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(+)-alpha-pinene + putidaredoxin + O2
(+)-cis-verbenol + (+)-myrtenol + (+)-verbenone + oxidized putidaredoxin + H2O
-
-
-
?
(+)-alpha-pinene + putidaredoxin + O2
? + oxidized putidaredoxin + H2O
-
-
-
-
?
(+)-camphor + O2 + reduced putidaredoxin
(+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
(+)-camphor + O2 + reduced putidaredoxin
exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
(+)-camphor + putidaredoxin + O2
(R)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
(+)-camphor + reduced ferredoxin + O2
(+)-exo-5-hydroxycamphor + oxidized ferredoxin + H2O
(+)-camphor + reduced putidaredoxin + NADH + H+ + O2
(+)-exo-5-hydroxycamphor + oxidized putidaredoxin + NAD+ + H2O
(+)-camphor + reduced putidaredoxin + O2
(+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
(+)-camphor + reduced putidaredoxin + O2
(R)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
-
-
?
(+)-camphor + reduced putidaredoxin + O2
borneol + oxidized putidaredoxin + H2O
(+)-exo-5-hydroxycamphor + reduced putidaredoxin + O2
5-oxocamphor + oxidized putidaredoxin + H2O
-
-
-
-
?
(1R)-(+)-camphor + O2 + reduced putidaredoxin
(+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
-
-
-
?
(1R)-(+)-camphor + reduced putidaredoxin + O2
(+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
(1R)-5,5-difluorocamphor + putidaredoxin + O2
? + oxidized putidaredoxin + H2O
-
-
-
?
(1R)-5-exo-methoxycamphor + putidaredoxin + O2
? + oxidized putidaredoxin + H2O
-
-
-
?
(1R)-5-methylenylcamphor + putidaredoxin + O2
? + oxidized putidaredoxin + H2O
-
-
-
?
(1R)-camphor + putidaredoxin + O2
5-exo-(1R)-hydroxycamphor + oxidized putidaredoxin + H2O
(1R)-camphor + putidaredoxin + O2
? + oxidized putidaredoxin + H2O
(1R)-camphor enol ether + putidaredoxin + O2
? + oxidized putidaredoxin + H2O
-
-
-
-
?
(1R)-camphor N-methyl imine + putidaredoxin + O2
? + oxidized putidaredoxin + H2O
-
-
-
-
?
(1R)-camphor oxime + putidaredoxin + O2
? + oxidized putidaredoxin + H2O
-
-
-
-
?
(1R)-endo-borneol allyl ether + putidaredoxin + O2
? + oxidized putidaredoxin + H2O
-
-
-
-
?
(1R)-endo-borneol methyl ether + putidaredoxin + O2
? + oxidized putidaredoxin + H2O
-
-
-
-
?
(1R)-endo-borneol propyl ether + putidaredoxin + O2
? + oxidized putidaredoxin + H2O
-
-
-
-
?
(1R)-iso-borneol methyl ether + putidaredoxin + O2
? + oxidized putidaredoxin + H2O
-
-
-
-
?
(1R)-norcamphor + putidaredoxin + O2
? + oxidized putidaredoxin + H2O
-
-
-
?
(1S)-camphor + putidaredoxin + O2
? + oxidized putidaredoxin + H2O
-
-
-
?
(4S)-limonene + putidaredoxin + O2
?
-
the 7-position is the major site of hydroxylation by P450cam
-
-
?
(R)-2-ethylhexanol + reduced putidaredoxin + O2
(R)-2-ethylhexanoic acid + oxidized putidaredoxin + H2O
-
-
-
-
r
(R)-3-ethylhexanol + putidaredoxin + O2
2-ethylhexanoic acid + 2-ethyl-1,2-hexanediol + 2-ethyl-1,3-hexanediol + 2-ethyl-1,4-hexanediol + oxidized putidaredoxin + H2O
-
-
ratio: 50:13:15:8
?
(R)-exo-5-hydroxycamphor + O2 + reduced putidaredoxin
2,5-diketocamphane + oxidized putidaredoxin + H2O
-
-
-
-
?
(S)-2-ethylhexanol + reduced putidaredoxin + O2
(S)-2-ethylhexanoic acid + oxidized putidaredoxin + H2O
-
-
-
-
r
(S)-3-ethylhexanol + putidaredoxin + O2
2-ethylhexanoic aicd + 2-ethyl-1,2-hexanediol + 2-ethyl-1,3-hexanediol + 2-ethyl-1,4-hexanediol + oxidized putidaredoxin + H2O
-
the (S)-isomer is turned over 1.4times faster than the (R)-isomer
ratio: 15:53:28:10
?
1,2,4,5-tetrachlorobenzene + putidaredoxin + O2
2,3,5,6-tetrachlorophenol + oxidized putidaredoxin + H2O
-
-
-
?
1,2-campholide + putidaredoxin + O2
5-exo-hydroxy-1,2-campholide + oxidized putidaredoxin + H2O
1,2-dibromo-3-chloropropane + O2 + reduced putidaredoxin
1-bromo-3-chloroacetone + allyl chloride + H2O + putidaredoxin + Br-
-
dehalogenation, bromochloroacetone is the major conversion product when the incubation medium is saturated with oxygen, while allyl chloride is the sole product in the absence of oxygen
a number of bromochloropropene are also formed to a minor extent by an elimination mechanism, product determination
-
?
1,2-dichlorobenzene + putidaredoxin + O2
2,3-dichlorophenol + 3,4-dichlorophenol + oxidized putidaredoxin + H2O
-
-
-
?
1,3,5-trichlorobenzene + putidaredoxin + O2
2,4,6-trichlorophenol + oxidized putidaredoxin + H2O
-
-
-
?
1,3,5-trichlorobenzene + putidaredoxin + O2
? + oxidized putidaredoxin + H2O
-
-
-
-
?
1,3-dichlorobenzene + putidaredoxin + O2
2,6-dichlorophenol + 2,4-dichlorophenol + 2,5-dichlorophenol + 2,3-dichlorophenol + oxidized putidaredoxin + H2O
-
-
-
?
1,4-dichlorobenzene + putidaredoxin + O2
2,5-dichlorophenol + oxidized putidaredoxin + H2O
-
-
-
?
1-dehydrocamphor + putidaredoxin + O2
exo-5,6-epoxycamphor + oxidized putidaredoxin + H2O
-
-
-
?
1-ethyl-2-methylbenzene + reduced putidaredoxin + O2
? + oxidized putidaredoxin + H2O
the S-enantiomer is preferred by the wild-type enzyme
-
-
?
1-ethyl-3-methylbenzene + reduced putidaredoxin + O2
? + oxidized putidaredoxin + H2O
the S-enantiomer is preferred by the wild-type enzyme
-
-
?
1-ethyl-4-methylbenzene + reduced putidaredoxin + O2
? + oxidized putidaredoxin + H2O
the S-enantiomer is preferred by the wild-type enzyme
-
-
?
1-methylimidazole + O2 + reduced putidaredoxin
?
-
-
-
?
2-adamantanone + O2 + reduced putidaredoxin
5-hydroxy-2-adamantanone + oxidized putidaredoxin + H2O
3 2-methylpentane + 3 reduced ferreredoxin + O2
2-methyl-pentan-2-ol + 2-methyl-pentan-3-ol + 2-methyl-pentan-4-ol + 3 oxidized ferredoxin
-
52.5% 2-methyl-pentan-2-ol + 13% 2-methyl-pentan-3-ol, and 3% 2-methyl-pentan-4-ol for the wild-type enzyme, 5% + 12% + 30% for the mutant Y96A
-
?
3-chloroindole + O2 + reduced putidaredoxin
isatin + H2O + Cl- + oxidized putidaredoxin + ?
no substrate of wild-type, substrate of mutants E156G/V247F/V253G/F256S, T56A/N116H/D297N and G60S/Y75H
-
-
?
3-chloroperbenzoic acid + O2 + reduced putidaredoxin
?
5,5-difluorocamphor + O2 + reduced putidaredoxin
?
-
-
-
-
?
5,5-difluorocamphor + putidaredoxin + O2
5,5-difluoro-9-hydroxycamphor + oxidized putidaredoxin + H2O
-
-
-
?
5-exo-bromocamphor + putidaredoxin + O2
5-ketocamphor + oxidized putidaredoxin + Br- + H2O
-
(+)- and (-)-enantiomer
-
?
5-methylenyl-camphor + O2 + reduced putidaredoxin
?
adamantane + reduced ferreredoxin + O2
1-adamantol + 2-adamantol + oxidized ferredoxin + H2O
-
98% 1-adamantol + 2% 2-adamantol for the wild-type enzyme, 97% + 3% for the mutant Y96A
-
?
adamantanone + O2 + reduced putidaredoxin
?
-
-
-
-
?
adamantanone + putidaredoxin + O2
? + oxidized putidaredoxin + H2O
-
-
-
?
adamantenone + reduced putidaredoxin + O2
?
