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Literature summary extracted from

  • Goo, K.S.; Chua, C.S.; Sim, T.S.
    Relevant double mutations in bioengineered Streptomyces clavuligerus deacetoxycephalosporin C synthase result in higher binding specificities which improve penicillin bioconversion (2008), Appl. Environ. Microbiol., 74, 1167-1175.
    View publication on PubMedView publication on EuropePMC

Protein Variants

EC Number Protein Variants Comment Organism
1.14.20.1 C281F substrate ampicillin, 191%, penicillin G, 264%, phenethicillin, 300%, and carbenicillin, 518% of wild-type activity Streptomyces clavuligerus
1.14.20.1 C281Y substrate ampicillin, 166%, penicillin G, 251%, phenethicillin, 280%, and carbenicillin, 572% of wild-type activity Streptomyces clavuligerus
1.14.20.1 C281Y/I305M substrate ampicillin, 491%, penicillin G, 495%, phenethicillin, 1109%, and carbenicillin, 1347% of wild-type activity Streptomyces clavuligerus
1.14.20.1 C281Y/N304A substrate ampicillin, 294%, penicillin G, 383%, phenethicillin, 714%, and carbenicillin, 911% of wild-type activity Streptomyces clavuligerus
1.14.20.1 C281Y/N304K substrate ampicillin, 376%, penicillin G, 428%, phenethicillin, 948%, and carbenicillin, 1180% of wild-type activity Streptomyces clavuligerus
1.14.20.1 C281Y/N304L substrate ampicillin, 330%, penicillin G, 211%, phenethicillin, 471%, and carbenicillin, 1001% of wild-type activity Streptomyces clavuligerus
1.14.20.1 C281Y/N304M substrate ampicillin, 225%, penicillin G, 277%, phenethicillin, 428%, and carbenicillin, 485% of wild-type activity Streptomyces clavuligerus
1.14.20.1 C281Y/N304R substrate ampicillin, 430%, penicillin G, 441%, phenethicillin, 1041%, and carbenicillin, 1309% of wild-type activity Streptomyces clavuligerus
1.14.20.1 C281Y/R306L substrate ampicillin, 394%, penicillin G, 191%, phenethicillin, 555%, and carbenicillin, 1010% of wild-type activity Streptomyces clavuligerus
1.14.20.1 C281Y/R307L substrate ampicillin, 274%, penicillin G, 334%, phenethicillin, 520%, and carbenicillin, 786% of wild-type activity Streptomyces clavuligerus
1.14.20.1 I305M substrate ampicillin, 313%, penicillin G, 234%, phenethicillin, 318%, and carbenicillin, 450% of wild-type activity Streptomyces clavuligerus
1.14.20.1 N304A substrate ampicillin, 192%, penicillin G, 163%, phenethicillin, 163%, and carbenicillin, 242% of wild-type activity Streptomyces clavuligerus
1.14.20.1 N304A/R306L substrate ampicillin, 214%, penicillin G, 114%, phenethicillin, 319%, and carbenicillin, 393% of wild-type activity Streptomyces clavuligerus
1.14.20.1 N304K substrate ampicillin, 270%, penicillin G, 237%, phenethicillin, 211%, an carbenicillin, 404% of wild-type activity Streptomyces clavuligerus
1.14.20.1 N304K/R306L substrate ampicillin, 245%, penicillin G, 80%, phenethicillin, 380%, and carbenicillin, 482% of wild-type activity Streptomyces clavuligerus
1.14.20.1 N304L substrate ampicillin, 163%, penicillin G, 129%, phenethicillin, 173%, and carbenicillin, 235% of wild-type activity Streptomyces clavuligerus
1.14.20.1 N304L/R306L substrate ampicillin, 145%, penicillin G, 40%, phenethicillin, 185%, and carbenicillin, 218% of wild-type activity Streptomyces clavuligerus
1.14.20.1 N304M substrate ampicillin, 120%, penicillin G,92%, phenethicillin, 127%, and carbenicillin, 161% of wild-type activity Streptomyces clavuligerus
1.14.20.1 N304M/R306L substrate ampicillin, 194%, penicillin G, 72%, phenethicillin, 245%, and carbenicillin, 232% of wild-type activity Streptomyces clavuligerus
1.14.20.1 N304R substrate ampicillin, 346%, penicillin G, 263%, phenethicillin, 360%, and carbenicillin, 398% of wild-type activity Streptomyces clavuligerus
1.14.20.1 N304R/R306L substrate ampicillin, 256%, penicillin G, 92%, phenethicillin, 344%, and carbenicillin, 514% of wild-type activity Streptomyces clavuligerus
1.14.20.1 R306L substrate ampicillin, 186%, penicillin G, 105%, phenethicillin, 173%, and carbenicillin, 283% of wild-type activity Streptomyces clavuligerus
1.14.20.1 R307L substrate ampicillin, 133%, penicillin G, 124%, phenethicillin, 122%, and carbenicillin, 108% of wild-type activity Streptomyces clavuligerus
1.14.20.1 V275I substrate ampicillin, 129%, penicillin G, 157%, phenethicillin, 150%, and carbenicillin, 227% of wild-type activity Streptomyces clavuligerus
