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1.21.3.1: isopenicillin-N synthase

This is an abbreviated version!
For detailed information about isopenicillin-N synthase, go to the full flat file.

Word Map on EC 1.21.3.1

Reaction

N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine
+
O2
=
isopenicillin N
 + 2 H2O

Synonyms

IPN cyclase, IPN synthase, IPNS, isopenicillin N synthase, isopenicillin N synthase (cyclase), isopenicillin N synthetase, isopenicillin N-synthase, isopenicillin-N synthase, isopenicillin-N-synthase, More, PcbC, synthetase, isopenicillin N (9Cl)

ECTree

     1 Oxidoreductases
         1.21 Catalysing the reaction X-H + Y-H = X-Y
             1.21.3 With oxygen as acceptor
                1.21.3.1 isopenicillin-N synthase

Engineering

Engineering on EC 1.21.3.1 - isopenicillin-N synthase

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PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C106S
-
site-directed mutagenesis, 63% reduced activity, 14fold increased Km for N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine
C106S/C255S
-
site-directed mutagenesis, 63% reduced activity, 14fold increased Km for N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine
C255S
-
site-directed mutagenesis, 33% reduced activity, 1.4fold increased Km for N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine
H116L
-
site-directed mutagenesis, reduced activity
H126L
-
site-directed mutagenesis, reduced activity
H137L
-
site-directed mutagenesis, reduced activity
H262L
-
site-directed mutagenesis, complete loss of activity
H272L
-
site-directed mutagenesis, complete loss of activity
H49L
-
site-directed mutagenesis, complete loss of activity
H64L
-
site-directed mutagenesis, reduced activity
P285L
-
site-directed mutagenesis, complete loss of activity, increased soluble expression in Escherichia coli host
Q227L
-
site-directed mutagenesis, 55.10% remaining activity compared to wild-type
Q337L
-
site-directed mutagenesis, slightly reduced activity compared to wild-type
Q230L
-
mutant of IPNS shows diminished enzyme activity, residue is highly conserved among dioxygenases and proximal to the active site, is the fourth ligand for the Fe2+ atom
D216E
-
mutant, retains 1% activity
K98E
the mutant shows about 8% of wild type activity
L223A
-
reduced activity with N-[(5S)-5-amino-5-carboxypentanoyl]-L-cysteinyl-D-valine, delta-(L-alpha-aminoadipoyl)-L-cysteinyl-D-alpha-aminobutyrate is a poor substrate
L223I
L223V
L231I
the mutant shows about 35% of wild type activity
L231V
the mutant shows about 25% of wild type activity
P283A
the mutant shows about 12% of wild type activity
P283I
the mutant shows about 10% of wild type activity
P283V
the mutant shows about 9% of wild type activity
V272T
the mutant shows about 27% of wild type activity
L223I
-
the mutant shows about 33% of wild type activity
-
L223V
-
the mutant shows about 33% of wild type activity
-
L231I
-
the mutant shows about 35% of wild type activity
-
L231V
-
the mutant shows about 25% of wild type activity
-
P283L
-
inactive
-
C104S
-
single-strand-site-directed mutagenesis, loss of more than 96% activity
C142S
-
single-strand-site-directed mutagenesis, loss of 24% activity
C251S
-
single-strand-site-directed mutagenesis, loss of 47.7% activity
C37S
-
single-strand-site-directed mutagenesis, loss of 18.3% activity
C37S/C142S/C251S
-
triple mutant, conformationally different from wild-type, prepared by recombining fragments of IPNS-encoding gene pcbC from each of the single mutants, loss of more than 90% activity
D214C
-
site-directed mutagenesis, active site mutant, complete loss of activity
D214E
-
site-directed mutagenesis, active site mutant, retains 1% of activity compared to wild-type
D214H
H212D
-
site-directed mutagenesis, active site mutant, complete loss of activity
H212N
-
site-directed mutagenesis, active site mutant, complete loss of activity
H212Q
-
site-directed mutagenesis, active site mutant, complete loss of activity
H268D
-
site-directed mutagenesis, active site mutant, complete loss of activity
H268N
-
site-directed mutagenesis, active site mutant, complete loss of activity
H268Q
-
site-directed mutagenesis, active site mutant, complete loss of activity
additional information