-
-
-
-
?
benzo[a]pyrene + putidaredoxin + O2
3-hydroxybenzo[a]pyrene + oxidized putidaredoxin + H2O
-
-
-
-
?
beta-ionone + O2 + reduced putidaredoxin
4-hydroxy-beta-ionone + oxidized putidaredoxin + H2O
camphane + reduced putidaredoxin + O2
? + oxidized putidaredoxin + H2O
-
-
-
?
cyclooctane + reduced ferreredoxin + O2
cyclooctanol + cyclooctanone + oxidized ferredoxin + H2O
-
99% cyclooctanol + 1% cyclooctanone for the wild-type enzyme, 97% + 3% for the mutant Y96A
-
?
DL-camphor + reduced putidaredoxin + NADH + H+ + O2
exo-5-hydroxycamphor + oxidized putidaredoxin + NAD+ + H2O
-
-
-
?
ethylbenzene + putidaredoxin + O2
1-phenylethanol + oxidized putidaredoxin + H2O
-
at 5% of the reaction with (+)-camphor
ratio of (R)- to (S)-1-phenylethanol produced depends on mutant form
?
fluoranthene + putidaredoxin + O2
3-fluoranthol + oxidized putidaredoxin + H2O
-
-
-
-
?
hexane + reduced ferreredoxin + O2
hexan-2-ol + hexan-3-ol + oxidized ferredoxin + H2O
-
56% hexan-2-ol + 44% hexan-3-ol for the wild-type enzyme and mutant Y96A
-
?
imidazole + O2 + reduced putidaredoxin
?
-
-
-
?
indole + O2 + reduced putidaredoxin
3-hydroxyindole + oxidized putidaredoxin + H2O
-
no substrate of the wild-type enzyme, but a good substrate for Y96 mutants, mutant screening, overview
3-hydroxyindole undergoes spontaneous air oxidation to produce the insoluble dye indigo
-
?
isoborneol + reduced putidaredoxin + O2
? + oxidized putidaredoxin + H2O
-
-
-
?
linalool + O2 + reduced putidaredoxin
8-hydroxy-linalool + oxidized putidaredoxin + H2O
norcamphor + O2 + reduced putidaredoxin
?
-
-
-
-
?
norcamphor + reduced putidaredoxin + O2
?
-
-
-
-
?
norcamphor + reduced putidaredoxin + O2
? + oxidized putidaredoxin + H2O
-
-
-
?
peracetic acid + O2 + reduced putidaredoxin
?
phenanthrene + putidaredoxin + O2
1-phenanthrol + 2-phenanthrol + 3-phenanthrol + 4-phenanthrol + oxidized putidaredoxin + H2O
-
-
-
-
?
pyrene + putidaredoxin + O2
1-pyrenol + 2-pyrenol + 1,6-pyrenequinone + 1,8-pyrenequinone + oxidized putidaredoxin + H2O
-
-
-
-
?
thiocamphor + reduced putidaredoxin + O2
? + oxidized putidaredoxin + H2O
-
-
-
?
additional information
?
-
(+)-camphor + O2 + reduced putidaredoxin
(+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
CYP101D1, CYP101D2
-
-
?
(+)-camphor + O2 + reduced putidaredoxin
(+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
CYP101D1, CYP101D2
-
-
?
(+)-camphor + O2 + reduced putidaredoxin
(+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
-
671535, 672100, 672115, 672742, 672760, 673037, 673071, 674152, 674438, 674474, 674516, 674959, 675234, 676280 -
-
?
(+)-camphor + O2 + reduced putidaredoxin
(+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
-
-
?
(+)-camphor + O2 + reduced putidaredoxin
(+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
-
-
-
?
(+)-camphor + O2 + reduced putidaredoxin
(+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
-
-
?
(+)-camphor + O2 + reduced putidaredoxin
(+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
terminal monooxygenase in a three-component camphor-hydroxylating system from Pseudomonas putida, the reaction cycle requires two distinct electron transfer processes from the [2Fe-2S] containing putidaredoxin to P450cam
-
-
?
(+)-camphor + O2 + reduced putidaredoxin
(+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
analysis of protein-protein interactions between enzyme and cofactor, overview
-
-
?
(+)-camphor + O2 + reduced putidaredoxin
(+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
binding of camphor is strongly dependent on the concentration of alcohols, alcohol expels camphor out of the heme cavity of the enzyme by affecting tertiary structure of Cyt P450cam as well as by modifying the solubility properties of camphor in aqueous medium, overview
-
-
?
(+)-camphor + O2 + reduced putidaredoxin
(+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
determination of appearance of transient intermediates at 3°C by double mixing rapid scanning stopped-flow spectroscopy, electron transfer from reduced putidaredoxin gives high spin deoxyferrous P-450, substrate-free enzyme also binds the cofactor, but the cofactor does not deliver the electrons to the substrate-free oxyferrous enzyme, binding structure of camphor-bound oxyferrous P450-CAM with reduced putidaredoxin, functional implications of the formation of the perturbed oxyferrous intermediate, overview
-
-
?
(+)-camphor + O2 + reduced putidaredoxin
(+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
mechanism of camphor hydroxylation incorporating an NADH-regeneration system, overview
-
-
?
(+)-camphor + O2 + reduced putidaredoxin
(+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
regio- and stereoselective C-H bond hydroxylation, rebound mechanism, overview
-
-
?
(+)-camphor + O2 + reduced putidaredoxin
(+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
regio- and stereospecific hydroxylation
-
-
?
(+)-camphor + O2 + reduced putidaredoxin
(+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
second reductive step of the mechanism of interaction and electron transfer, overview
-
-
?
(+)-camphor + O2 + reduced putidaredoxin
(+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
wild-type dioxygen complex structure: high occupancy and a ordered structure of the iron-linked dioxygen and two 'catalytic' water molecules that form part of a proton relay system to the iron-linked dioxygen, Thr252 accepts a hydrogen bond from the hydroperoxy (Fe(III)-OOH) intermediate that promotes the second protonation on the distal oxygen atom, leading to O-O bond cleavage and compound I formation
-
-
?
(+)-camphor + O2 + reduced putidaredoxin
(+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
wild-type enzyme, but not mutant T252A
-
-
?
(+)-camphor + O2 + reduced putidaredoxin
(+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
the reduced enzyme exhibits lower-amplitude motions of secondary structural features than the oxidized enzyme on all of the time scales accessible, and these differences are more pronounced in regions of the enzyme involved in substrate access to the active site (B' helix and beta3 and beta5 sheets) and binding of putidaredoxin (C and L helices), the iron-sulfur protein that acts as the effector and reductant of CYP101 in vivo
-
-
?
(+)-camphor + O2 + reduced putidaredoxin
(+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
though the active site of the enzyme resides deep inside the protein matrix, the substrate is recognized at the surface of the enzyme and directed towards the active site through the access channel. The threonine 192 that resides on the F-G loop and directed towards the putative substrate access channel of the enzyme, plays an important role in recognition of the substrate at the surface of the enzyme
-
-
?
(+)-camphor + O2 + reduced putidaredoxin
exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
-
-
-
?
(+)-camphor + O2 + reduced putidaredoxin
exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
-
-
?
(+)-camphor + O2 + reduced putidaredoxin
exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
conformational change in CYP101 upon binding of putidaredoxin that re-orients bound camphor appropriately for hydroxylation
-
-
?
(+)-camphor + O2 + reduced putidaredoxin
exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
ferric hydroperoxo complex, elusive reactive species of cytochrome P450cam, and the hydroxo intermediate (formed during camphor hydroxylation) in the catalytic cycle of cytochrome P450cam all have a doublet ground state which have a pronounced multiconfigurational character in the case of compound I and the hydroxo intermediate
-
-
?
(+)-camphor + O2 + reduced putidaredoxin
exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
the 7-propionate side chain plays a role in maintaining the high affinity of cytochrome P450cam for its substrate, the Asp297 and Gln322 residues are capable of undergoing a 7-propionate-associated conformational change in the protein interior
-
-
?
(+)-camphor + O2 + reduced putidaredoxin
exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
the Cpd II-like species is ineffective at hydroxylating camphor, but can be readily reduced by ascorbate to ferric P450cam, which can then bind camphor to form the high-spin heme
-
-
?
(+)-camphor + putidaredoxin + O2
(R)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
-
-
-
?
(+)-camphor + putidaredoxin + O2
(R)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
-
i.e. (+)-exo-5-hydroxycamphor
?
(+)-camphor + putidaredoxin + O2
(R)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
-
i.e. (+)-exo-5-hydroxycamphor
?
(+)-camphor + putidaredoxin + O2
(R)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
-
i.e. (+)-exo-5-hydroxycamphor
?
(+)-camphor + putidaredoxin + O2
(R)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
-
i.e. (+)-exo-5-hydroxycamphor
?
(+)-camphor + putidaredoxin + O2
(R)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
-
i.e. (+)-exo-5-hydroxycamphor
?
(+)-camphor + putidaredoxin + O2
(R)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
-
i.e. (+)-exo-5-hydroxycamphor
?
