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Literature summary for 3.5.2.6 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
Search for more literature for 3.5.2.6

Cloned(Commentary)

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

Protein Variants

Protein Variants Comment Organism
K104E thermal stability is increased, mutant enzyme loses activity against oxyimino-cephgalosporins, aztreonam, and ampicillin Providencia stuartii
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
S164E thermal stability is increased, mutant enzyme loses activity against ceftazidime Providencia stuartii

Inhibitors

Inhibitors Comment Organism Structure
beta-iodopenicillanate
-
 Providencia stuartii
tazobactam
-
 Providencia stuartii

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0017
-
 penicillin G mutant enzyme K104E Providencia stuartii
0.0042
-
 ampicillin mutant enzyme K104E Providencia stuartii
0.009
-
 penicillin G wild-type enzyme Providencia stuartii
0.009
-
 ampicillin wild-type enzyme Providencia stuartii
0.015
-
 penicillin G mutant enzyme S164R Providencia stuartii
0.018
-
 cephaloridine mutant enzyme P51L Providencia stuartii
0.02
-
 nitrocefin wild-type enzyme Providencia stuartii
0.028
-
 cephaloridine wild-type enzyme Providencia stuartii
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
0.041
-
 nitrocefin mutant enzyme K104E Providencia stuartii
0.055
-
 aztreonam wild-type enzyme Providencia stuartii
0.059
-
 ceftazidime wild-type enzyme Providencia stuartii
0.065
-
 nitrocefin mutant enzyme S164R Providencia stuartii
0.07
-
 cephaloridine mutant enzyme K104E Providencia stuartii
0.08
-
 ceftazidime mutant enzyme P51L Providencia stuartii
0.156
-
 cephaloridine mutant enzyme S164R Providencia stuartii
0.18
-
 nitrocefin mutant enzyme P51L Providencia stuartii
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
0.335
-
 ampicillin mutant enzyme S164R Providencia stuartii
0.45
-
 ampicillin mutant enzyme P51L Providencia stuartii
0.575
-
 penicillin G mutant enzyme P51L Providencia stuartii
0.66
-
 aztreonam mutant enzyme P51L Providencia stuartii
1.66
-
 aztreonam mutant enzyme S164R Providencia stuartii

Organism

Organism UniProt Comment Textmining
Providencia stuartii
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
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
-
 ?
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
-
 ?
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
-
 ?
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
-
 ?
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
-
 ?
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
-
 ?
penicillin G + H2O
-
 Providencia stuartii (2R,4S)-2-[(R)-carboxy[(phenylacetyl)amino]methyl]-5,5-dimethyl-1,3-thiazolidine-4-carboxylic acid
-
 ?

Synonyms

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

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
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]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.45
-
 cefotaxime mutant enzyme S164R Providencia stuartii
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
1.5
-
 cefotaxime wild-type enzyme Providencia stuartii
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
3.2
-
 ampicillin mutant enzyme P51L Providencia stuartii
5
-
 nitrocefin wild-type enzyme Providencia stuartii
8
-
 nitrocefin mutant enzyme P51L Providencia stuartii
9
-
 aztreonam wild-type enzyme Providencia stuartii
12
-
 ceftazidime wild-type enzyme Providencia stuartii
15
-
 cephaloridine wild-type enzyme Providencia stuartii
17
-
 aztreonam mutant enzyme P51L Providencia stuartii
18
-
 ampicillin wild-type enzyme Providencia stuartii
22
-
 cephaloridine mutant enzyme P51L Providencia stuartii
23
-
 penicillin G mutant enzyme P51L Providencia stuartii
26
-
 penicillin G wild-type enzyme Providencia stuartii
33
-
 penicillin G mutant enzyme K104E Providencia stuartii
60
-
 ceftazidime mutant enzyme P51L Providencia stuartii
127
-
 cephaloridine mutant enzyme K104E Providencia stuartii
146
-
 ampicillin mutant enzyme S164R Providencia stuartii
150
-
 aztreonam mutant enzyme S164R Providencia stuartii
176
-
 nitrocefin mutant enzyme K104E Providencia stuartii
200
-
 penicillin G mutant enzyme S164R Providencia stuartii
590
-
 cephaloridine mutant enzyme S164R Providencia stuartii
600
-
 nitrocefin mutant enzyme S164R Providencia stuartii

pI Value

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