Information on EC 3.5.1.11 - penicillin amidase and Organism(s) Escherichia coli and UniProt Accession P06875

-

-

?

D-phenylglycine methyl ester + 7-aminodesacetoxymethyl-3-chlorocephalosporanic acid

cephaclor + ?

-

-

?

diethyl phenylmalonate + H2O

(+)-ethyl phenylmalonate + ethanol

-

the enantiomeric excess is higher than 98%, producing mainly the (+)-ethyl phenylmalonate ester

686450

-

?

dimethyl phenylmalonate + H2O

(+)-methyl phenylmalonate + methanol

-

dimethyl phenylmalonate is fully hydrolyzed to methylphenylmalonate in only 2 h, but even after 10 h phenylmalonic acid is not detected. The enantiomeric excess is higher than 98%, producing mainly the (+)-methyl phenylmalonate ester

686450

-

?

DL-alpha-hydroxybenzylpenicillin + H2O

6-aminopenicillanate + DL-alpha-hydroxyphenylacetic acid

-

-

?

ethyl 2-phenylacetate + H2O

2-phenylacetate + ethanol

-

-

?

glutaryl-7-aminocephalosporanic acid + H2O

glutarate + 7-aminocephalosporanic acid

-

-

?

hydroxyphenylglycineamide + 7-aminodeacetoxycephalosporanic acid

cefadroxil + NH3

-

-

?

isobutoxymethylpenicillin + H2O

6-aminopenicillanate + carbonic acid isopropyl ester

-

-

?

N-(5-nitro-2-pyridyl)-phenylacetamide + H2O

phenylacetate + 5-nitropyridin-2-amine

-

-

?

N-(phenylacetyl)glycine + H2O

phenylacetic acid + glycine

-

-

?

N-phenylacetyl-alpha-homophenylalanine + H2O

homophenylalanine + phenylacetate

experimentally determined E-value (enantioselectivity) towards racemic mixtures of alpha- and beta-amino acids: 80

-

?

N-phenylacetyl-alpha-isoleucine + H2O

isoleucine + phenylacetate

experimentally determined E-value (enantioselectivity) towards racemic mixtures of alpha- and beta-amino acids: 80

-

?

N-phenylacetyl-alpha-leucine + H2O

leucine + phenylacetate

experimentally determined E-value (enantioselectivity) towards racemic mixtures of alpha- and beta-amino acids: 80

-

?

N-phenylacetyl-alpha-phenylalanine + H2O

phenylalanine + phenylacetate

experimentally determined E-value (enantioselectivity) towards racemic mixtures of alpha- and beta-amino acids: 90

-

?

N-phenylacetyl-alpha-tert-leucine + H2O

tert-leucine + phenylacetate

experimentally determined E-value (enantioselectivity) towards racemic mixtures of alpha- and beta-amino acids: 75

-

?

N-phenylacetyl-Asp + H2O

phenylacetic acid + Asp

-

-

?

N-phenylacetyl-Asp-Phe methyl ester + H2O

phenylacetic acid + Asp-Phe methyl ester

-

-

?

N-phenylacetyl-beta-homoleucine + H2O

beta-homoleucine + phenylacetate

experimentally determined E-value (enantioselectivity) towards racemic mixtures of alpha- and beta-amino acids: 55

-

?

N-phenylacetyl-beta-leucine + H2O

beta-leucine + phenylacetate

experimentally determined E-value (enantioselectivity) towards racemic mixtures of alpha- and beta-amino acids: 60

-

?

N-phenylacetyl-DL-leucine + H2O

?

-

-

?

N-phenylacetyl-Glu + H2O

phenylacetic acid + Glu

-

-

?

N-phenylacetyl-L-alpha-aminophenylacetic acid + H2O

?

-

-

?

N-phenylacetyl-Leu + H2O

phenylacetic acid + Leu

-

-

?

N-phenylacetyl-Phe + H2O

phenylacetic acid + Phe

-

-

?

N-phenylacetylglycine + H2O

?

-

-

?

n-propoxymethylpenicillin + H2O

6-aminopenicillanate + carbonic acid monopropyl ester

-

-

?

N2-phenylacetyl-2'-deoxyguanosine + H2O

phenylacetic acid + 2'-deoxyguanosine

N6-phenylacetyl-2'-deoxyadenosine + H2O

phenylacetic acid + 2'-deoxyadenosine

p-hydroxybenzylpenicillin + H2O

6-aminopenicillanate + p-hydroxyphenylacetic acid

-

-

?

penicillin G + H2O

6-aminopenicillanate + phenylacetic acid

11 entries

penicillin G + H2O

6-aminopenicillanic acid + phenyl acetic acid

penicillin G + H2O

6-aminopenicillanic acid + phenylacetic acid

penicillin G + H2O

phenyl acetic acid + 6-aminopenicillanate

-

-

?

penicillin G + H2O

phenylacetate + 6-aminopenicillanate

-

734341, 734463

-

?

penicillin G + H2O

phenylacetic acid + 6-aminopenicillanate

penicillin G + H2O

phenylacetic acid + 6-aminopenicillanic acid

penicillin-G + H2O

6-aminopenicillanic acid + phenylacetic acid

-

658320

-

?

phenylacetamide + 6-aminopenicillanic acid

?

-

-

r

phenylacetamide + H2O

?

-

-

?

phenylacetamide + H2O

phenylacetate + NH3

-

667115, 668459

-

?

phenylacetate 4-nitroanilide + H2O

phenylacetic acid + 4-nitrolaniline

-

171891, 668459

-

?

phenylacetic acid + 6-aminopenicillanic acid

penicillin G

phenylacetic hydrazide + H2O

phenylacetate + hydrazine

-

667115

-

?

phenylacetyl-2-naphthylamide + H2O

phenylacetate + 2-naphthylamine

-

-

?

phenylacetyl-4-aminobenzoic acid

4-aminobenzoic acid + phenylacetic acid

-

colometric assay

-

?

phenylacetyl-7-amido-4-methylcoumarin + H2O

phenylacetate + 7-amino-4-methylcoumarin

-

-

?

phenylacetyl-Gly + H2O

phenylacetic acid + glycine

-

-

?

phenylacetyl-L-asparagine + H2O

L-asparagine + phenylacetic acid

-

-

?

phenylacetylanthranilic acid + H2O

phenylacetate + 2-aminobenzoic acid

-

colometric assay

-

?

phenylacetylaspartame + H2O

phenylacetic acid + aspartame

-

-

?

phenylacetylglycine + 6-aminopenicillanic acid

penicillin G + glycine

phenylglycine methyl ester + aminopenicillanic acid

ampicillin + methanol

-

-

?

phenylglycineamide + 7-aminodeacetoxycephalosporanic acid

cefaclor + NH3

-

-

?

phenylglycinemethylester + 7-aminodeacetoxymethyl-3-chlorocephalosporanic acid

cephalexin + methanol

-

-

?

additional information

?

-

penicillin G + H2O

6-aminopenicillanic acid + phenyl acetic acid

D5M8S0

penicillin G acylase is a type II penicillin acylase

710789

-

?

penicillin G + H2O

6-aminopenicillanic acid + phenylacetic acid

-

171881, 171882, 171883

-

?

penicillin G + H2O

phenylacetate + 6-aminopenicillanate

-

734341, 734463

-

?

penicillin G + H2O

phenylacetic acid + 6-aminopenicillanate

-

710906, 711120

-

?

additional information

?

