3.5.1.11: penicillin amidase
This is an abbreviated version!
For detailed information about penicillin amidase, go to the full flat file.
Word Map on EC 3.5.1.11
-
3.5.1.11
-
6-aminopenicillanic
-
phenylacetic
-
biocatalyst
-
anandamide
-
acylases
-
cannabinoids
-
cephalosporin
-
synthesis
-
binuclear
-
alcaligenes
-
cephalexin
-
megaterium
-
endocannabinoids
-
semi-synthetic
-
3.5.1.1
-
kluyvera
-
l-asparagine
-
dihydroorotase
-
phenylacetylated
-
allantoinase
-
dihydropyrimidinase
-
lactonase
-
amoxicillin
-
rettgeri
-
eupergit
-
multipoint
-
acyl-enzyme
-
asparaginase
-
hydantoinase
-
phenoxyacetic
-
aminoacylase
-
7-aminocephalosporanic
-
phosphotriesterase
-
sphaericus
-
hydantoin
-
providencia
-
n-acylethanolamines
-
penicillanic
-
dihydrouracil
-
cleas
-
pharmacology
-
industry
- 3.5.1.11
-
6-aminopenicillanic
-
phenylacetic
-
biocatalyst
- anandamide
- acylases
- cannabinoids
- cephalosporin
- synthesis
-
binuclear
- alcaligenes
- cephalexin
- megaterium
-
endocannabinoids
-
semi-synthetic
-
3.5.1.1
-
kluyvera
- l-asparagine
- dihydroorotase
-
phenylacetylated
- allantoinase
- dihydropyrimidinase
- lactonase
- amoxicillin
- rettgeri
-
eupergit
-
multipoint
- acyl-enzyme
- asparaginase
- hydantoinase
-
phenoxyacetic
-
aminoacylase
-
7-aminocephalosporanic
- phosphotriesterase
- sphaericus
- hydantoin
- providencia
- n-acylethanolamines
-
penicillanic
- dihydrouracil
-
cleas
- pharmacology
- industry
Reaction
Synonyms
ACPGA001 PGA, AfPGA, alpha-acylamino-beta-lactam acylhydrolase, amidase, amidohydrolase, ampicillin acylase, AuAAC, benzylpenicillin acylase, BmPGA, Eca3205, KcPGA, maPGA, More, novozym 217, PA, PAC, penicillin acylase, penicillin amidase, Penicillin amidohydrolase, penicillin G acylase, Penicillin G amidase, Penicillin G amidohydrolase, penicillin V acylase, Penicillin V amidase, penicillin-G acylase, PGA, PGA650, PVA, semacylase, Sm-PVA, YxeI
ECTree
3.5.1.11 penicillin amidaseAdvanced search results
results (
results (Engineering
Engineering on EC 3.5.1.11 - penicillin amidase
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
top
PROTEIN VARIANTS 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
Q3C/P751C
-
temperature optimum is three degrees higher than for wild-type. Half-life of mutant at 55°C is increased by 50%
T206G
-
specific activity of the mutant enzyme is 60% higher compared to the wild-type enzyme, pI is 5.5
T206G/G213G
-
mutant enzyme with 1.9fold increased specific activity compared to completely processed wild-type enzyme, mutation stabilizes the precursor form, pI value is 5.6
T206G/S213G
-
construction of a mutant with extended C-terminus of the A-chain comprising parts of the connecting linker peptide showing almost 2fold increased activity and 3fold higher specificity compared to the wild-type enzyme, overview
T206G/S213G/T219G
-
mutant enzyme with 2.3fold increased specific activity compared to completely processed wild-type enzyme
T206P
-
mutant undergoes normal proteolytic processing leading to a completely processed enzyme with pI 5.3
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
alphaF146Y
betaF24A
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
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
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
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
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
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
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
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
S1C
replacement of serine of the beta-subunit with cysteine results in a fully processed but inactive enzyme
S1G
the second mutant in which the N-terminal serine is replaced by glycine remains in the unprocessed and inactive form
S290G
C1A
-
no catalytic activity, precursor form pre-C1S is not processed
C1S
-
no catalytic activity, precursor form pre-C1S is not processed
N175A
-
no catalytic activity, about 50% of precursor form pre-N175A is processed
N198Y
the mutant enzyme shows 1.36fold higher specific activity compared to wild-type enzyme
N198Y/S110C
the mutant enzyme shows 2.26fold higher specific activity compared to wild-type enzyme
T63S
the mutant enzyme shows 11.14fold higher specific activity compared to wild-type enzyme
T63S/N198Y
the mutant enzyme shows 4.6fold higher specific activity compared to wild-type enzyme
T63S/S110C
the mutant enzyme shows 3.97fold higher specific activity compared to wild-type enzyme
T63S/S110C/N198Y
the mutant enzyme shows 12.4fold higher specific activity and 11.3fold higher catalytic efficiency compared to wild-type enzyme. The mutant enzyme has a potential for large-scale industrial application for 6-aminopenicillanic acid production
D22E
the specific activity is 10.9% compared to wild-type enzyme. Protein expression yields is 34 mg/ml compared to 262 mg/ml for the native enzyme
