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3.5.1.93: glutaryl-7-aminocephalosporanic-acid acylase

This is an abbreviated version!
For detailed information about glutaryl-7-aminocephalosporanic-acid acylase, go to the full flat file.

Word Map on EC 3.5.1.93

Reaction

(7R)-7-(4-carboxybutanamido)cephalosporanate
+
H2O
=
(7R)-7-aminocephalosporanate
 +
Glutarate

Synonyms

7beta-(4-carboxybutanamido)cephalosporanic acid acylase, AcyI, acylase ACY 1 proenzyme, adipyl-cephalosporin acylase, antibiotic acylase, CA, CCA, cephalorin acylase, cephalosporin acylase, cephalosporin acylase II, cephalosporin C acylase, cephalosporin-C acylase, CephC acylase, class III acylase, class III GA, class III glutaryl acylase, CPC acylase, CPCAcy, GA, GAR, GCA, GL-7-ACA acylase, GL-7ACA acylase, Gl7ACA acylase, GLA, glutaryl 7-amino cephalosporanic acid acylase, glutaryl 7-aminocephalosporanic acid acylase, glutaryl acylase, glutaryl-7-(7-aminocephalosporanic acid) acylase, glutaryl-7-ACA acylase, glutaryl-7-amino cephalosporanic acid acylase, glutaryl-7-aminocephalosporanic acid acylase, glutaryl-7-aminocephalosporic acid acylase, J1 acylase, N176 cephalosporin C acylase, sCPCAcy, VAC, VAC acylase

ECTree

     3 Hydrolases
         3.5 Acting on carbon-nitrogen bonds, other than peptide bonds
             3.5.1 In linear amides
                3.5.1.93 glutaryl-7-aminocephalosporanic-acid acylase

Purification

Purification on EC 3.5.1.93 - glutaryl-7-aminocephalosporanic-acid acylase

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PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
a modified chitin-binding domain-GLA fusion protein is affinity purified on chitin column by changing the salt concentration of binding and elution buffer
-
acetone precipitation and Sepharose CM column chromatography
-
ammonium sulfate precipitation, chitin granula column chromatography, and Co2+-iminodiacetic acid-Sepharose column chromatography
-
ammonium sulfate precipitation, DEAE-Sepharose column chromatography, TEAE-cellulose chromatography, Toyopearl HW65F column chromatography, CM Toyopearl 650 column chromatography, Sephadex G-200 gel filtration, Q-Sepharose column chromatography, hydroxyapatite column chromatography, SP-Trisacryl gel filtration, and Sephacryl S-300gel filtration
-
ammonium sulfate precipitation, Toyopearl HW65F column chromatography, and CM Toyopearl 650 column chromatography
by HiTrap chelating affinity chromatography
-
Cu2+-immobilized iminodiacetic acid column chromatography
-
Ni2+-HiTrap chelating column chromatography and Cu2+-immobilized iminodiacetic acid affinity chromatography
overexpressed mutant protein
recombinant enzyme
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and desalting gel filtration
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli by nickel affinity chromatography
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and dialysis
recombinant protein, to high purity by immobilized metal affinity chromatography, 56% recovery
-
recombinant wild-type and mutant enzymes from Escherichia coli strain BL21(DE3)pLysS
-
several chemical treatment techniques commonly used for protein extraction are investigated for recovering glutaryl-7-aminocephalosporanic acid acylase (GLA) from recombinant Escherichia coli cells, best results are obtained by the combined use of 0.5% w/v cetyltrimethylammonium bromide (CTAB) and 0.3 M KCl at 10°C for 12 h, method evaluation and optimization of all extraction parameters, detailed overview
TALON affinity resin column chromatography