-

(R)-alpha-ethylphenylacetic acid

-

(R)-alpha-hydroxyphenylacetic acid

-

(R)-alpha-methylphenylacetic acid

-

(R)-phenylglycinamide

-

(R)-phenylglycine

-

(S)-alpha-ethylphenylacetic acid

-

(S)-alpha-hydroxyphenylacetic acid

-

(S)-alpha-methylphenylacetic acid

-

2-thienylacetic acid

-

competitive inhibitor

6-aminopenicillanic acid

7-amino-3-desacetoxycephalosporanic acid

-

non-competitive inhibitor

benzothiophene-2-boronic acid

-

fast reversible competitive inhibitor

benzyl penicillin

-

171882, 171883, 171887

Benzylisocyanate

-

penicillin G

the enzyme is covalently immobilized on aminopropyl functionalized mesocellular foam silica and is further cross-linked using glutaraldehyde without deactivation and upto 95% efficiency. The resulting biocatalyst demonstrates improved resistance to the substrate and product inhibition

phenylacetamide

-

phenylacetic acid

phenylmethanesulfonic acid N-hydroxysuccinimide ester

-

reactivation by incubation with 6-aminopenicillanic acid or proteins from E. coli

Phenylmethylsulfonyl azide

-

reactivation by incubation with 6-aminopenicillanic acid or proteins from E. coli

Phenylmethylsulfonyl chloride

-

reactivation by incubation with 6-aminopenicillanic acid or proteins from E. coli

phenylmethylsulfonyl fluoride

PMSF

-

irreversible, used for active site titrations of the cross-linked enzyme aggregates

712778

[bmim]dca

-

the enzyme activity in 25% [bmim]dca is 1.6fold than activity in water. The enzyme activity is decreased in higher concentration of [bmim]dca

additional information

-

no product inhibition by hydrazine

667115

-

[bmim]dca

-

the enzyme activity in 25% [bmim]dca is 1.6fold than activity in water. The enzyme activity is decreased in higher concentration of [bmim]dca

2.5

2-benzoxazolon-3-yl-acetic acid methyl ester

-

pH 6.8, 25°C

649527

0.081

2-nitro-5-phenylacetamidobenzoic acid

-

pH 7.5, 25°C

0.00614 - 0.03

2-nitro-5-[(phenyl-acetyl)amino]benzoic acid

0.0005 - 0.797

2-nitro-5-[(phenylacetyl)amino]-benzoic acid

9 entries

0.00063

2-phenylacetamidobenzoic acid

-

25°C, pH 7.0, 10% dimethylsulfoxide

0.0004

3-phenylacetamidobenzoic acid

-

25°C, pH 7.0, 10% dimethylsulfoxide

0.114

4-Hydroxyphenylacetamide

pH 7.0, 30°C

0.031

4-nitrophenyl acetate

-

0.0003

4-phenylacetamidobenzoic acid

-

25°C, pH 7.0, 10% dimethylsulfoxide

0.07 - 0.23

5-nitro-3-[(phenylacetyl)amino]benzoic acid

0.02

6-nitro-3-phenylacetamidobenzoic acid

-

pH 7.5, 25°C

650681

0.000015

6-nitro-3-[(phenylacetyl)amino]benzoic acid

-

25°C, pH 7.0, 10% dimethylsulfoxide

0.01

7-phenylacetamidodeacetoxycephalosporanic acid

-

1.07

amoxicillin

pH 7.0, 30°C

1.9 - 5.2

ampicillin

6 entries

0.0031 - 0.8

benzylpenicillin

7 entries

2

benzylpenicilloic acid

-

0.646

cefadroxil

pH 7.0, 30°C

1.5 - 2.1

cephalexin

0.1

cephaloridine

-

0.042

cephalothin

-

3.2

D-(-)-alpha-aminophenylacetic acid p-nitroanilide

-

1.3

D-2-nitro-5-[(phenylacetyl)amino]benzoic acid

pH 7.0, 30°C

12.2

D-4-hydroxyphenylglycine amide

pH 7.0, 30°C

2 - 30

D-phenylglycine amide

0.045

ethyl 2-phenylacetate

-

-

0.00051

N-(5-nitro-2-pyridyl)-phenylacetamide

-

25°C, pH 7.0, 10% dimethylsulfoxide

0.8

N2-phenylacetyl-2'-deoxyguanosine

-

pH 7.8, 25°C

0.46

N6-phenylacetyl-2'-deoxyadenosine

-

pH 7.8, 25°C

0.00001 - 87.08

penicillin G

10 entries

0.0002 - 0.156

phenylacetamide

0.0004

phenylacetate 4-nitroanilide

-

25°C, pH 7.0, 10% dimethylsulfoxide

0.097

phenylacetate p-nitroanilide

-

0.0002

phenylacetyl-2-naphthylamide

-

25°C, pH 7.0, 10% dimethylsulfoxide

0.4

phenylacetyl-3-aminobenzoic acid

-

0.18

phenylacetyl-4-aminobenzoic acid

-

0.000012

phenylacetyl-7-amido-4-methylcoumarin

-

25°C, pH 7.0, 10% dimethylsulfoxide

0.95

phenylacetyl-anthranilic acid

-

5.2

phenylacetyl-Gly

-

pH 7.8, 25°C

0.67

phenylacetylaspartame

-

pH 7.8, 25°C

0.08

phenylacetylglycine

-

additional information

-

1.2

(R) -2-(4-hydroxyphenyl)glycine amide

-

1.01 - 6.08

(S)-N-acetylphenylglycine

0.1

2-benzoxazolon-3-yl-acetic acid methyl ester

-

pH 6.8, 25°C

649527

0.204 - 15

2-nitro-5-[(phenyl-acetyl)amino]benzoic acid

18

2-nitro-5-[(phenylacetyl)amino]-benzoic acid

pH 7.0, 30°C, analysis of complex kinetic parameters alpha, beta, gamma

2.3

2-phenylacetamidobenzoic acid

-

25°C, pH 7.0, 10% dimethylsulfoxide

2.5

3-phenylacetamidobenzoic acid

-

25°C, pH 7.0, 10% dimethylsulfoxide

47

4-Hydroxyphenylacetamide

pH 7.0, 30°C, analysis of complex kinetic parameters alpha, beta, gamma

170

4-nitrophenyl acetate

-

3

4-phenylacetamidobenzoic acid

-

25°C, pH 7.0, 10% dimethylsulfoxide

0.9 - 1.2

5-nitro-3-[(phenylacetyl)amino]benzoic acid

20

6-nitro-3-phenylacetamidobenzoic acid

-

pH 7.5, 25°C

650681

16 - 27

6-nitro-3-[(phenylacetyl)amino]benzoic acid

50

7-phenylacetamidoacetoxycephalosporanic acid

-

17

amoxicillin

pH 7.0, 30°C

11 - 37

ampicillin

6 entries

48 - 50

benzylpenicillin

40

benzylpenicilloic acid

-

32

cefadroxil

pH 7.0, 30°C

29 - 54

cephalexin

33

cephaloridine

-

25

cephalothin

-

0.54

D-(-)-alpha-aminophenylacetic acid 4-nitroanilide

-

14

D-2-nitro-5-[(phenylacetyl)amino]benzoic acid

pH 7.0, 30°C, analysis of complex kinetic parameters alpha, beta, gamma

2 - 8

D-4-hydroxyphenylglycine amide

pH 7.0, 30°C, analysis of complex kinetic parameters alpha, beta, gamma

2.7 - 57

D-phenylglycine amide

170

ethyl 2-phenylacetate

-

-

7.8

N-(5-nitro-2-pyridyl)-phenylacetamide

-

25°C, pH 7.0, 10% dimethylsulfoxide

12000

N-phenylacetyl-Glu

-

18

N2-phenylacetyl-2'-deoxyguanosine

-

pH 7.8, 25°C

2

N6-phenylacetyl-2'-deoxyadenosine

-

pH 7.8, 25°C

39 - 50

penicillin G

46 - 50

phenylacetamide

20 - 55

phenylacetate 4-nitroanilide

1.9

phenylacetyl-2-naphthylamide

-

25°C, pH 7.0, 10% dimethylsulfoxide

3.2

phenylacetyl-7-amido-4-methylcoumarin

-

25°C, pH 7.0, 10% dimethylsulfoxide

47

phenylacetyl-Gly

-

pH 7.8, 25°C

35

phenylacetylaspartame

-

pH 7.8, 25°C

additional information

-

temperature dependence of turnover-numer for R- and S-p-hydroxyphenylglycineamide

650679

-

20 - 500

2-nitro-5-[(phenyl-acetyl)amino]benzoic acid

1700

6-nitro-3-(phenylacetamido)-benzoic acid

pH and temperature not specified in the publication

753335

10000

penicillin G

pH and temperature not specified in the publication

753335

0.276 - 5.7

(R)-alpha-ethylphenylacetic acid

3.5 - 31

(R)-alpha-hydroxyphenylacetic acid

0.076 - 0.47

(R)-alpha-methylphenylacetic acid

2 - 40

(R)-phenylglycinamide

0.37 - 2.1

(S)-alpha-ethylphenylacetic acid

4.05 - 10

(S)-alpha-hydroxyphenylacetic acid

0.36 - 1.5

(S)-alpha-methylphenylacetic acid

0.0086

benzothiophene-2-boronic acid

-

pH 7.5, 25°C

0.05 - 2

phenylacetamide

0.03 - 1.1

phenylacetic acid

0.049

substrate 2-nitro-5-[(phenylacetyl)amino]-benzoic acid, alphaF146Q mutant protein, pH 7.5, 25°C