(+)-camphor + putidaredoxin + O2
(R)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
-
i.e. (+)-exo-5-hydroxycamphor
?
(+)-camphor + putidaredoxin + O2
(R)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
-
i.e. (+)-exo-5-hydroxycamphor
?
(+)-camphor + putidaredoxin + O2
(R)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
-
i.e. (+)-exo-5-hydroxycamphor
?
(+)-camphor + putidaredoxin + O2
(R)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
-
i.e. (+)-exo-5-hydroxycamphor
?
(+)-camphor + putidaredoxin + O2
(R)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
-
i.e. (+)-exo-5-hydroxycamphor
?
(+)-camphor + putidaredoxin + O2
(R)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
-
i.e. (+)-exo-5-hydroxycamphor
?
(+)-camphor + putidaredoxin + O2
(R)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
-
i.e. (+)-exo-5-hydroxycamphor
?
(+)-camphor + putidaredoxin + O2
(R)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
-
i.e. (+)-exo-5-hydroxycamphor
?
(+)-camphor + putidaredoxin + O2
(R)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
-
i.e. (+)-exo-5-hydroxycamphor
?
(+)-camphor + putidaredoxin + O2
(R)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
-
i.e. (+)-exo-5-hydroxycamphor
?
(+)-camphor + putidaredoxin + O2
(R)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
the (+)- and (-)-enantiomers serve as substrates
i.e. (+)-exo-5-hydroxycamphor
?
(+)-camphor + putidaredoxin + O2
(R)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
when deuterated at either 5-exo- or 5-endo-position, only 5-exo-hydroxycamphor is the product
i.e. (+)-exo-5-hydroxycamphor
?
(+)-camphor + putidaredoxin + O2
(R)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
-
i.e. (+)-exo-5-hydroxycamphor
?
(+)-camphor + putidaredoxin + O2
(R)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
-
i.e. (+)-exo-5-hydroxycamphor
?
(+)-camphor + reduced ferredoxin + O2
(+)-exo-5-hydroxycamphor + oxidized ferredoxin + H2O
-
-
-
?
(+)-camphor + reduced ferredoxin + O2
(+)-exo-5-hydroxycamphor + oxidized ferredoxin + H2O
-
99% 5-exo-hydroxycamphor by wild-type enzyme and 92% by Y96A mutant
-
?
(+)-camphor + reduced ferredoxin + O2
(+)-exo-5-hydroxycamphor + oxidized ferredoxin + H2O
substrate of CYP101D1 and CYP101D2
-
-
?
(+)-camphor + reduced ferredoxin + O2
(+)-exo-5-hydroxycamphor + oxidized ferredoxin + H2O
-
-
-
?
(+)-camphor + reduced ferredoxin + O2
(+)-exo-5-hydroxycamphor + oxidized ferredoxin + H2O
substrate of CYP101D1 and CYP101D2
-
-
?
(+)-camphor + reduced putidaredoxin + NADH + H+ + O2
(+)-exo-5-hydroxycamphor + oxidized putidaredoxin + NAD+ + H2O
-
-
-
?
(+)-camphor + reduced putidaredoxin + NADH + H+ + O2
(+)-exo-5-hydroxycamphor + oxidized putidaredoxin + NAD+ + H2O
the catalytic cycle of P450cam requires two electrons, both of which are donated by putidaredoxin (Pdx), a ferredoxin containing a [2Fe-2S] cluster, structures of the Pdx-P450cam complex, overview
-
-
?
(+)-camphor + reduced putidaredoxin + NADH + H+ + O2
(+)-exo-5-hydroxycamphor + oxidized putidaredoxin + NAD+ + H2O
-
-
-
?
(+)-camphor + reduced putidaredoxin + O2
(+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
-
-
-
?
(+)-camphor + reduced putidaredoxin + O2
(+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
-
-
-
r
(+)-camphor + reduced putidaredoxin + O2
(+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
-
-
?
(+)-camphor + reduced putidaredoxin + O2
(+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
the enzyme shows regio- and stereospecific hydroxylation. Activation and cleavage of the oxygen molecule in the P450cam catalytic cycle is accompanied by two electron transfers from putidaredoxin. Ferric P450cam can accept the first electron from diverse chemical reductants and putidaredoxin homologues, but the second requires putidaredoxin as donor
-
-
?
(+)-camphor + reduced putidaredoxin + O2
(+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
-
-
?
(+)-camphor + reduced putidaredoxin + O2
(+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
-
-
-
?
(+)-camphor + reduced putidaredoxin + O2
(+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
-
-
-
?
(+)-camphor + reduced putidaredoxin + O2
borneol + oxidized putidaredoxin + H2O
-
under low oxygen conditions borneol is formed instead of 5-ketocamphor
-
-
?
(+)-camphor + reduced putidaredoxin + O2
borneol + oxidized putidaredoxin + H2O
-
the hydroxylation reaction proceeds via a catalytic cycle in which the reduction of dioxygen is coupled to the oxidation of the substrate. A key intermediate in the catalytic cycle is the iron-oxo species (Fe(IV)=O)
-
-
?
(+)-camphor + reduced putidaredoxin + O2
borneol + oxidized putidaredoxin + H2O
-
under low oxygen conditions
-
-
?
(+)-camphor + reduced putidaredoxin + O2
borneol + oxidized putidaredoxin + H2O
-
under low oxygen conditions borneol is formed instead of 5-ketocamphor
-
-
?
(+)-camphor + reduced putidaredoxin + O2
borneol + oxidized putidaredoxin + H2O
-
the hydroxylation reaction proceeds via a catalytic cycle in which the reduction of dioxygen is coupled to the oxidation of the substrate. A key intermediate in the catalytic cycle is the iron-oxo species (Fe(IV)=O)
-
-
?
(+)-camphor + reduced putidaredoxin + O2
borneol + oxidized putidaredoxin + H2O
-
under low oxygen conditions
-
-
?
(1R)-(+)-camphor + reduced putidaredoxin + O2
(+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
-
-
?
(1R)-(+)-camphor + reduced putidaredoxin + O2
(+)-exo-5-hydroxycamphor + oxidized putidaredoxin + H2O
-
-
-
?
(1R)-camphor + putidaredoxin + O2
5-exo-(1R)-hydroxycamphor + oxidized putidaredoxin + H2O
-
putidaredoxin transfers electrons from NADH to P450cam in a coupled assay method
-
-
?
(1R)-camphor + putidaredoxin + O2
5-exo-(1R)-hydroxycamphor + oxidized putidaredoxin + H2O
-
putidaredoxin transfers electrons from NADH to P450cam
-
-
?
(1R)-camphor + putidaredoxin + O2
5-exo-(1R)-hydroxycamphor + oxidized putidaredoxin + H2O
-
putidaredoxin transfers electrons from NADH to P450cam in a coupled assay method
-
-
?
(1R)-camphor + putidaredoxin + O2
5-exo-(1R)-hydroxycamphor + oxidized putidaredoxin + H2O
-
putidaredoxin transfers electrons from NADH to P450cam
-
-
?
(1R)-camphor + putidaredoxin + O2
? + oxidized putidaredoxin + H2O
-
-
-
-
?
(1R)-camphor + putidaredoxin + O2
? + oxidized putidaredoxin + H2O
-
-
-
?
1,2-campholide + putidaredoxin + O2
5-exo-hydroxy-1,2-campholide + oxidized putidaredoxin + H2O
-
-
-
?
1,2-campholide + putidaredoxin + O2
5-exo-hydroxy-1,2-campholide + oxidized putidaredoxin + H2O
-
-
-
?
1,2-campholide + putidaredoxin + O2
5-exo-hydroxy-1,2-campholide + oxidized putidaredoxin + H2O
-
-
-
?
1,2-campholide + putidaredoxin + O2
5-exo-hydroxy-1,2-campholide + oxidized putidaredoxin + H2O
-
-
-
?
2-adamantanone + O2 + reduced putidaredoxin
5-hydroxy-2-adamantanone + oxidized putidaredoxin + H2O
-
CYP101D1 and CYP101D2
-
-
?
2-adamantanone + O2 + reduced putidaredoxin
5-hydroxy-2-adamantanone + oxidized putidaredoxin + H2O
-
CYP101D1 and CYP101D2
-
-
?
3-chloroperbenzoic acid + O2 + reduced putidaredoxin
?
compound I, ferryl iron plus a porphyrin pi-cation radical (Fe(IV)=O/Por(+)), and compound ES, Fe(IV)=O/Tyr(), in reactions of substrate-free ferric enzyme with 3-chloroperbenzoic acid, compound ES arises by intramolecular electron transfer from nearby tyrosines to the porphyrin pi-cation radical of compound I, active site changes influence electron transfer from nearby tyrosines and affect formation of intermediates, the tyrosyl radical is assigned to Tyr96 for wild type or to Tyr75 for the Y96F variant, overview
-
-
?
3-chloroperbenzoic acid + O2 + reduced putidaredoxin
?
