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Literature summary for 3.5.2.6 extracted from

  • Lim, D.; Sanschagrin, F.; Passmore, L.; De Castro, L.; Levesque, R.C.; Strynadka, N.C.
    Insights into the molecular basis for the carbenicillinase activity of PSE-4 beta-lactamase from crystallographic and kinetic studies (2001), Biochemistry, 40, 395-402.
    View publication on PubMed
Search for more literature for 3.5.2.6

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type and mutant R234K in Escherichia coli BL21(DE3) Pseudomonas aeruginosa

Crystallization (Commentary)

Crystallization (Comment) Organism
0.003 ml 10 mg/ml purified recombinant wild-type enzyme or mutant R234K, mixed with 0.001 ml of well solution containing 2 M ammonium sulfate, 50 mM MOPS pH 6.4, and 0.1 M acetate pH 4.5, respectively, 100 mM MgCl2, 1-2 weeks at room temperature, X-ray diffraction structure determination and analysis at 1.75 A resolution Pseudomonas aeruginosa

Protein Variants

Protein Variants Comment Organism
R234K site-directed replacement mutagenesis, about 4fold reduced catalytic efficiency compared to the wild-type enzyme Pseudomonas aeruginosa

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.033
-
 ampicillin pH 7.0, 30°C, wild-type enzyme Pseudomonas aeruginosa
0.057
-
 penicillin G pH 7.0, 30°C, wild-type enzyme Pseudomonas aeruginosa
0.068
-
 carbenicillin pH 7.0, 30°C, wild-type enzyme Pseudomonas aeruginosa
0.074
-
 ampicillin pH 7.0, 30°C, mutant R234K Pseudomonas aeruginosa
0.1
-
 penicillin G pH 7.0, 30°C, mutant R234K Pseudomonas aeruginosa
0.129
-
 carbenicillin pH 7.0, 30°C, mutant R234K Pseudomonas aeruginosa

Organism

Organism UniProt Comment Textmining
Pseudomonas aeruginosa P16897 class A enzyme PSE-4, the active site structure involves Ser70, Lys73, Ser130, Glu166, Asn170, and Arg234 as catalytic residues
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant R234K from Escherichia coli BL21(DE3) Pseudomonas aeruginosa

Reaction

Reaction Comment Organism Reaction ID
a beta-lactam + H2O = a substituted beta-amino acid Arg234 is involved in catalysis Pseudomonas aeruginosa
a

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ampicillin + H2O
-
 Pseudomonas aeruginosa (2R,4S)-2-[(R)-[[(2R)-2-amino-2-phenylacetyl]amino](carboxy)methyl]-5,5-dimethyl-1,3-thiazolidine-4-carboxylic acid
-
 ?
carbenicillin + H2O
-
 Pseudomonas aeruginosa (2R,4S)-2-{(R)-carboxy[2-carboxy(phenyl)acetamido]methyl}-5,5-dimethyl-1,3-thiazolidine-4-carboxylic acid
-
 ?
nitrocefin + H2O
-
 Pseudomonas aeruginosa (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
-
 ?
penicillin G + H2O
-
 Pseudomonas aeruginosa (2R,4S)-2-[(R)-carboxy[(phenylacetyl)amino]methyl]-5,5-dimethyl-1,3-thiazolidine-4-carboxylic acid
-
 ?

Subunits

Subunits Comment Organism
More enzyme consists of an alpha/beta domain and a predominantly alpha-helical domain Pseudomonas aeruginosa

Synonyms

Synonyms Comment Organism
Carbenicillinase
-
 Pseudomonas aeruginosa
PSE-4
-
 Pseudomonas aeruginosa

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
 assay at Pseudomonas aeruginosa

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
57
-
 carbenicillin pH 7.0, 30°C, mutant R234K Pseudomonas aeruginosa
430
-
 penicillin G pH 7.0, 30°C, mutant R234K Pseudomonas aeruginosa
840
-
 ampicillin pH 7.0, 30°C, mutant R234K Pseudomonas aeruginosa
890
-
 penicillin G pH 7.0, 30°C, wild-type enzyme Pseudomonas aeruginosa
1170
-
 ampicillin pH 7.0, 30°C, wild-type enzyme Pseudomonas aeruginosa
1200
-
 carbenicillin pH 7.0, 30°C, wild-type enzyme Pseudomonas aeruginosa

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
 assay at Pseudomonas aeruginosa