Literature summary for 3.5.1.11 extracted from

Chand, D.; Varshney, N.; Ramasamy, S.; Panigrahi, P.; Brannigan, J.A.; Wilkinson, A.J.; Suresh, C.G.
Structure mediation in substrate binding and post-translational processing of penicillin acylases Information from mutant structures of Kluyvera citrophila penicillin G acylase (2015), Protein Sci., 24, 1660-1670 .

Cloned(Commentary)

Cloned (Comment)	Organism
expression in Escherichia coli strain BL21 pLys (DE3)	Kluyvera cryocrescens

Crystallization (Commentary)

Crystallization (Comment)	Organism
crystals of both mutants S1C and S1G belong to space group P1 with four molecules in the asymmetric unit. The three-dimensional structures of these mutants are refined at resolutions 2.8 A and 2.5 A, respectively. Comparison of the structures with similar structures of Escherichia coli PGA (EcPGA) reveals various conformational changes that lead to autocatalytic processing and consequent removal of the spacer peptide	Kluyvera cryocrescens

Crystallization (Comment)

Organism

crystals of both mutants S1C and S1G belong to space group P1 with four molecules in the asymmetric unit. The three-dimensional structures of these mutants are refined at resolutions 2.8 A and 2.5 A, respectively. Comparison of the structures with similar structures of Escherichia coli PGA (EcPGA) reveals various conformational changes that lead to autocatalytic processing and consequent removal of the spacer peptide

Kluyvera cryocrescens

Protein Variants

Protein Variants	Comment	Organism
S1C	replacement of serine of the beta-subunit with cysteine results in a fully processed but inactive enzyme	Kluyvera cryocrescens
S1G	the second mutant in which the N-terminal serine is replaced by glycine remains in the unprocessed and inactive form	Kluyvera cryocrescens

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
2.7	-	penicillin G	pH 7.5, 50°C	Kluyvera cryocrescens

Organism

Organism	UniProt	Comment	Textmining
Kluyvera cryocrescens	P07941	-	-

Posttranslational Modification

Posttranslational Modification	Comment	Organism
proteolytic modification	the enzyme is expressed as a 93000 Da inactive precursor in the cytoplasm which is directed to the periplasmic space by the 26 amino acid residue signal peptide. Subsequently, the spacer peptide is cleaved off by autocatalytic processing to generate the mature and active enzyme	Kluyvera cryocrescens

Purification (Commentary)

Purification (Comment)	Organism
-	Kluyvera cryocrescens

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
62.5	-	pH 7.5, 50°C	Kluyvera cryocrescens

Substrates and Products (Substrate)

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

Subunits

Subunits	Comment	Organism
heterodimer	1 * 23000 (alpha-subunit) + 1 * 63000 (beta-subunit)	Kluyvera cryocrescens

Synonyms

Synonyms	Comment	Organism
penicillin G acylase	-	Kluyvera cryocrescens

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
50	-	assay at	Kluyvera cryocrescens

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
60	-	30 min, complete loss of activity	Kluyvera cryocrescens

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.5	-	-	Kluyvera cryocrescens

pH Range

pH Minimum	pH Maximum	Comment	Organism
7	8	the enzyme is active over the pH range 7.0-8.0	Kluyvera cryocrescens

pH Stability

pH Stability	pH Stability Maximum	Comment	Organism
4	10	stable	Kluyvera cryocrescens
10	-	23°C, 8 h, the enzyme retains 80% of its activity	Kluyvera cryocrescens