-
reaction mechanism of substrate-free ferric cytochrome P450cam, via FeIV-O plus porphyrin Pi-cation radical, overview
-
-
?
5-methylenyl-camphor + O2 + reduced putidaredoxin
?
-
-
-
-
?
5-methylenyl-camphor + O2 + reduced putidaredoxin
?
-
wild-type enzyme and mutant T252A, in absence of the primary oxidant species of P450, the precursor species FeOOH can effect double bond activation of 5-methylenyl-camphor initiated by a homolytic cleavage of the O-O-bond and formation of an OH radical bound to the secondary oxidant by hydrogen bonding interaction, overview
-
-
?
beta-ionone + O2 + reduced putidaredoxin
4-hydroxy-beta-ionone + oxidized putidaredoxin + H2O
-
CYP101C1 and CYP101B1. CYP101C1 oxidizes beta-ionone to 4-hydroxy-beta-ionone (75%) with one other, unidentified product. This latter compound is the major product of beta-ionone oxidation by CYP101B1 (90%) where 4-hydroxy-beta-ionone is the minor product (10%)
-
-
?
beta-ionone + O2 + reduced putidaredoxin
4-hydroxy-beta-ionone + oxidized putidaredoxin + H2O
-
CYP101C1 and CYP101B1. CYP101C1 oxidizes beta-ionone to 4-hydroxy-beta-ionone (75%) with one other, unidentified product. This latter compound is the major product of beta-ionone oxidation by CYP101B1 (90%) where 4-hydroxy-beta-ionone is the minor product (10%)
-
-
?
linalool + O2 + reduced putidaredoxin
8-hydroxy-linalool + oxidized putidaredoxin + H2O
-
CYP111A2 and CYP111A1
-
-
?
linalool + O2 + reduced putidaredoxin
8-hydroxy-linalool + oxidized putidaredoxin + H2O
-
CYP111A2 and CYP111A1
-
-
?
peracetic acid + O2 + reduced putidaredoxin
?
-
-
-
-
?
peracetic acid + O2 + reduced putidaredoxin
?
-
reaction mechanism of substrate-free ferric cytochrome P450cam, via FeIV-O plus tyrosyl radical, overview
-
-
?
additional information
?
-
substrate binding and activity data for wild-type CYP101D2 and variants with different substrates, gas chromatography analysis of products, overview
-
-
?
additional information
?
-
-
substrate binding and activity data for wild-type CYP101D2 and variants with different substrates, gas chromatography analysis of products, overview
-
-
?
additional information
?
-
-
-
-
-
?
additional information
?
-
-
-
-
-
?
additional information
?
-
effects of heme environment on the hydrogen abstraction reaction of camphor in catalysis, overview
-
-
?
additional information
?
-
-
modelling of the hydroxylation of camphor using crystal structure, PDB code 1DZ9, and combined quantum mechanical/molecular mechanical method, heme propionate side chains are not involved in catalysis, Asp297 is important for the reaction mechanism, overview
-
-
?
additional information
?
-
-
relative stability of dibromochloropropane and products, overview
-
-
?
additional information
?
-
release of the substrate is caused both due to increased solubility of the substrate in solution in presence of alcohol and due to change in the tertiary structure of the active site of the enzyme, addition of alcohols to cytochrome P450cam causes a small change in the secondary structural elements but a significant change in the tertiary structural organization of the enzyme
-
-
?
additional information
?
-
-
the properties and reactivity of the oxyheme and of both the primary and the annealed intermediates are modulated by a bound substrate, including alterations in the properties of the heme center, the presence of any alternative substrate increases the lifetime of hydroperoxoferri-P450cam no less than about 20fold, especially 5-methylenyl-camphor
-
-
?
additional information
?
-
-
substrate specificities of wild-type and mutant enzymes, overview
-
-
?
additional information
?
-
-
under conditions of low oxygen, Pseudomonas putida cells and the isolated P450cam reduce camphor to borneol, product analysis by GC-MS
-
-
?
additional information
?
-
an additional coupling pathway transpires during H2O2 shunting of the cycle of wild-type P450cam and in mutant T252A. The reaction starts with the FeIII(O2H2) intermediate, which transforms to Cpd I via a O-O homolysis/H-abstraction mechanism. The substrate 5-methylenylcamphor prevents H2O2 release, while the protein controls the FeIII(O2H2) conversion to Cpd I by nailing through hydrogen-bonding interactionsthe conformation of the HO radical produced during O-O homolysis. This conformation prevents HO readical attack on the porphyrins meso position, as in heme oxygenase, and prefers H-abstraction from FeIVOH thereby generating H2O + Cpd I. Cpd I then performs substrate oxidations
-
-
?
additional information
?
-
analysis of the molecular substrate recognition of wild-type and mutant P450cams by infrared (IR) spectroscopy, differing conformational heterogeneity in the active site of the P450cam variants and changes in heterogeneity upon binding of different substrates likely contribute to their variable affinities via a conformational selection mechanism, overview. Although P450cam is relatively specific for camphor, it also recognizes a number of small, camphor-like substrates, albeit with variable affinity. Substrate binding and active site structure, overview
-
-
?
additional information
?
-
catalytic turnover in P450cam requires the enzymes putidaredoxin (Pdx) and putidaredoxin reductase (Pdr), which mediate electron transfer from NADH to heme, the process is tightly coupled to substrate hydroxylation
-
-
?
additional information
?
-
comparison of substrate binding and catalytic ability of wild-type enzyme with putidareductase and putidaredoxin (ternary complex), and recombinant fusion enzyme P450cam-putidareductase with putidaredoxin, overview. The wild-type and mutant systems show comparable activity with camphor. Further oxidation of 5-exo-hydroxycamphor to 5-oxo-camphor by the fusion enzyme is 39% lower than for the native system
-
-
?
additional information
?
-
enzyme 450cam binds camphor and converts from the closed to open conformation upon binding putidaredoxin, the binding thermodynamics of Pdx differ when the conformation of P450cam is held in different states, thermodynamic analysis, overview
-
-
?
additional information
?
-
enzyme P450cam catalyzes the regioselective hydroxylation of d-camphor to 5-exo-hydroxycamphor. P450cam hydroxylates thiocamphor with a regioselectivity lower than camphor but higher than norcamphor. The high regioselectivity of camphor hydroxylation, the hydrogen bond (H-bond) between the camphor ketone and the side chain of active site residue Y96 influences the local electrostatics of the active site but has little effect on either the relative populations of conformational states or the nature of the energy landscapes within the conformations. Infrared spectrometric analysis of enzyme-substrate intercations, overview
-
-
?
additional information
?
-
enzyme P450cam produces benzyl alcohols with only moderate enantioselectivities, enantioselective benzylic hydroxylation catalysed by P450 monooxygenases, enantioselectivity of a library of active-site mutants of chimeric P450cam-RhFRed towards the benzylic hydroxylation of structurally related regioisomers of ethylmethylbenzene, molecular dynamics simulations and computational molecular modelling, overview
-
-
?
additional information
?
-
putidaredoxin (Pdx) and putidaredoxin reductase (PdR) are expressed in Escherichia coli strain BL21 from a pMG211 plasmid as C-terminally His6-tagged proteins, and purified by nickel affinity chromatography and gel filtration
-
-
?
additional information
?
-
-
the P450FeIII-OOH intermediate must be the oxidizing species. The mechanism of hydrogen peroxide binding to the substrate-free form of P450cam is mainly governed by the ability of H2O2 to undergo deprotonation at the hydroxo ligand coordinated to the iron(III) center under conditions of pH > pKP450
-
-
?
additional information
?
-
putidaredoxin (Pdx) and putidaredoxin reductase (PdR) are expressed in Escherichia coli strain BL21 from a pMG211 plasmid as C-terminally His6-tagged proteins, and purified by nickel affinity chromatography and gel filtration
-
-
?
additional information
?
-
catalytic turnover in P450cam requires the enzymes putidaredoxin (Pdx) and putidaredoxin reductase (Pdr), which mediate electron transfer from NADH to heme, the process is tightly coupled to substrate hydroxylation
-
-
?
additional information
?
-
-
under conditions of low oxygen, Pseudomonas putida cells and the isolated P450cam reduce camphor to borneol, product analysis by GC-MS
-
-
?
additional information
?
-
-
substrate specificities of wild-type and mutant enzymes, overview
-
-
?
