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Literature summary for 3.5.1.93 extracted from

  • Volonte, F.; Marinelli, F.; Gastaldo, L.; Sacchi, S.; Pilone, M.S.; Pollegioni, L.; Molla, G.
    Optimization of glutaryl-7-aminocephalosporanic acid acylase expression in E. coli (2008), Protein Expr. Purif., 61, 131-137.
    View publication on PubMed

Application

Application Comment Organism
industry the huge production of engineered GLA produced in Escherichia coli under optimized conditions makes this enzyme an economic tool to be used in bioconversion processes at the industrial level, e.g., in 7-ACA production Pseudomonas sp.

Cloned(Commentary)

Cloned (Comment) Organism
into the vector pET24 for expression in Escherichia coli BL21DE3 pLysS cells Pseudomonas sp.

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
26000
-
alpha-subunit, determined by SDS-PAGE Pseudomonas sp.
58000
-
beta-subunit, determined by SDS-PAGE Pseudomonas sp.

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
(7R)-7-(4-carboxybutanamido)cephalosporanate + H2O Pseudomonas sp.
-
(7R)-7-aminocephalosporanate + glutarate
-
?

Organism

Organism UniProt Comment Textmining
Pseudomonas sp.
-
-
-

Purification (Commentary)

Purification (Comment) Organism
by HiTrap chelating affinity chromatography Pseudomonas sp.

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.28
-
LB Miller broth medium Pseudomonas sp.
3.7
-
purification step crude extract Pseudomonas sp.
11.4
-
optimized conditions, SB3 medium, 12.5 g NaCl/l, 0.6 mM IPTG Pseudomonas sp.
125
-
purification step HiTrap chelating chromatography Pseudomonas sp.
133
-
purification step PD10 desalting Pseudomonas sp.

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(7R)-7-(4-carboxybutanamido)cephalosporanate + H2O
-
Pseudomonas sp. (7R)-7-aminocephalosporanate + glutarate
-
?
glutaryl-7-aminocephalosporanic acid + H2O activity assay Pseudomonas sp. 7-aminocephalosporanic acid + glutarate
-
?

Subunits

Subunits Comment Organism
heterodimer
-
Pseudomonas sp.

Synonyms

Synonyms Comment Organism
glutaryl-7-aminocephalosporanic acid acylase
-
Pseudomonas sp.