2.7.7.23: UDP-N-acetylglucosamine diphosphorylase
This is an abbreviated version!
For detailed information about UDP-N-acetylglucosamine diphosphorylase, go to the full flat file.
Word Map on EC 2.7.7.23
Reaction
Synonyms
acetylglucosamine 1-phosphate uridylyltransferase, AGX-1, AGX1, AGX2, amino-sugar-1-phosphate acetyltransferase, bifunctional N-acetyltransferase/uridylyltransferase, BmUAP, CL6EHI_021200, CL6EHI_039830, EcGlmU, EnhiA.01126.a, EnhiA.01126.b, GlcNAc-1-P uridyltransferase, GlcNAc-1-P UTase, GlcNAc-1-phosphate nucleotidylyltransferase, GLCNAC1PUT1, GLCNAC1PUT2, GlmU, GlmUMTB, hiGlmU, LmUAP1, LmUAP2, LOC_Os08g10600, Mmy, More, mummy, N-acetyl-D-glucosamine-1-phosphate uridylyltransferase, N-acetyl-glucosamine-1-phosphate uridyltransferase, N-acetylglucosamine 1-phosphate uridyltransferase, N-acetylglucosamine-1-phosphate uridyltransferase, N-acetylglucosamine-1-phosphate uridylyltransferase, N-acetylglucosamine-1-phosphate-uridyltransferase/glucosamine-1-phosphate-acetyltransferase, Os08g10600, Rv1018c, spl29, spotted leaf 29, ST0452, ST0452 protein, STK_04520, Tb11.02.0120, UAP, UAP enzyme, UAP1, UAP2, UDP N-acetylglucosamine pyrophosphorylases 1, UDP N-acetylglucosamine pyrophosphorylases 2, UDP-GlcNAc pyrophosphorylase, UDP-GlcNAc pyrophosphorylase (UAP), UDP-HexNAc pyrophosphorylase, UDP-N-acetylgalactosamine pyrophosphorylase, UDP-N-acetylglucosamine diphosphorylase 1, UDP-N-acetylglucosamine diphosphorylase 2, UDP-N-acetylglucosamine pyrophosphorylase, UDP-N-acetylglucosamine pyrophosphorylase (UAP), UDP-N-acetylglucosamine pyrophosphorylase 1, UDP-N-acetylglucosamine pyrophosphorylase/glucosamine-1-phosphate N-acetyltransferase, UDP-N-acetylglucosamine-diphosphorylase, UDP-N-acetylhexosamine pyrophosphorylase, UDPacetylglucosamine pyrophosphorylase, UNAcP, uridine diphosphate N-acetylglucosamine pyrophosphorylase, uridine diphosphate-N-acetylglucosamine pyrophosphorylase, uridine diphosphoacetylglucosamine phosphorylase, uridine diphosphoacetylglucosamine pyrophosphorylase, uridine-diphospho-N-acetylglucosamine pyrophosphorylase, UTP:2-acetamido-2-deoxy-alpha-D-glucose-1-phosphate uridylyltransferase, UTP:N-acetylglucosamine-1-P uridylyltransferase, Ydl103c protein
ECTree
2.7.7.23 UDP-N-acetylglucosamine diphosphorylaseAdvanced search results
results (
results (Engineering
Engineering on EC 2.7.7.23 - UDP-N-acetylglucosamine diphosphorylase
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
top
PROTEIN VARIANTS 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
K421A
the activity of the mutant is very low (less than 5%) in comparison to the wild type enzyme
D105A
site-directed mutagenesis, the mutant shows 50% reduced activity compared to the wild-type enzyme
E154D
site-directed mutagenesis in the GlcNAc-binding region, the mutant shows about 80% reduced activity compared to the wild-type enzyme
E154K
site-directed mutagenesis in the GlcNAc-binding region, the mutant shows about 90% reduced activity compared to the wild-type enzyme
E154L
site-directed mutagenesis in the GlcNAc-binding region, the mutant shows about 70% reduced activity compared to the wild-type enzyme
N169A
site-directed mutagenesis in the GlcNAc-binding region, the mutant shows about 50% reduced activity compared to the wild-type enzyme
N169D
site-directed mutagenesis in the GlcNAc-binding region, the mutant shows unaltered activity compared to the wild-type enzyme
N169Q
site-directed mutagenesis in the GlcNAc-binding region, the mutant shows unaltered activity compared to the wild-type enzyme
N169R
site-directed mutagenesis in the GlcNAc-binding region, the mutant shows 1.4fold increased activity compared to the wild-type enzyme. The N169R mutant caused a slightly secondary structure changes, thus facilitating GlcNAc-1-phosphate to enter the active pocket through the additional interaction with N-acetyl arm of GlcNAc moiety
Q76A
site-directed mutagenesis in the uridine-binding region, the mutant has a catalytic activity to convert CTP and GlcNAc-1P into unnatural sugar nucleotide CDP-GlcNAc which is distinct from the wild-type, altered nucleotide specificity compared to wild-type, overview
