Literature summary for 3.2.2.27 extracted from

Xia, B.; Liu, Y.; Guevara, J.; Li, J.; Jilich, C.; Yang, Y.; Wang, L.; Dominy, B.N.; Cao, W.
Correlated mutation in the evolution of catalysis in uracil DNA glycosylase superfamily (2017), Sci. Rep., 7, 45978 .

Search for more literature for 3.2.2.27

Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Escherichia coli BL21(DE3) cells	Thermus thermophilus

Protein Variants

Protein Variants	Comment	Organism
E41A/G42D	the mutant shows severely reduced activity compared to the wild type enzyme	Thermus thermophilus
E41Q	the mutant shows 888fold reduced activity compared to the wild type enzyme	Thermus thermophilus
E41Q/G42D	the mutant shows 5.3fold reduced activity compared to the wild type enzyme	Thermus thermophilus
E47A	the mutant shows severely reduced activity compared to the wild type enzyme	Thermus thermophilus
F54A	the mutant shows severely reduced activity compared to the wild type enzyme	Thermus thermophilus
G42D	the mutant shows 89fold reduced activity compared to the wild type enzyme	Thermus thermophilus
H155S	the mutant shows reduced activity compared to the wild type enzyme	Thermus thermophilus
N80A	the mutant shows severely reduced activity compared to the wild type enzyme	Thermus thermophilus
N89A	the mutant shows reduced activity compared to the wild type enzyme	Thermus thermophilus

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.00017	-	double stranded DNA containing G-U mismatch	mutant enzyme E41Q/G42D, at pH 7.6 and 60°C	Thermus thermophilus
0.00074	-	double stranded DNA containing G-U mismatch	mutant enzyme N89A, at pH 7.6 and 60°C	Thermus thermophilus
0.00076	-	double stranded DNA containing G-U mismatch	mutant enzyme H155S, at pH 7.6 and 60°C	Thermus thermophilus
0.00097	-	double stranded DNA containing G-U mismatch	wild type enzyme, at pH 7.6 and 60°C	Thermus thermophilus

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
uracil-containing single-stranded DNA + H2O	Thermus thermophilus	-	uracil + single-stranded DNA with abasic site	-	?
uracil-mismatched double-stranded DNA + H2O	Thermus thermophilus	-	uracil + double-stranded DNA with abasic site	-	?

Organism

Organism	UniProt	Comment	Textmining
Thermus thermophilus	Q5SKC5	-	-

Purification (Commentary)

Purification (Comment)	Organism
-	Thermus thermophilus

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
double stranded DNA containing G-U mismatch + H2O	-	Thermus thermophilus	uracil + double-stranded DNA with abasic site	-	?
additional information	the enzyme does not show any detectable activity on other deaminated bases than uracil	Thermus thermophilus	?	-	?
uracil-containing single-stranded DNA + H2O	-	Thermus thermophilus	uracil + single-stranded DNA with abasic site	-	?
uracil-mismatched double-stranded DNA + H2O	-	Thermus thermophilus	uracil + double-stranded DNA with abasic site	-	?

Synonyms

Synonyms	Comment	Organism
family 4 UDGa	-	Thermus thermophilus
UDG	-	Thermus thermophilus

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
0.000072	-	double stranded DNA containing G-U mismatch	mutant enzyme H155S, at pH 7.6 and 60°C	Thermus thermophilus
0.0075	-	double stranded DNA containing G-U mismatch	mutant enzyme E41Q/G42D, at pH 7.6 and 60°C	Thermus thermophilus
0.025	-	double stranded DNA containing G-U mismatch	mutant enzyme N89A, at pH 7.6 and 60°C	Thermus thermophilus
0.23	-	double stranded DNA containing G-U mismatch	wild type enzyme, at pH 7.6 and 60°C	Thermus thermophilus

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
0.0092	-	double stranded DNA containing G-U mismatch	mutant enzyme H155S, at pH 7.6 and 60°C	Thermus thermophilus
35	-	double stranded DNA containing G-U mismatch	mutant enzyme N89A, at pH 7.6 and 60°C	Thermus thermophilus
45	-	double stranded DNA containing G-U mismatch	mutant enzyme E41Q/G42D, at pH 7.6 and 60°C	Thermus thermophilus
240	-	double stranded DNA containing G-U mismatch	wild type enzyme, at pH 7.6 and 60°C	Thermus thermophilus

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Release 2023.1 (January 2023)