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Literature summary 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

Cloned(Commentary)

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

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.5.2.6 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

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

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.5.2.6 0.033
-
 ampicillin pH 7.0, 30°C, wild-type enzyme Pseudomonas aeruginosa
3.5.2.6 0.057
-
 penicillin G pH 7.0, 30°C, wild-type enzyme Pseudomonas aeruginosa
3.5.2.6 0.068
-
 carbenicillin pH 7.0, 30°C, wild-type enzyme Pseudomonas aeruginosa
3.5.2.6 0.074
-
 ampicillin pH 7.0, 30°C, mutant R234K Pseudomonas aeruginosa
3.5.2.6 0.1
-
 penicillin G pH 7.0, 30°C, mutant R234K Pseudomonas aeruginosa
3.5.2.6 0.129
-
 carbenicillin pH 7.0, 30°C, mutant R234K Pseudomonas aeruginosa

Organism

EC Number Organism UniProt Comment Textmining
3.5.2.6 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)

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

Reaction

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

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.5.2.6 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
-
 ?
3.5.2.6 carbenicillin + H2O
-
 Pseudomonas aeruginosa (2R,4S)-2-{(R)-carboxy[2-carboxy(phenyl)acetamido]methyl}-5,5-dimethyl-1,3-thiazolidine-4-carboxylic acid
-
 ?
3.5.2.6 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
-
 ?
3.5.2.6 penicillin G + H2O
-
 Pseudomonas aeruginosa (2R,4S)-2-[(R)-carboxy[(phenylacetyl)amino]methyl]-5,5-dimethyl-1,3-thiazolidine-4-carboxylic acid
-
 ?

Subunits

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

Synonyms

EC Number Synonyms Comment Organism
3.5.2.6 Carbenicillinase
-
 Pseudomonas aeruginosa
3.5.2.6 PSE-4
-
 Pseudomonas aeruginosa

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.5.2.6 30
-
 assay at Pseudomonas aeruginosa

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.5.2.6 57
-
 carbenicillin pH 7.0, 30°C, mutant R234K Pseudomonas aeruginosa
3.5.2.6 430
-
 penicillin G pH 7.0, 30°C, mutant R234K Pseudomonas aeruginosa
3.5.2.6 840
-
 ampicillin pH 7.0, 30°C, mutant R234K Pseudomonas aeruginosa
3.5.2.6 890
-
 penicillin G pH 7.0, 30°C, wild-type enzyme Pseudomonas aeruginosa
3.5.2.6 1170
-
 ampicillin pH 7.0, 30°C, wild-type enzyme Pseudomonas aeruginosa
3.5.2.6 1200
-
 carbenicillin pH 7.0, 30°C, wild-type enzyme Pseudomonas aeruginosa

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.5.2.6 7
-
 assay at Pseudomonas aeruginosa