Escherichia coli
Streptomyces fradiae
2.4.1.285
expressed as a His-tagged fusion protein in Escherichia coli
Streptomyces fradiae
2.4.1.285
expressed in Escherichia coli BL21(DE3) cells
Streptomyces sp.
3.5.1.112
expressed as a His-tagged fusion protein in Escherichia coli
Streptomyces fradiae
3.5.1.113
expressed as a His-tagged fusion protein in Escherichia coli
Streptomyces fradiae
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
2.4.1.285
Ca2+
about 55% residual activity in the presence of 2 mM
Streptomyces sp.
2.4.1.285
Co2+
about 90% residual activity in the presence of 2 mM
Streptomyces sp.
2.4.1.285
Cu2+
inhibits activity
Streptomyces fradiae
2.4.1.285
Cu2+
complete inhibition in the presence of 2 mM
Streptomyces sp.
2.4.1.285
EDTA
about 20% residual activity in the presence of 10 mM
Streptomyces sp.
2.4.1.285
Ni2+
about 80% residual activity in the presence of 2 mM
Streptomyces sp.
2.4.1.285
Zn2+
inhibits activity
Streptomyces fradiae
2.4.1.285
Zn2+
complete inhibition in the presence of 2 mM
Streptomyces sp.
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
2.4.1.285
Mg2+
-
Streptomyces fradiae
2.4.1.285
Mg2+
100% activity in the presence of 2 mM
Streptomyces sp.
2.4.1.285
Mn2+
-
Streptomyces fradiae
2.4.1.285
Mn2+
105% activity in the presence of 2 mM Mn2+ compared to Mg2+
Streptomyces sp.
Organism
EC Number
Organism
UniProt
Commentary
Textmining
2.4.1.283
Streptomyces fradiae
-
-
-
2.4.1.283
Streptomyces sp.
-
-
-
2.4.1.285
Streptomyces fradiae
-
-
-
2.4.1.285
Streptomyces sp.
-
-
-
3.5.1.112
Streptomyces fradiae
-
-
-
3.5.1.113
Streptomyces fradiae
-
-
-
Purification (Commentary)
EC Number
Purification (Commentary)
Organism
2.4.1.283
a poor amount is isolated using Ni-NTA chromatography. The cell-free extract of Escherichia coli expressing NeoD is used for enzymatic assays
Streptomyces fradiae
2.4.1.285
DEAE Sepharose column chromatography, Superdex Hi-Load 200 gel filtration, and Mono Q column chromatography
Streptomyces sp.
2.4.1.285
purified to homogeneity by DEAE Sepharose column chromatography, gel filtration, and MonoQ column chromatography
Streptomyces fradiae
3.5.1.112
purified with TALON cobalt affinity resin
Streptomyces fradiae
3.5.1.113
purified with TALON cobalt affinity resin
Streptomyces fradiae
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
2.4.1.283
additional information
other possible glycosyl donor, UDP-glucose, and other possible glycosyl acceptors, paromamine, neamine, and ribostamycin are not accepted
719295
Streptomyces fradiae
?
-
-
-
-
2.4.1.283
additional information
the enzyme is unable to catalyze the glycosylation of ribostamycin
719295
Streptomyces sp.
?
-
-
-
-
2.4.1.283
UDP-N-acetyl-alpha-D-glucosamine + 2-deoxystreptamine
-
719295
Streptomyces fradiae
UDP + 2'-N-acetylparomamine
-
-
-
?
2.4.1.283
UDP-N-acetyl-alpha-D-glucosamine + 2-deoxystreptamine
about 80% of 2-deoxystreptamine is conversed
719295
Streptomyces sp.
UDP + 2'-N-acetylparomamine
-
-
-
?
2.4.1.285
UDP-N-acetyl-alpha-D-glucosamine + ribostamycin
-
719295
Streptomyces sp.
UDP + 2'''-acetyl-6'''-hydroxyneomycin C
-
-
-
?
2.4.1.285
UDP-N-acetyl-alpha-D-glucosamine + ribostamycin
-
719295
Streptomyces fradiae
UDP + 2'''-acetyl-6'''-hydroxyneomycin C
-
-
-
?
3.5.1.112
N-acetylparomamine + H2O
-
719295
Streptomyces fradiae
paromamine + acetate
-
-
-
?
3.5.1.113
2'''-N-acetyl-6'''-deamino-6'''-hydroxyneomycin C + H2O
-
719295
Streptomyces fradiae
6'''-deamino-6'''-hydroxyneomycin C
-
-
-
?
Synonyms
EC Number
Synonyms
Commentary
Organism
2.4.1.283
neoD
-
Streptomyces fradiae
2.4.1.283
neoD
-
Streptomyces sp.
2.4.1.285
Neo15
-
Streptomyces sp.
2.4.1.285
neoK
-
Streptomyces fradiae
2.4.1.285
neoK
-
Streptomyces sp.
3.5.1.112
neoL
-
Streptomyces fradiae
3.5.1.113
neoL
-
Streptomyces fradiae
Temperature Optimum [C]
EC Number
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
2.4.1.283
28
-
assay at
Streptomyces fradiae
2.4.1.285
28
-
assay at
Streptomyces fradiae
3.5.1.112
28
-
assay at
Streptomyces fradiae
3.5.1.113
28
-
assay at
Streptomyces fradiae
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.4.1.283
7
-
assay at
Streptomyces fradiae
2.4.1.285
7
-
assay at
Streptomyces fradiae
3.5.1.112
7
-
assay at
Streptomyces fradiae
3.5.1.113
7
-
assay at
Streptomyces fradiae
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
2.4.1.283
co-expressed with molecular chaperone GroES and GroEL in Escherichia coli BL21(DE3) cells
Streptomyces sp.
