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Literature summary extracted from

  • Caporale, B.; Franceschini, N.; Perilli, M.; Segatore, B.; Rossolini, G.M.; Amicosante, G.
    Biochemical characterization of laboratory mutants of extended-spectrum beta-lactamase TEM-60 (2004), Antimicrob. Agents Chemother., 48, 3579-3582.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.5.2.6 Escherichia coli strain HB101 is used as the host for the recombinant plasmids Providencia stuartii

Protein Variants

EC Number Protein Variants Comment Organism
3.5.2.6 K104E thermal stability is increased, mutant enzyme loses activity against oxyimino-cephgalosporins, aztreonam, and ampicillin Providencia stuartii
3.5.2.6 P51L thermal stability is greatly decreased, mutant enzyme loses activity against cefotaxime and exhibits a reduced catalytic efficiency with penicillins and aztreonam, no inhibition by beta-odopenicillanate Providencia stuartii
3.5.2.6 S164E thermal stability is increased, mutant enzyme loses activity against ceftazidime Providencia stuartii

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.5.2.6 beta-iodopenicillanate
-
 Providencia stuartii
3.5.2.6 tazobactam
-
 Providencia stuartii

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.5.2.6 0.0017
-
 penicillin G mutant enzyme K104E Providencia stuartii
3.5.2.6 0.0042
-
 ampicillin mutant enzyme K104E Providencia stuartii
3.5.2.6 0.009
-
 penicillin G wild-type enzyme Providencia stuartii
3.5.2.6 0.009
-
 ampicillin wild-type enzyme Providencia stuartii
3.5.2.6 0.015
-
 penicillin G mutant enzyme S164R Providencia stuartii
3.5.2.6 0.018
-
 cephaloridine mutant enzyme P51L Providencia stuartii
3.5.2.6 0.02
-
 nitrocefin wild-type enzyme Providencia stuartii
3.5.2.6 0.028
-
 cephaloridine wild-type enzyme Providencia stuartii
3.5.2.6 0.031
-
 (6R,7R)-3-[(acetyloxy)methyl]-7-[[(2E)-2-(2-amino-1,3-thiazol-4-yl)-2-(methoxyimino)acetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid wild-type enzyme Providencia stuartii
3.5.2.6 0.041
-
 nitrocefin mutant enzyme K104E Providencia stuartii
3.5.2.6 0.055
-
 aztreonam wild-type enzyme Providencia stuartii
3.5.2.6 0.059
-
 ceftazidime wild-type enzyme Providencia stuartii
3.5.2.6 0.065
-
 nitrocefin mutant enzyme S164R Providencia stuartii
3.5.2.6 0.07
-
 cephaloridine mutant enzyme K104E Providencia stuartii
3.5.2.6 0.08
-
 ceftazidime mutant enzyme P51L Providencia stuartii
3.5.2.6 0.156
-
 cephaloridine mutant enzyme S164R Providencia stuartii
3.5.2.6 0.18
-
 nitrocefin mutant enzyme P51L Providencia stuartii
3.5.2.6 0.21
-
 (6R,7R)-3-[(acetyloxy)methyl]-7-[[(2E)-2-(2-amino-1,3-thiazol-4-yl)-2-(methoxyimino)acetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid mutant enzyme S164R Providencia stuartii
3.5.2.6 0.335
-
 ampicillin mutant enzyme S164R Providencia stuartii
3.5.2.6 0.45
-
 ampicillin mutant enzyme P51L Providencia stuartii
3.5.2.6 0.575
-
 penicillin G mutant enzyme P51L Providencia stuartii
3.5.2.6 0.66
-
 aztreonam mutant enzyme P51L Providencia stuartii
3.5.2.6 1.66
-
 aztreonam mutant enzyme S164R Providencia stuartii