Q76E
site-directed mutagenesis in the uridine-binding region, the mutant has a catalytic activity to convert CTP and GlcNAc-1P into unnatural sugar nucleotide CDP-GlcNAc which is distinct from the wild-type, altered nucleotide specificity compared to wild-type, overview
Q76P
site-directed mutagenesis in the uridine-binding region, the mutant has a catalytic activity to convert CTP and GlcNAc-1P into unnatural sugar nucleotide CDP-GlcNAc which is distinct from the wild-type, altered nucleotide specificity compared to wild-type, overview
T82G
site-directed mutagenesis in the GlcNAc-binding region, the mutant shows about 65% reduced activity compared to the wild-type enzyme
T82Q
site-directed mutagenesis in the GlcNAc-binding region, the mutant shows about 80% reduced activity compared to the wild-type enzyme
T82S
site-directed mutagenesis in the GlcNAc-binding region, the mutant shows about 80% reduced activity compared to the wild-type enzyme
Y103A
Y103F
site-directed mutagenesis of the residue located nearby the uridyltransferase active pocket,the mutant shows increased activity compared to the wild-type enzyme
Y103N
Y103V
mutant protein shows a 30% decrease in activity compared to that of the wild-type enzyme
Y103A
Y103N
Y103V
-
mutant protein shows a 30% decrease in activity compared to that of the wild-type enzyme
-
G108A
dramatically reduced activity, significant increase in melting temperature
G210A
dramatically reduced activity, significant decrease in melting temperature
A229T
naturally occuring enzyme AGX1 mutation, and site-directed mutagenesis, the A229T mutation causes a reduction of protein thermal stability compared to wild-type AGX1, and AGX1A229T has lower activity in producing UDP-GlcNA. In diploid organisms, haploinsufficiency is a phenomenon in which a single copy of a functional gene is not sufficient to produce the normal/wild-type phenotype. The patient is only heterozygous for the UAP1 A229T missense mutation. The UAP1 gene is potentially haploinsufficient and LoF intolerant, and the heterozygous UAP1 A229T mutation is potentially pathogenic. The recombinant mutant enzyme shows a reduction of the melting temperature (Tm) by approximately 5.3°C compared to wild-type. The A229T mutation induces structural changes. The R228-E44 interaction is abolished in the AGX1A229T structure caused by the position shift of R228. The pushing effect is likely due to the bulkier side chain of threonine compared to that of alanine. Along with the conformational change of the N-terminal domain in the AGX1A229T structure, is M218 shifted by 0.8 A away from R169, weakening the Q112-R169-M218 interaction
G222A
-
traces of activity in forward and reverse reaction with N-acetyl-D-glucosamine 1-phosphate and N-acetyl-D-galactosamine 1-phosphate
G224A
-
low activity in forward and reverse reaction with N-acetyl-D-glucosamine 1-phosphate and N-acetyl-D-galactosamine 1-phosphate
P220A
-
only slight changes in activity with N-acetyl-D-glucosamine 1-phosphate and N-acetyl-D-galactosamine 1-phosphate
Y227A
-
only slight changes in activity with N-acetyl-D-glucosamine 1-phosphate and N-acetyl-D-galactosamine 1-phosphate
H374A
site-directed mutagenesis, the acetyltransferase active site mutant shows 1.7% of acetyltransferase activity and 96.7% of uridinyltransferase activity compared to the wild-type
K464A
site-directed mutagenesis, the mutant still shows acetyltransferase activity, the mutant shows 105.6% acetyltransferase activity and 97.9% of uridinyltransferase activity compared to the wild-type
N239A
the activity of the mutant is very low (less than 10%) in comparison to the wild type enzyme
N397A
site-directed mutagenesis, the acetyltransferase active site mutant shows 5.2% of acetyltransferase activity and 113.6% of uridinyltransferase activity compared to the wild-type
S416A
site-directed mutagenesis, the acetyltransferase active site mutant shows 100.9% of acetyltransferase activity and 96.4% of uridinyltransferase activity compared to the wild-type
T418A