D22N
the specific activity is 5.1% compared to wild-type enzyme. Protein expression yields is 2.5 mg/ml compared to 262 mg/ml for the native enzyme
R19H
the specific activity is 6% compared to wild-type enzyme. Protein expression yields is 121 mg/ml compared to 262 mg/ml for the native enzyme
R215L
the specific activity is 7.4% compared to wild-type enzyme. Protein expression yields is 5.6 mg/ml compared to 262 mg/ml for the native enzyme
W23F
the specific activity is 4% compared to wild-type enzyme. Protein expression yields is 6.9 mg/ml compared to 262 mg/ml for the native enzyme
W23F/W87Y
the specific activity is 0.13% compared to wild-type enzyme. Protein expression yields is 3.3 mg/ml compared to 262 mg/ml for the native enzyme
W23I
the specific activity is 0.9% compared to wild-type enzyme. Protein expression yields is 7.1 mg/ml compared to 262 mg/ml for the native enzyme
W23I/W87N
the specific activity is 0.02% compared to wild-type enzyme. Protein expression yields is 3.6 mg/ml compared to 262 mg/ml for the native enzyme
W87N
the specific activity is 81.2% compared to wild-type enzyme. Protein expression yields is 196.8 mg/ml compared to 262 mg/ml for the native enzyme
W87Y
the specific activity is 1.5% compared to wild-type enzyme. Protein expression yields is 6.7 mg/ml compared to 262 mg/ml for the native enzyme
R19H
-
the specific activity is 6% compared to wild-type enzyme. Protein expression yields is 121 mg/ml compared to 262 mg/ml for the native enzyme
-
R215L
-
the specific activity is 7.4% compared to wild-type enzyme. Protein expression yields is 5.6 mg/ml compared to 262 mg/ml for the native enzyme
-
W23F
-
the specific activity is 4% compared to wild-type enzyme. Protein expression yields is 6.9 mg/ml compared to 262 mg/ml for the native enzyme
-
W87Y
-
the specific activity is 1.5% compared to wild-type enzyme. Protein expression yields is 6.7 mg/ml compared to 262 mg/ml for the native enzyme
-
F145A
-
alpha chain, no formation of cephalexin, no activity with nitrophenylacetylaminobenzoic acid
F145L
-
alpha chain, no formation of cephalexin, no activity with nitrophenylacetylaminobenzoic acid
F145Y
-
alpha chain, slightly increased formation of cephalexin, 32% of nitrophenylacetylaminobenzoic acid hydrolyzing activity
K427A
K430A
K430A/K427A
-
mutation in beta-chain. Mutant shows a good stability to solvent and thermostability
V24F
-
beta chain, no effect on the formation of cephalexin, 11% of nitrophenylacetylaminobenzoic acid hydrolyzing activity
Y144R
-
alpha chain, reduced formation of cephalexin, 72% of nitrophenylacetylaminobenzoic acid hydrolyzing activity
Y144R/V24F
-
Y144R on alpha chain, V24F on beta chain, reduced formation of cephalexin, 4% of nitrophenylacetylaminobenzoic acid hydrolyzing activity
N272D
mutation at beta-subunit results in a drastic decrease of activity
N272V
mutation at beta-subunit results in a drastic decrease of activity
S1A
mutation at beta-subunit results in an inactive enzyme
S1C
mutation at beta-subunit results in an inactive enzyme
S1D
mutation at beta-subunit results in an inactive enzyme
S1H
mutation at beta-subunit results in an inactive enzyme
S1K
mutation at beta-subunit results in an inactive enzyme
V70A
mutation at beta-subunit results in a drastic decrease of activity
V70D
mutation at beta-subunit results in a drastic decrease of activity
N272D
Streptomyces lavendulae subsp. lavendulae ATCC 13664
-
mutation at beta-subunit results in a drastic decrease of activity
-
S1A
Streptomyces lavendulae subsp. lavendulae ATCC 13664
-
mutation at beta-subunit results in an inactive enzyme
-
S1C
Streptomyces lavendulae subsp. lavendulae ATCC 13664
-
mutation at beta-subunit results in an inactive enzyme
-
V70A
Streptomyces lavendulae subsp. lavendulae ATCC 13664
-
mutation at beta-subunit results in a drastic decrease of activity
-
V70D
Streptomyces lavendulae subsp. lavendulae ATCC 13664
-
mutation at beta-subunit results in a drastic decrease of activity
-
I57F
site-directed mutagenesis, the substrate specificity of the mutant shifts from penicillin K to penicillin G, Km for penicillin G is 4.5fold, while the catalytic efficiency remains at wild-type value
L188F
site-directed mutagenesis, the substrate specificity of the mutant shifts from penicillin K to penicillin G, penicillin G binding is 10fold improved compared to the wild-type enzyme
L188R
site-directed mutagenesis, the substrate specificity of the mutant shifts from penicillin K to penicillin G
L188R/S189F
site-directed mutagenesis, the substrate specificity of the mutant shifts from penicillin K to penicillin G
L24F
site-directed mutagenesis, the substrate specificity of the mutant shifts from penicillin K to penicillin G, penicillin G binding is 12fold improved compared to the wild-type enzyme
L24F/I57F
