1.14.11.26: deacetoxycephalosporin-C hydroxylase
This is an abbreviated version!
For detailed information about deacetoxycephalosporin-C hydroxylase, go to the full flat file.
Word Map on EC 1.14.11.26
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1.14.11.26
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clavuligerus
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penicillin
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dithiothreitol
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ascorbate
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acremonium
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alpha-ketoglutarate
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chrysogenum
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ring-expansion
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n-ethylmaleimide
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expandase
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cephalosporium
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synthesis
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biotechnology
- 1.14.11.26
- clavuligerus
- penicillin
- dithiothreitol
- ascorbate
- acremonium
- alpha-ketoglutarate
- chrysogenum
-
ring-expansion
- n-ethylmaleimide
-
expandase
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cephalosporium
- synthesis
- biotechnology
Reaction
Synonyms
cefF, Cephalosporin biosynthesis expandase/hydroxylase, DACS, DAOC hydroxylase, DAOC synthase, DAOC/DAC synthase, DAOCS, deacetoxycephalosporin C hydroxylase, deacetoxycephalosporin C synthase, deacetylcephalosporin C synthase
ECTree
Advanced search results
Engineering
Engineering on EC 1.14.11.26 - deacetoxycephalosporin-C hydroxylase
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M306I
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hydroxylation reaction of deacetoxycephalosporin C, EC 1.14.11.26, is abolished, 59% of wild-type ring expansion activity
N305L
R308L
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improved ability to convert penicillin analogs in ring expansion reaction of EC 1.14.20.1
W82A
W82S
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44% of wild-type ring expansion activity, 18% of wild-type hydroxylation activity
A106T
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80% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
A177V
random mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
C155Y
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90% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
C155Y/Y184H/V275I/C281Y
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580% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
C281Y
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200% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
E16G/T90A/T304A
E209Q
random mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
E82D
random mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
F267L
random mutagenesis, active site mutation, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
G300V
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410% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
G79E
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90% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
H244Q
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140% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
I193V
random mutagenesis, active site mutation, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
I305L
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230% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
I305M
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380% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
L236V
random mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
L277Q
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270% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
M184I
random mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
M188I
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90% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
M188V
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150% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
M73T
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180% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
N304K
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220% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
P186L
random mutagenesis and site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
P72L
random mutagenesis and site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
R182S
random mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
R182W
random mutagenesis, active site mutation, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
S251F
random mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
S260G
random mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
T90A/A311V
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
T90A/P72L
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
T90A/P72L/A311V
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
T90A/P72L/A311V/A40V/M229I/T273A
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
T90A/P72L/A311V/A40V/M229I/T273A/M184I/I193V/F267L
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
T90A/P72L/A311V/A40V/M229I/T273A/R182S
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
T90A/P72L/A311V/A40V/M229I/T273A/S251F
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
T90A/P72L/A311V/A40V/M229I/T273A/S260G
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
T90A/P72L/A311V/A40V/M229I/T273A/V171L/F267L
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L
site-directed mutagenesis, the mutant shows highly increased activity with cephalosporin G compared to the wild-type enzyme
T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L/A241V/V307A
site-directed mutagenesis, the mutant shows highly increased activity with cephalosporin G compared to the wild-type enzyme
T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L/G108D
site-directed mutagenesis, the mutant shows highly increased activity with cephalosporin G compared to the wild-type enzyme
T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L/N313D
site-directed mutagenesis, the mutant shows highly increased activity with cephalosporin G compared to the wild-type enzyme
T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L/R91G
site-directed mutagenesis, the mutant shows highly increased activity with cephalosporin G compared to the wild-type enzyme
T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L/R91G/A241V/V307A
site-directed mutagenesis, the mutant shows highly increased activity with cephalosporin G compared to the wild-type enzyme
T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L/T96S/A241V/V307A
site-directed mutagenesis, the mutant shows highly increased activity with cephalosporin G compared to the wild-type enzyme
T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L/T96S/G255D/A280S
site-directed mutagenesis, the mutant shows highly increased activity with cephalosporin G compared to the wild-type enzyme
T90A/P72L/A311V/A40V/M229I/T273A/V171L/R182W/F267L/V226I
site-directed mutagenesis, the mutant shows highly increased activity with cephalosporin G compared to the wild-type enzyme
T90A/P72L/A311V/A40V/M229I/T273A/V221P
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
T90A/P72L/A311V/F195L
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
T90A/P72L/A311V/P7L/A237V
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
T90A/P72L/A311V/P7L/T273A
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
T90A/P72L/A311V/R250L
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
T90A/P72L/A311V/V206I/A210V
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
T90A/P72L/A311V/V206I/A210V/T273A
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
T91A
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110% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
V171L
random mutagenesis, active site mutation, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
V171M
random mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
V221H
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
V221P
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
V221T
random mutagenesis and site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
V249I
random mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
V275I
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270% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
V275I/I305M
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500% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
Y184H
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200% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate
A177V
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random mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
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E82D
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random mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
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additional information
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107% of wild-type ring expansion activity, 85% of wild-type hydroxylation activity
N305L
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improved ability to convert penicillin analogs in ring expansion reaction of EC 1.14.20.1
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5.5% of wild-type ring expansion activity, 71% of wild-type hydroxylation activity
random mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
E16G/T90A/T304A
site-directed mutagenesis, the mutant shows increased activity with cephalosporin G compared to the wild-type enzyme
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truncation of C-terminus to residue 310, 2fold enhancement of ring expansion reaction of penicillin G. Double mutant with truncation at residue 310 and M306I, selective catalyzation of ring expansion. Triple mutant with truncation at residue 310, M306I and N305L, selective catalization of ring expansion with improved kinetic parameters
additional information
mutagenic cefF library creation by random mutagenesis and screening of the mutant deacetylcephalosporin C synthase enzyme library, homology modeling of DACS structure and mapping of mutant positions. Process-level biotransformation reaction of cephalosporin G to deacetylcephalosporin G by mutants of deacetylcephalosporin C synthase. Deacetylcephalosporin G can be converted completely into hydroxymethyl-7-amino-cephalosporanic acid (HACA) in about 30 min by a subsequent reaction, thus facilitating scalability toward commercialization. Directed-evolution strategies such as random, semirational, rational, and computational methods are used for systematic engineering of DACS for improved activity with cephalosporin G
additional information
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mutagenic cefF library creation by random mutagenesis and screening of the mutant deacetylcephalosporin C synthase enzyme library, homology modeling of DACS structure and mapping of mutant positions. Process-level biotransformation reaction of cephalosporin G to deacetylcephalosporin G by mutants of deacetylcephalosporin C synthase. Deacetylcephalosporin G can be converted completely into hydroxymethyl-7-amino-cephalosporanic acid (HACA) in about 30 min by a subsequent reaction, thus facilitating scalability toward commercialization. Directed-evolution strategies such as random, semirational, rational, and computational methods are used for systematic engineering of DACS for improved activity with cephalosporin G
additional information
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mutagenic cefF library creation by random mutagenesis and screening of the mutant deacetylcephalosporin C synthase enzyme library, homology modeling of DACS structure and mapping of mutant positions. Process-level biotransformation reaction of cephalosporin G to deacetylcephalosporin G by mutants of deacetylcephalosporin C synthase. Deacetylcephalosporin G can be converted completely into hydroxymethyl-7-amino-cephalosporanic acid (HACA) in about 30 min by a subsequent reaction, thus facilitating scalability toward commercialization. Directed-evolution strategies such as random, semirational, rational, and computational methods are used for systematic engineering of DACS for improved activity with cephalosporin G
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