site-directed mutagenesis, the acetyltransferase activity of mutant is severely compromised as compared with GlmUMtb wild-type, the mutant shows 2.4% acetyltransferase activity and 100.4% of uridinyltransferase activity compared to the wild-type
T418E
site-directed mutagenesis, the acetyltransferase activity of the T418E mutant that mimics a phosphorylated Thr, is severely compromised as compared with GlmUMtb wild-type, the mutant shows 2.2% acetyltransferase activity and 109.2% of uridinyltransferase activity compared to the wild-type
T418S
site-directed mutagenesis, the acetyltransferase activity of the mutant is compromised as compared with GlmUMtb wild-type, the mutant shows 19% acetyltransferase activity and 108.8% of uridinyltransferase activity compared to the wild-type
W460A
site-directed mutagenesis, the mutant displays almost complete loss in acetyltransferase activity, the mutant shows 8.4% acetyltransferase activity and 99.8% of uridinyltransferase activity compared to the wild-type
W460A/K64A
site-directed mutagenesis, the mutant shows 7.8% acetyltransferase activity and 104.7% of uridinyltransferase activity compared to the wild-type
N239A
-
the activity of the mutant is very low (less than 10%) in comparison to the wild type enzyme
-
D208A
E146A
G9A
G9A/K147A
G9A/T80A
G9A/Y97A
G9A/Y97F
K147A
R13A
T80A
T80A/K147A
T80A/Y97A
T80A/Y97F
T80C
site-directed mutagenesis, the mutant shows slightly reduced GlcNAc-1-P UTase activity compared to wild-type
T80D
site-directed mutagenesis, the mutant shows increased Glc-1-P UTase activity and reduced GlcNAc-1-P UTase activity compared to wild-type
T80E
site-directed mutagenesis, the mutant shows no Glc-1-P UTase activity and highly reduced GlcNAc-1-P UTase activity
T80F
site-directed mutagenesis, the mutant shows no and Glc-1-P UTase and GlcNAc-1-P UTase activity
T80G
site-directed mutagenesis, the mutant shows increased GlcNAc-1-P and Glc-1-P UTase activity compared to wild-type
T80H
site-directed mutagenesis, the mutant shows increased Glc-1-P UTase activity and reduced GlcNAc-1-P UTase activity compared to wild-type
T80I
site-directed mutagenesis, the mutant shows no Glc-1-P UTase and GlcNAc-1-P UTase activity
T80K
site-directed mutagenesis, the mutant shows no Glc-1-P UTase activity and highly reduced GlcNAc-1-P UTase activity
T80L
T80M
site-directed mutagenesis, the mutant shows reduced GlcNAc-1-P UTase activity and no Glc-1-P UTase activity
T80N
site-directed mutagenesis, the mutant shows increased GlcNAc-1-P UTase activity compared to wild-type
T80P
site-directed mutagenesis, the mutant shows slightly reduced GlcNAc-1-P UTase activity compared to wild-type
T80Q
site-directed mutagenesis, the mutant shows increased GlcNAc-1-P and Glc-1-P UTase activity compared to wild-type
T80R
site-directed mutagenesis, the mutant shows no Glc-1-P UTase activity and highly reduced GlcNAc-1-P UTase activity
T80S
site-directed mutagenesis, the mutant shows the mutant shows increased GlcNAc-1-P UTase activity compared to wild-type
T80S/Y97N
T80V
site-directed mutagenesis, the mutant shows slightly reduced GlcNAc-1-P UTase activity compared to wild-type
T80W
site-directed mutagenesis, the mutant shows no Glc-1-P UTase activity and highly reduced GlcNAc-1-P UTase activity
T80Y
site-directed mutagenesis, the mutant shows no Glc-1-P UTase activity and highly reduced GlcNAc-1-P UTase activity
Y97A
Y97A/K147A
Y97C
Y97D
Y97E
Y97F
Y97F/K147A
Y97G
Y97H
Y97I
Y97K
Y97L
Y97M
Y97N
Y97P
Y97Q
Y97R
Y97S
Y97T
Y97V
E146A
-
exhibits slightly weaker UDP-N-acetylglucosamine diphosphorylase activity than wild-type enzyme
-
G9A
-
enhanced UDP-N-acetylglucosamine diphosphorylase activity under optimal conditions
-
K147A
-
enhanced UDP-N-acetylglucosamine diphosphorylase activity under optimal conditions
-
T80A
-
site-directed mutagenesis, the mutant shows increased GlcNAc-1-P and Glc-1-P UTase activity compared to wild-type
-
T80D
-
site-directed mutagenesis, the mutant shows increased Glc-1-P UTase activity and reduced GlcNAc-1-P UTase activity compared to wild-type
-
T80P
-
site-directed mutagenesis, the mutant shows slightly reduced GlcNAc-1-P UTase activity compared to wild-type
-
T80S
-
