Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary extracted from

  • Hazra, S.; Xu, H.; Blanchard, J.S.
    Tebipenem, a new carbapenem antibiotic, is a slow substrate that inhibits the beta-lactamase from Mycobacterium tuberculosis (2014), Biochemistry, 53, 3671-3678.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.5.2.6 gene blaC, DNA and amino acid sequence determination and analysis, recombinant expression of His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Mycobacterium tuberculosis

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.5.2.6 purified recombinant detagged wild-type and mutant enzymes in complex with inhibitor tebipenem, hanging drop vapor diffusion, mixing of 12 mg/ml protein with 0.1 M HEPES and 2 M NH4H2PO4, pH 4.1, in a 1:1 ratio, 10°C, microseeding for the mutant enzyme, X-ray diffraction structure determination and analysis at 1.90 A and 1.75 A resolution Mycobacterium tuberculosis

Protein Variants

EC Number Protein Variants Comment Organism
3.5.2.6 K73A site-directed mutagenesis, inactive mutant, substrate/inhibitor binding structure Mycobacterium tuberculosis

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.5.2.6 additional information carbapenems such as meropenem, doripenem, and ertapenem react with the enzyme to form enzyme-drug covalent complexes that are hydrolyzed extremely slowly Mycobacterium tuberculosis
3.5.2.6 tebi-pivoxil binding structure analysis, overview Mycobacterium tuberculosis
3.5.2.6 tebipenem Michaelis-Menten complex, exhibits slow tight-binding inhibition at low micromolar concentrations versus the chromogenic substrate nitrocefin, binding structure analysis, overview. TBPM-PI is a prodrug that is quickly hydrolyzed to tebipenem (TEBI). Active site of BlaC-tebipenem Michaelis-Menten complex showing interactions and interatomic distances between tebipenem and active site residues, overview Mycobacterium tuberculosis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.5.2.6 0.0002
-
 ertapenem pH 6.5, 22°C, recombinant enzyme Mycobacterium tuberculosis
3.5.2.6 0.0002
-
 doripenem pH 6.5, 22°C, recombinant enzyme Mycobacterium tuberculosis
3.5.2.6 0.0008
-
 tebipenem pH 6.5, 22°C, recombinant enzyme Mycobacterium tuberculosis
3.5.2.6 0.0034
-
 meropenem pH 6.5, 22°C, recombinant enzyme Mycobacterium tuberculosis

