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Literature summary for 3.5.2.6 extracted from

  • Perilli, M.; Celenza, G.; De Santis, F.; Pellegrini, C.; Forcella, C.; Rossolini, G.M.; Stefani, S.; Amicosante, G.
    E240V substitution increases catalytic efficiency toward ceftazidime in a new natural TEM-type extended-spectrum beta -lactamase, TEM-149, from Enterobacter aerogenes and Serratia marcescens clinical isolates (2008), Antimicrob. Agents Chemother., 52, 915-919.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.5.2.6

Cloned(Commentary)

Cloned (Comment) Organism
cloned into pBC-SK and expressed in Escherichia coli HB101 Klebsiella aerogenes

Protein Variants

Protein Variants Comment Organism
E104K side directed mutagenesis Klebsiella aerogenes
E240V side directed mutagenesis Klebsiella aerogenes
E240V substitution increases catalytic efficiency toward ceftazidime Klebsiella aerogenes
M182T side directed mutagenesis Klebsiella aerogenes
R164S side directed mutagenesis Klebsiella aerogenes

Inhibitors

Inhibitors Comment Organism Structure
clavulanic acid
-
 Klebsiella aerogenes
Sulbactam
-
 Klebsiella aerogenes
tazobactam
-
 Klebsiella aerogenes

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0006
-
 benzylpenicillin 100 microM nitrocefin as the reporter substrate, TEM-149 wild-type, pH 7.0, 25°C Klebsiella aerogenes
0.0007
-
 benzylpenicillin 100 microM nitrocefin as the reporter substrate, TEM-149 mutant T182M, pH 7.0, 25°C Klebsiella aerogenes
0.0014
-
 piperacillin 100 microM nitrocefin as the reporter substrate, TEM-149 wild-type, pH 7.0, 25°C Klebsiella aerogenes
0.0034
-
 piperacillin 100 microM nitrocefin as the reporter substrate, TEM-149 mutant T182M, pH 7.0, 25°C Klebsiella aerogenes
0.005
-
 ampicillin 100 microM nitrocefin as the reporter substrate, TEM-149 wild-type, pH 7.0, 25°C Klebsiella aerogenes
0.016
-
 cefepime TEM-149 wild-type, pH 7.0, 25°C Klebsiella aerogenes
0.019
-
 ampicillin 100 microM nitrocefin as the reporter substrate, TEM-149 mutant T182M, pH 7.0, 25°C Klebsiella aerogenes
0.019
-
 ceftazidime TEM-149 wild-type, pH 7.0, 25°C Klebsiella aerogenes
0.02
-
 nitrocefin TEM-149 wild-type, pH 7.0, 25°C Klebsiella aerogenes
0.025
-
 nitrocefin TEM-149 mutant T182M, pH 7.0, 25°C Klebsiella aerogenes
0.026
-
 ceftazidime TEM-149 mutant T182M, pH 7.0, 25°C Klebsiella aerogenes
0.027
-
 aztreonam TEM-149 mutant T182M, pH 7.0, 25°C Klebsiella aerogenes
0.03
-
 cefepime TEM-149 mutant T182M, pH 7.0, 25°C Klebsiella aerogenes
0.032
-
 cefotaxime TEM-149 mutant T182M, pH 7.0, 25°C Klebsiella aerogenes
0.035
-
 cefazolin TEM-149 mutant T182M, pH 7.0, 25°C Klebsiella aerogenes
0.043
-
 cefotaxime TEM-149 wild-type, pH 7.0, 25°C Klebsiella aerogenes
0.051
-
 aztreonam TEM-149 wild-type, pH 7.0, 25°C Klebsiella aerogenes
0.068
-
 cefazolin TEM-149 wild-type, pH 7.0, 25°C Klebsiella aerogenes

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
28600
-
-
 Klebsiella aerogenes

Organism

Organism UniProt Comment Textmining
Klebsiella aerogenes Q1KY81 SS-13
-
Klebsiella aerogenes SS-13 Q1KY81 SS-13
-

Purification (Commentary)