site-directed mutagenesis, the mutant shows the mutant shows increased GlcNAc-1-P UTase activity compared to wild-type
-
Y97F
-
enhanced UDP-N-acetylglucosamine diphosphorylase activity under optimal conditions
-
G9A
G9A/T80A
-
specific GlcNAc-1-P UTase activity of the mutant enzyme is 35fold lower compared to specific activity of the wild-type enzyme
-
K147A
-
GlcNAc-1-P UTase activity is 25% of the activity of wild-type ST0452 protein
-
T80A
-
site-directed mutagenesis, the mutant shows increased GlcNAc-1-P and Glc-1-P UTase activity compared to wild-type
-
T80D
-
site-directed mutagenesis, the mutant shows increased Glc-1-P UTase activity and reduced GlcNAc-1-P UTase activity compared to wild-type
-
T80P
-
site-directed mutagenesis, the mutant shows slightly reduced GlcNAc-1-P UTase activity compared to wild-type
-
T80S
-
site-directed mutagenesis, the mutant shows the mutant shows increased GlcNAc-1-P UTase activity compared to wild-type
-
T80S/Y97N
-
the mutant enzyme shows 6.5times-higher activity, compared to that of the wild-type ST0452 protein, revealing that these two substituted residues function cooperatively to increase N-acetylglucosamine-1-phosphate uridyltransferase activity
-
Y97A
-
1.82fold increase in GlcNAc-1-P UTase activity compared to that of the wild-type enzyme
-
Y97F
Y97N
Y97V
-
3.56fold increase in GlcNAc-1-P UTase activity compared to that of the wild-type enzyme
-
T80A
-
site-directed mutagenesis, the mutant shows increased GlcNAc-1-P and Glc-1-P UTase activity compared to wild-type
-
T80D
-
site-directed mutagenesis, the mutant shows increased Glc-1-P UTase activity and reduced GlcNAc-1-P UTase activity compared to wild-type
-
T80P
-
site-directed mutagenesis, the mutant shows slightly reduced GlcNAc-1-P UTase activity compared to wild-type
-
T80S
-
site-directed mutagenesis, the mutant shows the mutant shows increased GlcNAc-1-P UTase activity compared to wild-type
-
T80A
-
site-directed mutagenesis, the mutant shows increased GlcNAc-1-P and Glc-1-P UTase activity compared to wild-type
-
T80D
-
site-directed mutagenesis, the mutant shows increased Glc-1-P UTase activity and reduced GlcNAc-1-P UTase activity compared to wild-type
-
T80P
-
site-directed mutagenesis, the mutant shows slightly reduced GlcNAc-1-P UTase activity compared to wild-type
-
T80S
-
site-directed mutagenesis, the mutant shows the mutant shows increased GlcNAc-1-P UTase activity compared to wild-type
-
C307S
additional information
Y103A
mutant protein shows a 24% increase in activity compared to that of the wild-type enzyme
Y103N
mutant protein shows a 32% increase in activity compared to that of the wild-type enzyme
Y103A
-
mutant protein shows a 24% increase in activity compared to that of the wild-type enzyme
-
Y103N
-
mutant protein shows a 32% increase in activity compared to that of the wild-type enzyme
-
D208A
enhanced UDP-N-acetylglucosamine diphosphorylase activity under optimal conditions
D208A
exhibits slightly weaker UDP-N-acetylglucosamine diphosphorylase activity than wild-type enzyme
D208A
GlcNAc-1-P UTase activity is 10% of the activity of wild-type ST0452 protein
E146A
exhibits slightly weaker UDP-N-acetylglucosamine diphosphorylase activity than wild-type enzyme
G9A
enhanced UDP-N-acetylglucosamine diphosphorylase activity under optimal conditions
G9A
site-directed mutagenesis, the mutant exhibits increased GlcNAc-1-P UTase activity compared to wild-type
G9A/K147A
specific GlcNAc-1-P UTase activity of the mutant enzyme is 71.8fold lower compared to specific activity of the wild-type enzyme
G9A/K147A
activity is 71.8fold lower than that of the wild-type enzyme
G9A/K147A
site-directed mutagenesis, the mutant shows highly reduced activity compared to wild-type enzyme
G9A/T80A
specific GlcNAc-1-P UTase activity of the mutant enzyme is 35fold lower compared to specific activity of the wild-type enzyme
G9A/T80A
activity is 35fold lower than that of the wild-type enzyme
G9A/T80A
site-directed mutagenesis, the mutant shows highly reduced activity compared to wild-type enzyme
G9A/Y97F
