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Literature summary for 3.5.2.6 extracted from

  • Garrity, J.D.; Carenbauer, A.L.; Herron, L.R.; Crowder, M.W.
    Metal binding Asp-120 in metallo-beta-lactamase L1 from Stenotrophomonas maltophilia plays a crucial role in catalysis (2004), J. Biol. Chem., 279, 920-927.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type and mutant enzymes in Escherichia coli Stenotrophomonas maltophilia

Protein Variants

Protein Variants Comment Organism
D120C site-directed mutagenesis, reduced activity and metal binding compared to the wild-type enzyme Stenotrophomonas maltophilia
D120N site-directed mutagenesis, reduced activity and metal binding compared to the wild-type enzyme Stenotrophomonas maltophilia
D120S site-directed mutagenesis, reduced activity and metal binding compared to the wild-type enzyme Stenotrophomonas maltophilia

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0011
-
cefoxitin wild-type enzyme, pH 7.0, 25°C Stenotrophomonas maltophilia
0.0041
-
nitrocefin wild-type enzyme, pH 7.0, 25°C Stenotrophomonas maltophilia
0.0089
-
cephalothin wild-type enzyme, pH 7.0, 25°C Stenotrophomonas maltophilia
0.013
-
cefaclor wild-type enzyme, pH 7.0, 25°C Stenotrophomonas maltophilia
0.015
-
meropenem wild-type enzyme, pH 7.0, 25°C Stenotrophomonas maltophilia
0.032
-
biapenem wild-type enzyme, pH 7.0, 25°C Stenotrophomonas maltophilia
0.033
-
cephalothin mutant D120C, pH 7.0, 25°C Stenotrophomonas maltophilia
0.038
-
penicillin G wild-type enzyme, pH 7.0, 25°C Stenotrophomonas maltophilia
0.045
-
biapenem mutant D120C, pH 7.0, 25°C Stenotrophomonas maltophilia
0.046
-
cephalothin mutant D120S, pH 7.0, 25°C Stenotrophomonas maltophilia
0.053
-
cefoxitin mutant D120S, pH 7.0, 25°C Stenotrophomonas maltophilia
0.055
-
cefoxitin mutant D120C, pH 7.0, 25°C Stenotrophomonas maltophilia
0.055
-
ampicillin wild-type enzyme, pH 7.0, 25°C Stenotrophomonas maltophilia
0.057
-
Imipenem wild-type enzyme, pH 7.0, 25°C Stenotrophomonas maltophilia
0.06
-
meropenem mutant D120C, pH 7.0, 25°C Stenotrophomonas maltophilia
0.065
-
Imipenem mutant D120C, pH 7.0, 25°C Stenotrophomonas maltophilia
0.068
-
nitrocefin mutant D120N, pH 7.0, 25°C Stenotrophomonas maltophilia
0.069
-
nitrocefin mutant D120C, pH 7.0, 25°C Stenotrophomonas maltophilia
0.07
-
meropenem mutant D120N, pH 7.0, 25°C Stenotrophomonas maltophilia
0.08
-
cefaclor mutant D120S, pH 7.0, 25°C Stenotrophomonas maltophilia
0.081
-
biapenem mutant D120S, pH 7.0, 25°C Stenotrophomonas maltophilia
0.094
-
Imipenem mutant D120S, pH 7.0, 25°C Stenotrophomonas maltophilia
0.097
-
meropenem mutant D120S, pH 7.0, 25°C Stenotrophomonas maltophilia
0.098
-
cefaclor mutant D120C, pH 7.0, 25°C Stenotrophomonas maltophilia
0.098
-
cefoxitin mutant D120N, pH 7.0, 25°C Stenotrophomonas maltophilia
0.103
-
nitrocefin mutant D120S, pH 7.0, 25°C Stenotrophomonas maltophilia
0.128
-
biapenem mutant D120N, pH 7.0, 25°C Stenotrophomonas maltophilia
0.15
-
cephalothin mutant D120N, pH 7.0, 25°C Stenotrophomonas maltophilia
0.31
-
Imipenem mutant D120N, pH 7.0, 25°C Stenotrophomonas maltophilia
0.384
-
penicillin G mutant D120N, pH 7.0, 25°C Stenotrophomonas maltophilia
0.55
-
cefaclor mutant D120N, pH 7.0, 25°C Stenotrophomonas maltophilia
1.33
-
ampicillin mutant D120N, pH 7.0, 25°C Stenotrophomonas maltophilia

