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

BRENDA support

Literature summary for 3.5.1.103 extracted from

  • Newton, G.L.; Ko, M.; Ta, P.; Av-Gay, Y.; Fahey, R.C.
    Purification and characterization of Mycobacterium tuberculosis 1D-myo-inosityl-2-acetamido-2-deoxy-alpha-D-glucopyranoside deacetylase, MshB, a mycothiol biosynthetic enzyme (2006), Protein Expr. Purif., 47, 542-550.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
1,7-phenanthroline 1 mM, slight enhancement of activity Mycobacterium tuberculosis

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Mycobacterium tuberculosis
Rv1170 is cloned to contain a C-terminal His-6 tag to facilitate purifiation, expression in Escherichia coli Mycobacterium tuberculosis

Inhibitors

Inhibitors Comment Organism Structure
1,10-phenanthroline 0.1 mM, 82% inhibition of the enzyme isolated on the Ni-affinity column. 0.1mM 1,10-phenanthroline produces no signifiant inhibition of the enzyme isolated on the Zn-affinity column and 10% activity remains after treatment with 1 mM 1,10-phenanthroline. MshB activity lost by incubation of the Ni enzyme with 1,10-phenanthroline can be restored following removal of 1,10-phenanthroline by incubation with 0.1 mM Zn2+, Ni2+, Mn2+, or Co2+, the latter promoting the highest activity, but Ca2+ and Mg2+ produce no restoration of activity; inhibition of the Zn enzyme by 1,10-phenanthroline is slower or less complete than inhibition of the Ni enzyme. MshB activity lost by incubation of the Ni enzyme with 1,10-phenanthroline can be restored following removal of 1,10-phenanthroline by incubation with 0.1 mM Zn2+, Ni2+, Mn2+, or Co2+, the latter promoting the highest activity. Ca2+ and Mg2+ produce no restoration of activity Mycobacterium tuberculosis
additional information no inhibition is observed with 0.1 M 1,7-phenanthroline Mycobacterium tuberculosis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.34
-
1-(2-acetamido-2-deoxy-alpha-D-glucopyranosyl)-1D-myo-inositol pH 7.5, 37┬░C Mycobacterium tuberculosis
0.34
-
N-deacetyl-N-formylmycothiol-monobromobimane conjugate pH 7.4, 37┬░C Mycobacterium tuberculosis

Metals/Ions

Metals/Ions Comment Organism Structure
Co2+ MshB activity lost by incubation of the Ni enzyme with 1,10-phenanthroline can be restored following removal of 1,10-phenanthroline by incubation with 0.1 mM Zn2+, Ni2+, Mn2+, or Co2+, the latter promoting the highest activity. Ca2+ and Mg2+ produce no restoration of activity Mycobacterium tuberculosis
Co2+ MshB contains a divalent transition metal ion essential for activity. MshB isolated on the Ni-affinity column contains 0.36 equivalent of Ca and 0.82 equivalent of Ni, and less than 0.08 equivalent of Zn per subunit. MshB isolated on the Zn-affinity column contains less than 0.1 equivalent of Ca and 2.3 equivalents of Zn, and less than 0.08 equivalent of Ni per subunit. MshB activity lost by incubation of the Ni enzyme with 1,10-phenanthroline can be restored following removal of 1,10-phenanthroline by incubation with 0.1 mM Zn2+, Ni2+, Mn2+, or Co2+, the latter promoting the highest activity, but Ca2+ and Mg2+ produce no restoration of activity Mycobacterium tuberculosis
Mn2+ MshB activity lost by incubation of the Ni enzyme with 1,10-phenanthroline can be restored following removal of 1,10-phenanthroline by incubation with 0.1 mM Zn2+, Ni2+, Mn2+, or Co2+, the latter promoting the highest activity. Ca2+ and Mg2+ produce no restoration of activity Mycobacterium tuberculosis
Mn2+ MshB contains a divalent transition metal ion essential for activity. MshB isolated on the Ni-affinity column contains 0.36 equivalent of Ca and 0.82 equivalent of Ni, and less than 0.08 equivalent of Zn per subunit. MshB isolated on the Zn-affinity column contains less than 0.1 equivalent of Ca and 2.3 equivalents of Zn, and less than 0.08 equivalent of Ni per subunit. MshB activity lost by incubation of the Ni enzyme with 1,10-phenanthroline can be restored following removal of 1,10-phenanthroline by incubation with 0.1 mM Zn2+, Ni2+, Mn2+, or Co2+, the latter promoting the highest activity, but Ca2+ and Mg2+ produce no restoration of activity Mycobacterium tuberculosis
Ni2+ MshB activity lost by incubation of the Ni enzyme with 1,10-phenanthroline can be restored following removal of 1,10-phenanthroline by incubation with 0.1 mM Zn2+, Ni2+, Mn2+, or Co2+, the latter promoting the highest activity. Ca2+ and Mg2+ produce no restoration of activity Mycobacterium tuberculosis
Ni2+ MshB contains a divalent transition metal ion essential for activity. MshB isolated on the Ni-affinity column contains 0.36 equivalent of Ca and 0.82 equivalent of Ni, and less than 0.08 equivalent of Zn per subunit. MshB isolated on the Zn-affinity column contains less than 0.1 equivalent of Ca and 2.3 equivalents of Zn, and less than 0.08 equivalent of Ni per subunit. MshB activity lost by incubation of the Ni enzyme with 1,10-phenanthroline can be restored following removal of 1,10-phenanthroline by incubation with 0.1 mM Zn2+, Ni2+, Mn2+, or Co2+, the latter promoting the highest activity, but Ca2+ and Mg2+ produce no restoration of activity Mycobacterium tuberculosis
Zn2+ contains an active site divalent transition metal. MshB activity lost by incubation of the Ni enzyme with 1,10-phenanthroline can be restored following removal of 1,10-phenanthroline by incubation with 0.1 mM Zn2+, Ni2+, Mn2+, or Co2+, the latter promoting the highest activity. Ca2+ and Mg2+ produce no restoration of activity Mycobacterium tuberculosis
Zn2+ MshB contains a divalent transition metal ion essential for activity. MshB isolated on the Ni-affinity column contains 0.36 equivalent of Ca and 0.82 equivalent of Ni, and less than 0.08 equivalent of Zn per subunit. MshB isolated on the Zn-affinity column contains less than 0.1 equivalent of Ca and 2.3 equivalents of Zn, and less than 0.08 equivalent of Ni per subunit. MshB activity lost by incubation of the Ni enzyme with 1,10-phenanthroline can be restored following removal of 1,10-phenanthroline by incubation with 0.1 mM Zn2+, Ni2+, Mn2+, or Co2+, the latter promoting the highest activity, but Ca2+ and Mg2+ produce no restoration of activity Mycobacterium tuberculosis

