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Literature summary for 1.14.11.26 extracted from

  • Wei, C.; Yang, Y.; Deng, C.; Liu, W.; Hsu, J.; Lin, Y.; Liaw, S.; Tsai, Y.
    Directed evolution of Streptomyces clavuligerus deacetoxycephalosporin C synthase for enhancement of penicillin G expansion (2005), Appl. Environ. Microbiol., 71, 8873-8880.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli Tuner(DE3) and BL21(DE3) cells Streptomyces clavuligerus

Protein Variants

Protein Variants Comment Organism
A106T 80% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate Streptomyces clavuligerus
C155Y 90% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate Streptomyces clavuligerus
C155Y/Y184H/V275I/C281Y 580% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate Streptomyces clavuligerus
C281Y 200% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate Streptomyces clavuligerus
G300V 410% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate Streptomyces clavuligerus
G79E 90% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate Streptomyces clavuligerus
H244Q 140% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate Streptomyces clavuligerus
I305L 230% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate Streptomyces clavuligerus
I305M 380% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate Streptomyces clavuligerus
L277Q 270% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate Streptomyces clavuligerus
M188I 90% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate Streptomyces clavuligerus
M188V 150% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate Streptomyces clavuligerus
M73T 180% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate Streptomyces clavuligerus
N304K 220% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate Streptomyces clavuligerus
T91A 110% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate Streptomyces clavuligerus
V275I 270% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate Streptomyces clavuligerus
V275I/I305M 500% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate Streptomyces clavuligerus
Y184H 200% relative activity compared to the wild type enzyme using 1 mM penicillin G as substrate Streptomyces clavuligerus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.004
-
Penicillin N mutant enzyme N304K Streptomyces clavuligerus
0.006
-
Penicillin N mutant enzyme C281Y Streptomyces clavuligerus
0.006
-
Penicillin N mutant enzyme I305L Streptomyces clavuligerus
0.006
-
Penicillin N mutant enzyme M73T Streptomyces clavuligerus
0.009
-
Penicillin N mutant enzyme G79E Streptomyces clavuligerus
0.009
-
Penicillin N mutant enzyme H244Q Streptomyces clavuligerus
0.009
-
Penicillin N mutant enzyme T91A Streptomyces clavuligerus
0.011
-
Penicillin N mutant enzyme L277Q Streptomyces clavuligerus
0.011
-
Penicillin N mutant enzyme M188I Streptomyces clavuligerus
0.012
-
Penicillin N mutant enzyme I305M Streptomyces clavuligerus
0.012
-
Penicillin N mutant enzyme V275I Streptomyces clavuligerus
0.013
-
Penicillin N mutant enzyme A106T Streptomyces clavuligerus
0.013
-
Penicillin N mutant enzyme V275I/I305M Streptomyces clavuligerus
0.014
-
Penicillin N wild type enzyme Streptomyces clavuligerus
0.016
-
Penicillin N mutant enzyme C155Y Streptomyces clavuligerus
0.047
-
Penicillin N mutant enzyme M188V Streptomyces clavuligerus
0.092
-
Penicillin N mutant enzyme C155Y/Y184/V275I/C281Y Streptomyces clavuligerus
0.19
-
penicillin G mutant enzyme C155Y/Y184/V275I/C281Y Streptomyces clavuligerus
0.22
-
penicillin G mutant enzyme N304K Streptomyces clavuligerus
0.25
-
penicillin G mutant enzyme V275I/I305M Streptomyces clavuligerus
0.295
-
Penicillin N mutant enzyme Y184H Streptomyces clavuligerus
0.66
-
penicillin G mutant enzyme I305L Streptomyces clavuligerus
0.68
-
penicillin G mutant enzyme C281Y Streptomyces clavuligerus
0.74
-
penicillin G mutant enzyme M73T Streptomyces clavuligerus
0.75
-
penicillin G mutant enzyme G79E Streptomyces clavuligerus
0.75
-
penicillin G mutant enzyme I305M Streptomyces clavuligerus
0.76
-
penicillin G mutant enzyme M188V Streptomyces clavuligerus
0.95
-
penicillin G mutant enzyme Y184H Streptomyces clavuligerus
1.02
-
penicillin G mutant enzyme L277Q Streptomyces clavuligerus
1.36
-
penicillin G mutant enzyme T91A Streptomyces clavuligerus
1.39
-
penicillin G mutant enzyme H244Q Streptomyces clavuligerus
1.68
-
penicillin G mutant enzyme V275I Streptomyces clavuligerus
1.76
-
penicillin G mutant enzyme C155Y Streptomyces clavuligerus
1.77
-
penicillin G mutant enzyme A106T Streptomyces clavuligerus
1.96
-
penicillin G mutant enzyme M188I Streptomyces clavuligerus
2.58
-
penicillin G wild type enzyme Streptomyces clavuligerus

Metals/Ions

Metals/Ions Comment Organism Structure
Fe2+ Fe2+-dependent Streptomyces clavuligerus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
penicillin N + 2-oxoglutarate + O2 Streptomyces clavuligerus
-
? + succinate + CO2
-
?

