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Literature summary for 6.3.3.6 extracted from

  • Gerratana, B.; Stapon, A.; Townsend, C.A.
    Inhibition and alternate substrate studies on the mechanism of carbapenam synthetase from Erwinia carotovora (2003), Biochemistry, 42, 7836-7847.
    View publication on PubMed
Search for more literature for 6.3.3.6

Cloned(Commentary)

Cloned (Comment) Organism
overexpression of the plasmid pET24a/carA transformed in Escherichia coli BL21(DE3) cells Pectobacterium carotovorum

Inhibitors

Inhibitors Comment Organism Structure
(3S,5S)-carbapenam 3-carboxylate
-
 Pectobacterium carotovorum
AMP uncompetitive inhibition versus both substrates Pectobacterium carotovorum
diphosphate the competitive inhibition exhibited by diphosphate versus ATP allows the assignment of diphosphate as the last product released Pectobacterium carotovorum
L-proline the uncompetitive nature of the inhibition of the dead-end inhibitor, L-proline, versus ATP is consistent with a mechanism with ordered substrate binding where ATP binds first followed by (2S,5S)-carboxymethylproline Pectobacterium carotovorum

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.11
-
 ATP pH 7.8, 22°C Pectobacterium carotovorum
0.23
-
 (2S,5S)-5-carboxymethylproline pH 7.8, 22°C Pectobacterium carotovorum
4.6
-
 (2R,5R)-5-carboxymethylproline pH 7.8, 22°C Pectobacterium carotovorum
7
-
 (2S,5R)-5-carboxymethylproline pH 7.8, 22°C Pectobacterium carotovorum

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
56000
-
 2 * 56000, calculated from sequence Pectobacterium carotovorum
119000
-
 native PAGE Pectobacterium carotovorum

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + (2S,5S)-5-carboxymethylproline Pectobacterium carotovorum the enzyme is involved in the biosynthesis of the carbapenem beta-lactam antibiotic (5R)-carbapen-2-em-3-carboxylate AMP + diphosphate + (3S,5S)-carbapenam 3-carboxylate
-
 ?

Organism

Organism UniProt Comment Textmining
Pectobacterium carotovorum Q9XB61
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Pectobacterium carotovorum

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + (2R,5R)-5-carboxymethylproline low activity, kcat/Km is 2% compared to the value for (2S,5S)-5-carboxymethylproline Pectobacterium carotovorum ?
-
 ?
ATP + (2S,5R)-5-carboxymethylproline low activity, kcat/Km is 2% compared to the value for (2S,5S)-5-carboxymethylproline Pectobacterium carotovorum ?
-
 ?
ATP + (2S,5S)-5-carboxymethylproline the enzyme is involved in the biosynthesis of the carbapenem beta-lactam antibiotic (5R)-carbapen-2-em-3-carboxylate Pectobacterium carotovorum AMP + diphosphate + (3S,5S)-carbapenam 3-carboxylate
-
 ?
ATP + (2S,5S)-5-carboxymethylproline the kinetic mechanism is bi-ter where ATP is the first substrate to bind followed by (2S,5S)-5-carboxymethyl proline and diphosphate is the last product released. Low activity with (2S,5R)-5-carboxymethylproline or (2R,5R)-5-carboxymethylproline Pectobacterium carotovorum AMP + diphosphate + (3S,5S)-carbapenam 3-carboxylate
-
 ?

Subunits

Subunits Comment Organism
homodimer 2 * 56000, calculated from sequence Pectobacterium carotovorum

Synonyms

Synonyms Comment Organism
CarA
-
 Pectobacterium carotovorum
carbapenam 3-carboxylate synthase
-
 Pectobacterium carotovorum

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
22
-
 assay at Pectobacterium carotovorum

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.122
-
 (2R,5R)-5-carboxymethylproline pH 7.8, 22°C Pectobacterium carotovorum
0.2
-
 (2S,5R)-5-carboxymethylproline pH 7.8, 22°C Pectobacterium carotovorum
0.28
-
 ATP pH 7.8, 22°C Pectobacterium carotovorum
0.28
-
 (2S,5S)-5-carboxymethylproline pH 7.8, 22°C Pectobacterium carotovorum

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.8
-
 assay at Pectobacterium carotovorum

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.005
-
 diphosphate pH 7.8, 22°C, variable substrate: (2S,5R)-carboxymethylproline, Ki(slope) Pectobacterium carotovorum
0.012
-
 diphosphate pH 7.8, 22°C, variable substrate: ATP, Ki(slope) Pectobacterium carotovorum
0.019
-
 diphosphate pH 7.8, 22°C, variable substrate: (2S,5R)-carboxymethylproline, Ki(intercept) Pectobacterium carotovorum
0.17
-
 AMP pH 7.8, 22°C, variable substrate: ATP, Ki(intercept) Pectobacterium carotovorum
0.28
-
 AMP pH 7.8, 22°C, variable substrate: (2S,5R)-carboxymethylproline, Ki(intercept) Pectobacterium carotovorum
1.1
-
 (3S,5S)-carbapenam 3-carboxylate pH 7.8, 22°C, variable substrate: (2S,5R)-carboxymethylproline, Ki(intercept) Pectobacterium carotovorum
1.3
-
 (3S,5S)-carbapenam 3-carboxylate pH 7.8, 22°C, variable substrate: (2S,5R)-carboxymethylproline, Ki(slope) Pectobacterium carotovorum
1.3
-
 (3S,5S)-carbapenam 3-carboxylate pH 7.8, 22°C, variable substrate: ATP, Ki(slope) Pectobacterium carotovorum
90
-
 L-proline pH 7.8, 22°C, variable substrate: (2S,5R)-carboxymethylproline, Ki(slope) Pectobacterium carotovorum
207
-
 L-proline pH 7.8, 22°C, variable substrate: ATP, Ki(intercept) Pectobacterium carotovorum

General Information

General Information Comment Organism
physiological function the enzyme is involved in the biosynthesis of the carbapenem beta-lactam antibiotic (5R)-carbapen-2-em-3-carboxylate Pectobacterium carotovorum

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.026
-
 (2R,5R)-5-carboxymethylproline pH 7.8, 22°C Pectobacterium carotovorum
0.027
-
 (2S,5R)-5-carboxymethylproline pH 7.8, 22°C Pectobacterium carotovorum
1.2
-
 (2S,5S)-5-carboxymethylproline pH 7.8, 22°C Pectobacterium carotovorum
2.5
-
 ATP pH 7.8, 22°C Pectobacterium carotovorum