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
32000
-
2 * 32000, SDS-PAGE Mycobacterium tuberculosis
32000
-
2 * 32000, the enzyme appears to behave as a dimer at normal ionic strength but may associate to a tetramer at low ionic strength, SDS-PAGE Mycobacterium tuberculosis
32000
-
4 * 32000, the enzyme appears to behave as a dimer at normal ionic strength but may associate to a tetramer at low ionic strength, SDS-PAGE Mycobacterium tuberculosis
33423
-
2 * 33423, the enzyme appears to behave as a dimer at normal ionic strength but may associate to a tetramer at low ionic strength, calculated from sequence Mycobacterium tuberculosis
33423
-
4 * 33423, the enzyme appears to behave as a dimer at normal ionic strength but may associate to a tetramer at low ionic strength, calculated from sequence Mycobacterium tuberculosis
79000
-
dimer, gel filtration Mycobacterium tuberculosis
79000
-
dimer, the enzyme behaves as a dimer at normal ionic strength but may associate to a tetramer at low ionic strength, gel filtration Mycobacterium tuberculosis
158000
-
tetramer, gel filtration Mycobacterium tuberculosis
158000
-
tetramer, the enzyme behaves as a dimer at normal ionic strength but may associate to a tetramer at low ionic strength, gel filtration Mycobacterium tuberculosis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1-(2-acetamido-2-deoxy-alpha-D-glucopyranosyl)-1D-myo-inositol + H2O Mycobacterium tuberculosis mycothiol biosynthesis 1-(2-amino-2-deoxy-alpha-D-glucopyranoside)-1D-myo-inositol + acetate
-
?
1-(2-acetamido-2-deoxy-alpha-D-glucopyranosyl)-1D-myo-inositol + H2O Mycobacterium tuberculosis H37Rv mycothiol biosynthesis 1-(2-amino-2-deoxy-alpha-D-glucopyranoside)-1D-myo-inositol + acetate
-
?
1-D-myo-inosityl-2-acetamido-2-deoxy-alpha-D-glucopyranoside + H2O Mycobacterium tuberculosis key enzyme in mycothiol biosynthesis. Mycothiol is the major low molecular weight thiol in actinomycetes and is essential for growth of Mycobacterium tuberculosis 1-(2-amino-2-deoxy-alpha-D-glucopyranoside)-1D-myo-inositol + acetate
-
?

Organism

Organism UniProt Comment Textmining
Mycobacterium tuberculosis P9WJN3
-
-
Mycobacterium tuberculosis H37Rv P9WJN3
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Mycobacterium tuberculosis

Specific Activity [micromol/min/mg]