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L253V
site-directed mutagenesis, the mutant shows 95% reduced activity compared to the wild-type enzyme, crystal structure analysis of mutant enzyme with bound substrate
M98F
site-directed mutagensis, the mutant shows 85% reduced activity compared to the wild-type enzyme, crystal structure analysis of mutant enzyme with bound substrate
Y96A
site-directed mutagensis, the mutant shows increased affinity for hydrocarbon substrates including adamantane, cyclooctane, hexane and 2-methylpentane, the monooxygenase activity of the mutant towards alkane substrates is enhanced compared to the wild-type enzyme, crystal structure analysis of mutant enzyme with bound substrate
C136A
-
altered NADH turnover rate
C136S
site-directed mutagenesis, putidaredoxin binding compared to wild-type
C148A
-
altered NADH turnover rate
C285S
site-directed mutagenesis, putidaredoxin binding compared to wild-type
C357M
-
site-directed mutagenesis, comparison of the mutant structure to the wild-type one
C357U/R365L/E366Q
site-directed mutagenesis, structural, electronic, and catalytic properties of cytochrome P450cam are subtly altered when the cysteine that coordinates to the heme iron is replaced with a selenocysteine, mapping of the effects of the sulfur-to-selenium substitution on the individual steps of the catalytic cycle. The more electron-donating selenolate ligand has only negligible effects on substrate, product, and oxygen binding, electron transfer, catalytic turnover, and coupling efficiency. Off-pathway reduction of oxygen to give superoxide is the only step significantly affected by the mutation. Incorporation of selenium accelerates this uncoupling reaction approximately 50fold compared to sulfur, but because the second electron transfer step is much faster, the impact on overall catalytic turnover is minimal. Quantum mechanical calculations, overview. Steady-state kinetic analysis revealed that the selenocysteine substitution has essentially no effect on the specific catalytic activity or the binding interaction with the electron donor Pdx, as both kcat and KM,Pdx are very similar for wild-type and mutant enzymes
C58A
-
altered NADH turnover rate
C58S
site-directed mutagenesis, putidaredoxin binding compared to wild-type
C85A
-
altered NADH turnover rate
C85S
site-directed mutagenesis, putidaredoxin binding compared to wild-type
D125A
site-directed mutagenesis
D251N
-
site-directed mutagenesis, the mutant shows altered conformation of the I helix groove and misses the catalytically important water molecules in the dioxygen complex leading to lower catalytic activity and slower proton transfer to the dioxygen ligand compared to the wild-type enzyme
D38A
-
site-directed mutagenesis, the mutant shows altered electron transfer activity with higher Kd values for ferric P450cam and about 20% of the first electron transferring ability compared to the wild-type enzyme, the mutant forms a complex with 1,3-dimethoxy-5-methyl-1,4-benzoquinone
D38N
-
site-directed mutagenesis, the mutant shows altered electron transfer activity with higher Kd values for ferric P450cam and about 20% of the first electron transferring ability compared to the wild-type enzyme
D97F/P122L/Q183L/L244Q
mutant isolated by Sequence Saturation Mutagenesis, converts 3-chloroindole to isatin
E14C/S29C/C85S/C73S
site-directed mutagenesis, putidaredoxin binding compared to wild-type
E156G/V247F/V253G/F256S
mutant isolated by Sequence Saturation Mutagenesis, shows the highest maximal velocity in the conversion of 3-chloroindole to isatin
E195C/A199C/C334A
site-directed mutagenesis, substrate and cofactor binding of the mutant compared to the wild-type, overview
E366Q
site-directed mutagenesis
F87W/Y96F
enhanced binding and oxidation of (+)-alpha-pinene
F87W/Y96F/L244A
enhanced binding and oxidation of (+)-alpha-pinene, production of 86% (+)-cis-verbenol + 5% (+)-verbenone
F87W/Y96F/L244A/V247L
enhanced binding and oxidation of (+)-alpha-pinene
G120A/Y179H/G248S/D297H
mutant isolated by Sequence Saturation Mutagenesis, converts 3-chloroindole to isatin
G248D
low catalytic activity
G248E
low catalytic activity, incubation with camphor, putidaredoxin reductase, and NADH results in partial covalent binding of heme to protein, pronase digestion of heme-bound protein releases 5-hydroxyheme
G326A
-
site-directed mutagenesis in order to decrease the flexibility of the polypeptide at that point, spin state fractions with different substrates and compared to the wild-type enzyme. The mutant shows 40% reduced activity compared to the wild-type enzyme
G60S/Y75H
mutant isolated by Sequence Saturation Mutagenesis, shows highest Km/kcat values for the conversion of 3-chloroindole to isatin
G93C/K314R/L319M
mutant isolated by Sequence Saturation Mutagenesis, converts 3-chloroindole to isatin
H352A
site-directed mutagenesis
H361A
site-directed mutagenesis
I396A
-
site-directed mutagenesis, the substrate specificity is altered compared to the wild-type enzyme
I396G
-
site-directed mutagenesis, the substrate specificity is altered compared to the wild-type enzyme
I396V
-
site-directed mutagenesis, the substrate specificity is altered compared to the wild-type enzyme
K344C
-
altered NADH turnover rate
L244A/C334A
site-directed mutagenesis, mutation C334A prevents adventitious dimerization to facilitate crystallization but has no further effect on structure or activity of the enzyme, the L244A mutation leads to a highly increased Km and reduced activity for imidazole, but not for for 1-methylimidazole, and altered binding of imidazole to the active site and the active site heme involving residue Val247, overview
L244F/V247L
site-directed mutagenesis, the mutant exhibits moderate to high R-selectivity toward ethylmethylbenzene substrates and shows a narrow width of the binding pocket
L244N/V247L
site-directed mutagenesis, the mutant displays the highest S-selectivity toward substrates 1-ethyl-2-methylbenzene and 1-ethyl-3-methylbenzene, and low R-selectivity toward 1-ethyl-4-methylbenzene and shows a narrow width of the binding pocket
M184V/T185F
site-directed mutagenesis, the mutation introduces changes above the heme plane, prefers S-orientation of 1-ethyl-4-methylbenzene in the binding pocket of mutant, enantioselectivities of 1-ethyl-2-methylbenzene and 1-ethyl-3-methylbenzene are similar to the wild-type enzyme
M395I
-
site-directed mutagenesis, the substrate specificity is altered compared to the wild-type enzyme
M96Y
-
site-directed mutagenesis, the substrate specificity is altered compared to the wild-type enzyme
N244L
-
site-directed mutagenesis, the substrate specificity is altered compared to the wild-type enzyme
P89I
-
yields a mixture of both bound camphor orientations, that seen in putidaredoxin-free and that seen in putidaredoxin-bound CYP101. A mutation in CYP101 that destabilizes the cis conformer of the Ile-88-Pro-89 amide bond results in weaker binding of putidaredoxin
R112C
-
altered NADH turnover rate
R364C
-
altered NADH turnover rate
R365L
site-directed mutagenesis
R66A
-
site-directed mutagenesis, reduced mutant electron transfer activity and increased Kd values for ferric P450cam compared to the wild-type enzyme
R66E
-
site-directed mutagenesis, reduced mutant electron transfer activity and increased Kd values for ferric P450cam compared to the wild-type enzyme
R72C
-
altered NADH turnover rate
S190D
does not show any significant change in the rate constants of the substrate association, has almost no effect on the activation energy of substrate binding to the enzyme
T101M
-
ratio of (R)- to (S)-1-phenylethanol produced from ethylbenzene is 89:11
T101M/T185F/V247M
-
ratio of (R)- to (S)-1-phenylethanol produced from ethylbenzene is 87:13
T101V
site-directed mutagenesis, the mutant shows decreased thermal stability of the heme active site and reaction intermediates in the reaction, equilibrium unfolding compared to the wild-type enzyme
T185F
-
ratio of (R)- to (S)-1-phenylethanol produced from ethylbenzene is 78:22
T185L
-
ratio of (R)- to (S)-1-phenylethanol produced from ethylbenzene is 80:20
T185V
-
ratio of (R)- to (S)-1-phenylethanol produced from ethylbenzene is 74:26
T192E
rate constants of the substrate association is much lower compared to the wild-type, activation energy for the substrate association is significantly higher in the T192E mutant compared to the S190D mutant or the wild-type enzyme
T252I
-
10% of wild-type activity
T252N
has comparable turnover number but higher Km value relative to the wild-type enzyme, due to a decrease in the camphor binding affinity, non-productive H2O2 generation is negligible
T252N/V253T
has comparable turnover number but higher Km value relative to the wild-type enzyme, due to a decrease in the camphor binding affinity, non-productive H2O2 generation is negligible
T297D
-
site-directed mutagenesis, the substrate specificity is altered compared to the wild-type enzyme
T56A/N116H/D297N
mutant isolated by Sequence Saturation Mutagenesis, shows highest Km/kcat values for the conversion of 3-chloroindole to isatin
V247A
-
ratio of (R)- to (S)-1-phenylethanol produced from ethylbenzene is 87:13
V247L
-
increased turnover rate for NADH
V247M
-
ratio of (R)- to (S)-1-phenylethanol produced from ethylbenzene is 83:17
V295I
-
