1.1.1.22: UDP-glucose 6-dehydrogenase

This is an abbreviated version, for detailed information about UDP-glucose 6-dehydrogenase, go to the full flat file.

Reaction

UDP-alpha-D-glucose
+ 2 NAD+ +
H2O
=
UDP-alpha-D-glucuronate
+ 2 NADH + 2 H+

Synonyms

ADH-like UDP-glucose dehydrogenase, AglM, BceC, dehydrogenase, uridine diphosphoglucose, dual specificity UDP-glucose dehydrogenase, GbUGD, HasB, HasB2, HVO_1531, PA2022, PA3559, sqv-4, Sugarless protein, UDG1, UDP-alpha-D-glucose:NAD oxidoreductase, UDP-D-glucose dehydrogenase, UDP-GDH, UDP-Glc dehydrogenase, UDP-Glc DH, UDP-GlcDH, UDP-GlcDHase, UDP-glucose dehydrogenase, UDPDH, UDPG dehydrogenase, UDPG-DH, UDPG:NAD oxidoreductase, UDPGD, UDPGDH, UDPGDH-A, UDPGDH-B, UDPGlc dehydrogenase, UDPglucose dehydrogenase, UDPglucose:NAD+ oxidoreductase, Ugd, UGD1, UgdG, UGDH, uridine diphosphate D-glucose dehydrogenase, uridine diphosphate glucose dehydrogenase, uridine diphosphate glucose-6-dehydrogenase, uridine diphosphate-glucose dehydrogenase, uridine diphosphoglucose dehydrogenase, uridine-5'-diphosphoglucose dehydrogenase, uridyl phosphate dehydrogenase, VNG1048G, VNG_1048G

ECTree

     1 Oxidoreductases
         1.1 Acting on the CH-OH group of donors
             1.1.1 With NAD+ or NADP+ as acceptor
                1.1.1.22 UDP-glucose 6-dehydrogenase

Engineering

Engineering on EC 1.1.1.22 - UDP-glucose 6-dehydrogenase

Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Y10F
mutation in GXGYXG consensus motif, 9% residual activity. Tyr10 plays a catalytic role in the final hydrolysis step. Upon release of NADH after the second oxidation step, Tyr10 may work as a proton conveyer from the aqueous hydrogen-bonding proton wire system to the hydrolytic site
Y10K
mutation in GXGYXG consensus motif, 2% residual activity
Y10S
mutation in GXGYXG consensus motif, 3% residual activity
K323A
-
site-directed mutagenesis, the mutant shows altered kinetics compared to the wild-type enzyme
R324A
-
site-directed mutagenesis, the mutant purifies in much lower amounts relative to wild-type and is prone to degradation and has negligible activity
Y71F
-
site-directed mutagenesis, the mutant shows unaltered catalytic activity
A222Q/S233G
C276E
-
activity is not measurable at pH 8.7, 22°C
C276G
-
activity is not measurable at pH 8.7, 22°C
C276K
-
activity is not measurable at pH 8.7, 22°C
C276L
-
activity is not measurable at pH 8.7, 22°C
C276Y
-
activity is not measurable at pH 8.7, 22°C
D280A
-
extremely poor enzymic activity
D280E
-
site-directed mutagenesis, 3-fold increase in Km for UDP-glucose and a 2-fold reduced Vmax relative to that of the wild type
DELTA132
-
deletion of residue Val132 from the Thr131 loop to approximate an intermediate state in the allosteric transition. The crystal structure of the deletion construct reveals an open conformation that relaxes steric constraints and facilitates repacking of the protein core. The open conformation stabilizes the construct as a hexamer with point group symmetry 32, similar to that of the active complex. The DELTA132 and UDP-alpha-D-xylose-inhibited structures have similar hexamer-building interfaces
E110A
-
site-directed mutagenesis, the mutant, although dimeric in the apo form, exhibits only about 50% reduction in Vmax, but is highly unstable in solution and in cultured cells so it cannot be evaluated unambiguously
E161Q
-
hydrolysis step becomes completely rate-limiting so that a thioester enzyme intermediate accumulates at steady state. Crystallization of E161Q in the presence of 5 mM UDP-glucose and 2 mM NAD results in trapping a thiohemiacetal enzyme intermediate
G13E
normal expression and stability of mutant, no enzymic activity, no photoaffinity labeling with nicotinamide 2-azidoadenosine dinucleotide
K220H
-
site-directed mutagenesis, putative active site residue, mutation severly impairs enzyme function
K220R
-
site-directed mutagenesis, putative active site residue, mutation severly impairs enzyme function
N224A
-
steady-state kinetic parameters are within an order of magnitude of the native enzyme
T131S
-
steady-state kinetic parameters are within an order of magnitude of the native enzyme
T325A
-
site-directed mutagenesis, the mutant occurs as dimeric species that can be induced to form hexamers in the ternary complex with substrate and cofactor. The inducible hexamer shows that upon increasing enzyme concentration, which favors the hexameric species, activity is modestly decreased and exhibits cooperativity. The T325A mutant is significantly less efficient in promoting downstream hyaluronan production by HEK293 cells than the wild-type. The activity of the T325A mutant is the most labile, with a half-life of only 24 h that is not extended significantly by substrate and cofactor addition
T325D
-
site-directed mutagenesis, the mutant yields exclusively dimeric species. The T325D mutant is significantly less efficient in promoting downstream hyaluronan production by HEK293 cells than the wild-type. UGDH T325D retains its activity similarly to the wild-type enzyme but does not exhibit increased stability in the abortive ternary complex
C260A
-
no oxidation of UDP-glucose to glucuronic acid, but capable of both reducing the aldehyde intermediate and oxidizing the hydrated form of the aldehyde intermediate, protein is expressed in inclusion bodies
E141Q
-
kcat-value 10fold lower than wild-type
E145Q
-
kcat-value 10fold lower than wild-type
T118A
-
160fold reduction of kcat value
additional information