Literature summary for 3.5.2.6 extracted from

Ellerby, L.M.; Escobar, W.A.; Fink, A.L.; Mitchinson, C.; Wells, J.A.
The role of lysine-234 in beta-lactamase catalysis probed by site directed mutagenesis (1990), Biochemistry, 29, 5797-5806.

Search for more literature for 3.5.2.6

Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Bacillus subtilis	Bacillus licheniformis

Protein Variants

Protein Variants	Comment	Organism
K234A	no significant structural change, mutant with decreased Km 1-2 orders of magnitude and kcat 2-3 orders of magnitude	Bacillus licheniformis
K234E	no significant structural change, mutant with decreased Km 1-2 orders of magnitude and kcat 2-3 orders of magnitude	Bacillus licheniformis

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.041	-	nitrocefin	wild type	Bacillus licheniformis
0.1	-	phenoxymethylpenicillin	wild type	Bacillus licheniformis
0.12	-	benzylpenicillin	wild type	Bacillus licheniformis
0.14	-	7-(thienyl-2-acetamido)-3-[2-(4-,N,N-dimethylaminophenylazo)pyridinium-methyl]-3-cephem-4-carboxylic acid	wild type	Bacillus licheniformis
0.21	-	ampicillin	wild type	Bacillus licheniformis

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
extracellular	-	Bacillus licheniformis	-	-

Organism

Organism	UniProt	Comment	Textmining
Bacillus licheniformis	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
-	Bacillus licheniformis

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
7-(thienyl-2-acetamido)-3-[2-(4-N,N-dimethylaminophenylazo)pyridinium-methyl]-3-cephem-4-carboxylic acid + H2O	i.e. PADAC	Bacillus licheniformis	(2R)-2-[(R)-carboxy{(E)-[1-hydroxy-2-(thiophen-2-yl)ethylidene]amino}methyl]-5-{[(2E)-2-{[4-(dimethyliminio)cyclohexa-2,5-dien-1-ylidene]hydrazinylidene}pyridin-1(2H)-yl]methyl}-3,6-dihydro-2H-1,3-thiazine-4-carboxylate	-	?
ampicillin + H2O	-	Bacillus licheniformis	(2R,4S)-2-[(R)-[[(2R)-2-amino-2-phenylacetyl]amino](carboxy)methyl]-5,5-dimethyl-1,3-thiazolidine-4-carboxylic acid	-	?
benzylpenicillin + H2O	-	Bacillus licheniformis	(2R,4S)-2-[(R)-carboxy(2-phenylacetamido)methyl]-5,5-dimethyl-1,3-thiazolidine-4-carboxylic acid	-	?
nitrocefin + H2O	-	Bacillus licheniformis	(2R)-2-{(R)-carboxy[2-(thiophen-2-yl)acetamido]methyl}-5-[(E)-2-(2,4-dinitrophenyl)ethenyl]-3,6-dihydro-2H-1,3-thiazine-4-carboxylic acid	-	?
phenoxymethylpenicillin + H2O	-	Bacillus licheniformis	(2R,4S)-2-[(R)-carboxy(2-phenoxyacetamido)methyl]-5,5-dimethyl-1,3-thiazolidine-4-carboxylic acid	-	?

Temperature Range [°C]

Temperature Minimum [°C]	Temperature Maximum [°C]	Comment	Organism
50	75	pH 7: stable up to 58, 62, 68°C for K234E, wild type and K234A respectively	Bacillus licheniformis

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
667	-	7-(thienyl-2-acetamido)-3-[2-(4-,N,N-dimethylaminophenylazo)pyridinium-methyl]-3-cephem-4-carboxylic acid	-	Bacillus licheniformis
1090	-	nitrocefin	-	Bacillus licheniformis
2240	-	ampicillin	-	Bacillus licheniformis
2550	-	phenoxymethylpenicillin	-	Bacillus licheniformis
2650	-	benzylpenicillin	-	Bacillus licheniformis

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
6.5	-	substrate phenoxymethylpenicillin	Bacillus licheniformis

pH Range

pH Minimum	pH Maximum	Comment	Organism
4	9	substrate phenoxymethylpenicillin: kcat 1/6 at pH 9 compared to pH 6.5	Bacillus licheniformis

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Release 2023.1 (January 2023)