ratio of (R)- to (S)-1-phenylethanol produced from ethylbenzene is 76:24
V87F
-
site-directed mutagenesis, the substrate specificity is altered compared to the wild-type enzyme
W106A
-
site-directed mutagenesis, the mutant shows altered electron transfer activity with higher Kd values for ferric P450cam and about 20% of the first electron transferring ability compared to the wild-type enzyme
W106F
-
site-directed mutagenesis, reduced mutant electron transfer activity and increased Kd values for ferric P450cam compared to the wild-type enzyme
Y179H
mutant isolated by Sequence Saturation Mutagenesis, converts 3-chloroindole to isatin
Y29F
-
the cis conformer is destabilized by the absence of the hydrogen bond between the carbonyl oxygen of Ile-88 and the Tyr-29 hydroxyl group
Y33A
-
site-directed mutagenesis, reduced mutant electron transfer activity and increased Kd values for ferric P450cam compared to the wild-type enzyme
Y33F
-
site-directed mutagenesis, reduced mutant electron transfer activity and increased Kd values for ferric P450cam compared to the wild-type enzyme
Y96A
-
site-directed mutagenesis, the mutant gains the ability to hydroxylate indole to 3-hydroxyindole
Y96C
-
site-directed mutagenesis, the mutant gains the ability to hydroxylate indole to 3-hydroxyindole
Y96C/C334A
site-directed mutagenesis, substrate recognition and binding compared to the wild-type, conformational selection mechanism
Y96F/C334A
site-directed mutagenesis, substrate recognition and binding compared to the wild-type, conformational selection mechanism
Y96F/L244A/V247L
enhanced binding and oxidation of (+)-alpha-pinene, production of 55% (+)-cis-verbenol + 32% (+)-verbenone
Y96F/V247L
enhanced binding and oxidation of (+)-alpha-pinene
Y96G
-
site-directed mutagenesis, the mutant gains the ability to hydroxylate indole to 3-hydroxyindole
Y96M
-
site-directed mutagenesis, the mutant gains the ability to hydroxylate indole to 3-hydroxyindole
Y96N/C334A
site-directed mutagenesis, substrate recognition and binding compared to the wild-type, conformational selection mechanism
Y96Q
-
site-directed mutagenesis, the mutant gains the ability to hydroxylate indole to 3-hydroxyindole
Y96S
-
site-directed mutagenesis, the mutant gains the ability to hydroxylate indole to 3-hydroxyindole
Y96T
-
site-directed mutagenesis, the mutant gains the ability to hydroxylate indole to 3-hydroxyindole
Y96Y
-
altered product spectrum
C357U/R365L/E366Q
-
site-directed mutagenesis, structural, electronic, and catalytic properties of cytochrome P450cam are subtly altered when the cysteine that coordinates to the heme iron is replaced with a selenocysteine, mapping of the effects of the sulfur-to-selenium substitution on the individual steps of the catalytic cycle. The more electron-donating selenolate ligand has only negligible effects on substrate, product, and oxygen binding, electron transfer, catalytic turnover, and coupling efficiency. Off-pathway reduction of oxygen to give superoxide is the only step significantly affected by the mutation. Incorporation of selenium accelerates this uncoupling reaction approximately 50fold compared to sulfur, but because the second electron transfer step is much faster, the impact on overall catalytic turnover is minimal. Quantum mechanical calculations, overview. Steady-state kinetic analysis revealed that the selenocysteine substitution has essentially no effect on the specific catalytic activity or the binding interaction with the electron donor Pdx, as both kcat and KM,Pdx are very similar for wild-type and mutant enzymes
-
E366Q
-
site-directed mutagenesis
-
R365L
-
site-directed mutagenesis
-
M96Y
-
site-directed mutagenesis, the substrate specificity is altered compared to the wild-type enzyme
-
N244L
-
site-directed mutagenesis, the substrate specificity is altered compared to the wild-type enzyme
-
T297D
-
site-directed mutagenesis, the substrate specificity is altered compared to the wild-type enzyme
-
V87F
-
site-directed mutagenesis, the substrate specificity is altered compared to the wild-type enzyme
-
C334A
-
identical with the wild-type monomer in terms of optical spectra, camphor-binding and turnover
C334A
spectroscopically and enzymatically identical to wild-type, but does not form dimers in solution
C334A
-
is identical spectroscopically and enzymatically to wild-type but does not dimerize in solution at high concentrations
C334A
-
mutant, that is spectroscopically and enzymatically identical to wild-type CYP101 enzyme, but does not form dimers in solution, and so is more suitable for solution NMR studies than wild-type enzyme
C334A
the mutation of P450cam increases the protein stability compared to the wild-type enzyme
C334A
-
the mutation of P450cam increases the protein stability compared to the wild-type enzyme
C334A
-
site-directed mutagenesis, the C334A mutant is spectroscopically and enzymatically identical to the wild type but does not form dimers in solution, and so is more suitable for NMR structure analysis than the wild type enzyme
C334A
site-directed mutagenesis, the mutation reduces protein aggregation, but has no effect on catalytic activity
C357U
site-directed mutagenesis, selenocysteine increases the affinity for oxygen 3-4-fold and accelerates the formation of superoxide 50fold, but the net effect of the C357U mutation on substrate hydroxylation is minimal because the second electron transfer step is much faster than superoxide formation under normal turnover conditions. As a consequence, selenocysteine is an excellent surrogate for the proximal cysteine in P450cam, maintaining both high monooxygenase activity and coupling efficiency
C357U
site-directed mutagenesis, the engineered gene contains the requisite UGA codon for selenocysteine, the sulfur-to-selenium substitution subtly modulates the structural, electronic, and catalytic properties of the enzyme. Catalytic activity decreases only 2fold, whereas substrate oxidation becomes partially uncoupled from electron transfer. The structure of mutant C357U, including the active site, is very similar to that of wild-type enzyme and mutant R365L/E366Q. The specific activity of the selenoenzyme mutant C357U is approximately half that of the mutant R365L/E366Q, which is 2fold less active than the wild-type enzyme
F87A/Y96F
-
altered product spectrum
F87A/Y96F
enhanced binding and oxidation of (+)-alpha-pinene
F87L/Y96F
-
altered product spectrum
F87L/Y96F
enhanced binding and oxidation of (+)-alpha-pinene
F87W/Y96F/V247L
-
enhanced activity for oxidation of 1,3,5-trichlorobenzene or (+)-alpha-pinene, compared to wild-type, analysis of active-site structure, crystallization
F87W/Y96F/V247L
enhanced binding and oxidation of (+)-alpha-pinene
L358P
-
stereo- and regioselectivity for d-camphor hydroxylation unchanged
L358P
-
in absence of putidaredoxin, mutant shows ring-current signals typical for wild-type enzyme in presence of putidaredoxin, heme-environment of mutant mimics that of the putidaredoxin-bound wild-type, mutant accepts nonphysiological electron donors dithionite and ascorbic acid
L358P
-
site-directed mutagenesis, spin-state equilibrium in the L358P mutant is more sensitive to K+ than the wild-type enzyme
Q227C
mutation used for double electron-electron resonance studies
Q227C
site-directed mutagenesis, putidaredoxin binding compared to wild-type
Q272C
mutation used for double electron-electron resonance studies
Q272C
site-directed mutagenesis, putidaredoxin binding compared to wild-type
R365L/E366Q
site-directed mutagenesis
R365L/E366Q
site-directed mutagenesis, the structure of mutant R365L/E366Q is very similar to that of wild-type enzyme and mutant C357U. The specific activity of the selenoenzyme mutant C357U is approximately half that of the mutant R365L/E366Q, which is 2fold less active than the wild-type enzyme
S190C
mutation used for double electron-electron resonance studies. Residues S48C and S190C are at opposite ends of the substrate access channel to provide a longer distance measurement
S190C
site-directed mutagenesis, putidaredoxin binding compared to wild-type
S48C
mutation used for double electron-electron resonance studies. Residues S48C and S190C are at opposite ends of the substrate access channel to provide a longer distance measurement
S48C
site-directed mutagenesis, putidaredoxin binding compared to wild-type
T252A
about 5% of wild-type activity, similar spectra for oxyferrous mutant and wild-type except for Soret band position blue shifts. Epoxidation substrate 5-methylenylcamphor has a anomalous binding mode for the mutant
T252A
-
site-directed mutagenesis, the mutant does not show altered conformation of the I helix groove and the catalytically important water molecules in the dioxygen complex
T252A
-
site-sirected mutagenesis, comparison of substrate binding properties to the wild-type enzyme, overview
T252A
-
the mutant can epoxidize olefins like 5-methylenyl-camphor, but is ineffective in camphor hydroxylation
T252A
non-productive H2O2 generation is dominant, does not oxidize camphor, substrate binding affinity is similar to that of the wild-type enzyme
T252A
mutant displays an additional coupling pathway responsible for the epoxidation of 5-methylenylcamphor. During the reaction, camphor cannot prevent H2O2 release and hence the T252A mutant does not oxidize camphor
Y75F
