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Literature summary 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

Cloned(Commentary)

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

Inhibitors

EC Number Inhibitors Comment Organism Structure
6.3.3.6 (3S,5S)-carbapenam 3-carboxylate
-
 Pectobacterium carotovorum
6.3.3.6 AMP uncompetitive inhibition versus both substrates Pectobacterium carotovorum
6.3.3.6 diphosphate the competitive inhibition exhibited by diphosphate versus ATP allows the assignment of diphosphate as the last product released Pectobacterium carotovorum
6.3.3.6 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]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
6.3.3.6 0.11
-
 ATP pH 7.8, 22°C Pectobacterium carotovorum
6.3.3.6 0.23
-
 (2S,5S)-5-carboxymethylproline pH 7.8, 22°C Pectobacterium carotovorum
6.3.3.6 4.6
-
 (2R,5R)-5-carboxymethylproline pH 7.8, 22°C Pectobacterium carotovorum
6.3.3.6 7
-
 (2S,5R)-5-carboxymethylproline pH 7.8, 22°C Pectobacterium carotovorum

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
6.3.3.6 56000
-
 2 * 56000, calculated from sequence Pectobacterium carotovorum
6.3.3.6 119000
-
 native PAGE Pectobacterium carotovorum

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
6.3.3.6 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

EC Number Organism UniProt Comment Textmining
6.3.3.6 Pectobacterium carotovorum Q9XB61
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
6.3.3.6
-
 Pectobacterium carotovorum

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6.3.3.6 ATP + (2R,5R)-5-carboxymethylproline low activity, kcat/Km is 2% compared to the value for (2S,5S)-5-carboxymethylproline Pectobacterium carotovorum ?
-
 ?
6.3.3.6 ATP + (2S,5R)-5-carboxymethylproline low activity, kcat/Km is 2% compared to the value for (2S,5S)-5-carboxymethylproline Pectobacterium carotovorum ?
-
 ?
6.3.3.6 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
-
 ?
6.3.3.6 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

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

Synonyms

EC Number Synonyms Comment Organism
6.3.3.6 CarA
-
 Pectobacterium carotovorum
6.3.3.6 carbapenam 3-carboxylate synthase
-
 Pectobacterium carotovorum

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
6.3.3.6 22
-
 assay at Pectobacterium carotovorum

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
6.3.3.6 0.122
-
 (2R,5R)-5-carboxymethylproline pH 7.8, 22°C Pectobacterium carotovorum
6.3.3.6 0.2
-
 (2S,5R)-5-carboxymethylproline pH 7.8, 22°C Pectobacterium carotovorum
6.3.3.6 0.28
-
 ATP pH 7.8, 22°C Pectobacterium carotovorum
6.3.3.6 0.28
-
 (2S,5S)-5-carboxymethylproline pH 7.8, 22°C Pectobacterium carotovorum

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
6.3.3.6 7.8
-
 assay at Pectobacterium carotovorum

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
6.3.3.6 0.005
-
 diphosphate pH 7.8, 22°C, variable substrate: (2S,5R)-carboxymethylproline, Ki(slope) Pectobacterium carotovorum
6.3.3.6 0.012
-
 diphosphate pH 7.8, 22°C, variable substrate: ATP, Ki(slope) Pectobacterium carotovorum
6.3.3.6 0.019
-
 diphosphate pH 7.8, 22°C, variable substrate: (2S,5R)-carboxymethylproline, Ki(intercept) Pectobacterium carotovorum
6.3.3.6 0.17
-
 AMP pH 7.8, 22°C, variable substrate: ATP, Ki(intercept) Pectobacterium carotovorum
6.3.3.6 0.28
-
 AMP pH 7.8, 22°C, variable substrate: (2S,5R)-carboxymethylproline, Ki(intercept) Pectobacterium carotovorum
6.3.3.6 1.1
-
 (3S,5S)-carbapenam 3-carboxylate pH 7.8, 22°C, variable substrate: (2S,5R)-carboxymethylproline, Ki(intercept) Pectobacterium carotovorum
6.3.3.6 1.3
-
 (3S,5S)-carbapenam 3-carboxylate pH 7.8, 22°C, variable substrate: (2S,5R)-carboxymethylproline, Ki(slope) Pectobacterium carotovorum
6.3.3.6 1.3
-
 (3S,5S)-carbapenam 3-carboxylate pH 7.8, 22°C, variable substrate: ATP, Ki(slope) Pectobacterium carotovorum
6.3.3.6 90
-
 L-proline pH 7.8, 22°C, variable substrate: (2S,5R)-carboxymethylproline, Ki(slope) Pectobacterium carotovorum
6.3.3.6 207
-
 L-proline pH 7.8, 22°C, variable substrate: ATP, Ki(intercept) Pectobacterium carotovorum

General Information

EC Number General Information Comment Organism
6.3.3.6 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]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
6.3.3.6 0.026
-
 (2R,5R)-5-carboxymethylproline pH 7.8, 22°C Pectobacterium carotovorum
6.3.3.6 0.027
-
 (2S,5R)-5-carboxymethylproline pH 7.8, 22°C Pectobacterium carotovorum
6.3.3.6 1.2
-
 (2S,5S)-5-carboxymethylproline pH 7.8, 22°C Pectobacterium carotovorum
6.3.3.6 2.5
-
 ATP pH 7.8, 22°C Pectobacterium carotovorum