site-directed mutagenesis, the substrate specificity of the mutant shifts from penicillin K to penicillin G
L24F/I57F/L188F
site-directed mutagenesis, the substrate specificity of the mutant shifts from penicillin K to penicillin G
S189F
site-directed mutagenesis, the substrate specificity of the mutant shifts from penicillin K to penicillin G, penicillin G binding is 9fold improved compared to the wild-type enzyme
additional information
F146L
-
mutation in alpha-subunit,3fold increase in transferase/hydrolase ratio using 6-aminopenicillanic acid as nucleophile
F146L
-
mutation in alpha-subunit. 169% of ampicillin synthesis activity compared to wild-type
F146W
-
mutation in alpha-subunit, 40fold decrease in transferase/hydrolase ratio using 6-aminopenicillanic acid as nucleophile
F146W
-
mutation in alpha-subunit. 10% of ampicillin synthesis activity compared to wild-type
F146Y
-
mutation in alpha-subunit, 40fold decrease in transferase/hydrolase ratio using 6-aminopenicillanic acid as nucleophile
F24A
-
beta-subunit, increased affinity for Calpha-substituted substrates, 20fold reduced affinity for phenylacetic acid
F24A
-
mutation in beta-subunit, 3fold increase in transferase/hydrolase ratio using 6-aminopenicillanic acid as nucleophile
F24A
-
mutation in the beta-subunit produces a protein with a higher synthesis/hydrolysis ratio, increased acylase activity, and more resistance to inhibition by phenyl acetic acid
F24A
-
suppressed hydrolysis rate of N-phenylacetyl-L-Gln and N-phenylacetyl-L-Glu, respectively, compared to wild-type enzyme
V56R
-
mutation of beta-subunit, 12% of wild-type activity with substrate penicillin G, 230% of wild-type activity with cephalosporin acylase substrate glutaryl-7-aminocephalosporanic acid
V56R
-
mutation of beta-subunit, plus F146Y in alpha-subunit, 5% of wild-type activity with substrate penicillin G, 280% of wild-type activity with cephalosporin acylase substrate glutaryl-7-aminocephalosporanic acid
S290G
site-directed mutagenesis of the internal proteolytic cleavage site of the pro-enzyme
S290G
-
site-directed mutagenesis of the internal proteolytic cleavage site of the pro-enzyme
-
K427A
-
mutation in beta-chain. Half-life is improved by 60% compared to wild-type enzyme
K427A
-
mutation in beta-chain. Mutant shows a great stability to organic solvents
K430A
-
mutation in beta-chain. Half-life is improved by 166% compared to wild-type enzyme
K430A
-
mutation in beta-chain. Mutant shows a good stability to solvent and thermostability
additional information
-
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. In comparison with the two wild-type enzymes the hybrid enzyme has a higher turnover-number for benzylpenicillin, ampicillin and 6-nitro-3-phenylacetamido-benzoic acid. The KM-values are between the values of the wild-type enzymes or close ro that of K. citrophila
additional information
-
addition of a tag of three lysine residues alternating with three glycines to C-terminus of enzyme results in improvement of the immobilization efficiency on glyoxyl agarose and the catalytic protperties of immobilized enzyme, but impairs posttranslational steps of overexpressed protein maturation
additional information
-
conjugation of Saccharomyces cerevisiae mannan with enzyme yielding neoglycoproteins containing 42 to 67% saccharides. Significant increase in half-life at 37°C and 50°C of both free enzyme and concanavalin A-linked enzyme
additional information
construction hybrids of the penicillin acylase-encoding genes from Eacherichia coli and Kluyvera cryocrescens, with additional point mutations. The hybrid enzymes display a 40-90% increase in the relative rate of acyl transfer to the beta-lactam nucleus during ampicillin synthesis. This increase is not accompanied by a reduction of synthetic activity. Similar improvements in acyl transfer are obtained for the synthesis of amoxicillin, cephalexin and cefadroxil making the new hybrid enzymes interesting candidates for the biocatalytic synthesis of several beta-lactam antibiotics
additional information
-
the mutant enzyme contains 85 exposed Glu-plus-Asp residues, while the native enzyme exposed in the surface only 77 acidic residues. Such an increase in the number of negative charges reduces the isoelectric point of the mutant enzyme from 6.4 to 4.3. 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, suggesting that these conditions allow the penetration of the enzyme inside the polymeric beds, thus becoming fully covered with the polymer. After the enzyme inactivation, it can be desorbed to reuse the support
additional information
-
construction of a fusion protein, consisting of Pseudomonas Sec- or Tat-specific signal peptides, the elastase propeptide and the mature penicillin G acylase, termed a TatProPGA hybrid. 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
additional information
-
