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Information on EC 3.5.1.103 - N-acetyl-1-D-myo-inositol-2-amino-2-deoxy-alpha-D-glucopyranoside deacetylase and Organism(s) Mycobacterium tuberculosis and UniProt Accession P9WJN3

for references in articles please use BRENDA:EC3.5.1.103
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EC Tree
IUBMB Comments
This enzyme is considered the key enzyme and rate limiting step in the mycothiol biosynthesis pathway . In addition to acetylase activity, the enzyme possesses weak activity of EC 3.5.1.115, mycothiol S-conjugate amidase, and shares sequence similarity with that enzyme . The enzyme requires a divalent transition metal ion for activity, believed to be Zn2+ .
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This record set is specific for:
Mycobacterium tuberculosis
UNIPROT: P9WJN3
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Word Map
The taxonomic range for the selected organisms is: Mycobacterium tuberculosis
The expected taxonomic range for this enzyme is: Bacteria, Archaea
Synonyms
rv1170, glcnac-ins deacetylase, acgi deacetylase, n-acetyl-1-d-myo-inosityl-2-amino-2-deoxy-alpha-d-glucopyranoside deacetylase, 1-d-myo-inosityl 2-n-acetamido-2-deoxy-alpha-d-glucopyranoside deacetylase, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
1-D-myo-inosityl 2-acetamido-2-deoxy-alpha-D-glucopyranoside deacetylase
-
1-D-myo-inosityl 2-N-acetamido-2-deoxy-alpha-D-glucopyranoside deacetylase
-
1D-myo-inosityl-2-acetamido-2-deoxy-alpha-D-glucopyranoside deacetylase
-
N-acetyl-1-D-myo-inosityl-2-amino-2-deoxy-alpha-D-glucopyranoside deacetylase
-
N-acetyl-1-D-myo-inosityl-2-amino-2-deoxy-D-glucopyranoside deacetylase
-
N-acetyl-1-D-myo-inosityl-2-deoxy-alpha-D-glucopyranoside deacetylase
-
GlcNAc-Ins-deacetylase
-
-
N-acetylglucosaminylinositol-deacetylase
-
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
1-O-(2-acetamido-2-deoxy-alpha-D-glucopyranosyl)-1D-myo-inositol + H2O = 1-O-(2-amino-2-deoxy-alpha-D-glucopyranosyl)-1D-myo-inositol + acetate
show the reaction diagram
PATHWAY SOURCE
PATHWAYS
-
-
SYSTEMATIC NAME
IUBMB Comments
1-(2-acetamido-2-deoxy-alpha-D-glucopyranosyl)-1D-myo-inositol acetylhydrolase
This enzyme is considered the key enzyme and rate limiting step in the mycothiol biosynthesis pathway [1]. In addition to acetylase activity, the enzyme possesses weak activity of EC 3.5.1.115, mycothiol S-conjugate amidase, and shares sequence similarity with that enzyme [2]. The enzyme requires a divalent transition metal ion for activity, believed to be Zn2+ [3].
CAS REGISTRY NUMBER
COMMENTARY hide
340703-87-9
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
1-(2-acetamido-2-deoxy-alpha-D-glucopyranosyl)-1D-myo-inositol + H2O
1-(2-amino-2-deoxy-alpha-D-glucopyranoside)-1D-myo-inositol + acetate
show the reaction diagram
1-D-myo-inosityl-2-acetamido-2-deoxy-alpha-D-glucopyranoside + H2O
1-(2-amino-2-deoxy-alpha-D-glucopyranoside)-1D-myo-inositol + acetate
show the reaction diagram
key enzyme in mycothiol biosynthesis. Mycothiol is the major low molecular weight thiol in actinomycetes and is essential for growth of Mycobacterium tuberculosis
-
-
?
1-O-(2-acetamido-2-deoxy-alpha-D-glucopyranosyl)-1D-myo-inositol + H2O
1-O-(2-amino-2-deoxy-alpha-D-glucopyranosyl)-1D-myo-inositol + acetate
show the reaction diagram
-
-
-
?
1-O-(2-acetamido-2-deoxy-alpha-D-glucopyranosyl)-D-myo-inositol + H2O
1-O-(2-amino-2-deoxy-alpha-D-glucopyranosyl)-D-myo-inositol + acetate
show the reaction diagram
-
-
-
?