Organism

Organism UniProt Comment Textmining
Streptomyces clavuligerus
-
-
-

Purification (Commentary)

Purification (Comment) Organism
anion-exchange chromatography and gel filtration Streptomyces clavuligerus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
deacetoxycephalosporin C + 2-oxoglutarate + O2
-
Streptomyces clavuligerus deacetylcephalosporin C + succinate + CO2
-
?
penicillin G + 2-oxoglutarate + O2
-
Streptomyces clavuligerus ? + succinate + CO2
-
?
penicillin N + 2-oxoglutarate + O2
-
Streptomyces clavuligerus ? + succinate + CO2
-
?
phenylacetyl-7-aminodeacetoxycephalosporanic acid + 2-oxoglutarate + O2
-
Streptomyces clavuligerus phenylacetyl-7-aminodeacetylcephalosporin + succinate + CO2
-
?

Synonyms

Synonyms Comment Organism
DAOCS
-
Streptomyces clavuligerus
deacetoxycephalosporin C synthase
-
Streptomyces clavuligerus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.0315
-
penicillin G mutant enzyme G79E Streptomyces clavuligerus
0.0453
-
penicillin G wild type enzyme Streptomyces clavuligerus
0.0453
-
penicillin G mutant enzyme A106T Streptomyces clavuligerus
0.0456
-
penicillin G mutant enzyme M188V Streptomyces clavuligerus
0.0476
-
penicillin G mutant enzyme C155Y Streptomyces clavuligerus
0.048
-
Penicillin N mutant enzyme C155Y/Y184/V275I/C281Y Streptomyces clavuligerus
0.048
-
Penicillin N mutant enzyme Y184H Streptomyces clavuligerus
0.0502
-
penicillin G mutant enzyme V275I Streptomyces clavuligerus
0.0507
-
penicillin G mutant enzyme M188I Streptomyces clavuligerus
0.0522
-
penicillin G mutant enzyme T91A Streptomyces clavuligerus
0.0564
-
penicillin G mutant enzyme N304K Streptomyces clavuligerus
0.0627
-
penicillin G mutant enzyme M73T Streptomyces clavuligerus
0.0698
-
penicillin G mutant enzyme H244Q Streptomyces clavuligerus
0.0744
-
penicillin G mutant enzyme C281Y Streptomyces clavuligerus
0.0759
-
penicillin G mutant enzyme I305L Streptomyces clavuligerus
0.0798
-
penicillin G mutant enzyme Y184H Streptomyces clavuligerus
0.084
-
Penicillin N mutant enzyme M188V Streptomyces clavuligerus
0.1036
-
penicillin G mutant enzyme L277Q Streptomyces clavuligerus
0.1398
-
penicillin G mutant enzyme C155Y/Y184/V275I/C281Y Streptomyces clavuligerus
0.1452
-
penicillin G mutant enzyme I305M Streptomyces clavuligerus
0.1458
-
penicillin G mutant enzyme V275I/I305M Streptomyces clavuligerus
0.178
-
Penicillin N mutant enzyme G79E Streptomyces clavuligerus
0.235
-
Penicillin N mutant enzyme C155Y Streptomyces clavuligerus
0.239
-
Penicillin N mutant enzyme M73T Streptomyces clavuligerus
0.247
-
Penicillin N mutant enzyme T91A Streptomyces clavuligerus
0.252
-
Penicillin N mutant enzyme V275I Streptomyces clavuligerus
0.26
-
Penicillin N mutant enzyme H244Q Streptomyces clavuligerus
0.273
-
Penicillin N mutant enzyme C281Y Streptomyces clavuligerus
0.277
-
Penicillin N mutant enzyme A106T Streptomyces clavuligerus
0.277
-
Penicillin N mutant enzyme M188I Streptomyces clavuligerus
0.284
-
Penicillin N mutant enzyme I305L Streptomyces clavuligerus
0.297
-
Penicillin N mutant enzyme L277Q Streptomyces clavuligerus
0.307
-
Penicillin N wild type enzyme Streptomyces clavuligerus
0.31
-
Penicillin N mutant enzyme I305M Streptomyces clavuligerus
0.316
-
Penicillin N mutant enzyme V275I/I305M Streptomyces clavuligerus
0.366
-
Penicillin N mutant enzyme N304K Streptomyces clavuligerus