the mutant shows an altered active site structure influencing catalysis
Y75F
-
reaction with meta-chloroperbenzoic acid at 25°C, pH 8.0, is similar to that with the Y96F variant, although slightly more Cpd I (and possibly some Cpd ES) is present
Y96F
site-directed mutagenesis
Y96F
-
100fold increase of activity
Y96F
-
altered product spectrum
Y96F
enhanced binding and oxidation of (+)-alpha-pinene
Y96F
the mutant shows an altered active site structure influencing catalysis
Y96F
-
reaction with peracetic acid at pH 8.0, 25°C, is similar to that with meta-chloroperbenzoic acid, except that even with 2.4 mM peracetic acid, all steps are slower than those with 0.150 mM meta-chloroperbenzoic acid
Y96F
site-directed mutagenesis, population and dynamics of the conformational states are largely unaltered by the Y96F mutation compared to the wild-type enzyme
Y96F/Y75F
the mutant shows an altered active site structure influencing catalysis
Y96F/Y75F
-
mutants produce changes in hydrogen bonding patterns and increase hydrophobicity that affect the ratio of heterolytic to homolytic pathways in reactions with cumene hydroperoxide, resulting in a shift of this ratio from 84/16 for wild-type to 72/28 for the Y96F/Y75F double mutant
C357U
-
site-directed mutagenesis, selenocysteine increases the affinity for oxygen 3-4-fold and accelerates the formation of superoxide 50fold, but the net effect of the C357U mutation on substrate hydroxylation is minimal because the second electron transfer step is much faster than superoxide formation under normal turnover conditions. As a consequence, selenocysteine is an excellent surrogate for the proximal cysteine in P450cam, maintaining both high monooxygenase activity and coupling efficiency
-
C357U
-
site-directed mutagenesis, the engineered gene contains the requisite UGA codon for selenocysteine, the sulfur-to-selenium substitution subtly modulates the structural, electronic, and catalytic properties of the enzyme. Catalytic activity decreases only 2fold, whereas substrate oxidation becomes partially uncoupled from electron transfer. The structure of mutant C357U, including the active site, is very similar to that of wild-type enzyme and mutant R365L/E366Q. The specific activity of the selenoenzyme mutant C357U is approximately half that of the mutant R365L/E366Q, which is 2fold less active than the wild-type enzyme
-
R365L/E366Q
-
site-directed mutagenesis
-
R365L/E366Q
-
site-directed mutagenesis, the structure of mutant R365L/E366Q is very similar to that of wild-type enzyme and mutant C357U. The specific activity of the selenoenzyme mutant C357U is approximately half that of the mutant R365L/E366Q, which is 2fold less active than the wild-type enzyme
-
additional information
substitutions at Tyr96, Met98 and Leu253 in CYP101D2 analogously to closely related CYP101A1 from Pseudomonas putida, reduce both the spin state shift on camphor binding and the camphor oxidation activity
additional information
-
substitutions at Tyr96, Met98 and Leu253 in CYP101D2 analogously to closely related CYP101A1 from Pseudomonas putida, reduce both the spin state shift on camphor binding and the camphor oxidation activity
additional information
-
a facile way to significantly enhance the catalytic efficiency of the P450cam system by the coupling of its native electron transfer system with enzymatic NADH regeneration catalyzed by glycerol dehydrogenase in Escherichia coli whole cell biocatalysts, production of (+)-exo-5-hydroxycamphor and 5-keto-camphor, overview
additional information
-
construction of a catalytically active recombinant Escherichia coli whole cell biocatalyst harboring a cytochrome P450cam monooxygenase system from Pseudomonas putida coupled with enzymatic cofactor putidaredoxin mutant C73S/C85S regeneration, the cofactor mutant is 2fold more effective than the wild-type putidaredoxin
additional information
-
construction of the deletion mutant DELTA106, the mutant shows reduced electron transfer activity and increased Kd values for ferric P450cam compared to the wild-type enzyme
additional information
-
establishment of a functional CYP101-system in a cell-like aqueous compartment build by a stable water-oil emulsion with the non-ioninc surfactant tetraethylene glycol dodecyl ether in micro-scale, the enzyme is not active in an organic-aqueous biphasic system, e.g. with etahnol or glycol, incorporating an NADH-regeneration system using recombinant His-tagged bacterial glycerol dehydrogenase, method optimization, overview
additional information
-
in contrast to the wild-type, the more hydrophobic sites of the Tyr-to-Phe variants, compared to that of wild-type, favor homolysis more strongly and lead to considerable fractions of the Cpd II like species in reactions with peracids, especially at higher pH
additional information
removal of heme-7-propionate decreases the (+)-camphor affinity by approximately 3fold, but exerts less of an influence on the other steps of the catalytic cycle of the monooxygenation reaction catalyzed by P450cam, does not exert an influence on the structure and electronic properties of the heme
additional information
-
establishment of an in vitro screening system for P450cam selecting variants by the activity on NADH
additional information
-
generation of a G327 insertion mutant in order to determine whether the length of the helix played a role in the sensitivity of the K' helix to substrate, the insertion mutant fails to express
additional information
construction of a fusion enzyme composed of enzyme P450cam and putidaroxin reductase, i.e. P450cam-PdR, kinetic model based on two-site binding of putidaredoxin by P450cam-PdR and inactive dimer formation of the fusion protein. Fusion co-expression with putidaredoxin results in a functional system with in vivo camphor oxidation activity comparable to the wild-type system , but P450cam-PdR is a class I P450 fusion protein that exhibits significantly more favorable catalytic behavior than that of the wild-type system. Further oxidation of 5-exo-hydroxycamphor to 5-oxo-camphor by the fusion enzyme is 39% lower than for the native system
additional information
enantioselectivity of a library of active-site mutants of chimeric P450cam-RhFRed towards the benzylic hydroxylation of structurally related regioisomers of ethylmethylbenzene, computational molecular modeling, overview
additional information
interaction analysis of enzyme wild-type and mutant (D125A, H352A, and H361A) proteins with putidaredoxxin wild-type and mutant (Y33A, S42A, and S44A) proteins, kinetics, overview
additional information
-
establishment of an in vitro screening system for P450cam selecting variants by the activity on NADH
-
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Pseudomonas putida
brenda
Michizoe, J.; Ichinose, H.; Kamiya, N.; Maruyama, T.; Goto, M.
Functionalization of the cytochrome P450cam monooxygenase system in the cell-like aqueous compartments of water-in-oil emulsions
J. Biosci. Bioeng.
99
12-17
2005
Pseudomonas putida
brenda
Spolitak, T.; Dawson, J.H.; Ballou, D.P.
Rapid kinetics investigations of peracid oxidation of ferric cytochrome P450cam: nature and possible function of compound ES
J. Inorg. Biochem.
100
2034-2044
2006
Pseudomonas putida (P00183)
brenda
Hirao, H.; Kumar, D.; Shaik, S.
On the identity and reactivity patterns of the "second oxidant" of the T252A mutant of cytochrome P450cam in the oxidation of 5-methylenenylcamphor
J. Inorg. Biochem.
100
2054-2068
2006
Pseudomonas putida
brenda
Zurek, J.; Foloppe, N.; Harvey, J.N.; Mulholland, A.J.
Mechanisms of reaction in cytochrome P450: hydroxylation of camphor in P450cam
Org. Biomol. Chem.
4
3931-3937
2006
Pseudomonas putida
brenda
Verras, A.; Alian, A.; de Montellano, P.R.
Cytochrome P450 active site plasticity: attenuation of imidazole binding in cytochrome P450(cam) by an L244A mutation
Protein Eng. Des. Sel.
19
491-496
2006
Pseudomonas putida (P00183)
brenda
Sakurai, K.; Shimada, H.; Hayashi, T.; Tsukihara, T.
Substrate binding induces structural changes in cytochrome P450cam
Acta Crystallogr. Sect. F
65
80-83
2009
Pseudomonas putida (P00183)
brenda
Bell, S.G.; Dale, A.; Rees, N.H.; Wong, L.L.
A cytochrome P450 class I electron transfer system from Novosphingobium aromaticivorans
Appl. Microbiol. Biotechnol.
86
163-175
2010
Novosphingobium aromaticivorans
brenda
Kim, D.; Heo, Y.S.; Ortiz de Montellano, P.R.
Efficient catalytic turnover of cytochrome P450(cam) is supported by a T252N mutation
Arch. Biochem. Biophys.
474
150-156
2008
Pseudomonas putida (P00183)
brenda
Pochapsky, S.S.; Dang, M.; OuYang, B.; Simorellis, A.K.; Pochapsky, T.C.
Redox-dependent dynamics in cytochrome P450cam
Biochemistry
48
4254-4261
2009
Pseudomonas putida
brenda
Behera, R.K.; Mazumdar, S.
Roles of two surface residues near the access channel in the substrate recognition by cytochrome P450cam
Biophys. Chem.
135
1-6
2008
Pseudomonas putida (P00183)
brenda
Ryan, J.; Clark, D.
P450cam biocatalysis in surfactant-stabilized two-phase emulsions
Biotechnol. Bioeng.
99
1311-1319
2008
Pseudomonas putida
brenda
Kim, D.; Ortiz de Montellano, P.R.
Tricistronic overexpression of cytochrome P450cam, putidaredoxin, and putidaredoxin reductase provides a useful cell-based catalytic system
Biotechnol. Lett.
31
1427-1431
2009
Pseudomonas putida
brenda
Hayashi, T.; Harada, K.; Sakurai, K.; Shimada, H.; Hirota, S.