N-acetyl-1-D-myo-inosityl-2-amino-2-deoxy-alpha-D-glucopyranoside + H2O
1-D-myo-inosityl-2-amino-2-deoxy-alpha-D-glucopyranoside + acetate
show the reaction diagram
-
-
-
?
N-acetyl-1-D-myo-inosityl-2-amino-2-deoxy-D-glucopyranoside + H2O
1-O-(2-amino-2-deoxy-alpha-D-glucopyranosyl)-1D-myo-inositol + acetate
show the reaction diagram
molecular docking analysis of the MshB active site binding pocket, overview
-
-
?
N-acetyl-D-glucosamine + H2O
acetate + D-glucosamine
show the reaction diagram
N-acetyl-D-glucosamine + H2O
D-glucosamine + acetate
show the reaction diagram
N-deacetyl-N-formylmycothiol-monobromobimane conjugate + H2O
?
show the reaction diagram
-
-
-
?
1-(2-acetamido-2-deoxy-alpha-D-glucopyranosyl)-1D-myo-inositol + H2O
1-(2-amino-2-deoxy-alpha-D-glucopyranoside)-1D-myo-inositol + acetate
show the reaction diagram
cyclohexyl-2-acetamido-2-deoxy-1-thio-alpha-D-glucopyranoside + H2O
cyclohexyl-2-amino-2-deoxy-1-thio-alpha-D-glucopyranoside + acetate
show the reaction diagram
-
6% of the activity with 1-(2-acetamido-2-deoxy-alpha-D-glucopyranosyl)-1D-myo-inositol
-
-
?
cyclohexyl-2-acetamido-2-deoxy-alpha-D-glucopyranoside + H2O
cyclohexyl-2-amino-2-deoxy-alpha-D-glucopyranoside + acetate
show the reaction diagram
-
6% of the activity with 1-(2-acetamido-2-deoxy-alpha-D-glucopyranosyl)-1D-myo-inositol
-
-
?
phenyl 2-acetamido-2-deoxy-1-thio-alpha-D-glucopyranoside + H2O
phenyl 2-amino-2-deoxy-1-thio-alpha-D-glucopyranoside + acetate
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
1-(2-acetamido-2-deoxy-alpha-D-glucopyranosyl)-1D-myo-inositol + H2O
1-(2-amino-2-deoxy-alpha-D-glucopyranoside)-1D-myo-inositol + acetate
show the reaction diagram
1-D-myo-inosityl-2-acetamido-2-deoxy-alpha-D-glucopyranoside + H2O
1-(2-amino-2-deoxy-alpha-D-glucopyranoside)-1D-myo-inositol + acetate
show the reaction diagram
key enzyme in mycothiol biosynthesis. Mycothiol is the major low molecular weight thiol in actinomycetes and is essential for growth of Mycobacterium tuberculosis
-
-
?
1-O-(2-acetamido-2-deoxy-alpha-D-glucopyranosyl)-D-myo-inositol + H2O
1-O-(2-amino-2-deoxy-alpha-D-glucopyranosyl)-D-myo-inositol + acetate
show the reaction diagram
-
-
-
?
N-acetyl-1-D-myo-inosityl-2-amino-2-deoxy-alpha-D-glucopyranoside + H2O
1-D-myo-inosityl-2-amino-2-deoxy-alpha-D-glucopyranoside + acetate
show the reaction diagram
-
-
-
?