0.053

substrate 2-nitro-5-[(phenylacetyl)amino]-benzoic acid, alphaF146R mutant protein, pH 7.5, 25°C

0.073

substrate 2-nitro-5-[(phenylacetyl)amino]-benzoic acid, alphaF146A mutant protein, pH 7.5, 25°C

0.15

-

substrate glutaryl-7-aminocephalosporanic acid, wild-type, pH 8.0

0.6

substrate 2-nitro-5-[(phenylacetyl)amino]-benzoic acid, betaF24S mutant protein, pH 7.5, 25°C

0.7

substrate penicillin G, betaF24A/alphaF146Y mutant protein, pH 8.0, 37°C

1

substrate 2-nitro-5-[(phenylacetyl)amino]-benzoic acid, betaF24A mutant protein, pH 7.5, 25°C

1.3

substrate 2-nitro-5-[(phenylacetyl)amino]-benzoic acid, alphaF146Y mutant protein, pH 7.5, 25°C

1.64

-

penicillin G

11

substrate penicillin G, betaF24A mutant protein, pH 8.0, 37°C

12

16

-

substrate 6-nitro-3-[(phenylacetyl)amino]benzoic acid, wild-type, pH 8.5, 25°C

16.3

-

purified recombinant enzyme, pH 8.0, 37°C

17.3

substrate 2-nitro-5-[(phenylacetyl)amino]-benzoic acid, wild type protein, pH 7.5, 25°C

18

-

substrate penicillin G, mutant with three lysine residues alternating with three glycines at C-terminus, pH 8.5, 25°C

2.35

-

cephalosporin G

2.9

substrate 2-nitro-5-[(phenylacetyl)amino]-benzoic acid, betaF24P mutant protein, pH 7.5, 25°C

22240

-

30°C, pH 7.0

670783

3 - 8

substrate penicillin G, wild type protein, pH 8.0, 37°C

33

-

substrate penicillin G, wild-type, pH 8.5, 25°C

38.5

-

40°C

669820

4.6

-

6-aminopenicillanic acid, pH 8, 27°C

48

-

6.3

-

6-aminopenicillanic acid, pH 7

6.6

-

6-aminopenicillanic acid, pH 7.5

666

-

substrate penicillin G, wild-type, pH 8.0

7.1

-

6-aminopenicillanic acid, pH 8-8.5

additional information

-

further values of mutants

10

-

acylation of beta-phenylalanine

5 - 6

-

6.5 - 7.6

-

7

7 - 7.5

-

assay at

7 - 8.5

-

7.2

7.5

7.6

-

8

8 - 8.5

-

8.15

-

8.2

-

additional information

-

overview on values for similar organisms

3 - 10

-

activity range of the immobilized enzyme, profile overview

5 - 8.5

-

6 - 9

-

6.5 - 8.5

7 - 8.5

-

8 - 11

-

pH 8: about 50% of maximal activity, pH 11: about 60% of maximal activity

685738

25

-

assay at

37

40

45

-

50

50 - 60

soluble enzyme

55

-

benzylpenicillin

60

-

171909

additional information

-

overview on values for similar organisms

20 - 50

-

activity range of the free enzyme, profile overview

710906

20 - 60

-

activity range of the immobilized enzyme, profile overview

30 - 35

-

30 - 60

30 - 70

4.3

-

wild-type enzyme

6.4

-

mutant enzyme

6.7

7

8.5

-

-

668439, 685724, 686434, 696160, 718676, 720964, 753335, 754436, 754707

Uniprot

cytosol

-

the mature protein, expressed from a TatProPGA hybrid, is not only found in the extracellular medium and the periplasm, but also in the cytoplasm as assessed by comparison to the reporter beta-lactamase protein

-

-

physiological function

P06875 pBLAST

PAC_ECOLX

846

0

94643

Swiss-Prot

-

20500

22100

-

alpha,beta, x * 22100 + x * 63600, SDS-PAGE

171907

23000

-

1 * 23000 + 1 * 62000, alphabeta, SDS-PAGE

62000

-

1 * 23000 + 1 * 62000, alphabeta, SDS-PAGE

63600

-

alpha,beta, x * 22100 + x * 63600, SDS-PAGE

69000

70000

71000

-

sedimentation equilibrium

?

heterodimer

additional information

-

penicillin G acylase is a 86-kDa heterodimeric protein produced by intein-mediated auto-splicing of a 92-kDa precursor

proteolytic modification

8 entries

both wild-type and mutants F146Y, F24Y, F146Y/F24Y

-

crystals of space group P1 diffracted to beyond 2.3 A

-

171911

hanging-drop vapour diffusion at 18°C, crystal structure of a slowly processing precursor mutant T263G

-

A149Q

activity is too low to measure thermal stability

A305D

mutation in beta-subunit. Half-life at 50°C is 1.6fold higher than wild-type value. Activity is about 60% of wild-type value

A545K

mutation in beta-subunit. Half-life at 50°C is about 80% of wild-type value. Activity is about 80% of wild-type value

A80R

mutation in alpha-subunit. Half-life at 50°C is 2.6fold higher than wild-type value. Activity is 1.6fold higher than wild-type value

A84P

mutation in beta-subunit. Half-life at 50°C is 1.3fold higher than wild-type value. Activity is identical to wild-type value

alphaF146A

mutant selective for (S)-ampicillin synthesis

alphaF146Q

mutant selective for (S)-ampicillin synthesis

alphaF146R

mutant selective for (S)-ampicillin synthesis

alphaF146Y

betaF24A

betaF24A/alphaF146Y

very low amidase activity

betaF24C

mutant selective for (R)-ampicillin synthesis

betaF24P

mutant selective for (R)-ampicillin synthesis

betaF24S

mutant selective for (R)-ampicillin synthesis

betaF24T

used as negative control because of the negative effect on synthetic and hydrolytic activities

betaF24T/alphaF146Y

used as negative control because of the negative effect on synthetic and hydrolytic activities

C290C

-

20% decrease in production of penicillin G acylase activity

D13K

-

mutation in beta-subunit, no change in enzyme stability or kinetic properties, but improved stability after immobilization on glyoxyl-agarose

E130T

mutation in alpha-subunit. Half-life at 50°C is about 60% of wild-type value. Activity is about 1.2fold higher than wild-type value

E272K

-

mutation in beta-subunit, no change in enzyme stability or kinetic properties, but improved stability after immobilization on glyoxyl-agarose

F146A

-

mutation in alpha-subunit. 99% of ampicillin synthesis activity compared to wild-type

F146C

-

mutation in alpha-subunit. 241% of ampicillin synthesis activity compared to wild-type

F146D

-

mutation in alpha-subunit. No ampicillin synthesis activity

F146E

-

mutation in alpha-subunit. 13% of ampicillin synthesis activity compared to wild-type

F146G

-

mutation in alpha-subunit. 61% of ampicillin synthesis activity compared to wild-type

F146H

-

mutation in alpha-subunit. 174% of ampicillin synthesis activity compared to wild-type

F146I

-

mutation in alpha-subunit. 135% of ampicillin synthesis activity compared to wild-type

F146K

-

mutation in alpha-subunit. 120% of ampicillin synthesis activity compared to wild-type

F146L

F146M

-

mutation in alpha-subunit. 85% of ampicillin synthesis activity compared to wild-type

F146N

-

mutation in alpha-subunit. 151% of ampicillin synthesis activity compared to wild-type

F146P

-

mutation in alpha-subunit. 112% of ampicillin synthesis activity compared to wild-type

F146Q

-

mutation in alpha-subunit. 114% of ampicillin synthesis activity compared to wild-type

F146R

-

mutation in alpha-subunit. 3% of ampicillin synthesis activity compared to wild-type

F146S

-

mutation in alpha-subunit. 238% of ampicillin synthesis activity compared to wild-type

F146T

-

mutation in alpha-subunit. 376% of ampicillin synthesis activity compared to wild-type

F146V

-

mutation in alpha-subunit. 153% of ampicillin synthesis activity compared to wild-type

F146W

F146Y

F146Y/F24A

-

mutation F24Y in beta-, F146Y in alpha-subunit, increased affinity for Calpha-substituted substrates

F24A

F71C

-

mutation in B-subunit shows a 100fold increase in kcat/Km towards glutaryl-L-leucine

F71L

-

mutation in B-subunit shows a 100fold increase in kcat/Km towards glutaryl-L-leucine

