Literature summary for 1.14.20.1 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.

Search for more literature for 1.14.20.1

Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Escherichia coli	Streptomyces clavuligerus

Protein Variants

Protein Variants	Comment	Organism
A106T	80% relative activity with 1 mM substrate and 170% relative activity with 10 mM penicillin G compared to wild-type activity detected with 1 mM penicillin G, 1.5fold reduction in Km, penicillin G as substrate	Streptomyces clavuligerus
A106T/C155Y	epPCR random mutagenesis	Streptomyces clavuligerus
A106T/M188V	epPCR random mutagenesis	Streptomyces clavuligerus
A11V/T91A/C281Y/I305L	220% relative activity with 1 mM and 200% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G	Streptomyces clavuligerus
C155Y	90% relative activity with 1 mM and 180% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G, 1.5fold reduction in Km, penicillin G as substrate	Streptomyces clavuligerus
C155Y/Y184H/V275I/C281Y	580% relative activity with 1 mM and 670% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G	Streptomyces clavuligerus
C281R	activity similar to wild-type enzyme	Streptomyces clavuligerus
C281Y	epPCR random mutagenesis	Streptomyces clavuligerus
D107G	activity similar to wild-type enzyme	Streptomyces clavuligerus
D107G/L277Q	epPCR random mutagenesis	Streptomyces clavuligerus
D53H/C281Y	epPCR random mutagenesis	Streptomyces clavuligerus
E144K/M188I/A198T/V275I/C281Y	160% relative activity with 1 mM and 200% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G	Streptomyces clavuligerus
G300V	410% relative activity with 1 mM and 370% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G	Streptomyces clavuligerus
G300V	substrate inhibition, epPCR random mutagenesis	Streptomyces clavuligerus
H244Q	140% relative activity with 1 mM and 270% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G, 1.9fold reduction in Km, penicillin G as substrate, epPCR random mutagenesis	Streptomyces clavuligerus
I305L	epPCR random mutagenesis	Streptomyces clavuligerus
L277Q	270% relative activity with 1 mM and 410% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G, 2.5fold reduction in Km, penicillin G as substrate	Streptomyces clavuligerus
L277Q/I305M	610% relative activity with 1 mM and 520% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G	Streptomyces clavuligerus
M188I	90% relative activity with 1 mM and 190% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G, epPCR random mutagenesis	Streptomyces clavuligerus
M188V	150% relative activity with 1 mM and 190% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G, epPCR random mutagenesis	Streptomyces clavuligerus
M188V/V275I/G300V	440% relative activity with 1 mM and 310% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G	Streptomyces clavuligerus
M73T	180% relative activity with 1 mM and 250% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G, 3.5fold reduction in Km for penicillin N and 2.3fold reduction in Km for penicillin G, epPCR random mutagenesis	Streptomyces clavuligerus
M73T/C281Y	610% relative activity with 1 mM and 530% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G	Streptomyces clavuligerus
M73T/G300V/I305L	640% relative activity with 1 mM and 380% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G	Streptomyces clavuligerus
M73T/I118V/A140V/H244Q/C281Y	450% relative activity with 1 mM and 350% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G	Streptomyces clavuligerus
M73T/I277Q	350% relative activity with 1 mM and 350% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G	Streptomyces clavuligerus
M73T/P145L	epPCR random mutagenesis	Streptomyces clavuligerus
R135Q/C155Y/R179Q/M188V/I305M	360% relative activity with 1 mM and 290% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G	Streptomyces clavuligerus
T91A	110% relative activity with 1 mM and 200% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G, 1.9fold reduction in Km, penicillin G as substrate, epPCR random mutagenesis	Streptomyces clavuligerus
V275I/C281Y	260% relative activity with 1 mM and 310% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G	Streptomyces clavuligerus
V275I/C281Y/G300V	620% relative activity with 1 mM and 380% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G	Streptomyces clavuligerus
V275I/I305L	310% relative activity with 1 mM and 330% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G	Streptomyces clavuligerus
Y184H	200% relative activity with 1 mM and 330% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G	Streptomyces clavuligerus
Y184H/C281R	epPCR random mutagenesis	Streptomyces clavuligerus
Y184H/H244Q/T259I/L277Q	320% relative activity with 1 mM and 310% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G	Streptomyces clavuligerus
Y184H/M188I/C281Y/I305L	610% relative activity with 1 mM and 460% relative activity with 10 mM substrate penicillin G compared to wild-type activity detected with 1 mM penicillin G	Streptomyces clavuligerus

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.006	-	Penicillin N	mutant M73T	Streptomyces clavuligerus
0.009	-	Penicillin N	mutant H244Q	Streptomyces clavuligerus
0.009	-	Penicillin N	mutant T91A	Streptomyces clavuligerus
0.011	-	Penicillin N	mutant L277Q	Streptomyces clavuligerus
0.011	-	Penicillin N	mutant M188I	Streptomyces clavuligerus
0.013	-	Penicillin N	mutant A106T	Streptomyces clavuligerus
0.014	-	Penicillin N	wild-type	Streptomyces clavuligerus
0.016	-	Penicillin N	mutant C155Y	Streptomyces clavuligerus
0.047	-	Penicillin N	mutant M188V	Streptomyces clavuligerus
0.092	-	Penicillin N	mutant C155Y/Y184H/V275I/C281Y	Streptomyces clavuligerus
0.19	-	penicillin G	mutant C155Y/Y184H/V275I/C281Y	Streptomyces clavuligerus
0.295	-	Penicillin N	mutant Y184H	Streptomyces clavuligerus
0.74	-	penicillin G	mutant M73T	Streptomyces clavuligerus
0.76	-	penicillin G	mutant M188V	Streptomyces clavuligerus
0.95	-	penicillin G	mutant Y184H	Streptomyces clavuligerus
1.02	-	penicillin G	mutant L277Q	Streptomyces clavuligerus
1.36	-	penicillin G	mutant T91A	Streptomyces clavuligerus
1.39	-	penicillin G	mutant H244Q	Streptomyces clavuligerus
1.76	-	penicillin G	mutant C155Y	Streptomyces clavuligerus
1.77	-	penicillin G	mutant A106T	Streptomyces clavuligerus
1.96	-	penicillin G	mutant M188I	Streptomyces clavuligerus
2.58	-	penicillin G	wild-type	Streptomyces clavuligerus

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
penicillin G + 2-oxoglutarate + O2	Streptomyces clavuligerus	-	phenylacetyl-7-aminodeacetoxycephalosporanic acid + succinate + CO2 + H2O	-	?

Organism

Organism	UniProt	Comment	Textmining
Streptomyces clavuligerus	P18548	-	-

Purification (Commentary)

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

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
additional information	-	160% relative activity with 10 mM substrate penicillin G compared to the activity detected with 1 mM penicillin G	Streptomyces clavuligerus

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
penicillin G + 2-oxoglutarate + O2	-	Streptomyces clavuligerus	phenylacetyl-7-aminodeacetoxycephalosporanic acid + succinate + CO2 + H2O	-	?
penicillin N + 2-oxoglutarate + O2	-	Streptomyces clavuligerus	deacetoxycephalosporin C + succinate + CO2 + H2O	-	?

Synonyms

Synonyms	Comment	Organism
DAOCS	-	Streptomyces clavuligerus