2.4.1.283
expressed as a His-tagged fusion protein in Escherichia coli
Streptomyces fradiae
2.4.1.285
expressed as a His-tagged fusion protein in Escherichia coli
Streptomyces fradiae
2.4.1.285
expressed in Escherichia coli BL21(DE3) cells
Streptomyces sp.
3.5.1.112
expressed as a His-tagged fusion protein in Escherichia coli
Streptomyces fradiae
3.5.1.113
expressed as a His-tagged fusion protein in Escherichia coli
Streptomyces fradiae
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
2.4.1.285
Ca2+
about 55% residual activity in the presence of 2 mM
Streptomyces sp.
2.4.1.285
Co2+
about 90% residual activity in the presence of 2 mM
Streptomyces sp.
2.4.1.285
Cu2+
inhibits activity
Streptomyces fradiae
2.4.1.285
Cu2+
complete inhibition in the presence of 2 mM
Streptomyces sp.
2.4.1.285
EDTA
about 20% residual activity in the presence of 10 mM
Streptomyces sp.
2.4.1.285
Ni2+
about 80% residual activity in the presence of 2 mM
Streptomyces sp.
2.4.1.285
Zn2+
inhibits activity
Streptomyces fradiae
2.4.1.285
Zn2+
complete inhibition in the presence of 2 mM
Streptomyces sp.
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
2.4.1.285
Mg2+
-
Streptomyces fradiae
2.4.1.285
Mg2+
100% activity in the presence of 2 mM
Streptomyces sp.
2.4.1.285
Mn2+
-
Streptomyces fradiae
2.4.1.285
Mn2+
105% activity in the presence of 2 mM Mn2+ compared to Mg2+
Streptomyces sp.
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
2.4.1.283
a poor amount is isolated using Ni-NTA chromatography. The cell-free extract of Escherichia coli expressing NeoD is used for enzymatic assays
Streptomyces fradiae
2.4.1.285
DEAE Sepharose column chromatography, Superdex Hi-Load 200 gel filtration, and Mono Q column chromatography
Streptomyces sp.
2.4.1.285
purified to homogeneity by DEAE Sepharose column chromatography, gel filtration, and MonoQ column chromatography
Streptomyces fradiae
3.5.1.112
purified with TALON cobalt affinity resin
Streptomyces fradiae
3.5.1.113
purified with TALON cobalt affinity resin
Streptomyces fradiae
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
2.4.1.283
additional information
other possible glycosyl donor, UDP-glucose, and other possible glycosyl acceptors, paromamine, neamine, and ribostamycin are not accepted
719295
Streptomyces fradiae
?
-
-
-
-
2.4.1.283
additional information
the enzyme is unable to catalyze the glycosylation of ribostamycin
719295
Streptomyces sp.
?
-
-
-
-
2.4.1.283
UDP-N-acetyl-alpha-D-glucosamine + 2-deoxystreptamine
-
719295
Streptomyces fradiae
UDP + 2'-N-acetylparomamine
-
-
-
?
2.4.1.283
UDP-N-acetyl-alpha-D-glucosamine + 2-deoxystreptamine
about 80% of 2-deoxystreptamine is conversed
719295
Streptomyces sp.
UDP + 2'-N-acetylparomamine
-
-
-
?
2.4.1.285
UDP-N-acetyl-alpha-D-glucosamine + ribostamycin
-
719295
Streptomyces sp.
UDP + 2'''-acetyl-6'''-hydroxyneomycin C
-
-
-
?
2.4.1.285
UDP-N-acetyl-alpha-D-glucosamine + ribostamycin
-
719295
Streptomyces fradiae
UDP + 2'''-acetyl-6'''-hydroxyneomycin C
-
-
-
?
3.5.1.112
N-acetylparomamine + H2O
-
719295
Streptomyces fradiae
paromamine + acetate
-
-
-
?
3.5.1.113
2'''-N-acetyl-6'''-deamino-6'''-hydroxyneomycin C + H2O
-
719295
Streptomyces fradiae
6'''-deamino-6'''-hydroxyneomycin C
-
-
-
?
Temperature Optimum [C] (protein specific)
EC Number
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
2.4.1.283
28
-
assay at
Streptomyces fradiae
2.4.1.285
28
-
assay at
Streptomyces fradiae
3.5.1.112
28
-
assay at
Streptomyces fradiae
3.5.1.113
28
-
assay at
Streptomyces fradiae
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
2.4.1.283
7
-
assay at
Streptomyces fradiae
2.4.1.285
7
-
assay at
Streptomyces fradiae
3.5.1.112
7
-
assay at
Streptomyces fradiae
3.5.1.113
7
-
assay at
Streptomyces fradiae
General Information
EC Number
General Information
Commentary
Organism
2.4.1.285
metabolism
NeoK is a glycosyltransferase that catalyzes the retaining glycosylation of ribostamycin with UDP-GlcNAc as a sugar donor
Streptomyces fradiae
General Information (protein specific)
EC Number
General Information
Commentary
Organism
2.4.1.285
metabolism
NeoK is a glycosyltransferase that catalyzes the retaining glycosylation of ribostamycin with UDP-GlcNAc as a sugar donor
Streptomyces fradiae