H26G

activity is too low to measure thermal stability

K299H

-

mutant shows very low processing and 90% loss of activity

K299Q

-

mutant enzyme shows no processing

K299S

-

mutant enzyme shows no processing

L100E

mutation in beta-subunit. Half-life at 50°C is 1.2fold higher than wild-type value. Activity is about 50% of wild-type value

M90R

mutation in alpha-subunit. Half-life at 50°C is nearly identical to wild-type value. Activity is about 70% of wild-type value

N241G

-

site-directed mutagensis of subunit B residue, leads to reduced activity compared to the wild-type enzyme

N241S

-

site-directed mutagensis of subunit B residue, leads to reduced activity compared to the wild-type enzyme

N348D

mutation in beta-subunit. Half-life at 50°C is 1.5fold higher than wild-type value. Activity is nearly identical to wild-type value

Q118E

activity is too low to measure thermal stability

R145A

-

mutation in alpha-subunit. 154% of ampicillin synthesis activity compared to wild-type

R145C

-

mutation in alpha-subunit. 169% of ampicillin synthesis activity compared to wild-type

R145D

-

mutation in alpha-subunit. 15% of ampicillin synthesis activity compared to wild-type

R145E

-

mutation in alpha-subunit. 6% of ampicillin synthesis activity compared to wild-type

R145F

-

mutation in alpha-subunit. 131% of ampicillin synthesis activity compared to wild-type

R145G

-

mutation in alpha-subunit. 256% of ampicillin synthesis activity compared to wild-type. Due to increased tendency of the acyl-enzyme intermediate to react with beta-lactam nucleophile instead of water, the mutant demonstrates an enhanced synthetic yield over wild-type penicillin acylase at high substrate concentrations. This is accompanied by an increased conversion of 6-aminopenicillanic acid to ampicillin as well as a decreased undesirable hydrolysis of the acyl donor

R145H

-

mutation in alpha-subunit. 78% of ampicillin synthesis activity compared to wild-type

R145I

-

mutation in alpha-subunit. 120% of ampicillin synthesis activity compared to wild-type

R145K

-

mutation in alpha-subunit. 145% of ampicillin synthesis activity compared to wild-type

R145L

-

mutation in alpha-subunit. 237% of ampicillin synthesis activity compared to wild-type. Due to increased tendency of the acyl-enzyme intermediate to react with beta-lactam nucleophile instead of water, the mutant demonstrates an enhanced synthetic yield over wild-type penicillin acylase at high substrate concentrations. This is accompanied by an increased conversion of 6-aminopenicillanic acid to ampicillin as well as a decreased undesirable hydrolysis of the acyl donor

R145M

-

mutation in alpha-subunit. 129% of ampicillin synthesis activity compared to wild-type

R145N

-

mutation in alpha-subunit. 173% of ampicillin synthesis activity compared to wild-type

R145P

-

mutation in alpha-subunit. 137% of ampicillin synthesis activity compared to wild-type

R145Q

-

mutation in alpha-subunit. 158% of ampicillin synthesis activity compared to wild-type

R145S

-

mutation in alpha-subunit. 192% of ampicillin synthesis activity compared to wild-type. Due to increased tendency of the acyl-enzyme intermediate to react with beta-lactam nucleophile instead of water, the mutant demonstrates an enhanced synthetic yield over wild-type penicillin acylase at high substrate concentrations. This is accompanied by an increased conversion of 6-aminopenicillanic acid to ampicillin as well as a decreased undesirable hydrolysis of the acyl donor

R145T

-

mutation in alpha-subunit. 127% of ampicillin synthesis activity compared to wild-type

R145V

-

mutation in alpha-subunit. 127% of ampicillin synthesis activity compared to wild-type

R145W

-

mutation in alpha-subunit. 103% of ampicillin synthesis activity compared to wild-type

R145Y

-

mutation in alpha-subunit. 37% of ampicillin synthesis activity compared to wild-type

R276K

-

mutation in beta-subunit, no change in enzyme stability or kinetic properties, but improved stability after immobilization on glyoxyl-agarose

S374T

mutation in beta-subunit. Half-life at 50°C is about 90% of wild.type value. Activity about 50% of wild-type value

S9E

activity is too low to measure thermal stability

T121D

mutation in alpha-subunit. Half-life at 50°C is about 70% of wild-type value. Activity is about 40% of wild-type value

T150N

mutation in alpha-subunit. Half-life at 50°C is 1.5fold higher than wild-type value. Activity is about 50% of wild-type value

T263G

-

slowly processing mutant enzyme

650295

T289C

-

production of penicillin G acylase activity is 92% improved

T289G

-

production of penicillin G acylase activity is 20% improved

T289S

-

production of penicillin G acylase activity is 85% improved

T311P/Q312A

mutation in beta-subunit. Half-life at 50°C is 2fold higher than wild-type value. Activity is about 90% of wild-type value

V184K

mutation in beta-subunit. Half-life at 50°C is about 60% of wild-type value. Activity is about 60% of wild-type value

V359L

mutation in beta-subunit. Half-life at 50°C is 1.4fold higher than wild-type value. Activity is about 60% of wild-type value

V400L

mutation in beta-subunit. Half-life at 50°C is 1.8fold higher than wild-type value. Activity is about 75% of wild-type value

V56R

V56R/T32Y

-

mutation of beta-subunit, plus F146Y in alpha-subunit, 2.3% of wild-type activity with substrate penicillin G, 460% of wild-type activity with cephalosporin acylase substrate glutaryl-7-aminocephalosporanic acid

V56R/T32Y/I177Y

-

mutation of beta-subunit, plus F146Y in alpha-subunit, 1.8% of wild-type activity with substrate penicillin G, 490% of wild-type activity with cephalosporin acylase substrate glutaryl-7-aminocephalosporanic acid

V56R/T32Y/I177Y/P49Q

-

mutation of beta-subunit, plus F146Y in alpha-subunit, 1.3% of wild-type activity with substrate penicillin G, 510% of wild-type activity with cephalosporin acylase substrate glutaryl-7-aminocephalosporanic acid

V56R/T32Y/I177Y/P49Q/W154Y

-

mutation of beta-subunit, plus F146Y in alpha-subunit, 1.2% of wild-type activity with substrate penicillin G, 600% of wild-type activity with cephalosporin acylase substrate glutaryl-7-aminocephalosporanic acid

V56R/T32Y/I177Y/P49Q/W154Y/F24L

-

mutation of beta-subunit, plus F146Y in alpha-subunit, 0.3% of wild-type activity with substrate penicillin G, 760% of wild-type activity with cephalosporin acylase substrate glutaryl-7-aminocephalosporanic acid

V97K

activity is too low to measure thermal stability

W25Y

mutation in alpha-subunit. Half-life at 50°C is 2.7fold higher than wild-type value. Activity is 1.4fold higher than wild-type value

additional information

10 entries

10

3 - 9.5

-

the enzyme maintains the folded structure in the pH range from pH 3.0 to 9.5

685398

4.5 - 4.8

-

high inactivation at pH 4.5, but stable at pH 4.8

711120

8

-

half-life at 25°C is 6 days

667754

9.1

-

half-life at 25°C is 9.1 h

667754

additional information

-

study on inactivation kinetics over a wide pH-range at 25°C and 50°C. Enzyme is very stable in neutral solutions and quickly loses its catalytic activity in acidic and alkaline solutions. Inactivation proceeds according to first order reaction kinetics

667754

40

-

denaturation temperature at pH 3.0 or 9.5

685398

5

-

stable during the whole working period

50

-

loss of stability

60

-

enzyme immobilized on epoxy polymer: 80% loss of activity after 30 min. Enzyme immobilized on amino polymers with short spacer arms: about 80% loss of activity after 60 min. Enzyme immobilized on amino polymers with long spacer arm: about 95% loss of activity after 5 min, about 80% loss of activity after 120 min. Native enzyme in solution: complete loss of activity after 5 min

684252

65

-

denaturation temperature at pH 6.0

685398

70

-

thermal stability of the enzyme immobilized on alginate/chitosan hybrid beads is much higher that that of the soluble enzyme

additional information

crosslinking with glutaraldehyde fails to protect the immobilized enzyme activity in case of proline, tryptophan, and casein acid hydrolysate

-

710906

enzyme immobilized on alginate/chitosan hybrid beads can be reused more times as compared to enzyme immobilized on alginate beads

-

enzyme immobilized on alginate/chitosan hybrid beads shows higher operational stability compared to enzyme immobilized on alginate beads

