column_label: Table 3; row AoPOx 1,4-Benzoquinone; columns Km, kcat, and kcat/Km; footnote a pH 4.5.

row_label: 1,4-Benzoquinone / AoPOx


enzyme_display_name: galactose oxidase

organism: Hypomyces rosellus

substrate_name: D-galactose

source_db: brenda

verification_status: unverified

km_display: Not reported

ki_display: Not reported

kcat_over_km_display: Not reported

source_license: CC BY 4.0

sequence_variant_note: Mutation context is present, but no sequence was resolved for this record.

mutation_type: substitution

enzyme: galactose oxidase

substrate: D-galactose

status: unverified

source: brenda


mutation_signature: Unknown

sequence_variant_note: No sequence was resolved for this record, so variant-aware sequence handling could not be applied.

proof_lines:
native wild-type enzyme
recombinant wild-type enzyme


enzyme_display_name: aryl-alcohol oxidase

organism: Pseudomonas nitroreducens

verification_status: corrected

kcat_over_km_origin: calculated

km_unit: mM

sequence_variant_note: Sequence is treated as wild-type because the source explicitly marked the record as wild-type.

assay_conditions_summary: pH 7

column_label: TABLE 3. Steady-state kinetic constants for the oxidation of vanillyl alcohol, HMF, DFF, and FDCA by HMFOs; row FFCA / 5-formylfuran-2-carboxylate / PseniHMFOHis; columns kcat, Km, kcat/Km; kcat raw_value=0.05 +/- 0.01 raw_unit=min^-1; Km raw_value=0.6 +/- 0.3 raw_unit=mM; kcat/Km raw_value=0.09 +/- 0.05 raw_unit=mM^-1*min^-1

normalized_value_unit_evidence: kcat normalized_value=0.00083 normalized_unit=s^-1; Km normalized_value=0.6 normalized_unit=mM; kcat/Km normalized_value=0.0013833333333333334 normalized_unit=s^-1*mM^-1

data_origin: experimental

enzyme: aryl-alcohol oxidase

status: corrected


verification_status: verified

column_label: TABLE 3. Steady-state kinetic constants for the oxidation of vanillyl alcohol, HMF, DFF, and FDCA by HMFOs; row DFF / furan-2,5-dicarbaldehyde / PseniHMFOHis; columns kcat, Km, kcat/Km; kcat raw_value=1.0 +/- 0.1 raw_unit=min^-1; Km raw_value=4.8 +/- 1.9 raw_unit=mM; kcat/Km raw_value=0.2 +/- 0.08 raw_unit=mM^-1*min^-1

normalized_value_unit_evidence: kcat normalized_value=0.017 normalized_unit=s^-1; Km normalized_value=4.8 normalized_unit=mM; kcat/Km normalized_value=0.003541666666666667 normalized_unit=s^-1*mM^-1

status: verified


column_label: TABLE 3. Steady-state kinetic constants for the oxidation of vanillyl alcohol, HMF, DFF, and FDCA by HMFOs; row HMF / 5-(hydroxymethyl)furfural / PseniHMFOHis; columns kcat, Km, kcat/Km; kcat raw_value=14.3 +/- 0.9 raw_unit=min^-1; Km raw_value=5.1 +/- 1.1 raw_unit=mM; kcat/Km raw_value=2.8 +/- 0.6 raw_unit=mM^-1*min^-1

normalized_value_unit_evidence: kcat normalized_value=0.24 normalized_unit=s^-1; Km normalized_value=5.1 normalized_unit=mM; kcat/Km normalized_value=0.047058823529411764 normalized_unit=s^-1*mM^-1


enzyme_display_name: pyranose oxidase

organism: Pseudarthrobacter siccitolerans

substrate_name: D-glucose

proof_lines: with O2 as electron acceptor, at pH 6.5 and 37°C

proof_lines: with 1,4-benzoquinone as electron acceptor, at pH 6.5 and 37°C

enzyme: pyranose oxidase

substrate: D-glucose


substrate_name: L-arabinose

substrate: L-arabinose


proof_lines: with D-glucose as cosubstrate, at pH 6.5 and 37°C


enzyme_display_name: alcohol oxidase

organism: Ogataea angusta

substrate_name: formaldehyde

smiles: C=O

proof_lines:
pH 7.5, 30°C
cosubstrate: p-benzoquinone
cosubstrate: O2

enzyme: alcohol oxidase

substrate: formaldehyde


substrate_name: ethanol

smiles: CCO

proof_lines:
pH 7.5, 30°C
free enzyme
immobilized enzyme

substrate: ethanol


substrate_name: methanol

smiles: CO

substrate: methanol


organism: Paecilomyces variotii

kcat_display: Not reported


organism: Methylovorus sp. MP688

sequence_variant_note: Sequence identity was resolved, but no explicit wild-type or mutant call was captured.