1-(2-acetamido-2-deoxy-alpha-D-glucopyranosyl)-1D-myo-inositol + H2O
1-(2-amino-2-deoxy-alpha-D-glucopyranoside)-1D-myo-inositol + acetate
show the reaction diagram
-
biosynthesis of mycothiol and detoxification of xenobiotics as their mycothiol-S-conjugates
-
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Fe2+
the active site metal ion is coordinated to two histidine residues, His13 and His147, and an aspartate, Asp16
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1,10-phenanthroline
benzyl 2-deoxy-2-acetamido-1-thio-beta-D-glucopyranoside
-
0.5 mM, 13% inhibition
cyclohexyl (2''R),(2''S)-3,3''-anhydro-2-deoxy-2-C-(2'',3''-dihydroxypropyl)-alpha-D-glucopyranoside
-
0.5 mM, 6.6% inhibition
cyclohexyl 2-deoxy-2-C-(2'',3''-epoxypropyl)-alpha-D-glucopyranoside
-
0.5 mM, 19.7% inhibition
cyclohexyl 2-deoxy-2-C-(2''-hydroxypropyl)-alpha-D-glucopyranoside
-
0.5 mM, 11.4% inhibition
cyclohexyl 2-deoxy-2-C-(2''-oxopropyl)-a-D-glucopyranoside
-
0.5 mM, 6.7% inhibition
cyclohexyl-2-chloroacetamido-2-deoxy-1-thio-alpha-D-glucopyranoside
-
0.5 mM, 15% inhibition
phenyl-2-acetamido-2-deoxy-1-thio-alpha-D-glucopyranoside
-
0.5 mM, 6% inhibition
phenyl-2-deoxy-2-acetamido-1-thio-beta-D-glucopyranoside
-
0.5 mM, 15% inhibition
phenyl-2-deoxy-2-[3'-(8''-hydroxy-3''-methyl-1'',4''-dioxo-1'',4''-dihydronaphthalen-2''-yl)butanamido]-1-thio-alpha-D-glucopyranoside
-
0.5 mM, 81.6% inhibition
phenyl-2-deoxy-2-[3'-(8''-hydroxy-3''-methyl-1'',4''-dioxo-1'',4''-dihydronaphthalen-2''-yl)pentanamido]-1-thio-alpha-D-glucopyranoside
-
0.5 mM, 81.4% inhibition
phenyl-2-deoxy-2-[3'-(8''-hydroxy-3''-methyl-1'',4''-dioxo-1'',4''-dihydronaphthalen-2''-yl)propanamido]-1-thio-alpha-D-glucopyranoside
-
0.5 mM, 57.4% inhibition
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1,7-phenanthroline
1 mM, slight enhancement of activity
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.34
1-(2-acetamido-2-deoxy-alpha-D-glucopyranosyl)-1D-myo-inositol
pH 7.5, 37°C
0.34
1-O-(2-acetamido-2-deoxy-alpha-D-glucopyranosyl)-1D-myo-inositol
pH 7.5, 30°C
17.5 - 114
N-acetyl-D-glucosamine
0.34
N-deacetyl-N-formylmycothiol-monobromobimane conjugate
pH 7.4, 37°C
0.348
1-(2-acetamido-2-deoxy-alpha-D-glucopyranosyl)-1D-myo-inositol
-
pH 7.5, 37°C
1.695
phenyl-2-acetamido-2-deoxy-1-thio-alpha-D-glucopyranoside
-
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.49
1-(2-acetamido-2-deoxy-alpha-D-glucopyranosyl)-1D-myo-inositol
pH 7.5, 37°C
0.0048 - 1.05
N-acetyl-D-glucosamine
0.49
N-deacetyl-N-formylmycothiol-monobromobimane conjugate
pH 7.4, 37°C
0.24
1-(2-acetamido-2-deoxy-alpha-D-glucopyranosyl)-1D-myo-inositol
-
pH 7.5, 37°C
0.12
phenyl-2-acetamido-2-deoxy-1-thio-alpha-D-glucopyranoside
-
pH 7.5, 37°C
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.44
1-(2-acetamido-2-deoxy-alpha-D-glucopyranosyl)-1D-myo-inositol
pH 7.5, 37°C
0.00009 - 0.06
N-acetyl-D-glucosamine
0.68
1-(2-acetamido-2-deoxy-alpha-D-glucopyranosyl)-1D-myo-inositol
-
pH 7.5, 37°C
0.07
phenyl-2-acetamido-2-deoxy-1-thio-alpha-D-glucopyranoside
-
pH 7.5, 37°C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
GlcNAc-Ins is absent in mycothiol-deficient mutant strain 49. This strain can concentrate GlcNAc-Ins from the medium and convert it to mycothiol
metabolism
additional information
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
158000
32000
33423
79000
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dimer
tetramer
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
crystals prepared in the presence of beta-octylglucoside as a key additive, are suitable for high-resolution X-ray structural analysis. The crystals are orthorhombic, with unit-cell parameters a = 71.69, b = 83.74, c = 95.65 A, space group P2(1)2(1)2(1) and two molecules in the asymmetric unit. Collection of X-ray diffraction data to 1.9 A resolution
hanging-drop vapour-diffusion method, crystallized both in native and SeMet-substituted forms