-

immobilized enzyme, higher stability than soluble enzyme

-

the enzyme is covalently immobilized on aminopropyl functionalized mesocellular foam silica and is further cross-linked using glutaraldehyde without deactivation and upto 95% efficiency. The resulting biocatalyst has an activity of 1185 IU mg/l and demonstrates improved pH and thermal stability

754707

the enzyme is more stable in [bmim]PF6 than in conventional aqueous solution

-

the mutant enzyme adsorbed on DEAE is more stable than the enzyme covalently immobilized on CNBr agarose. The most stable preparations are those where the enzyme is adsorbed on polyethyleneimine

-

the native enzyme does not become significantly immobilized on any of the three supports (DEAE and two supports coated with polyethyleneimine of different sizes), while the mutant enzyme becomes fully immobilized on them. The use of restrictive conditions during the enzyme adsorption on anionic exchangers (pH 5 and high ionic strength) further increases the strength of adsorption and the enzyme stability in the presence of organic solvents

-

dioxane

-

investigation of inactivation of immobilized enzyme by 70% (v/v) dioxan

Methanol

-

in absence of methanol very stable, in 40% methanol, inactivation

171896

-20°C, freeze dried pure enzyme

-

171882, 171887

2°C, immobilized enzyme

-

171883

enzyme immobilized on alginate/chitosan hybrid beads shows higher storage stability compared to enzyme immobilized on alginate beads

-

-

commercial preparation

-

commercial preparation, Eupergit-immobilized

-

719296

His-tagged recombinant enzyme

-

670783

ion exchange chromatography (Q-Sepharose)

mixed ion exchange supports are useful ion exchangers for purification of penicillin G acylase

-

685491

mutant proenzyme T263G, expressed in Escherichia coli

-

650295

native enzyme 4.5-7.5fold from crude cell extract or homogenate by expanded bed adsorption method using a cationic resin, method comparisons show similar results for packed-bed and for expanded-bed modes, evaluation, overview

-

partially

-

periplasmic fraction

purification of His-tagged enzyme secreted into periplasmic psace by immobilized metal-ion affnity chromatography. Maximum specific activity of enzyme is 38.5 U/mg, yield is 70%

-

669820

recombinant enzyme from Escherichia coli strain 5KpHM12 culture supernatant by anion and cation exchange chromatographic high-throughput absorption membranes, method development and evaluation, binding dynamics, overview

-

734463

recombinant extracellular enzyme 3.2fold from Escherichia coli strain HB101 culture supernatant by anion exchange chromatography, single-step purification

-

-

a cross-species penicillin G amidase gene coding for an alpha-peptide and a linker peptide from K. citrophila and a beta-peptide from Escherichia coli constructed and cloned in Escherichia coli

-

650678

cloned into pUC 9 and transformed into Escherichia coli

-

171911

expressed in Escherichia coli HB101, sequence shows one mutation resulting in a betaVal148Leu substitution in the mature protein as compared with the Swiss-Prot entry P06875 derived from Escherichia coli ATC11105

expression in a phage display in Escherichia coli strain TG1

-

expression in Escherichia coli

-

expression levels in various Escherichia coli host cells. Intracellular proteolytic degradation of the newly synthesized penicillin amidase precursor and translocation through the plasma membrane are determined to be the main posttranslational processes limiting enzyme production. The production of mature active penicillin amidase is increased up to 10fold when the protease deficient strain Escherichia coli BL21 (DE3) is cultivated in medium without a proteinaceous substrate. Simultaneous coexpression of the OmpT pac gene with some proteins of the Sec export machinery of the cell results in up to threefold-enhanced penicillin amidase production

-

649380

expression of functional wild-type and mutant enzymes in Pseudomonas aeruginosa PAO1 and PAO1DELTAtatABC strains using the pMMB67EH shuttle vector, the mutant a TatProPGA hybrid only shows an active site, when the transformed bacterium is grown at 25°C or lower temperatures. Processing of recombinant PGA can also occur in the cytoplasm of Pseudomonas aeruginosa. The extracellular localization of the TatProPGA hybrid is not dependent on the tatABC-genes

-

712528

expresssion of the enzyme in Escherichia coli strain 5KpHM12

-

734463

gene pac, overexpression in Escherichia coli strain HB101 and secretion of the enzyme to the culture medium, method optimization, overview

-

PGA gene cloning isolated from environmental and clinical samples, DNA and amino acid sequence determination and analysis, subcloning in Escherichia coli strain DH5alpha, overexpression as C-terminally His-tagged protein in Escherichia coli strain BL21 under control of the T7 promoter using pGEM -T easy vector

D5M8S0

710789

production of penicillin amidase hybrid I and hybrid II in Escherichia coli. Hybrid A contains A-chain from K. citrophila and B-chain from Escherichia coli. Hybrid II contains A-chain from Escherichia coli and B-chain from Kluyvera citrophila

-

partially inactivated immobilized enzyme, by addition of e.g. ethyleneglycol and catalytic modulators (competitive (reduction in reactivation yield) and non-competitive (increase in reactivation yield) inhibitors)

-

synthesis

Bauer, K.; Kaufmann, W.; Ludwig, S.A.

A simplified determination of penicillin amidase from E. coli

Hoppe-Seyler's Z. Physiol. Chem.

352

1723-1724

1971

Escherichia coli

4947502

171882

Balasingham, K.; Warburton, O.; Dunnil, P.; Lilly, M.D.

The isolation and kinetics of penicillin amidase from Escherichia coli

Biochim. Biophys. Acta

276

250-256

1972

Escherichia coli

4340118

171883

Warburton, D.; Balasingham, K.; Dunnil, P.; Lilly, M.D.

The preparation and kinetics of immobilised penicillin amidase from Escherichia coli

Biochim. Biophys. Acta

284

278-284

1972

Escherichia coli

4342221

171884

Boeck, A.; Wirth, R.; Schmid, G.; Schumacher, G.; Lang, G.; Buckel, P.

The penicillin acylase from Escherichia coli ATCC 11105 consists of two dissimilar subunits

FEMS Microbiol. Lett.

20

135-139

1983

Escherichia coli

-

171885

Berezin, I.V.; Klibanov, A.M.; Klyosov, A.A.; Martinek, K.; Svedas, V.K.

The effect of ultrasound as a new method of studying conformational transitions in enzyme active sites. pH- and temperature-induced conformational transitions in the active centre of penicillin amidase

FEBS Lett.

49

325-328

1975

Escherichia coli

234079

171886

Cole, M.; Savidge, T.; vander Haeghe, H.

Penicillin acylase (assay)

Methods Enzymol.

43

698-705

1975

Bacillus subtilis, Escherichia coli

805897

Savidge, T.A.; Cole, M.

Penicillin acylase (bacterial)

Methods Enzymol.

43

705-721

1975

Priestia megaterium, Escherichia coli

237220

171889

Szewczuk, A.; Ziomek, E.; Mordarski, M.; Siewinski, M.; Wieczorek, J.

Properties of penicillin amidase immobilized by copolymerization with acrylamide

Biotechnol. Bioeng.

21

1543-1552

1979

Escherichia coli

387106

Szewczuk, A.; Siewinski, M.; Slowinska, R.

Colorimetric assay of penicillin amidase activity using phenylacetyl-aminobenzoic acid as substrate

Anal. Biochem.

103

166-169

1980

Escherichia coli

6990827

Morgolin, A.L.; Svedas, V.K.; Berezin, I.V.

Substrate specificity of penicillin amidase from E. coli

Biochim. Biophys. Acta

616

283-289

1980

Escherichia coli

7011386

171892

Svedas, V.K.; Margolin, A.L.; Borisov, I.L.; Berezin, I.V.

Kinetics of the enzymatic synthesis of benzylpenicillin

Enzyme Microb. Technol.

2

313-317

1980

Escherichia coli

-

171893

McDougall, B.; Dunnill, P.; Lilly, M.D.

Enzymatic acylation of 6-aminopenicillanic acid

Enzyme Microb. Technol.

4

114-115

1982

Escherichia coli

-

Siewinski, M.; Kuropatwa, M.; Szewczuk, A.

Phenylalkylsulfonyl derivatives as covalent inhibitors of penicillin amidase

Hoppe-Seyler's Z. Physiol. Chem.

365

829-837

1984

Escherichia coli

6386649

171895

Kasche, V.; Haufler, U.; Z�llner, R.

Kinetic studies on the mechanism of the penicillin amidase-catalysed synthesis of ampicillin and benzylpenicillin

Hoppe-Seyler's Z. Physiol. Chem.

