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Overview for MACiE Entry M0288

Version history

General Information

EC Number: 3.5.1.93 (A member of the Hydrolases, Acting on carbon-nitrogen bonds, other than peptide bonds, In linear amides)

Enzyme Name: glutaryl-7-aminocephalosporanic-acid acylase

Biological Species: Pseudomonas sp. (Baceria)

Catalytic Chain UniprotKB Accession Codes:

  • P07662 - Glutaryl-7-aminocephalosporanic-acid acylase subunit beta

Representative PDB Code: 3s8r - CRYSTAL STRUCTURES OF GLUTARYL 7-AMINOCEPHALOSPORANIC ACID ACYLASE:INSIGHT INTO AUTOPROTEOLYTIC ACTIVATION (Resolution = 2.50 Å).

Catalytic CATH Codes:

  • Unassigned Domain

Display structure information

Overall Reaction:

Image of water

Image of 7-glutarylaminocephalosporanate

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Image of 7-aminocephalosporanic acid

Image of glutarate

water
C00001
CHEBI:15377
7-glutarylaminocephalosporanate
C15666
CHEBI:41425
7-aminocephalosporanic acid
C07756
CHEBI:2255
glutarate
C00489
CHEBI:17859

Overall Comment: In order for the enzyme to be activated towards catalysing the hydrolysis of the glutaryl form of 7-aminocephalosporanic acid, autoproteolysis must occur at the nucleophilic Ser170, revealing an N terminal nucleophile capable of inducing nucleophilic attack at the substrate's glutaryl carbonyl. The process forms two descrete proteins chains. The crystal structure depicts the non-cleaved, precursor protein, whereas the active protein froms a heterotetramer from the two resulting chains.


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Stepwise Description of the Reaction

Step 1A water molecule acts as a proton carrier in the deprotonation of the Ser170A side chain hydroxyl by the N terminal amine group.
Step 2The side chain hydroxide of Ser170A attacks the substrate amide carbonyl, forming a tetrahedral intermediate.
Step 3The tetrahedral intermediate collapses, forming a serine linked glutarate intermediate and eliminating 7-Aminocephalosporanic acid.
Step 4The structurally conserved water molecule is activated by the non-protonated terminal amine of Ser170 to attack at the enzyme-substrate intermediate's carbonyl group, forming a second tetrahedral intermediate.
Step 5The tetrahedral anion intermediate collapses, releasing glutarate and the N terminal serine.
Step 6The anionic serine side chain undergoes an intamolecular deprotonation, removing a proton from the terminal amine group via a conserved water molecule.

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Catalytic Residues Involved

Type Number Chain Location of Function
Val 140 A Main Chain Amide
Asn 314 A Side Chain
Glu 525 A Side Chain
Ser 170 A Side Chain
Main Chain N Terminus
His 193 A Side Chain

References

  1. J. K. Kim et al. (2003), Biochemistry, 42, 4084-4093. Crystal structures of glutaryl 7-aminocephalosporanic acid acylase: insight into autoproteolytic activation.
    Medline: 12680762
  2. K. Fritz-Wolf et al. (2002), Protein Sci., 11, 92-103. Structure-based prediction of modifications in glutarylamidase to allow single-step enzymatic production of 7-aminocephalosporanic acid from cephalosporin C.
    Medline: 11742126

Homologue information for M0288 (3s8r)

MACiE Homologues (within the PDB)

MACiE Homologues (within UniprotKB/SwissProt)


Links to this entry in other databases

Link to EC-PDB-SUM Link to PDB-SUM Link to RCSB PDB Link to PDBe Link to CSA
Link to MetaCyc Link to KEGG Link to BRENDA Link to ExplorENZ

GOA logo
hydrolase activity (molecular function)
hydrolase activity, acting on carbon-nitrogen (but not peptide) bonds, in linear amides (molecular function)
antibiotic biosynthetic process (biological process)
glutaryl-7-aminocephalosporanic-acid acylase activity (molecular function)
periplasmic space (cellular component)
response to antibiotic (biological process)
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