Purification (Comment) Organism
-
 Klebsiella aerogenes

Source Tissue

Source Tissue Comment Organism Textmining
additional information isolated from Homo sapiens urine Klebsiella aerogenes
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ampicillin + H2O
-
 Klebsiella aerogenes (2R,4S)-2-[(R)-[[(2R)-2-amino-2-phenylacetyl]amino](carboxy)methyl]-5,5-dimethyl-1,3-thiazolidine-4-carboxylic acid
-
 ?
ampicillin + H2O
-
 Klebsiella aerogenes SS-13 (2R,4S)-2-[(R)-[[(2R)-2-amino-2-phenylacetyl]amino](carboxy)methyl]-5,5-dimethyl-1,3-thiazolidine-4-carboxylic acid
-
 ?
aztreonam + H2O
-
 Klebsiella aerogenes [(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
-
 ?
aztreonam + H2O
-
 Klebsiella aerogenes SS-13 [(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
-
 ?
benzylpenicillin + H2O
-
 Klebsiella aerogenes (2R,4S)-2-[(R)-carboxy(2-phenylacetamido)methyl]-5,5-dimethyl-1,3-thiazolidine-4-carboxylic acid
-
 ?
benzylpenicillin + H2O
-
 Klebsiella aerogenes SS-13 (2R,4S)-2-[(R)-carboxy(2-phenylacetamido)methyl]-5,5-dimethyl-1,3-thiazolidine-4-carboxylic acid
-
 ?
cefazolin + H2O
-
 Klebsiella aerogenes (2R)-2-[(R)-carboxy[(1H-tetrazol-1-ylacetyl)amino]methyl]-5-[[(5-methyl-1,3,4-thiadiazol-2-yl)sulfanyl]methyl]-3,6-dihydro-2H-1,3-thiazine-4-carboxylic acid
-
 ?
cefazolin + H2O
-
 Klebsiella aerogenes SS-13 (2R)-2-[(R)-carboxy[(1H-tetrazol-1-ylacetyl)amino]methyl]-5-[[(5-methyl-1,3,4-thiadiazol-2-yl)sulfanyl]methyl]-3,6-dihydro-2H-1,3-thiazine-4-carboxylic acid
-
 ?
cefepime + H2O
-
 Klebsiella aerogenes (2R)-2-[(R)-[[(2Z)-2-(2-amino-1,3-thiazol-4-yl)-2-(methoxyimino)acetyl]amino](carboxy)methyl]-5-[(1-methylpyrrolidinium-1-yl)methyl]-3,6-dihydro-2H-1,3-thiazine-4-carboxylate
-
 ?
cefotaxime + H2O
-
 Klebsiella aerogenes (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
-
 ?
cefotaxime + H2O
-
 Klebsiella aerogenes SS-13 (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 preferred and best substrate Klebsiella aerogenes (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
-
 ?
nitrocefin + H2O
-
 Klebsiella aerogenes (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
-
 ?
piperacillin + H2O
-
 Klebsiella aerogenes (2R,4S)-2-[(R)-carboxy[[(2R)-2-[[(4-ethyl-2,3-dioxopiperazin-1-yl)carbonyl]amino]-2-phenylacetyl]amino]methyl]-5,5-dimethyl-1,3-thiazolidine-4-carboxylic acid
-
 ?

Synonyms

Synonyms Comment Organism
beta-lactamase
-
 Klebsiella aerogenes
TEM-149
-
 Klebsiella aerogenes

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
25 55 thermal stability of TEM-149 wild-type and TEM-149T182M mutant is determined by measuring the residual activity after incubation of the enzymes at three different temperatures: 25°C, 40°C, and 55°C. TEM-149 wild-type exhibits half-lives of 90 min at 25°C, 70 min at 40°C, and 30 min at 55°C, whereas TEM-149T182M mutant shows half-lives greater than 120 min at 25°C and 40°C and 45 min at 55°C. Compared to the thermal stability of TEM-149, the thermal stability of the TEM-149T182M mutant is substantially increased Klebsiella aerogenes

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.07
-
 cefotaxime TEM-149 wild-type, pH 7.0, 25°C Klebsiella aerogenes
0.41
-
 cefotaxime TEM-149 mutant T182M, pH 7.0, 25°C Klebsiella aerogenes
0.43
-
 cefepime TEM-149 mutant T182M, pH 7.0, 25°C Klebsiella aerogenes
0.56
-
 cefazolin TEM-149 mutant T182M, pH 7.0, 25°C Klebsiella aerogenes
2.2
-
 cefazolin TEM-149 wild-type, pH 7.0, 25°C Klebsiella aerogenes
2.4
-
 ampicillin TEM-149 mutant T182M, pH 7.0, 25°C Klebsiella aerogenes
2.4
-
 cefepime TEM-149 wild-type, pH 7.0, 25°C Klebsiella aerogenes
2.7
-
 piperacillin TEM-149 wild-type, pH 7.0, 25°C Klebsiella aerogenes
3.5
-
 benzylpenicillin TEM-149 wild-type, pH 7.0, 25°C Klebsiella aerogenes
3.6
-
 aztreonam TEM-149 mutant T182M, pH 7.0, 25°C Klebsiella aerogenes
4.7
-
 aztreonam TEM-149 wild-type, pH 7.0, 25°C Klebsiella aerogenes
5
-
 piperacillin TEM-149 mutant T182M, pH 7.0, 25°C Klebsiella aerogenes
5.4
-
 ampicillin TEM-149 wild-type, pH 7.0, 25°C Klebsiella aerogenes
7.4
-
 nitrocefin TEM-149 wild-type, pH 7.0, 25°C Klebsiella aerogenes
8.3
-
 ceftazidime TEM-149 mutant T182M, pH 7.0, 25°C Klebsiella aerogenes
8.3
-
 ceftazidime TEM-149 wild-type, pH 7.0, 25°C Klebsiella aerogenes
10.5
-
 benzylpenicillin TEM-149 mutant T182M, pH 7.0, 25°C Klebsiella aerogenes
26
-
 nitrocefin TEM-149 mutant T182M, pH 7.0, 25°C Klebsiella aerogenes

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.000018
-
 tazobactam TEM-149 wild-type, pH 7.0, 25°C Klebsiella aerogenes
0.000026
-
 tazobactam TEM-149T182M mutant, pH 7.0, 25°C Klebsiella aerogenes
0.00011
-
 clavulanic acid TEM-149T182M mutant, pH 7.0, 25°C Klebsiella aerogenes
0.00015
-
 Sulbactam TEM-149T182M mutant, pH 7.0, 25°C Klebsiella aerogenes
0.00021
-
 Sulbactam TEM-149 wild-type, pH 7.0, 25°C Klebsiella aerogenes
0.00026
-
 clavulanic acid TEM-149 wild-type, pH 7.0, 25°C Klebsiella aerogenes

pI Value

Organism Comment pI Value Maximum pI Value
Klebsiella aerogenes and above 9.0, isoelectric focusing analysis
-
 5.8
Klebsiella aerogenes above, and at 5.8, isoelectric focusing analysis
-
 9