365

1435-1443

1984

Escherichia coli

6098545

171896

Kasche, V.

Ampicillin- and cephalhexin-synthesis catalyzed by E. coli penicillin amidase. yield increase due to substrate recycling.

Biotechnol. Lett.

7

877-882

1985

Escherichia coli

-

171897

Sudhakaran, V.K.; Shewale, J.G.

Hydrophobic interaction chromatography of penicillin amidase

Biotechnol. Lett.

9

539-542

1987

Escherichia coli

-

171898

Brandl, E.

Information on penicillin amidase

Hoppe-Seyler's Z. Physiol. Chem.

342

86

1965

Escherichia coli

4960666

171899

Self, D.A.; Kay, G.; Lilly, M.D.; Dunnill, P.

The conversion of benzyl penicillin to 6-aminopenicillanic acid using an insoluble derivative of penicillin amidase

Biotechnol. Bioeng.

11

337

1969

Escherichia coli

4898121

171900

Kutzbach, C.; Rauenbusch, E.

Preparation and general properties of crystalline penicillin acylase from Escherichia coli ATCC 11 105

Hoppe-Seyler's Z. Physiol. Chem.

355

45-53

1974

Escherichia coli

4611892

171901

Nys, P.S.; Kolygina, T.S.; Garaev, M.M.

Penicillin amidase from E. coli. A direct spectrophotometric method of determining the enzymes activity

Antibiotiki

22

211-216

1977

Escherichia coli

15505

171902

Margolin, A.L.; Izumrudov, V.A.; Svedas, V.K.; Zezin, A.B.; Kabanov, V.A.; Berezin, I.V.

Preparation and properties of penicillin amidase immobilized in polyelectrolyte complexes

Biochim. Biophys. Acta

660

359-365

1981

Escherichia coli

7025915

Niersbach, H.; Kuhne, A.; Tischer, W.; Weber, M.; Wedekind F.; Plapp, R.

Improvement of the catalytic properties of penicillin G acylase from Escherichia coli ATCC 11105 by selection of a new substrate specificity

Appl. Microbiol. Biotechnol.

43

679-684

1995

Escherichia coli

7546605

171907

Sriubolmas, N.; Panbangred, W.; Sriurairatana, S.; Meevootisom, V.

Localization and characterization of inclusion bodies in recombinant Escherichia coli cells overproducing penicillin G acylase

Appl. Microbiol. Biotechnol.

47

373-378

1974

Escherichia coli

9163951

171908

Fernandez-Lafuente, R.; Rosell, C.M.; Guisan, J.M.

Enzyme reaction engineering: synthesis of antibiotics catalysed by stabilized penicillin G acylase in the predence of organic solvents

Enzyme Microb. Technol.

13

898-905

1991

Escherichia coli, Kluyvera cryocrescens

1368000

171909

Erarslan, A.; Terzi, I.; Guray, A.; Bermek, E.

Purification and kinetics of penicillin G acylase from a mutant strain of Escherichia coli ATCC 11105

J. Chem. Technol. Biotechnol.

51

27-40

1991

Escherichia coli

-

171910

Erarlan, A.; Guray, A.

Kinetic investigation of penicillin G acylase from a mutant strain of Escheria coli ATCC 11105 immobilized on oxirane-acrylic beads

J. Chem. Technol. Biotechnol.

51

181-195

1991

Escherichia coli

1369308

171911

Hunt, P.D.; Tolley, S.P.; Ward, R.J.; Hill, C.P.; Dodson, G.G.

Expression, purification and crystallization of penicillin G acylase from Escherichia coli ATCC 11105

Protein Eng.

3

635-639

1990

Escherichia coli

2217137

171914

Azevedo, A.M.; Fonseca, L.P.; Prazeres, D.M.F.

Stability and stabilisation of penicillin acylase

J. Chem. Technol. Biotechnol.

74

1110-1116

1999

Escherichia coli

-

649380

Ignatova, Z.; Mahsunah, A.; Georgieva, M.; Kasche, V.

Improvement of posttranslational bottlenecks in the production of penicillin amidase in recombinant Escherichia coli strains

Appl. Environ. Microbiol.

69

1237-1245

2003

Escherichia coli

12571052

649527

Stambolieva, N.; Mincheva, Z.; Galunsky, B.

Kinetic comparison of penicillin amidase catalsed transfer of nonspecific and specific acyl moieties to 7-aminodeacetoxycephalosporanic acid

Biocatal. Biotransform.

16

225-232

1998

Escherichia coli

-

650295

Kasche, V.; Lummer, K.; Nurk, A.; Piotraschke, E.; Rieks, A.; Stoeva, S.; Voelter, W.

Intramolecular autoproteolysis initiates the maturation of penicillin amidase from Escherichia coli

Biochim. Biophys. Acta

1433

76-86

1999

Escherichia coli

10446361

Dineva, M.A.; Gatunsky, B.; Kasche, V.; Petkov, D.D.

Phenylacetyl group as enzyme-cleavable aminoprotection of purine nucleosides

Bioorg. Med. Chem. Lett.

3

2781-2784

1993

Escherichia coli

-

650678

Piotraschke, E.; Nurk, A.; Galunsky, B.; Kasche, V.

Genetic construction of catalytically active cross-species heterodimer penicillin G amidase

Biotechnol. Lett.

16

119-124

1994

Escherichia coli, Kluyvera cryocrescens

-

650679

Kasche, V.; Galunsky, B.; Nurk, E.; Piotraschke, E.; Rieks, A.

The dependency of the stereoselectivity of penicillin amidases � enzymes with R-specific S1 and S-specific S'1-subsites � on temperature and primary structure

Biotechnol. Lett.

18

455-460

1996

Escherichia coli

-

650681

Ignatova, Z.; Stoeva, S.; Galunsky, B.; H�rnle, C.; Nurk, A.; Piotraschke, E.; Voelter, W.; Kasche, V.

Proteolytic processing of penicillin amidase from Alcaligenes faecalis cloned in Escherichia coli yields several active forms

Biotechnol. Lett.

20

977-982

1998

Alcaligenes faecalis, Escherichia coli

-

650683

Ignatova, Z.; Taruttis, S.; Kasche, V.

Role of the intracellular proteolysis in the production of the periplasmic penicillin amidase in Escherichia coli

Biotechnol. Lett.

22

1727-1732

2000

Escherichia coli

-

651013

Ignatova, Z.; Enfors, S.O.; Hobbie, M.; Taruttis, S.; Vogt, C.; Kasche, V.

The relative importance of intracellular proteolysis and transport on the yield of the periplasmic enzyme penicillin amidase in Escherichia coli

Enzyme Microb. Technol.

26

165-170

2000

Escherichia coli

10689073

652830

Hewitt, L.; Kasche, V.; Lummer, K.; Lewis, R.J.; Murshudov, G.N.; Verma, C.S.; Dodson, G.G.; Wilson, K.S.

Structure of a slow processing precursor penicillin acylase from Escherichia coli reveals the linker peptide blocking the active-site cleft

J. Mol. Biol.

302

887-898

2000

Escherichia coli

10993730

Galunsky, B.; Lummer, K.; Kasche, V.

Comparative study of substrate- and stereospecificity of penicillin G amidases from different sources and hybrid isoenzymes

Monatsh. Chem.

131

623-632

2000

Alcaligenes faecalis, Rhizobium viscosum, Escherichia coli, Kluyvera cryocrescens

-

658320

Tramper, J.

Chemical versus biochemical conversion: when and how to use biocatalysts

Biotechnol. Bioeng.

52

290-295

1996

Escherichia coli

18629895

Hernandez-Justiz, O.; Fernandez-Lafuente, R.; Terrini, M.; Guisan, J.M.

Use of aqueous two-phase systems for in situ extraction of water soluble antibiotics during their synthesis by enzymes immobilized on porous supports

Biotechnol. Bioeng.

59

73-79

1998

Escherichia coli

10099316

667115

Basso, A.; Ebert, C.; Gardossi, L.; Linda, P.; Phuong, T.T.; Zhu, M.; Wessjohann, L.

Penicillin G amidase-catalysed hydrolysis of phenylacetic hydrazides on a solid phase: A new route to enzyme-cleavable linkers

Adv. Synth. Catal.

347

963-966

2005

Escherichia coli

-

Abian, O.; Grazu, V.; Hermoso, J.; Gonzalez, R.; Garcia, J.L.; Fernandez-Lafuente, R.; Guisan, J.M.