specific GlcNAc-1-P UTase activity of the mutant enzyme is 14.5fold lower compared to specific activity of the wild-type enzyme
G9A/Y97F
activity is 14.5fold lower than that of the wild-type enzyme
G9A/Y97F
site-directed mutagenesis, the mutant shows reduced activity compared to wild-type enzyme
K147A
enhanced UDP-N-acetylglucosamine diphosphorylase activity under optimal conditions
K147A
GlcNAc-1-P UTase activity is 25% of the activity of wild-type ST0452 protein
K147A
site-directed mutagenesis, the mutant exhibits increased GlcNAc-1-P UTase activity compared to wild-type
R13A
GlcNAc-1-P UTase activity is 10% of the activity of wild-type ST0452 protein
T80A
enhanced UDP-N-acetylglucosamine diphosphorylase activity under optimal conditions
T80A
GlcNAc-1-P UTase activity is 56% of the activity of wild-type ST0452 protein
T80A
site-directed mutagenesis, the mutant exhibits increased GlcNAc-1-P UTase activity compared to wild-type
T80A
site-directed mutagenesis, the mutant shows increased GlcNAc-1-P and Glc-1-P UTase activity compared to wild-type
T80A/K147A
specific GlcNAc-1-P UTase activity of the mutant enzyme is 11.09fold lower compared to specific activity of the wild-type enzyme
T80A/K147A
activity is 11.1fold lower than that of the wild-type enzyme
T80A/K147A
site-directed mutagenesis, the mutant shows reduced activity compared to wild-type enzyme
T80A/Y97A
specific GlcNAc-1-P UTase activity of the mutant enzyme is 31.4fold lower compared to specific activity of the wild-type enzyme
T80A/Y97A
activity is 31.4fold lower than that of the wild-type enzyme
T80A/Y97A
site-directed mutagenesis, the mutant shows highly reduced activity compared to wild-type enzyme
T80A/Y97F
specific GlcNAc-1-P UTase activity of the mutant enzyme is 25.1fold lower compared to specific activity of the wild-type enzyme
T80A/Y97F
activity is 25.1fold lower than that of the wild-type enzyme
T80A/Y97F
site-directed mutagenesis, the mutant shows highly reduced activity compared to wild-type enzyme
T80L
site-directed mutagenesis, the mutant shows reduced GlcNAc-1-P UTase activity and no Glc-1-P UTase activity
T80S/Y97N
the mutant enzyme shows 6.5times-higher activity, compared to that of the wild-type ST0452 protein, revealing that these two substituted residues function cooperatively to increase N-acetylglucosamine-1-phosphate uridyltransferase activity
T80S/Y97N
site-directed mutagenesis, the mutant shows 6.5times higher activity with N-acetylglucosamine-1-phosphate compared to that of the wild-type ST0452 protein
Y97A
enhanced UDP-N-acetylglucosamine diphosphorylase activity under optimal conditions
Y97A
1.82fold increase in GlcNAc-1-P UTase activity compared to that of the wild-type enzyme
Y97A
GlcNAc-1-P UTase activity is 66% of the activity of wild-type ST0452 protein
Y97A
activity is 1.83fold higher than that of the wild-type enzyme
Y97A
site-directed mutagenesis, the mutant exhibits increased GlcNAc-1-P UTase activity compared to wild-type
Y97A/K147A
specific GlcNAc-1-P UTase activity of the mutant enzyme is 11.9fold lower compared to specific activity of the wild-type enzyme
Y97A/K147A
activity is 11.9fold lower than that of the wild-type enzyme
Y97A/K147A
site-directed mutagenesis, the mutant shows reduced activity compared to wild-type enzyme
Y97C
activity is 1.4fold lower than that of the wild-type enzyme
Y97C
site-directed mutagenesis, the mutant shows reduced activity compared to wild-type enzyme
Y97D
activity is 1.3fold higher than that of the wild-type enzyme
Y97D
site-directed mutagenesis, the mutant exhibits increased GlcNAc-1-P UTase activity compared to wild-type
Y97E
activity is 2.1fold lower than that of the wild-type enzyme
Y97E
site-directed mutagenesis, the mutant shows reduced activity compared to wild-type enzyme
Y97F
enhanced UDP-N-acetylglucosamine diphosphorylase activity under optimal conditions
Y97F
GlcNAc-1-P UTase activity is similar to wild-type ST0452 protein
Y97F
activity is 1.2fold higher than that of the wild-type enzyme
Y97F
site-directed mutagenesis, the mutant exhibits increased GlcNAc-1-P UTase activity compared to wild-type
Y97F/K147A
