Literature summary extracted from

Varshney, N.K.; Ramasamy, S.; Brannigan, J.A.; Wilkinson, A.J.; Suresh, C.G.
Cloning, overexpression, crystallization and preliminary X-ray crystallographic analysis of a slow-processing mutant of penicillin G acylase from Kluyvera citrophila (2013), Acta Crystallogr. Sect. F, 69, 925-929.

Application

EC Number	Application	Comment	Organism
3.5.1.11	synthesis	the enzyme KcPGA is quite resilient to harsh conditions and is easier to immobilize for the industrial hydrolysis of natural penicillins to generate the 6-aminopenicillin nucleus, which is the starting material for semisynthetic antibiotic production	Kluyvera cryocrescens

Cloned(Commentary)

EC Number	Cloned (Comment)	Organism
3.5.1.11	gene pac, recombinant expression of N-terminally His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21 (DE3) pLysS	Kluyvera cryocrescens

Crystallization (Commentary)

EC Number	Crystallization (Comment)	Organism
3.5.1.11	purified recombinant enzyme, sitting drop vapour diffusion method, mixing of 300 nl of 45 mg/ml protein solution with 0.1 ml of precipitant solution containing 30% w/v PEG 4000, 50 mM sodium cacodylate pH 5.6, 0.5 M potassium thiocyanate, 3-4 days, mixing of 500 nl of protein and of precipitant solutions each results in crystals that grow in a week under a wide range of pH conditions using 50 mM sodium cacodylate buffer, method optimization, X-ray diffraction structure determination and analysis at 2.5-3.5 A resolution, molecular replacement method	Kluyvera cryocrescens

Protein Variants

EC Number	Protein Variants	Comment	Organism
3.5.1.11	S290G	site-directed mutagenesis of the internal proteolytic cleavage site of the pro-enzyme	Kluyvera cryocrescens

Localization

EC Number	Localization	Comment	Organism	GeneOntology No.	Textmining

Molecular Weight [Da]

EC Number	Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
3.5.1.11	92000	-	1 * 92000, pro-enzyme, SDS-PAGE	Kluyvera cryocrescens

Natural Substrates/ Products (Substrates)

EC Number	Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
3.5.1.11	penicillin G + H2O	Kluyvera cryocrescens	-	phenylacetate + 6-aminopenicillanate	-	?
3.5.1.11	penicillin G + H2O	Kluyvera cryocrescens DMSZ 2660	-	phenylacetate + 6-aminopenicillanate	-	?

Organism

EC Number	Organism	UniProt	Comment	Textmining
3.5.1.11	Kluyvera cryocrescens	P07941	gene pac	-
3.5.1.11	Kluyvera cryocrescens DMSZ 2660	P07941	gene pac	-

Posttranslational Modification

EC Number	Posttranslational Modification	Comment	Organism
3.5.1.11	proteolytic modification	the enzyme is synthesized as a single-chain inactive pro-enzyme, which upon autocatalytic processing becomes an active heterodimer of alpha and beta chains. Cleavage of the Thr289-Ser290 bond leads to the unveiling of the primary amine group of Serbeta1 (Ser290 of the precursor), creating the active centre in mature enzyme	Kluyvera cryocrescens

Purification (Commentary)

EC Number	Purification (Comment)	Organism
3.5.1.11	recombinant N-terminally His-tagged wild-type and mutant enzymes to homogeneity from Escherichia coli strain BL21 (DE3) pLysS by nickel affinity chromatography and gel filtration	Kluyvera cryocrescens

Substrates and Products (Substrate)

EC Number	Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
3.5.1.11	penicillin G + H2O	-	Kluyvera cryocrescens	phenylacetate + 6-aminopenicillanate	-	?
3.5.1.11	penicillin G + H2O	-	Kluyvera cryocrescens DMSZ 2660	phenylacetate + 6-aminopenicillanate	-	?

Subunits

EC Number	Subunits	Comment	Organism
3.5.1.11	heterodimer	-	Kluyvera cryocrescens
3.5.1.11	monomer	1 * 92000, pro-enzyme, SDS-PAGE	Kluyvera cryocrescens

Synonyms

EC Number	Synonyms	Comment	Organism
3.5.1.11	penicillin G acylase	-	Kluyvera cryocrescens
3.5.1.11	PGA	-	Kluyvera cryocrescens

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Release 2023.1 (January 2023)