1.14.20.1 V275I/I305M substrate ampicillin, 406%, penicillin G, 420%, phenethicillin, 685%, and carbenicillin, 1057% of wild-type activity Streptomyces clavuligerus
1.14.20.1 V275I/N304A substrate ampicillin, 265%, penicillin G, 233%, phenethicillin, 330%, and carbenicillin,721% of wild-type activity Streptomyces clavuligerus
1.14.20.1 V275I/N304K substrate ampicillin, 334%, penicillin G, 294%, phenethicillin, 635%, and carbenicillin, 867% of wild-type activity Streptomyces clavuligerus
1.14.20.1 V275I/N304L substrate ampicillin, 225%, penicillin G, 141%, phenethicillin, 245%, and carbenicillin, 524% of wild-type activity Streptomyces clavuligerus
1.14.20.1 V275I/N304M substrate ampicillin, 160%, penicillin G, 174%, phenethicillin, 261%, and carbenicillin, 230% of wild-type activity Streptomyces clavuligerus
1.14.20.1 V275I/N304R substrate ampicillin, 360%, penicillin G, 381%, phenethicillin, 698%, and carbenicillin,814% of wild-type activity Streptomyces clavuligerus
1.14.20.1 V275I/R306L substrate ampicillin, 343%, penicillin G, 168%, phenethicillin, 379%, and carbenicillin, 665% of wild-type activity Streptomyces clavuligerus
1.14.20.1 V275I/R307L substrate ampicillin, 172%, penicillin G, 169%, phenethicillin, 238%, and carbenicillin, 333% of wild-type activity Streptomyces clavuligerus
1.14.20.1 V275L substrate ampicillin, 85%, penicillin G, 130%, phenethicillin, 99%, and carbenicillin, 113% of wild-type activity Streptomyces clavuligerus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.14.20.1 0.05
-
ampicillin mutant C281Y7N304R, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.11
-
ampicillin mutant V275I/N304R, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.12
-
ampicillin mutant N304K/R306L, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.13
-
ampicillin mutant C281Y/N304K, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.15
-
ampicillin mutant C281Y/R306L, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.19
-
ampicillin mutant N304R/R306L, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.2
-
ampicillin mutant N304A/R306L, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.21
-
ampicillin mutant C281Y/I305M, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.26
-
ampicillin mutant N304R, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.34
-
ampicillin mutant C281Y/N304L, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.36
-
ampicillin mutant N304M/R306L, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.38
-
ampicillin mutant C281Y/N304A, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.38
-
ampicillin mutant V275I7N304K, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.4
-
ampicillin mutant N304L/R306L, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.51
-
ampicillin mutant V275I/R306L, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.54
-
ampicillin mutant N304K, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.68
-
ampicillin mutant V275I/N304A, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.7
-
ampicillin mutant V275I/I305M, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.8
-
ampicillin mutant C281Y/N304M, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 1
-
ampicillin mutant I305M, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 1.01
-
ampicillin mutant R306L, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 1.02
-
ampicillin mutant C281Y, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 1.04
-
ampicillin mutant V275I/N304L, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 1.09
-
ampicillin mutant N304A, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 1.2
-
ampicillin mutant C281Y/R307L, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 1.35
-
ampicillin mutant C281F, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 1.65
-
ampicillin mutant N304L, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 3.41
-
ampicillin mutant V275I, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 3.6
-
ampicillin mutant V275I/N304M, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 4.26
-
ampicillin mutant V275L, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 4.27
-
ampicillin mutant V275I/R307L, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 5.12
-
ampicillin mutant N304M, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 5.96
-
ampicillin mutant R307L, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 6.94
-
ampicillin wild type, pH 7.4, 30°C Streptomyces clavuligerus