Stabilization of penicillin G acylase from Escherichia coli: site-directed mutagenesis of the protein surface to increase multipoint covalent attachment

Appl. Environ. Microbiol.

70

1249-1251

2004

Escherichia coli

14766616

Scaramozzino, F.; Estruch, I.; Rossolillo, P.; Terreni, M.; Albertini, A.M.

Improvement of catalytic properties of Escherichia coli penicillin G acylase immobilized on glyoxyl agarose by addition of a six-amino-acid tag

Appl. Environ. Microbiol.

71

8937-8940

2005

Escherichia coli

16332894

Oh, B.; Kim, K.; Park, J.; Yoon, J.; Han, D.; Kim, Y.

Modifying the substrate specificity of penicillin G acylase to cephalosporin acylase by mutating active-site residues

Biochem. Biophys. Res. Commun.

319

486-492

2004

Escherichia coli

15178432

667754

Guranda, D.T.; Volovik, T.S.; Svedas, V.K.

pH Stability of penicillin acylase from Escherichia coli

Biochemistry (Moscow)

69

1386-1390

2004

Escherichia coli

15627395

668030

Masarova, J.; Mislovicova, D.; Mendichi, R.; Svitel, J.; Gemeiner, P.; Danielsson, B.

Mannan-penicillin G acylase neoglycoproteins and their potential applications in biotechnology

Biotechnol. Appl. Biochem.

39

285-291

2004

Escherichia coli

15154839

668035

Youshko, M.I.; Moody, H.M.; Bukhanov, A.L.; Boosten, W.H.; Svedas, V.K.

Penicillin acylase-catalyzed synthesis of beta-lactam antibiotics in highly condensed aqueous systems: beneficial impact of kinetic substrate supersaturation

Biotechnol. Bioeng.

85

323-329

2004

Escherichia coli

14748088

668062

Xu, Y.; Hsieh, M.Y.; Narayanan, N.; Anderson, W.A.; Scharer, J.M.; Moo-Young, M.; Chou, C.P.

Cytoplasmic overexpression, folding, and processing of penicillin acylase precursor in Escherichia coli

Biotechnol. Prog.

21

1357-1365

2005

Escherichia coli

16209538

668240

Wu, W.B.; Wang, N.; Xu, J.M.; Wu, Q.; Lin, X.F.

Penicillin G acylase catalyzed Markovnikov addition of allopurinol to vinyl ester

Chem. Commun. (Camb.)

2005

2348-2350

2005

Escherichia coli

15877124

Gabor, E.M.; de Vries, E.J.; Janssen, D.B.

A novel penicillin acylase from the environmental gene pool with improved synthetic properties

Enzyme Microb. Technol.

36

182-190

2005

Escherichia coli (P06875), uncultured Gammaproteobacteria bacterium (Q6PWR5)

-

668443

Terreni, M.; Ubiali, D.; Pagani, G.; Hernandez-Justiz, O.; Fernandez-Lafuente, R.; Guisan, J.M.

Penicillin G acylase catalyzed acylation of 7-ACA in aqueous two-phase systems using kinetically and thermodynamically controlled strategies: improved enzymatic synthesis of 7-[(1-hydroxy-1-phenyl)-acetamido]-3-acetoxymethyl-D3-cephem-4-carboxylic acid

Enzyme Microb. Technol.

36

672-679

2005

Escherichia coli

-

Guncheva, M.; Ivanov, I.; Galunsky, B.; Stambolieva, N.; Kaneti, J.

Kinetic studies and molecular modelling attribute a crucial role in the specificity and stereoselectivity of penicillin acylase to the pair ArgA145-ArgB263

Eur. J. Biochem.

271

2272-2279

2004

Escherichia coli

15153118

668619

Viegas, S.C.; Schmidt, D.; Kasche, V.; Arraiano, C.M.; Ignatova, Z.

Effect of the increased stability of the penicillin amidase mRNA on the protein expression levels

FEBS Lett.

579

5069-5073

2005

Escherichia coli

16137683

669820

Kim, J.; Kang, H.; Kim, E.; Kim, J.; Koo, Y.

One-step purification of poly-His tagged penicillin G acylase expressed in E. coli

J. Microbiol. Biotechnol.

14

231-236

2004

Escherichia coli

-

669900

Ribeiro, M.P.; Ferreira, A.L.; Giordano, R.L.; Giordano, R.C.

Selectivity of the enzymatic synthesis of ampicillin by E. coli PGA in the presence of high concentrations of substrates

J. Mol. Catal. B

33

81-86

2005

Escherichia coli

-

Alkema, W.B.; Hensgens, C.M.; Snijder, H.J.; Keizer, E.; Dijkstra, B.W.; Janssen, D.B.

Structural and kinetic studies on ligand binding in wild-type and active-site mutants of penicillin acylase

Protein Eng. Des. Sel.

17

473-480

2004

Escherichia coli

15254299

670783

Cheng, T.; Chen, M.; Zheng, H.; Wang, J.; Yang, S.; Jiang, W.

Expression and purification of penicillin G acylase enzymes from four different micro-organisms, and a comparative evaluation of their synthesis/hydrolysis ratios for cephalexin

Protein Expr. Purif.

46

107-113

2006

Alcaligenes faecalis, Escherichia coli, Kluyvera cryocrescens, Providencia rettgeri

16139515

684252

Basso, A.; Braiuca, P.; Cantone, S.; Ebert, C.; Linda, P.; Spizzo, P.; Caimi, P.; Hanefeld, U.; Degrassi, G.; Gardossi, L.

In silico analysis of enzyme surface and glycosylation effect as a tool for efficient covalent immobilisation of CalB and PGA on Sepabeads

Adv. Synth. Catal.

349

877-886

2007

Escherichia coli, Providencia rettgeri

-

Montes, T.; Grazu, V.; Lopez-Gallego, F.; Hermoso, J.A.; Garcia, J.L.; Manso, I.; Galan, B.; Gonzalez, R.; Fernandez-Lafuente, R.; Guisan, J.M.

Genetic modification of the penicillin G acylase surface to improve its reversible immobilization on ionic exchangers

Appl. Environ. Microbiol.

73

312-319

2007

Escherichia coli

17098917

685398

Grinberg, V.Y.; Burova, T.V.; Grinberg, N.V.; Shcherbakova, T.A.; Guranda, D.T.; Chilov, G.G.; Svedas, V.K.

Thermodynamic and kinetic stability of penicillin acylase from Escherichia coli

Biochim. Biophys. Acta

1784

736-746

2008

Escherichia coli

18314015

685491

Fuentes, M.; Batalla, P.; Grazu, V.; Pessela, B.C.; Mateo, C.; Montes, T.; Hermoso, J.A.; Guisan, J.M.; Fernandez-Lafuente, R.

Mixed ion exchange supports as useful ion exchangers for protein purification: purification of penicillin G acylase from Escherichia coli

Biomacromolecules

8

703-707

2007

Escherichia coli

17291095

Zhang, W.G.; Wei, D.Z.; Yang, X.P.; Song, Q.X.

Penicillin acylase catalysis in the presence of ionic liquids

Bioprocess Biosyst. Eng.

29

379-383

2006

Escherichia coli

17082915

Polizzi, K.M.; Chaparro-Riggers, J.F.; Vazquez-Figueroa, E.; Bommarius, A.S.

Structure-guided consensus approach to create a more thermostable penicillin G acylase

Biotechnol. J.

1

531-536

2006

Escherichia coli (P06875)

16892288

685738

Li, D.; Cheng, S.; Wei, D.; Ren, Y.; Zhang, D.

Production of enantiomerically pure (S)-beta-phenylalanine and (R)-beta-phenylalanine by penicillin G acylase from Escherichia coli in aqueous medium

Biotechnol. Lett.

29

1825-1830

2007

Escherichia coli

17657412

685811

Cecchini, D.A.; Serra, I.; Ubiali, D.; Terreni, M.; Albertini, A.M.

New active site oriented glyoxyl-agarose derivatives of Escherichia coli penicillin G acylase

BMC Biotechnol.

7

54

2007

Escherichia coli

17845725

686434

Jager, S.A.; Jekel, P.A.; Janssen, D.B.

Hybrid penicillin acylases with improved properties for synthesis of beta -lactam antibiotics

Enzyme Microb. Technol.

40

1335-1344

2007

Escherichia coli (P06875)

-

686450

Cabrera, Z.; Lopez-Gallego, F.; Fernandez-Lorente, G.; Palomo, J.M.; Montes, T.; Grazu, V.; Guisan, J.M.; Fernandez-Lafuente, R.