A role of the heme-7-propionate side chain in cytochrome P450cam as a gate for regulating the access of water molecules to the substrate-binding site
J. Am. Chem. Soc.
131
1398-1400
2009
Pseudomonas putida (P00183)
brenda
Spolitak, T.; Dawson, J.H.; Ballou, D.P.
Replacement of tyrosine residues by phenylalanine in cytochrome P450cam alters the formation of Cpd II-like species in reactions with artificial oxidants
J. Biol. Inorg. Chem.
13
599-611
2008
Pseudomonas putida
brenda
Altun, A.; Kumar, D.; Neese, F.; Thiel, W.
Multireference ab initio quantum mechanics/molecular mechanics study on intermediates in the catalytic cycle of cytochrome P450(cam)
J. Phys. Chem. A
112
12904-12910
2008
Pseudomonas putida
brenda
OuYang, B.; Pochapsky, S.S.; Dang, M.; Pochapsky, T.C.
A functional proline switch in cytochrome P450cam
Structure
16
916-923
2008
Pseudomonas putida
brenda
Lee, Y.T.; Wilson, R.F.; Rupniewski, I.; Goodin, D.B.
P450cam visits an open conformation in the absence of substrate
Biochemistry
49
3412-3419
2010
Pseudomonas putida (P00183), Pseudomonas putida
brenda
Asciutto, E.K.; Dang, M.; Pochapsky, S.S.; Madura, J.D.; Pochapsky, T.C.
Experimentally restrained molecular dynamics simulations for characterizing the open states of cytochrome P450cam
Biochemistry
50
1664-1671
2011
Pseudomonas putida (P00183), Pseudomonas putida
brenda
Hoffmann, G.; Boensch, K.; Greiner-Stoeffele, T.; Ballschmiter, M.
Changing the substrate specificity of P450cam towards diphenylmethane by semi-rational enzyme engineering
Protein Eng. Des. Sel.
24
439-446
2011
Pseudomonas putida, Pseudomonas putida PpG1
brenda
Bell, S.G.; Yang, W.; Dale, A.; Zhou, W.; Wong, L.L.
Improving the affinity and activity of CYP101D2 for hydrophobic substrates
Appl. Microbiol. Biotechnol.
97
3979-3990
2013
Novosphingobium aromaticivorans (Q2G8A2), Novosphingobium aromaticivorans
brenda
Yang, W.; Bell, S.; Wang, H.; Zhou, W.; Bartlam, M.; Wong, L.; Rao, Z.
The structure of CYP101D2 unveils a potential path for substrate entry into the active site
Biochem. J.
433
85-93
2011
Novosphingobium aromaticivorans (Q2G8A2), Novosphingobium aromaticivorans DSM 12444 (Q2G8A2)
brenda
Asciutto, E.K.; Young, M.J.; Madura, J.; Pochapsky, S.S.; Pochapsky, T.C.
Solution structural ensembles of substrate-free cytochrome P450(cam)
Biochemistry
51
3383-3393
2012
Pseudomonas putida
brenda
Hiruma, Y.; Gupta, A.; Kloosterman, A.; Olijve, C.; Olmez, B.; Hass, M.A.; Ubbink, M.
Hot-spot residues in the cytochrome p450cam-putidaredoxin binding interface
ChemBioChem
15
80-86
2014
Pseudomonas putida
brenda
Prasad, B.; Rojubally, A.; Plettner, E.
Identification of camphor oxidation and reduction products in Pseudomonas putida: new activity of the cytochrome P450cam system
J. Chem. Ecol.
37
657-667
2011
Pseudomonas putida, Pseudomonas putida ATCC 17453
brenda
Dang, M.; Pochapsky, S.; Pochapsky, T.
Spring-loading the active site of cytochrome P450 cam
Metallomics
3
339-343
2011
Pseudomonas putida
brenda
Kelly, P.; Eichler, A.; Herter, S.; Kranz, D.; Turner, N.; Flitsch, S.
Active site diversification of P450cam with indole generates catalysts for benzylic oxidation reactions
Beilstein J. Org. Chem.
11
1713-1720
2015
Pseudomonas putida (P00183)
brenda
Vandemeulebroucke, A.; Aldag, C.; Stiebritz, M.; Reiher, M.; Hilvert, D.
Kinetic consequences of introducing a proximal selenocysteine ligand into cytochrome P450cam
Biochemistry
54
6692-6703
2015
Pseudomonas putida (P00183), Pseudomonas putida ATCC 12633 (P00183)
brenda
Basom, E.; Manifold, B.; Thielges, M.
Conformational heterogeneity and the affinity of substrate molecular recognition by cytochrome P450cam
Biochemistry
56
3248-3256
2017
Pseudomonas putida (P00183)
brenda
Liou, S.H.; Myers, W.K.; Oswald, J.D.; Britt, R.D.; Goodin, D.B.
Putidaredoxin binds to the same site on cytochrome P450cam in the open and closed conformation
Biochemistry
56
4371-4378
2017
Pseudomonas putida (P00183)
brenda
Kammoonah, S.; Prasad, B.; Balaraman, P.; Mundhada, H.; Schwaneberg, U.; Plettner, E.
Selecting of a cytochrome P450cam SeSaM library with 3-chloroindole and endosulfan - Identification of mutants that dehalogenate 3-chloroindole
Biochim. Biophys. Acta
1866
68-79
2018
Pseudomonas putida (P00183), Pseudomonas putida
brenda
Johnson, E.O.; Wong, L.L.
Partial fusion of a cytochrome P450 system by carboxy-terminal attachment of putidaredoxin reductase to P450cam (CYP101A1)
Catal. Sci. Technol.
6
7549-7560
2016
Pseudomonas putida (P00183)
brenda
Hiruma, Y.; Gupta, A.; Kloosterman, A.; Olijve, C.; Oelmez, B.; Hass, M.; Ubbink, M.
Hot-spot residues in the cytochrome P450cam-putidaredoxin binding interface
ChemBioChem
15
80-86
2014
Pseudomonas putida (P00183)
brenda
Eichler, A.; Gricman, .; Herter, S.; Kelly, P.; Turner, N.; Pleiss, J.; Flitsch, S.
Enantioselective benzylic hydroxylation catalysed by P450 monooxygenases characterisation of a P450cam mutant library and molecular modelling
ChemBioChem
17
426-432
2016
Pseudomonas putida (P00183)
brenda
Franke, A.; van Eldik, R.
Spectroscopic and kinetic evidence for the crucial role of compound 0 in the P450cam-catalyzed hydroxylation of camphor by hydrogen peroxide
Chemistry
21
15201-15210
2015
Pseudomonas putida
brenda
Wang, B.; Li, C.; Dubey, K.D.; Shaik, S.
Quantum mechanical/molecular mechanical calculated reactivity networks reveal how cytochrome P450cam and its T252A mutant select their oxidation pathways
J. Am. Chem. Soc.
137
7379-7390
2015
Pseudomonas putida (P00183)
brenda
Basom, E.; Spearman, J.; Thielges, M.
Conformational landscape and the selectivity of cytochrome P450cam
J. Phys. Chem. B
119
6620-6627
2015
Pseudomonas putida (P00183)
brenda
Lai, R.; Li, H.
Hydrogen abstraction of camphor catalyzed by cytochrome P450cam A QM/MM study
J. Phys. Chem. B
120
12312-12320
2016
Pseudomonas putida (P00183)
brenda
Aldag, C.; Gromov, I.A.; Garcia-Rubio, I.; von Koenig, K.; Schlichting, I.; Jaun, B.; Hilvert, D.
Probing the role of the proximal heme ligand in cytochrome P450cam by recombinant incorporation of selenocysteine
Proc. Natl. Acad. Sci. USA
106
5481-5486
2009
Pseudomonas putida (P00183), Pseudomonas putida ATCC 12633 (P00183)
brenda
Skinner, S.; Liu, W.; Hiruma, Y.; Timmer, M.; Blok, A.; Hass, M.; Ubbink, M.
Delicate conformational balance of the redox enzyme cytochrome P450cam
Proc. Natl. Acad. Sci. USA
112
9022-9027
2015
Pseudomonas putida (P00183)
brenda
Rydzewski, J.; Nowak, W.
Thermodynamics of camphor migration in cytochrome P450cam by atomistic simulations
Sci. Rep.
7
7736
2017
Pseudomonas putida (P00183)
brenda
Murarka, V.C.; Batabyal, D.; Amaya, J.A.; Sevrioukova, I.F.; Poulos, T.L.
Unexpected differences between two closely related bacterial P450 camphor monooxygenases
Biochemistry
59
2743-2750
2020
Pseudomonas sp. TCU-HL1
brenda
Skinner, S.P.; Follmer, A.H.; Ubbink, M.; Poulos, T.L.; Houwing-Duistermaat, J.J.; Paci, E.
Partial opening of cytochrome P450cam (CYP101A1) is driven by allostery and putidaredoxin binding
Biochemistry
60
2932-2942
2021
Pseudomonas putida (P00183)
brenda