Specific Activity Minimum [┬Ámol/min/mg] Specific Activity Maximum [┬Ámol/min/mg] Comment Organism
0.19
-
-
Mycobacterium tuberculosis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1-(2-acetamido-2-deoxy-alpha-D-glucopyranosyl)-1D-myo-inositol + H2O
-
Mycobacterium tuberculosis 1-(2-amino-2-deoxy-alpha-D-glucopyranoside)-1D-myo-inositol + acetate
-
?
1-(2-acetamido-2-deoxy-alpha-D-glucopyranosyl)-1D-myo-inositol + H2O mycothiol biosynthesis Mycobacterium tuberculosis 1-(2-amino-2-deoxy-alpha-D-glucopyranoside)-1D-myo-inositol + acetate
-
?
1-(2-acetamido-2-deoxy-alpha-D-glucopyranosyl)-1D-myo-inositol + H2O
-
Mycobacterium tuberculosis H37Rv 1-(2-amino-2-deoxy-alpha-D-glucopyranoside)-1D-myo-inositol + acetate
-
?
1-(2-acetamido-2-deoxy-alpha-D-glucopyranosyl)-1D-myo-inositol + H2O mycothiol biosynthesis Mycobacterium tuberculosis H37Rv 1-(2-amino-2-deoxy-alpha-D-glucopyranoside)-1D-myo-inositol + acetate
-
?
1-D-myo-inosityl-2-acetamido-2-deoxy-alpha-D-glucopyranoside + H2O key enzyme in mycothiol biosynthesis. Mycothiol is the major low molecular weight thiol in actinomycetes and is essential for growth of Mycobacterium tuberculosis Mycobacterium tuberculosis 1-(2-amino-2-deoxy-alpha-D-glucopyranoside)-1D-myo-inositol + acetate
-
?
additional information MshB shows amidase activity with a wide range of substrates Mycobacterium tuberculosis ?
-
?
additional information the enzyme is unable to remove the acetyl residue from the acetylcysteinyl group of mycothiol or S-(2-oxo-2-phenylethyl)mycothiol, and it exhibits barely detectable amidase activity with mycothiol or mycothiol disulfide. It has significant amidase activity with S-(2-oxo-2-phenylethyl)mycothiol and even greater activity with N-deacetyl-N-formylmycothiol-monobromobimane conjugate, N-deacetyl-mycothiol-monobromobimane conjugate, formyl-CySmB-GlcN-Ins, and mycothiol-monobromobimane conjugate Mycobacterium tuberculosis ?
-
?
additional information MshB shows amidase activity with a wide range of substrates Mycobacterium tuberculosis H37Rv ?
-
?
additional information the enzyme is unable to remove the acetyl residue from the acetylcysteinyl group of mycothiol or S-(2-oxo-2-phenylethyl)mycothiol, and it exhibits barely detectable amidase activity with mycothiol or mycothiol disulfide. It has significant amidase activity with S-(2-oxo-2-phenylethyl)mycothiol and even greater activity with N-deacetyl-N-formylmycothiol-monobromobimane conjugate, N-deacetyl-mycothiol-monobromobimane conjugate, formyl-CySmB-GlcN-Ins, and mycothiol-monobromobimane conjugate Mycobacterium tuberculosis H37Rv ?
-
?
N-acetyl-D-glucosamine + H2O activity is 73fold lower than with 1-D-myo-inosityl-2-acetamido-2-deoxy-alpha-D-glucopyranoside Mycobacterium tuberculosis D-glucosamine + acetate
-
?
N-acetyl-D-glucosamine + H2O activity is 73fold lower than with 1-D-myo-inosityl-2-acetamido-2-deoxy-alpha-D-glucopyranoside Mycobacterium tuberculosis H37Rv D-glucosamine + acetate
-
?
N-deacetyl-N-formylmycothiol-monobromobimane conjugate + H2O
-
Mycobacterium tuberculosis ?
-
?

Subunits

Subunits Comment Organism
dimer 2 * 32000, SDS-PAGE Mycobacterium tuberculosis
dimer 2 * 32000, the enzyme appears to behave as a dimer at normal ionic strength but may associate to a tetramer at low ionic strength, SDS-PAGE Mycobacterium tuberculosis
dimer 2 * 33423, the enzyme appears to behave as a dimer at normal ionic strength but may associate to a tetramer at low ionic strength, calculated from sequence Mycobacterium tuberculosis
tetramer 4 * 32000, the enzyme appears to behave as a dimer at normal ionic strength but may associate to a tetramer at low ionic strength, SDS-PAGE Mycobacterium tuberculosis
tetramer 4 * 33423, the enzyme appears to behave as a dimer at normal ionic strength but may associate to a tetramer at low ionic strength, calculated from sequence Mycobacterium tuberculosis

Synonyms

Synonyms Comment Organism
1D-myo-inosityl-2-acetamido-2-deoxy-alpha-D-glucopyranoside deacetylase
-
Mycobacterium tuberculosis
GlcNAc-Ins deacetylase
-
Mycobacterium tuberculosis
MshB
-
Mycobacterium tuberculosis
Rv1170
-
Mycobacterium tuberculosis

Temperature Optimum [┬░C]

Temperature Optimum [┬░C] Temperature Optimum Maximum [┬░C] Comment Organism
37
-
assay at Mycobacterium tuberculosis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.49
-
1-(2-acetamido-2-deoxy-alpha-D-glucopyranosyl)-1D-myo-inositol pH 7.5, 37┬░C Mycobacterium tuberculosis
0.49
-
N-deacetyl-N-formylmycothiol-monobromobimane conjugate pH 7.4, 37┬░C Mycobacterium tuberculosis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.4
-
assay at Mycobacterium tuberculosis

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.44
-
1-(2-acetamido-2-deoxy-alpha-D-glucopyranosyl)-1D-myo-inositol pH 7.5, 37┬░C Mycobacterium tuberculosis