Organism

EC Number Organism UniProt Comment Textmining
1.14.20.1 Streptomyces clavuligerus P18548
-
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.14.20.1 0.76
-
substrate carbenicillin, wild-type, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 3.57
-
substrate phenethicillin, wild-type, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 5.67
-
substrate ampicillin, wild-type, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 6.72
-
substrate penicillin G, wild-type, pH 7.4, 30°C Streptomyces clavuligerus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.20.1 ampicillin + 2-oxoglutarate + O2
-
Streptomyces clavuligerus ? + succinate + CO2
-
?
1.14.20.1 carbenicillin + 2-oxoglutarate + O2
-
Streptomyces clavuligerus ? + succinate + CO2
-
?
1.14.20.1 penicillin G + 2-oxoglutarate + O2
-
Streptomyces clavuligerus ? + succinate + CO2
-
?
1.14.20.1 phenethicillin + 2-oxoglutarate + O2
-
Streptomyces clavuligerus ? + succinate + CO2
-
?

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.14.20.1 0.026
-
ampicillin mutant N304M/R306L, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.029
-
ampicillin mutant N304L/R306L, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.03
-
ampicillin mutant N304R/R306L, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.032
-
ampicillin mutant V275L, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.033
-
ampicillin mutant N304K/R306L, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.034
-
ampicillin mutant N304A/R306L, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.037
-
ampicillin mutant C281Y/R306L, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.037
-
ampicillin mutant V275I/R306L, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.038
-
ampicillin mutant N304K, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.039
-
ampicillin mutant V275I/N304R, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.04
-
ampicillin mutant N304A, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.04
-
ampicillin mutant V275I7N304K, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.041
-
ampicillin mutant N304L, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.042
-
ampicillin mutant C281F, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.042
-
ampicillin mutant V275I/N304A, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.043
-
ampicillin mutant C281Y/N304L, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.043
-
ampicillin mutant C281Y7N304R, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.043
-
ampicillin mutant V275I/N304L, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.044
-
ampicillin mutant C281Y, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.044
-
ampicillin mutant V275I, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.046
-
ampicillin mutant C281Y/N304K, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.046
-
ampicillin mutant C281Y/R307L, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.047
-
ampicillin mutant C281Y/N304M, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.048
-
ampicillin mutant V275I/N304M, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.049
-
ampicillin mutant V275I/R307L, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.05
-
ampicillin mutant R306L, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.05
-
ampicillin mutant R307L, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.052
-
ampicillin mutant C281Y/N304A, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.052
-
ampicillin mutant N304R, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.054
-
ampicillin mutant N304M, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.059
-
ampicillin wild type, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.104
-
ampicillin mutant I305M, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.105
-
ampicillin mutant V275I/I305M, pH 7.4, 30°C Streptomyces clavuligerus
1.14.20.1 0.122
-
ampicillin mutant C281Y/I305M, pH 7.4, 30°C Streptomyces clavuligerus