Literature summary for 4.1.1.7 extracted from

Yep, A.; Kenyon, G.L.; McLeish, M.J.
Saturation mutagenesis of putative catalytic residues of benzoylformate decarboxylase provides a challenge to the accepted mechanism (2008), Proc. Natl. Acad. Sci. USA, 105, 5733-5738.

Search for more literature for 4.1.1.7

Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Escherichia coli BL21(DE3) cells	Pseudomonas putida

Protein Variants

Protein Variants	Comment	Organism
H281A	mutant with 152fold decreased kcat value compared to the wild type enzyme	Pseudomonas putida
H281F	mutant with 4.9fold decrease in kcat value compared to the wild type enzyme	Pseudomonas putida
H281N	mutant with 17fold decrease in kcat value compared to the wild type enzyme	Pseudomonas putida
H281Q	mutant with 37fold decrease in kcat value compared to the wild type enzyme	Pseudomonas putida
H281T	mutant with 159fold decrease in kcat value compared to the wild type enzyme	Pseudomonas putida
H281W	mutant with 19fold decrease in kcat value compared to the wild type enzyme	Pseudomonas putida
H281Y	mutant with 46fold decrease in kcat value compared to the wild type enzyme	Pseudomonas putida
H70A	mutant exhibits 4000fold decreased catalytic activity compared to the wild type enzyme	Pseudomonas putida
H70F	mutant exhibits a 236fold decrease in kcat/Km compared to the wild type enzyme	Pseudomonas putida
H70L	mutant exhibits a 33fold decrease in kcat/Km compared to the wild type enzyme	Pseudomonas putida
H70S	mutant exhibits a 197fold decrease in kcat/Km compared to the wild type enzyme	Pseudomonas putida
H70T	mutant exhibits a 25fold decrease in kcat/Km compared to the wild type enzyme	Pseudomonas putida
S26A	mutant with 21fold decrease in kcat value compared to the wild type enzyme	Pseudomonas putida
S26L	mutant exhibits no significant loss of activity compared to the wild type enzyme (2fold decrease in kcat value)	Pseudomonas putida
S26M	mutant exhibits no significant loss of activity compared to the wild type enzyme (12fold decrease in kcat value)	Pseudomonas putida
S26T	mutant exhibits no significant loss of activity compared to the wild type enzyme (3fold decrease in kcat value)	Pseudomonas putida

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.27	-	benzoylformate	wild type enzyme, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C	Pseudomonas putida
0.29	-	benzoylformate	mutant enzyme H281Q, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C	Pseudomonas putida
0.29	-	benzoylformate	mutant enzyme H281W, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C	Pseudomonas putida
0.38	-	benzoylformate	mutant enzyme H70L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C	Pseudomonas putida
0.49	-	benzoylformate	mutant enzyme H281Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C	Pseudomonas putida
0.54	-	benzoylformate	mutant enzyme H281F, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C	Pseudomonas putida
0.8	-	benzoylformate	mutant enzyme S26M, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C	Pseudomonas putida
0.85	-	benzoylformate	mutant enzyme H70F, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C	Pseudomonas putida
0.85	-	benzoylformate	mutant enzyme S26T, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C	Pseudomonas putida
0.89	-	benzoylformate	mutant enzyme H70T, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C	Pseudomonas putida
0.9	-	benzoylformate	mutant enzyme H281T, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C	Pseudomonas putida
0.94	-	benzoylformate	mutant enzyme H70S, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C	Pseudomonas putida
1.2	-	benzoylformate	mutant enzyme H281A, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C	Pseudomonas putida
1.2	-	benzoylformate	mutant enzyme S26L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C	Pseudomonas putida
1.5	-	benzoylformate	mutant enzyme H70A, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C	Pseudomonas putida
2.9	-	benzoylformate	mutant enzyme H281N in 100 mM potassium phosphate buffer (pH 6.0), at 30°C	Pseudomonas putida
7.8	-	benzoylformate	mutant enzyme S26A, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C	Pseudomonas putida

Organism

Organism	UniProt	Comment	Textmining
Pseudomonas putida	P20906	-	-

Purification (Commentary)

Purification (Comment)	Organism
-	Pseudomonas putida

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
benzoylformate	-	Pseudomonas putida	benzaldehyde + CO2	-	?

Synonyms

Synonyms	Comment	Organism
BFDC	-	Pseudomonas putida

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
0.46	-	benzoylformate	mutant enzyme H70A, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C	Pseudomonas putida
2	-	benzoylformate	mutant enzyme H281T, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C	Pseudomonas putida
2.1	-	benzoylformate	mutant enzyme H281A, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C	Pseudomonas putida
4.5	-	benzoylformate	mutant enzyme H70F, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C	Pseudomonas putida
5.6	-	benzoylformate	mutant enzyme H70S, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C	Pseudomonas putida
6.9	-	benzoylformate	mutant enzyme H281Y, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C	Pseudomonas putida
8.6	-	benzoylformate	mutant enzyme H281Q, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C	Pseudomonas putida
14	-	benzoylformate	mutant enzyme H70L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C	Pseudomonas putida
15	-	benzoylformate	mutant enzyme S26A, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C	Pseudomonas putida
17	-	benzoylformate	mutant enzyme H281W, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C	Pseudomonas putida
19	-	benzoylformate	mutant enzyme H281N in 100 mM potassium phosphate buffer (pH 6.0), at 30°C	Pseudomonas putida
26	-	benzoylformate	mutant enzyme S26M, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C	Pseudomonas putida
42	-	benzoylformate	mutant enzyme H70T, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C	Pseudomonas putida
65	-	benzoylformate	mutant enzyme H281F, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C	Pseudomonas putida
94	-	benzoylformate	mutant enzyme S26T, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C	Pseudomonas putida
132	-	benzoylformate	mutant enzyme S26L, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C	Pseudomonas putida
320	-	benzoylformate	wild type enzyme, in 100 mM potassium phosphate buffer (pH 6.0), at 30°C	Pseudomonas putida

Cofactor

Cofactor	Comment	Organism	Structure
thiamine diphosphate	0.5 mM, required for activity	Pseudomonas putida

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Release 2023.1 (January 2023)