Organism

EC Number Organism UniProt Comment Textmining
3.5.2.6 Mycobacterium tuberculosis P9WKD3 gene blaC
-
3.5.2.6 Mycobacterium tuberculosis H37Rv P9WKD3 gene blaC
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.5.2.6 recombinant His6-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, tag cleavage by thrombin, and gel filtration Mycobacterium tuberculosis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.5.2.6 doripenem + H2O very low activity Mycobacterium tuberculosis (4R,5S)-5-[(1S,2R)-1-carboxy-2-hydroxypropyl]-4-methyl-3-([(3S,5S)-5-[(sulfamoylamino)methyl]pyrrolidin-3-yl]sulfanyl)-4,5-dihydro-1H-pyrrole-2-carboxylic acid
-
 ?
3.5.2.6 doripenem + H2O very low activity Mycobacterium tuberculosis H37Rv (4R,5S)-5-[(1S,2R)-1-carboxy-2-hydroxypropyl]-4-methyl-3-([(3S,5S)-5-[(sulfamoylamino)methyl]pyrrolidin-3-yl]sulfanyl)-4,5-dihydro-1H-pyrrole-2-carboxylic acid
-
 ?
3.5.2.6 ertapenem + H2O very low activity Mycobacterium tuberculosis (4R,5S)-5-[(1S,2R)-1-carboxy-2-hydroxypropyl]-3-([(3S,5S)-5-[(3-carboxyphenyl)carbamoyl]pyrrolidin-3-yl]sulfanyl)-4-methyl-4,5-dihydro-1H-pyrrole-2-carboxylic acid
-
 ?
3.5.2.6 ertapenem + H2O very low activity Mycobacterium tuberculosis H37Rv (4R,5S)-5-[(1S,2R)-1-carboxy-2-hydroxypropyl]-3-([(3S,5S)-5-[(3-carboxyphenyl)carbamoyl]pyrrolidin-3-yl]sulfanyl)-4-methyl-4,5-dihydro-1H-pyrrole-2-carboxylic acid
-
 ?
3.5.2.6 meropenem + H2O very low activity Mycobacterium tuberculosis (4R,5S)-5-[(1S,2R)-1-carboxy-2-hydroxypropyl]-3-[[(3S,5S)-5-(dimethylcarbamoyl)pyrrolidin-3-yl]sulfanyl]-4-methyl-4,5-dihydro-1H-pyrrole-2-carboxylic acid
-
 ?
3.5.2.6 meropenem + H2O very low activity Mycobacterium tuberculosis H37Rv (4R,5S)-5-[(1S,2R)-1-carboxy-2-hydroxypropyl]-3-[[(3S,5S)-5-(dimethylcarbamoyl)pyrrolidin-3-yl]sulfanyl]-4-methyl-4,5-dihydro-1H-pyrrole-2-carboxylic acid
-
 ?
3.5.2.6 additional information enzyme BlaC catalyzes the opening of the beta-lactam ring via nucleophilic attack by an active site serine residue (Ser70) to generate the acyl enzyme followed by hydrolysis of the ester bond to generate the inactive ring-opened product. Enzyme BlaC has an extremely broad spectrum of activity against penicillins and cephalosporins but weak activity against newer carbapenems. Carbapenems such as meropenem, doripenem, and ertapenem react with the enzyme to form enzyme-drug covalent complexes that are hydrolyzed extremely slowly Mycobacterium tuberculosis ?
-
 ?
3.5.2.6 additional information enzyme BlaC catalyzes the opening of the beta-lactam ring via nucleophilic attack by an active site serine residue (Ser70) to generate the acyl enzyme followed by hydrolysis of the ester bond to generate the inactive ring-opened product. Enzyme BlaC has an extremely broad spectrum of activity against penicillins and cephalosporins but weak activity against newer carbapenems. Carbapenems such as meropenem, doripenem, and ertapenem react with the enzyme to form enzyme-drug covalent complexes that are hydrolyzed extremely slowly Mycobacterium tuberculosis H37Rv ?
-
 ?
3.5.2.6 nitrocefin + H2O
-
 Mycobacterium tuberculosis (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 nitrocefin + H2O
-
 Mycobacterium tuberculosis H37Rv (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 tebipenem + H2O very low activity, active site of BlaC-tebipenem Michaelis-Menten complex showing interactions and interatomic distances between tebipenem and active site residues, overview Mycobacterium tuberculosis (4R,5S)-5-[(1S,2R)-1-carboxy-2-hydroxypropyl]-3-[[1-(4,5-dihydro-1,3-thiazol-2-yl)azetidin-3-yl]sulfanyl]-4-methyl-4,5-dihydro-1H-pyrrole-2-carboxylic acid
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.5.2.6 BlaC
-
 Mycobacterium tuberculosis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.5.2.6 22
-
 assay at room temperature Mycobacterium tuberculosis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.5.2.6 0.00033
-
 ertapenem pH 6.5, 22°C, recombinant enzyme Mycobacterium tuberculosis
3.5.2.6 0.00033
-
 doripenem pH 6.5, 22°C, recombinant enzyme Mycobacterium tuberculosis
3.5.2.6 0.0005
-
 tebipenem pH 6.5, 22°C, recombinant enzyme Mycobacterium tuberculosis
3.5.2.6 0.0013
-
 meropenem pH 6.5, 22°C, recombinant enzyme Mycobacterium tuberculosis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.5.2.6 6.5
-
 assay at Mycobacterium tuberculosis

General Information

EC Number General Information Comment Organism
3.5.2.6 evolution BlaC is a class A beta-lactamase Mycobacterium tuberculosis

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.5.2.6 0.3
-
 meropenem pH 6.5, 22°C, recombinant enzyme Mycobacterium tuberculosis
3.5.2.6 0.63
-
 tebipenem pH 6.5, 22°C, recombinant enzyme Mycobacterium tuberculosis
3.5.2.6 1.67
-
 ertapenem pH 6.5, 22°C, recombinant enzyme Mycobacterium tuberculosis
3.5.2.6 1.67
-
 doripenem pH 6.5, 22°C, recombinant enzyme Mycobacterium tuberculosis