Metals/Ions

Metals/Ions Comment Organism Structure
Zn2+ absolutely required, dinuclear zinc-metalloenzyme, metal-binding Asp, the wild-type conatins 1.9 mol of Zn2+ per mol of subunit, D120 mutant enzymes show reduced metal content, overview Stenotrophomonas maltophilia

Organism

Organism UniProt Comment Textmining
Stenotrophomonas maltophilia P52700 metallo-beta-lactamase L1
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant enzymes from Escherichia coli Stenotrophomonas maltophilia

Reaction

Reaction Comment Organism Reaction ID
a beta-lactam + H2O = a substituted beta-amino acid active site, Asp120 is important for catalysis, as well as substrate and Zn2+ binding Stenotrophomonas maltophilia

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ampicillin + H2O
-
Stenotrophomonas maltophilia (2R,4S)-2-[(R)-[[(2R)-2-amino-2-phenylacetyl]amino](carboxy)methyl]-5,5-dimethyl-1,3-thiazolidine-4-carboxylic acid
-
?
biapenem + H2O
-
Stenotrophomonas maltophilia (4R,5S)-5-[(1S,2R)-1-carboxy-2-hydroxypropyl]-3-[(6,7-dihydro-5H-pyrazolo[1,2-a][1,2,4]triazol-4-ium-6-yl)sulfanyl]-4-methyl-4,5-dihydro-1H-pyrrole-2-carboxylate
-
?
cefaclor + H2O
-
Stenotrophomonas maltophilia (2R)-2-[(R)-{[(2R)-2-amino-2-phenylacetyl]amino}(carboxy)methyl]-5-chloro-3,6-dihydro-2H-1,3-thiazine-4-carboxylic acid
-
?
cefoxitin + H2O
-
Stenotrophomonas maltophilia (2R)-5-[(carbamoyloxy)methyl]-2-[(S)-carboxy(methoxy)[(thiophen-2-ylacetyl)amino]methyl]-3,6-dihydro-2H-1,3-thiazine-4-carboxylic acid
-
?
cephalotin + H2O
-
Stenotrophomonas maltophilia (2R)-5-[(acetyloxy)methyl]-2-[(R)-carboxy[(thiophen-2-ylacetyl)amino]methyl]-3,6-dihydro-2H-1,3-thiazine-4-carboxylic acid
-
?
imipenem + H2O
-
Stenotrophomonas maltophilia (5R)-5-[(1S,2R)-1-carboxy-2-hydroxypropyl]-3-({2-[(iminomethyl)amino]ethyl}sulfanyl)-4,5-dihydro-1H-pyrrole-2-carboxylic acid
-
?
meropenem + H2O
-
Stenotrophomonas maltophilia (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
-
?
nitrocefin + H2O
-
Stenotrophomonas maltophilia (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
-
Stenotrophomonas maltophilia (2R,4S)-2-[(R)-carboxy[(phenylacetyl)amino]methyl]-5,5-dimethyl-1,3-thiazolidine-4-carboxylic acid
-
?