purified recombinant enzyme, from 0.1 M sodium acetate, 0.2 M ammonium sulfate, 25% PEG 4000 pH 4.6, with glycerol and the reaction product acetate in the active site, X-ray diffraction structure determination and analysis at 1.94 A resolution, modelling
structure, determined at 1.9 A resolution by X-ray crystallography
the structure, determined at 1.9 A resolution by X-ray crystallography, reveals an alpha/beta fold in which helices pack against a seven-stranded mostly parallel beta-sheet
vapor-diffusion method
vapor-diffusion method with a mother liquor consisting of 15% polyethylene glycol 4000, 50 mm Tris-HCl (pH 8.0), 0.1 m Mg(NO3)2, 6% 1,6-hexanediol, and 10% ethylene glycol. A 1:2 ratio of protein solution (6 mg/ml) to mother liquor is mixed and left for vapor equilibration. Triclinic crystals form after approximately 1 week at room temperature
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D146A
D146N
the D146N mutant is about 10fold higher than that of the D146A mutant, suggesting that the ability to accept a hydrogen bond at this position contributes to GlcNAc substrate specificity. Because there does not appear to be a direct contact between Asp146 and substrate, this effect is likely mediated via positioning of other catalytically important residues. The mutant enzyme shows 3.7% of the activity compared to the wild-type enzyme with the substrate N-acetyl-D-glucosamine
D15A
the mutant enzyme shows 0.5% of the activity compared to the wild-type enzyme with the substrate N-acetyl-D-glucosamine
D95A
inactive mutant enzyme
E47A
the mutant enzyme shows 300% of the activity compared to the wild-type enzyme with the substrate N-acetyl-D-glucosamine. Mutation decreases the value of KM GlcNAc (2-fold) and increases the value of kcat/KM GlcNAc (3-fold)
F216A
the mutant enzyme shows 110% of the activity compared to the wild-type enzyme with the substrate N-acetyl-D-glucosamine
L19A
the mutant enzyme shows 115% of the activity compared to the wild-type enzyme with the substrate N-acetyl-D-glucosamine
M98A
the mutant enzyme shows 15% of the activity compared to the wild-type enzyme with the substrate N-acetyl-D-glucosamine
R68A
inactive mutant enzyme
additional information
the enzyme is unable to catalyze the turnover of GlcNAc upon loss of the Arg68 or Asp95 side chains, consistent with the proposal that these side chains make critical hydrogen bonding interactions with substrate
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli
expression in Escherichia coli both in native and SeMet-substituted forms
recombinant enzyme expression
Rv1170 is cloned to contain a C-terminal His-6 tag to facilitate purifiation, expression in Escherichia coli
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
drug development
enzyme MshB is a target for the discovery of drugs to treat tuberculosis
medicine
antisense oligonucleotides to GlcNAc-Ins deacetylase (Rv1170) mRNA affect mycobacterial growth. The enzyme is sensitive to free radical generating antituberculosis drugs and may be a useful targets for new drug development
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Hayward, D.; Wiid, I.; van Helden, P.
Differential expression of mycothiol pathway genes: are they affected by antituberculosis drugs?
IUBMB Life
56
131-138
2004
Mycobacterium tuberculosis (P9WJN3), Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv (P9WJN3)
Manually annotated by BRENDA team
McCarthy, A.A.; Knijff, R.; Peterson, N.A.; Baker, E.N.
Crystallization and preliminary X-ray analysis of N-acetyl-1-D-myo-inosityl-2-deoxy-alpha-D-glucopyranoside deacetylase (MshB) from Mycobacterium tuberculosis
Acta Crystallogr. Sect. D
59
2316-2318
2003
Mycobacterium tuberculosis (P9WJN3), Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv (P9WJN3)
Manually annotated by BRENDA team
Nicholas, G.M.; Eckman, L.L.; Kovac, P.; Otero-Quintero, S.; Bewley, C.A.