specific GlcNAc-1-P UTaseactivity of the mutant enzyme is 5.7fold lower compared to specific activity of the wild-type enzyme
Y97F/K147A
activity is 5.4fold lower than that of the wild-type enzyme
Y97F/K147A
site-directed mutagenesis, the mutant shows reduced activity compared to wild-type enzyme
Y97G
activity is 1.2fold higher than that of the wild-type enzyme
Y97G
site-directed mutagenesis, the mutant exhibits slightly increased GlcNAc-1-P UTase activity compared to wild-type
Y97H
activity is 2.9fold higher than that of the wild-type enzyme
Y97H
site-directed mutagenesis, the mutant exhibits highly increased GlcNAc-1-P UTase activity compared to wild-type
Y97I
activity is 1.2fold higher than that of the wild-type enzyme
Y97I
site-directed mutagenesis, the mutant exhibits slightly increased GlcNAc-1-P UTase activity compared to wild-type
Y97K
activity is 1.35fold higher than that of the wild-type enzyme
Y97K
site-directed mutagenesis, the mutant exhibits increased GlcNAc-1-P UTase activity compared to wild-type
Y97L
activity is 1.5fold higher than that of the wild-type enzyme
Y97L
site-directed mutagenesis, the mutant exhibits increased GlcNAc-1-P UTase activity compared to wild-type
Y97M
activity is 1.3fold higher than that of the wild-type enzyme
Y97M
site-directed mutagenesis, the mutant exhibits slightly increased GlcNAc-1-P UTase activity compared to wild-type
Y97N
4.45fold increase in GlcNAc-1-P UTase activity compared to that of the wild-type enzyme
Y97N
the mutant enzyme exhibits over 4 times higher N-acetylglucosamine-1-phosphate uridyltransferase activity, compared with that of the wild-type ST0452 protein. The three-dimensional structure of the Y97N protein is not changed by this substitution but the interactions with the substrate are slightly modified, which might cause the activity to increase. The crystal structure of the Y97N protein shows that positions 146 (Glu) and 80 (Thr) form interactions with GlcNAc, and an engineering strategy is applied to these residues to increase activity
Y97N
activity is 4.5fold higher than that of the wild-type enzyme
Y97N
site-directed mutagenesis, the mutant exhibits highly increased GlcNAc-1-P UTase activity compared to wild-type
Y97N
naturally occuring mutation, the Y97N mutant of the ST0452 protein, isolated from Sulfolobus tokodaii, exhibits over 4times higher N-acetylglucosamine-1-phosphate (GlcNAc-1-P) uridyltransferase (UTase, EC 2.7.7.23) activity, compared with that of the wild-type ST0452 protein, three-dimensional structure analysis of the Y97N protein. The overall structure is almost identical to that of the wild-type ST0452 protein (PDB ID 2GGO), with residue 97 (Asn) interacting with the O-5 atom of N-acetylglucosamine (GlcNAc) in the complex without metal ions
Y97P
activity is 1.5fold higher than that of the wild-type enzyme
Y97P
site-directed mutagenesis, the mutant shows reduced activity compared to wild-type enzyme
Y97Q
activity is 2.3fold higher than that of the wild-type enzyme
Y97Q
site-directed mutagenesis, the mutant exhibits increased GlcNAc-1-P UTase activity compared to wild-type
Y97S
activity is 1.97fold higher than that of the wild-type enzyme
Y97S
site-directed mutagenesis, the mutant exhibits increased GlcNAc-1-P UTase activity compared to wild-type
Y97T
activity is 2.1fold higher than that of the wild-type enzyme
Y97T
site-directed mutagenesis, the mutant exhibits increased GlcNAc-1-P UTase activity compared to wild-type
Y97V
3.56fold increase in GlcNAc-1-P UTase activity compared to that of the wild-type enzyme
Y97V
activity is 3.6fold higher than that of the wild-type enzyme
Y97V
site-directed mutagenesis, the mutant exhibits highly increased GlcNAc-1-P UTase activity compared to wild-type
G9A
-
site-directed mutagenesis, the mutant exhibits increased GlcNAc-1-P UTase activity compared to wild-type
-
Y97F
-
activity is 1.2fold higher than that of the wild-type enzyme
-
Y97F
-
GlcNAc-1-P UTase activity is similar to wild-type ST0452 protein
-
Y97N
-
