Information on EC 3.5.1.70 - aculeacin-A deacylase

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The expected taxonomic range for this enzyme is: Actinoplanes utahensis

EC NUMBER
COMMENTARY hide
3.5.1.70
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RECOMMENDED NAME
GeneOntology No.
aculeacin-A deacylase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
hydrolysis of the amide bond in aculeacin A and related neutral lipopeptide antibiotics, releasing the long-chain fatty acid side-chain
show the reaction diagram
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of linear amides
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SYSTEMATIC NAME
IUBMB Comments
aculeacin-A amidohydrolase
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CAS REGISTRY NUMBER
COMMENTARY hide
121479-50-3
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ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
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enzyme hydrolyzes the acyl moieties of antifungal echinocandin antibiotics, efficiently hydrolyzes penicillin V and natural aliphatic penicillins to yield 6-aminopenicillanic acid
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
aculeacin A + H2O
? + palmitic acid
show the reaction diagram
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-
-
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?
aculeacin A + H2O
cyclo-hexapeptide + palmitic acid
show the reaction diagram
echinocandin A + H2O
cyclo-hexapeptide + linoleic acid
show the reaction diagram
echinocandin C + H2O
cyclo-hexapeptide + linoleic acid
show the reaction diagram
echinocandin D + H2O
cyclo-hexapeptide + linoleic acid
show the reaction diagram
penicillin dihydroF + H2O
6-aminopenicillanic acid + hexanoic acid
show the reaction diagram
-
-
-
-
?
penicillin dihydroF + H2O
?
show the reaction diagram
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-
-
-
?
penicillin F + H2O
6-aminopenicillanic acid + (3E)-hex-3-enoic acid
show the reaction diagram
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-
-
-
?
penicillin F + H2O
?
show the reaction diagram
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-
-
-
?
penicillin G + H2O
6-aminopenicillanic acid + phenylacetic acid
show the reaction diagram
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-
-
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?
penicillin K + H2O
6-aminopenicillanic acid + octanoic acid
show the reaction diagram
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-
-
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?
penicillin K + H2O
?
show the reaction diagram
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-
-
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?
penicillin V + H2O
6-aminopenicillanic acid + phenoxyacetic acid
show the reaction diagram
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-
-
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?
additional information
?
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NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
aculeacin A + H2O
? + palmitic acid
show the reaction diagram
-
-
-
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
KCl
-
1 M: 1.47fold activation
NaCl
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1 M: 1.69fold activation
additional information
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no significant effect, no activation, by Ca2+, Co2+, Cu2+, Mg2+, Mn2+ or Zn2+
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
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no inhibition by EDTA, up to 10 mM, no inhibition by Ca2+, Co2+, Cu2+, Mg2+, Mn2+ or Zn2+
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.53 - 6.3
aculeacin A
3.9 - 5.6
penicillin dihydroF
9.5 - 15.1
penicillin F
155.8
penicillin G
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pH 8.0, 45°C
0.45 - 1
penicillin K
6.45 - 15.4
Penicillin V
additional information
additional information
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kinetic data
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TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
4.2
penicillin dihydroF
Actinoplanes utahensis
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pH 8.0, 45°C
1.9
penicillin F
Actinoplanes utahensis
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pH 8.0, 45°C
2.2
penicillin G
Actinoplanes utahensis
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pH 8.0, 45°C
33.3
penicillin K
Actinoplanes utahensis
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pH 8.0, 45°C
70.3
Penicillin V
Actinoplanes utahensis
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pH 8.0, 45°C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
9.5 - 11.5
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from recombinant Streptomyces strains
11.1
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from recombinant strain Streptomyces lividans JT46
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8 - 8.5
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substrate: penicillin V
8.5
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recombinant immobilized enzyme, highest activity with penicillin V, above the activity decreases very quickly as a result of the denaturation of the enzyme, 40°C
additional information
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pI: above 10.25
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5 - 11
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ca. 17 U/g wet carrier at pH 6.5 and ca. 20 U/g wet carrier at pH 11
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
19000
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1 * 55000 + 1 * 19000, SDS-PAGE or gel filtration in the presence of 6 M guanidine hydrochloride, two dissimilar subunits
19100
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1 * 60300 + 1 * 19100, two dissimilar subunits, calculated from nucleotide sequence
38000
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HPLC gel filtration, the large discrepancy between the molecular weight estimated by HPLC and that estimated by SDS-PAGE is due to the particularly high pI of the enzyme: the molecular weight of acylase should be 74000 Da, the sum of the values of the two subunits obtained by SDS-PAGE
55000
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1 * 55000 + 1 * 19000, SDS-PAGE or gel filtration in the presence of 6 M guanidine hydrochloride, two dissimilar subunits
60300
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1 * 60300 + 1 * 19100, two dissimilar subunits, calculated from nucleotide sequence
84070
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precursor peptide of aculeacin A acylase, calculated from nucleotide sequence
87000
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precursor peptide of aculeacin A acylase, gel filtration and SDS-PAGE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
proteolytic modification
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
2
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4 h, 30°C: inactivation
209226
4 - 8
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4 h, 30°C: stable at
209226
6.5 - 11
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the immobilized enzyme is stable at the whole pH range from pH 6.5 to 11. Stability is tested for the storage of the biocatalyst for two hours at different pH values at 4°C
711493
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30
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4 h, inactivation at pH 2, stable at pH 4-8
45
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both soluble and immobilized enzyme apparently suffers no loss of enzymatic activity when stored at 45°C for at least 550 min
50
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20 min, stable up to
60
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24 h, 0.1 M phosphate buffer, pH 7, stable at
65
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the recombinant immobilized enzyme is highly thermostable since it suffers no loss of activity at 65°C and pH 8.0 during 3 h
70
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above, 0.1 M phosphate buffer, pH 7, rapid loss of activity
75
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the recombinant immobilized enzyme is completely deactivated after its incubation during two hours at temperatures above 75°C
82
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aculeacin A acylase is highly thermostable, with a midpoint transition temperature of 81.5°C
additional information
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half-life of the soluble enzyme at 65°C is estimate, t1/2: 477 min
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
the recombinant immobilized enzyme can be recycled for at least 30 consecutive batch reactions at pH 8.0 and 45°C without loss of catalytic activity. Both soluble and immobilized enzyme apparently suffers no loss of enzymatic activity when stored at 45°C for at least 550 min
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
enzyme precursor peptide
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from recombinant strain Streptomyces lividans JT46
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from recombinant strains: Streptomyces lividans JT46, Streptomyces avermitilis K139, Streptomyces albus J1074, Streptomyces ambofaciens ATCC151540, Streptomyces parvulus 2283 and Streptomyces griseus ATCC23345
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recombinant enzyme
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression in Streptomyces lividans
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expression in Streptomyces lividans JT46
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overexpression and overproduction in Streptomyces lividans JT46, Streptomyces albus J1074, Streptomyces ambofaciens ATCC151540, Streptomyces parvulus 2283, Streptomyces avermitilis K139 and Streptomyces griseus ATCC23345, using multi-copy vector pIJ702
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recombinant enzyme covalently immobilized onto several epoxy-activated supports in order to obtain a robust biocatalyst to be used in industrial bioreactors. The best biocatalyst is obtained by attaching the enzyme on Sepabeads EC-EP5
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
biotechnology
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recombinant enzyme covalently immobilized onto several epoxy-activated supports in order to obtain a robust biocatalyst to be used in industrial bioreactors. The best biocatalyst is obtained by attaching the enzyme on Sepabeads EC-EP5
pharmacology
synthesis