Organism

EC Number Organism UniProt Comment Textmining
3.5.2.6 Providencia stuartii
-
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.5.2.6 ampicillin + H2O
-
 Providencia stuartii (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 aztreonam + H2O
-
 Providencia stuartii [(1S,2S)-1-[[(2Z)-2-(2-ammonio-1,3-thiazol-4-yl)-2-[[(2-carboxypropan-2-yl)oxy]imino]acetyl]amino]-1-carboxypropan-2-yl]sulfamate
-
 ?
3.5.2.6 cefotaxime + H2O
-
 Providencia stuartii (2R)-5-[(acetyloxy)methyl]-2-[(R)-[[(2E)-2-(2-amino-1,3-thiazol-4-yl)-2-(methoxyimino)acetyl]amino](carboxy)methyl]-3,6-dihydro-2H-1,3-thiazine-4-carboxylic acid
-
 ?
3.5.2.6 ceftazidime + H2O
-
 Providencia stuartii (2R)-2-[(R)-{[(2Z)-2-(2-amino-1,3-thiazol-4-yl)-2-{[(2-carboxypropan-2-yl)oxy]imino}acetyl]amino}(carboxy)methyl]-5-[(pyridin-1-ium-1-yl)methyl]-3,6-dihydro-2H-1,3-thiazine-4-carboxylate
-
 ?
3.5.2.6 cephaloridine + H2O
-
 Providencia stuartii (2R)-2-[(R)-carboxy[(thiophen-2-ylacetyl)amino]methyl]-5-(pyridinium-1-ylmethyl)-3,6-dihydro-2H-1,3-thiazine-4-carboxylate
-
 ?
3.5.2.6 nitrocefin + H2O
-
 Providencia stuartii (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
-
 Providencia stuartii (2R,4S)-2-[(R)-carboxy[(phenylacetyl)amino]methyl]-5,5-dimethyl-1,3-thiazolidine-4-carboxylic acid
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.5.2.6 extended-spectrum beta-lactamase TEM-60
-
 Providencia stuartii

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.5.2.6 42
-
 30 min, mutant enzyme R146S is stable, mutant enzyme K104E loses 50% of maximal activity, mutant enzyme P51L loses 95% of maximal activity, wild-type enzyme loses 80% of maximal activity Providencia stuartii

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.5.2.6 0.45
-
 cefotaxime mutant enzyme S164R Providencia stuartii
3.5.2.6 0.45
-
 (6R,7R)-3-[(acetyloxy)methyl]-7-[[(2E)-2-(2-amino-1,3-thiazol-4-yl)-2-(methoxyimino)acetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid mutant enzyme S164R Providencia stuartii
3.5.2.6 1.5
-
 cefotaxime wild-type enzyme Providencia stuartii
3.5.2.6 1.5
-
 (6R,7R)-3-[(acetyloxy)methyl]-7-[[(2E)-2-(2-amino-1,3-thiazol-4-yl)-2-(methoxyimino)acetyl]amino]-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid wild-type enzyme Providencia stuartii
3.5.2.6 3.2
-
 ampicillin mutant enzyme P51L Providencia stuartii
3.5.2.6 5
-
 nitrocefin wild-type enzyme Providencia stuartii
3.5.2.6 8
-
 nitrocefin mutant enzyme P51L Providencia stuartii
3.5.2.6 9
-
 aztreonam wild-type enzyme Providencia stuartii
3.5.2.6 12
-
 ceftazidime wild-type enzyme Providencia stuartii
3.5.2.6 15
-
 cephaloridine wild-type enzyme Providencia stuartii
3.5.2.6 17
-
 aztreonam mutant enzyme P51L Providencia stuartii
3.5.2.6 18
-
 ampicillin wild-type enzyme Providencia stuartii
3.5.2.6 22
-
 cephaloridine mutant enzyme P51L Providencia stuartii
3.5.2.6 23
-
 penicillin G mutant enzyme P51L Providencia stuartii
3.5.2.6 26
-
 penicillin G wild-type enzyme Providencia stuartii
3.5.2.6 33
-
 penicillin G mutant enzyme K104E Providencia stuartii
3.5.2.6 60
-
 ceftazidime mutant enzyme P51L Providencia stuartii
3.5.2.6 127
-
 cephaloridine mutant enzyme K104E Providencia stuartii
3.5.2.6 146
-
 ampicillin mutant enzyme S164R Providencia stuartii
3.5.2.6 150
-
 aztreonam mutant enzyme S164R Providencia stuartii
3.5.2.6 176
-
 nitrocefin mutant enzyme K104E Providencia stuartii
3.5.2.6 200
-
 penicillin G mutant enzyme S164R Providencia stuartii
3.5.2.6 590
-
 cephaloridine mutant enzyme S164R Providencia stuartii
3.5.2.6 600
-
 nitrocefin mutant enzyme S164R Providencia stuartii

pI Value

EC Number Organism Comment pI Value Maximum pI Value
3.5.2.6 Providencia stuartii isoelectric focusing, mutant enzyme K104E
-
 5.4
3.5.2.6 Providencia stuartii isoelectric focusing, mutant enzyme P51L
-
 6
3.5.2.6 Providencia stuartii isoelectric focusing, mutant enzyme S164R
-
 6.1