directed evolution of penicillin acylase using phage display technology for extending its specificity. Fusion of the penicillin acylase to fd phage coat protein III and used pIII secretion signal sequence instead of penicillin acylase, which couples gene and enzyme on phage particle and can is useful for directed evolution of penicillin acylase. Penicillin acylase is functionally displayed on phage surface. Determination by site-directed mutagenesis of the effect of Ser B1 and Asn B241 variants on post-translational maturation of phage fused penicillin acylase, overview
additional information
-
effect of the degree of cross-linking on the properties of different cross-linked enzyme aggregates, CLEAs, of penicillin acylase, overview
additional information
-
immobilization of the enzyme, quantitative characteristic of the catalytic properties and microstructure of cross-linked enzyme aggregates of penicillin acylase under different conditions by confocal fluorescent microscopy, overview. Comparison of fresh aggregates and mature aggregates. The aggregate size might regulate the extent of covalent modification of the enzyme and thereby influence the catalytic properties of cross-linked enzyme aggregates
additional information
-
penicillin G acylase immobilization using highly porous cellulose-based polymeric membrane, immobilized enzyme specific activity is 145 U/g, and a 2.4fold increase in activity compared to the free enzyme. The immobilized enzyme retains almost 50% activity after 107 days and 50 cycles of operation, method evaluation and enzyme stability, overview. Effect of different ionic molecules/compounds as ligands coupled to membrane on enzyme immobilization, best is Brilliant green
additional information
-
penicillin G acylase immobilization using highly porous cellulose-based polymeric membrane, immobilized enzyme specific activity is 145 U/g, and a 2.4fold increase in activity compared to the free enzyme. The immobilized enzyme retains almost 50% activity after 107 days and 50 cycles of operation, method evaluation and enzyme stability, overview. Effect of different ionic molecules/compounds as ligands coupled to membrane on enzyme immobilization, best is Brilliant green
-
additional information
-
a cross-species penicillin G amidase gene coding for an alpha-peptide and a linker peptide from Kluyvera citrophila and a beta-peptide from Escherichia coli constructed and cloned in Escherichia coli. In comparison with the two wild-type enzymes the hybrid enzyme has a higher turnover-number for benzylpenicillin, ampicillin and 6-nitro-3-phenylacetamido-benzoic acid. The Km-values are between the values of the wild-type enzymes or close ro that of Kluyvera citrophila
additional information
-
transversion mutation of thymine to guanine at position 1163 results in 90% and 50% of activity loss in penicillin G acylase activity on penicillin G and phenylacetyl-L-leucine respectively
additional information
-
leader peptide was replaced by the Bacillus megaterium LipA counterpart, changing the signal peptide increases the amount of secreted enzyme
additional information
-
immobilization of the enzyme on mesocellular silica foams from 0.1 M phosphate buffer, pH 7.0, the immobilized enzyme activity depends on molecular crowding and applied to catalysis in non-aqueous medium, addition of dextran, method optimization, using different enzyme preparations, organic solvents and mesocellular silica foams-dextran mixtures, overview
additional information
-
immobilization of the recombinant enzyme by covalent linking to epoxy-activated beads in homogenous distribution to avoid enzyme aggregate formation, and usage as biocatalyst. The immobilized enzyme shows altered optimal reaction conditions and enlarged substrate specificity, as well as increased thermal stability compared to the soluble enzyme, it can be recycled 30times in consecutive batch reactions, overview
additional information
substrate specificity change through engineering: generation of a set of enzyme variants containing between one and four amino acid replacements, with altered catalytic properties in the hydrolyses of penicillins K and G. The introduction of a single phenylalanine residue in position alpha188, alpha189, or beta24 improves the Km for penicillin G between 9 and 12fold, and the catalytic efficiency of these variants for penicillin G was improved up to 6.6fold
additional information
-
substrate specificity change through engineering: generation of a set of enzyme variants containing between one and four amino acid replacements, with altered catalytic properties in the hydrolyses of penicillins K and G. The introduction of a single phenylalanine residue in position alpha188, alpha189, or beta24 improves the Km for penicillin G between 9 and 12fold, and the catalytic efficiency of these variants for penicillin G was improved up to 6.6fold