Synthesis of 1-D- and 1-L-myo-inosityl 2-N-acetamido-2-deoxy-alpha-D-glucopyranoside establishes substrate specificity of the Mycobacterium tuberculosis enzyme AcGI deacetylase
Bioorg. Med. Chem.
11
2641-2647
2003
Mycobacterium tuberculosis (P9WJN3), Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv (P9WJN3)
Manually annotated by BRENDA team
Gammon, D.W.; Steenkamp, D.J.; Mavumengwana, V.; Marakalala, M.J.; Mudzunga, T.T.; Hunter, R.; Munyololo, M.
Conjugates of plumbagin and phenyl-2-amino-1-thioglucoside inhibit MshB, a deacetylase involved in the biosynthesis of mycothiol
Bioorg. Med. Chem.
18
2501-2514
2010
Mycobacterium tuberculosis
Manually annotated by BRENDA team
Newton, G.L.; Av-Gay, Y.; Fahey, R.C.
N-Acetyl-1-D-myo-inosityl-2-amino-2-deoxy-alpha-D-glucopyranoside deacetylase (MshB) is a key enzyme in mycothiol biosynthesis
J. Bacteriol.
182
6958-6963
2000
Mycobacterium tuberculosis (P9WJN3), Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv (P9WJN3)
Manually annotated by BRENDA team
Maynes, J.T.; Garen, C.; Cherney, M.M.; Newton, G.; Arad, D.; Av-Gay, Y.; Fahey, R.C.; James, M.N.
The crystal structure of 1-D-myo-inosityl 2-acetamido-2-deoxy-alpha-D-glucopyranoside deacetylase (MshB) from Mycobacterium tuberculosis reveals a zinc hydrolase with a lactate dehydrogenase fold
J. Biol. Chem.
278
47166-47170
2003
Mycobacterium tuberculosis (P9WJN3), Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv (P9WJN3)
Manually annotated by BRENDA team
McCarthy, A.A.; Peterson, N.A.; Knijff, R.; Baker, E.N.
Crystallization and preliminary X-ray analysis of N-acetyl-1-D-myo-inosityl-2-deoxy-alpha-D-glucopyranoside deacetylase (MshB) from Mycobacterium tuberculosis
J. Mol. Biol.
335
1131-1141
2004
Mycobacterium tuberculosis (P9WJN3), Mycobacterium tuberculosis H37Rv (P9WJN3)
Manually annotated by BRENDA team
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
Protein Expr. Purif.
47
542-550
2006
Mycobacterium tuberculosis (P9WJN3), Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Rv (P9WJN3)
Manually annotated by BRENDA team
Broadley, S.G.; Gumbart, J.C.; Weber, B.W.; Marakalala, M.J.; Steenkamp, D.J.; Sewell, B.T.
A new crystal form of MshB from Mycobacterium tuberculosis with glycerol and acetate in the active site suggests the catalytic mechanism
Acta Crystallogr. Sect. D
68
1450-1459
2012
Mycobacterium tuberculosis (P9WJN3)
Manually annotated by BRENDA team
Huang, X.; Hernick, M.
Automated docking studies provide insights into molecular determinants of ligand recognition by N-acetyl-1-D-myo-inosityl-2-amino-2-deoxy-alpha-D-glucopyranoside deacetylase (MshB)
Biopolymers
101
406-417
2014
Mycobacterium tuberculosis (P9WJN3)
Manually annotated by BRENDA team
Lamprecht, D.A.; Muneri, N.O.; Eastwood, H.; Naidoo, K.J.; Strauss, E.; Jardine, A.
An enzyme-initiated Smiles rearrangement enables the development of an assay of MshB, the GlcNAc-Ins deacetylase of mycothiol biosynthesis
Org. Biomol. Chem.
10
5278-5288
2012
Mycobacterium tuberculosis (P9WJN3), Mycobacterium tuberculosis
Manually annotated by BRENDA team
Huang, X.; Hernick, M.
Molecular determinants of N-acetylglucosamine recognition and turnover by N-acetyl-1-D-myo-inosityl-2-amino-2-deoxy-alpha-D-glucopyranoside deacetylase (MshB)
Biochemistry
54
3784-3790
2015
Mycobacterium tuberculosis (P9WJN3), Mycobacterium tuberculosis ATCC 25618 (P9WJN3)
Manually annotated by BRENDA team