Asymmetric hydrolysis of dimethyl phenylmalonate by immobilized penicillin G acylase from E. coli

Enzyme Microb. Technol.

40

997-1000

2007

Escherichia coli

-

Chandel, A.K.; Rao, L.V.; Narasu, M.L.; Singh, O.V.

The realm of penicillin G acylase in beta-lactam antibiotics

Enzyme Microb. Technol.

42

199-207

2008

Priestia megaterium, Escherichia coli, Kluyvera cryocrescens

-

Jager, S.A.; Shapovalova, I.V.; Jekel, P.A.; Alkema, W.B.; Svedas, V.K.; Janssen, D.B.

Saturation mutagenesis reveals the importance of residues alphaR145 and alphaF146 of penicillin acylase in the synthesis of beta-lactam antibiotics

J. Biotechnol.

133

18-26

2008

Escherichia coli

17933411

690107

Carboni, C.; Kierkels, H.G.; Gardossi, L.; Tamiola, K.; Janssen, D.B.; Quaedflieg, P.J.

Preparation of D-amino acids by enzymatic kinetic resolution using a mutant of penicillin-G acylase from E. coli

Tetrahedron Asymmetry

17

245-251

2006

Escherichia coli

-

695688

Illanes, A.; Wilson, L.; Aguirre, C.

Synthesis of cephalexin in aqueous medium with carrier-bound and carrier-free penicillin acylase biocatalysts

Appl. Biochem. Biotechnol.

157

98-110

2009

Escherichia coli

18618085

696160

Chilov, G.G.; Stroganov, O.V.; Svedas, V.K.

Molecular modeling studies of substrate binding by penicillin acylase

Biochemistry (Moscow)

73

56-64

2008

Escherichia coli (P06875)

18294130

696877

Bergeron, L.M.; Tokatlian, T.; Gomez, L.; Clark, D.S.

Redirecting the inactivation pathway of penicillin amidase and increasing amoxicillin production via a thermophilic molecular chaperone

Biotechnol. Bioeng.

102

417-424

2009

Escherichia coli

18846552

696880

Romero, O.; Vergara, J.; Fernandez-Lafuente, R.; Guisan, J.M.; Illanes, A.; Wilson, L.

Simple strategy of reactivation of a partially inactivated penicillin g acylase biocatalyst in organic solvent and its impact on the synthesis of beta-lactam antibiotics

Biotechnol. Bioeng.

103

472-479

2009

Escherichia coli

19253933

696890

Bernardino, S.M.; Fernandes, P.; Fonseca, L.P.

A new biocatalyst: Penicillin G acylase immobilized in sol-gel micro-particles with magnetic properties

Biotechnol. J.

4

695-702

2009

Escherichia coli

19418472

699157

Massolini, G.; Temporini, C.; Calleri, E.

Penicillin G acylase as chiral selector in LC and CE: exploring the origins of enantioselectivity

J. Chromatogr. B Analyt. Technol. Biomed. Life Sci.

875

20-29

2008

Rhizobium viscosum, Priestia megaterium, Escherichia coli, Kluyvera cryocrescens

18579456

710789

Kafshnochi, M.; Farajnia, S.; Aboshof, R.; Babaei, H.; Aminolroayaee, M.

Cloning and over-expression of penicillin G acylase in Escherichia coli BL21

Afr. J. Biotechnol.

9

2697-2701

2010

Escherichia coli (D5M8S0)

-

710906

Adikane, H.V.; Thakar, D.M.

Studies of penicillin G acylase immobilization using highly porous cellulose-based polymeric membrane

Appl. Biochem. Biotechnol.

160

1130-1145

2010

Escherichia coli, Escherichia coli NCIM 2400

19484409

711120

Pinotti, L.; Fonseca, L.; Prazeres, D.; Rodrigues, D.; Nucci, E.; Giordano, R.

Recovery and partial purification of penicillin G acylase from E. coli homogenate and B. megaterium culture medium using an expanded bed adsorption column

Biochem. Eng. J.

44

111-118

2009

Priestia megaterium, Escherichia coli, Priestia megaterium ATCC 14945, Escherichia coli ATCC 9637

-

712528

Krzeslak, J.; Braun, P.; Voulhoux, R.; Cool, R.H.; Quax, W.J.

Heterologous production of Escherichia coli penicillin G acylase in Pseudomonas aeruginosa

J. Biotechnol.

142

250-258

2009

Escherichia coli

19481123

Shi, Y.F.; Soumillion, P.; Ueda, M.

Effects of catalytic site mutations on active expression of phage fused penicillin acylase

J. Biotechnol.

145

139-142

2010

Escherichia coli

19932137

712778

Pchelintsev, N.; Youshko, M.; Švedas, V.

Quantitative characteristic of the catalytic properties and microstructure of cross-linked enzyme aggregates of penicillin acylase

J. Mol. Catal. B

56

202-207

2009

Escherichia coli

-

713438

Wilson, L.; Illanes, A.; Soler, L.; Henri�quez, M.

Effect of the degree of cross-linking on the properties of different CLEAs of penicillin acylase

Process Biochem.

44

322-326

2009

Escherichia coli

-

Cecchini, D.A.; Pavesi, R.; Sanna, S.; Daly, S.; Xaiz, R.; Pregnolato, M.; Terreni, M.

Efficient biocatalyst for large-scale synthesis of cephalosporins, obtained by combining immobilization and site-directed mutagenesis of penicillin acylase

Appl. Microbiol. Biotechnol.

95

1491-1500

2012

Escherichia coli (P06875), Escherichia coli

22228258

719296

Blum, J.K.; Deaguero, A.L.; Perez, C.V.; Bommarius, A.S.

Ampicillin synthesis using a two-enzyme cascade with both alpha-amino ester hydrolase and penicillin G acylase

ChemCatChem

2

987-991

2010

Escherichia coli

22039394

Miranda, V.; Wilson, L.; Cardenas, C.; Illanes, A.

Reactivation of immobilized penicillin G acylase: Influence of cosolvents and catalytic modulators

J. Mol. Catal. B

68

77-82

2011

Escherichia coli

-

Deaguero, A.; Blum, J.; Bommarius, A.

Improving the diastereoselectivity of penicillin G acylase for ampicillin synthesis from racemic substrates

Protein Eng. Des. Sel.

25

135-144

2012

Escherichia coli (P06875)

22271751

Orr, V.; Scharer, J.; Moo-Young, M.; Honeyman, C.; Fenner, D.; Crossley, L.; Suen, S.; Chou, C.

Integrated development of an effective bioprocess for extracellular production of penicillin G acylase in Escherichia coli and its subsequent one-step purification

J. Biotechnol.

161

19-26

2012

Escherichia coli, Escherichia coli JE5505

22728392

734463

Moenster, A.; Villain, L.; Scheper, T.; Beutel, S.

One-step-purification of penicillin G amidase from cell lysate using ion-exchange membrane adsorbers

J. Membr. Sci.

444

359-364

2013

Escherichia coli

-

Adediran, S.A.; Pratt, R.F.

Penicillin acylase and O-aryloxycarbonyl hydroxamates Two acyl-enzymes, one leading to hydrolysis, the other to inactivation

Arch. Biochem. Biophys.

614

65-71

2017

Escherichia coli

28038956

753335

Avinash, V.; Pundle, A.; Ramasamy, S.; Suresh, C.

Penicillin acylases revisited Importance beyond their industrial utility

Crit. Rev. Biotechnol.

36

303-316

2016

Achromobacter sp., Alcaligenes faecalis, Achromobacter xylosoxidans, Bacillus badius, Bacillus subtilis, Lysinibacillus sphaericus, Fusarium oxysporum, Providencia rettgeri, Escherichia coli (P06875), Kluyvera cryocrescens (P07941), Rhizobium viscosum (P31956), Priestia megaterium (Q60136)

25430891

Grulich, M.; Brezovsky, J.; Stepanek, V.; Palyzova, A.; Kyslikova, E.; Kyslik, P.

Resolution of alpha/beta-amino acids by enantioselective penicillin G acylase from Achromobacter sp.

J. Mol. Catal. B

122

240-247

2015

Escherichia coli (P06875), Achromobacter sp. CCM 4824 (Q3ZEF0)

-

754707

Ayakar, S.; Yadav, G.

Development of novel support for penicillin acylase and its application in 6-aminopenicillanic acid production

Mol. Catal.

476

110484

2019

Escherichia coli (P06875)

-