Synonyms

Synonyms Comment Organism
L1
-
Stenotrophomonas maltophilia

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Stenotrophomonas maltophilia

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.01
-
penicillin G below, mutant D120C, pH 7.0, 25°C Stenotrophomonas maltophilia
0.01
-
ampicillin below, mutant D120C, pH 7.0, 25°C Stenotrophomonas maltophilia
0.01
-
penicillin G below, mutant D120S, pH 7.0, 25°C Stenotrophomonas maltophilia
0.01
-
ampicillin below, mutant D120S, pH 7.0, 25°C Stenotrophomonas maltophilia
0.028
-
nitrocefin mutant D120S, pH 7.0, 25°C Stenotrophomonas maltophilia
0.18
-
nitrocefin mutant D120C, pH 7.0, 25°C Stenotrophomonas maltophilia
0.28
-
cefaclor mutant D120S, pH 7.0, 25°C Stenotrophomonas maltophilia
0.29
-
meropenem mutant D120C, pH 7.0, 25°C Stenotrophomonas maltophilia
0.3
-
biapenem mutant D120C, pH 7.0, 25°C Stenotrophomonas maltophilia
0.3
-
cefaclor mutant D120C, pH 7.0, 25°C Stenotrophomonas maltophilia
0.31
-
cephalothin mutant D120C, pH 7.0, 25°C Stenotrophomonas maltophilia
0.34
-
cefoxitin mutant D120S, pH 7.0, 25°C Stenotrophomonas maltophilia
0.36
-
Imipenem mutant D120C, pH 7.0, 25°C Stenotrophomonas maltophilia
0.37
-
cephalothin mutant D120S, pH 7.0, 25°C Stenotrophomonas maltophilia
0.5
-
biapenem mutant D120S, pH 7.0, 25°C Stenotrophomonas maltophilia
0.56
-
cefoxitin mutant D120C, pH 7.0, 25°C Stenotrophomonas maltophilia
0.63
-
Imipenem mutant D120S, pH 7.0, 25°C Stenotrophomonas maltophilia
0.75
-
meropenem mutant D120S, pH 7.0, 25°C Stenotrophomonas maltophilia
1.53
-
cefoxitin mutant D120N, pH 7.0, 25°C Stenotrophomonas maltophilia
1.9
-
cefoxitin wild-type enzyme, pH 7.0, 25°C Stenotrophomonas maltophilia
2.54
-
nitrocefin mutant D120N, pH 7.0, 25°C Stenotrophomonas maltophilia
3.6
-
biapenem mutant D120N, pH 7.0, 25°C Stenotrophomonas maltophilia
6.3
-
cefaclor mutant D120N, pH 7.0, 25°C Stenotrophomonas maltophilia
8.1
-
cephalothin mutant D120N, pH 7.0, 25°C Stenotrophomonas maltophilia
10.6
-
meropenem mutant D120N, pH 7.0, 25°C Stenotrophomonas maltophilia
15
-
Imipenem mutant D120N, pH 7.0, 25°C Stenotrophomonas maltophilia
41
-
nitrocefin wild-type enzyme, pH 7.0, 25°C Stenotrophomonas maltophilia
42
-
cefaclor wild-type enzyme, pH 7.0, 25°C Stenotrophomonas maltophilia
45
-
penicillin G mutant D120N, pH 7.0, 25°C Stenotrophomonas maltophilia
77
-
ampicillin mutant D120N, pH 7.0, 25°C Stenotrophomonas maltophilia
82
-
cephalothin wild-type enzyme, pH 7.0, 25°C Stenotrophomonas maltophilia
134
-
biapenem wild-type enzyme, pH 7.0, 25°C Stenotrophomonas maltophilia
157
-
meropenem wild-type enzyme, pH 7.0, 25°C Stenotrophomonas maltophilia
370
-
Imipenem wild-type enzyme, pH 7.0, 25°C Stenotrophomonas maltophilia
520
-
ampicillin wild-type enzyme, pH 7.0, 25°C Stenotrophomonas maltophilia
600
-
penicillin G wild-type enzyme, pH 7.0, 25°C Stenotrophomonas maltophilia

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
assay at Stenotrophomonas maltophilia