the mutant enzyme exhibits over 4 times higher N-acetylglucosamine-1-phosphate uridyltransferase activity, compared with that of the wild-type ST0452 protein. The three-dimensional structure of the Y97N protein is not changed by this substitution but the interactions with the substrate are slightly modified, which might cause the activity to increase. The crystal structure of the Y97N protein shows that positions 146 (Glu) and 80 (Thr) form interactions with GlcNAc, and an engineering strategy is applied to these residues to increase activity
-
Y97N
-
4.45fold increase in GlcNAc-1-P UTase activity compared to that of the wild-type enzyme
-
additional information
KT282116
knockdown of LdUAP1 or combined knockdown of LdUAP1 and LdUAP2
additional information
KT282117
knockdown of LdUAP1 or combined knockdown of LdUAP1 and LdUAP2
additional information
KT282116
knockdown of LdUAP2 or combined knockdown of LdUAP1 and LdUAP2
additional information
KT282117
knockdown of LdUAP2 or combined knockdown of LdUAP1 and LdUAP2
additional information
construction of Mycobacterium tuberculosis mutant strains overproducing GlmU allows determination of the contribution of the protein to mycobacterial entry into human neutrophils
additional information
-
construction of Mycobacterium tuberculosis mutant strains overproducing GlmU allows determination of the contribution of the protein to mycobacterial entry into human neutrophils
-
additional information
-
construction of Mycobacterium tuberculosis mutant strains overproducing GlmU allows determination of the contribution of the protein to mycobacterial entry into human neutrophils
-
additional information
the lesion mimic mutant spl29, regenerated from the cultivar Zhonghua 11, exhibits spotted leaves and rapid leaf senescence from the seedling stage throughout the rest of its life cycle, and no enzymatic activity in the spl29 mutant. Functional complementation with LOC_Os08g10600 in the spl29 mutant
additional information
-
the lesion mimic mutant spl29, regenerated from the cultivar Zhonghua 11, exhibits spotted leaves and rapid leaf senescence from the seedling stage throughout the rest of its life cycle, and no enzymatic activity in the spl29 mutant. Functional complementation with LOC_Os08g10600 in the spl29 mutant
-
additional information
for industrial applications, activity needs to be increased without decreasing thermostability. To enhance this activity, mutations are introduced into the amino acid residues located within the predicted reaction centre by targeted mutagenesis. All 12 mutant ST0452 proteins show no decrease in thermostability. Among them, six mutant proteins are found to have increased UDP-N-acetylglucosamine diphosphorylase activity under optimal reaction conditions with sufficient substrates or an appropriate metal ion
additional information
construction of expression vectors encoding a series of ST0452 C-terminal deletion mutants with hexahistidine tags at their C-termini, designated pST0452(DC005)H, pST0452(DC011)H, pST0452(DC021)H, pST0452(DC031)H, pST0452(DC041) H, pST0452(DC051)H, pST0452(DC071)H, pST0452 (DC121)H and pST0452(DC171)H. The deletion mutants retain the same tertiary structures as the wild-type ST0452 protein
additional information
-
construction of expression vectors encoding a series of ST0452 C-terminal deletion mutants with hexahistidine tags at their C-termini, designated pST0452(DC005)H, pST0452(DC011)H, pST0452(DC021)H, pST0452(DC031)H, pST0452(DC041) H, pST0452(DC051)H, pST0452(DC071)H, pST0452 (DC121)H and pST0452(DC171)H. The deletion mutants retain the same tertiary structures as the wild-type ST0452 protein
additional information
N-acetyl-D-glucosamine-1-phosphate uridylyltransferase activity of the mutant enzyme deletion mutant lacking the 170-residue C-terminal domain remains in the truncated enzyme after 5 min of heating at 65 °C but is completely removed by treatment over 70°C
additional information
ST0452 proteins exhibiting a further increase in activity are created using a site saturation mutagenesis strategy at the 97th position. Kinetic analyses show that the increased activities of the mutant proteins are principally due to increased apparent kcat values. Nine double-mutant ST0452 proteins are generated with the intent of obtaining enzymes exhibiting a further increase in catalysis, but all show less than 15% of the wild-type N-acetyl-D-glucosamine-1-phosphate uridyltransferase (GlcNAc-1-P UTase) activity. The Y97A mutant exhibits the highest activity of the single-mutant proteins. Analysis of mutant ST0452 proteins generated using a site saturation mutagenesis strategy at the 97th position, overview
additional information
-
ST0452 proteins exhibiting a further increase in activity are created using a site saturation mutagenesis strategy at the 97th position. Kinetic analyses show that the increased activities of the mutant proteins are principally due to increased apparent kcat values. Nine double-mutant ST0452 proteins are generated with the intent of obtaining enzymes exhibiting a further increase in catalysis, but all show less than 15% of the wild-type N-acetyl-D-glucosamine-1-phosphate uridyltransferase (GlcNAc-1-P UTase) activity. The Y97A mutant exhibits the highest activity of the single-mutant proteins. Analysis of mutant ST0452 proteins generated using a site saturation mutagenesis strategy at the 97th position, overview
additional information
all proteins substituted at position 146 have drastically decreased activities, whereas several proteins substituted at position 80 show higher GlcNAc-1-P UTase activity, compared to that of the wild-type protein
additional information
-
all proteins substituted at position 146 have drastically decreased activities, whereas several proteins substituted at position 80 show higher GlcNAc-1-P UTase activity, compared to that of the wild-type protein
additional information
-
for industrial applications, activity needs to be increased without decreasing thermostability. To enhance this activity, mutations are introduced into the amino acid residues located within the predicted reaction centre by targeted mutagenesis. All 12 mutant ST0452 proteins show no decrease in thermostability. Among them, six mutant proteins are found to have increased UDP-N-acetylglucosamine diphosphorylase activity under optimal reaction conditions with sufficient substrates or an appropriate metal ion
-
additional information
-
all proteins substituted at position 146 have drastically decreased activities, whereas several proteins substituted at position 80 show higher GlcNAc-1-P UTase activity, compared to that of the wild-type protein
-
additional information
-
ST0452 proteins exhibiting a further increase in activity are created using a site saturation mutagenesis strategy at the 97th position. Kinetic analyses show that the increased activities of the mutant proteins are principally due to increased apparent kcat values. Nine double-mutant ST0452 proteins are generated with the intent of obtaining enzymes exhibiting a further increase in catalysis, but all show less than 15% of the wild-type N-acetyl-D-glucosamine-1-phosphate uridyltransferase (GlcNAc-1-P UTase) activity. The Y97A mutant exhibits the highest activity of the single-mutant proteins. Analysis of mutant ST0452 proteins generated using a site saturation mutagenesis strategy at the 97th position, overview
-
additional information
-
N-acetyl-D-glucosamine-1-phosphate uridylyltransferase activity of the mutant enzyme deletion mutant lacking the 170-residue C-terminal domain remains in the truncated enzyme after 5 min of heating at 65 °C but is completely removed by treatment over 70°C
-
additional information
-
all proteins substituted at position 146 have drastically decreased activities, whereas several proteins substituted at position 80 show higher GlcNAc-1-P UTase activity, compared to that of the wild-type protein
-
additional information
-
all proteins substituted at position 146 have drastically decreased activities, whereas several proteins substituted at position 80 show higher GlcNAc-1-P UTase activity, compared to that of the wild-type protein
-
additional information
-
all proteins substituted at position 146 have drastically decreased activities, whereas several proteins substituted at position 80 show higher GlcNAc-1-P UTase activity, compared to that of the wild-type protein
-