column_label: TABLE 3. Steady-state kinetic constants for the oxidation of vanillyl alcohol, HMF, DFF, and FDCA by HMFOs; row DFF / furan-2,5-dicarbaldehyde / MetspHMFO; columns kcat, Km, kcat/Km; kcat raw_value=6.1 +/- 0.2 raw_unit=min^-1; Km raw_value=6.8 +/- 0.6 raw_unit=mM; kcat/Km raw_value=0.9 +/- 0.1 raw_unit=mM^-1*min^-1

normalized_value_unit_evidence: kcat normalized_value=0.1 normalized_unit=s^-1; Km normalized_value=6.8 normalized_unit=mM; kcat/Km normalized_value=0.014705882352941178 normalized_unit=s^-1*mM^-1


column_label: TABLE 3. Steady-state kinetic constants for the oxidation of vanillyl alcohol, HMF, DFF, and FDCA by HMFOs; row HMF / 5-(hydroxymethyl)furfural / MetspHMFO; columns kcat, Km, kcat/Km; kcat raw_value=82.5 +/- 1.6 raw_unit=min^-1; Km raw_value=9.1 +/- 0.7 raw_unit=mM; kcat/Km raw_value=9.1 +/- 0.8 raw_unit=mM^-1*min^-1

normalized_value_unit_evidence: kcat normalized_value=1.38 normalized_unit=s^-1; Km normalized_value=9.1 normalized_unit=mM; kcat/Km normalized_value=0.15164835164835164 normalized_unit=s^-1*mM^-1


organism: Paenibacillus sp. AIU 311

substrate_name: glyceraldehyde

smiles: C(C(C=O)O)O

substrate: glyceraldehyde


substrate_name: glycolaldehyde

smiles: C(C=O)O

substrate: glycolaldehyde


enzyme_display_name: L-sorbose oxidase

organism: Trametes sanguinea

proof_lines:
acceptor O2
acceptor 2,6-dichlorophenolindophenol

enzyme: L-sorbose oxidase


substrate_name: L-sorbose

substrate: L-sorbose


enzyme_display_name: pyridoxine 4-oxidase

organism: Pseudomonas sp.

enzyme: pyridoxine 4-oxidase


substrate_name: pyridoxine

substrate: pyridoxine


organism: Mesorhizobium loti

sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature H460A to reconstruct the assayed variant sequence.

assay_conditions_summary: pH 8

proof_lines: pH 8.0, 30°C, recombinant His6-tagged mutant H460A


sequence_variant_note: Resolved sequence already matches the mutant residue(s), so it is treated as a source-provided variant sequence.

proof_lines: pH 8.0, 30°C, recombinant His6-tagged mutant H462A


proof_lines: pH 8.0, 30°C, recombinant His6-tagged wild-type enzyme


organism: Levilactobacillus brevis

substrate_name: Cyclohexanol

proof_lines: fusion protein with phenylacetone monooxygenase, pH 7.5, 25°C

substrate: Cyclohexanol


smiles: O=O


organism: Fungi

substrate_name: hexadecanol

smiles: CCCCCCCCCCCCCCCCO

substrate: hexadecanol


organism: Penicillium purpurescens

substrate_name: ethylene glycol

smiles: C(CO)O

substrate: ethylene glycol


organism: Thermoascus aurantiacus

proof_lines:
extracellular enzyme, pH 5.9, 30°C
intracellular enzyme, pH 5.9, 30°C


organism: Aspergillus terreus

substrate_name: n-butanol

smiles: CCCCO

substrate: n-butanol


substrate_name: n-Octanol

smiles: CCCCCCCCO

substrate: n-Octanol


organism: Aspergillus ochraceus


organism: Thermoanaerobacter brockii


protein_accession_database: UniProt


enzyme_display_name: cholesterol oxidase

organism: gamma proteobacterium Y-134

substrate_name: cholesterol

assay_conditions_summary: pH 6

enzyme: cholesterol oxidase

substrate: cholesterol


organism: Streptomyces aegyptia


organism: Schizophyllum commune


organism: Bordetella sp.


organism: Pleurotus ostreatus


organism: Komagataella pastoris

substrate_name: (R,S)-4-methoxybenzyl alcohol

proof_lines: protein fused to peroxidase, linker (GGGGS)12, pH 4, 25°C

proof_lines: protein fused to peroxidase, linker (GGGGS)9, pH 4, 25°C

proof_lines: protein fused to peroxidase, linker (AP)5(GGGGS)1, pH 4, 25°C

substrate: (R,S)-4-methoxybenzyl alcohol


organism: Lentinus sajor-caju

substrate_name: 3,4-dimethoxybenzyl alcohol

proof_lines:
enzyme form VAO II
enzyme form VAO I

substrate: 3,4-dimethoxybenzyl alcohol


organism: Bacillus subtilis


organism: Phanerodontia chrysosporium

substrate_name: Vanillyl alcohol

substrate: Vanillyl alcohol


organism: Phratora vitellinae

substrate_name: 2-hydroxybenzyl alcohol

substrate: 2-hydroxybenzyl alcohol


substrate_name: veratryl alcohol

substrate: veratryl alcohol


organism: Trametes versicolor

substrate_name: 4-methoxybenzyl alcohol

substrate: 4-methoxybenzyl alcohol


enzyme_display_name: L-gulonolactone oxidase

organism: Grifola frondosa

enzyme: L-gulonolactone oxidase


organism: Hypomyces rosellus

substrate_name: 1-O-methyl beta-D-galactopyranoside

proof_lines: Cu(II)-bound H2O is required in the protonated aquo state for catalysis and is responsible for the kinetic solvent isotope effect

substrate: 1-O-methyl beta-D-galactopyranoside


proof_lines: substrate 1-O-methyl-alpha-D-galactopyranoside, 2 H-atoms in position 6 are substituted by D-atoms

proof_lines: substrate oxygen at saturated 1-O-methyl-alpha-D-galactopyranoside concentration, 2 H-atoms in position 6 are substituted by D-atoms


organism: Aspergillus nidulans

substrate_name: O2; D-Galactose

source_license: SABIO-RK terms (free for academic use; see sabiork.h-its.org)

substrate: O2; D-Galactose


organism: Mycobacterium tuberculosis

assay_conditions_summary: cofactor FAD

proof_lines: the FAD-binding motif Rv1771 protein is a metalloenzyme that oxidizes L-gulono-1,4-lactone


substrate_name: D-galactose

substrate: D-galactose


organism: Fusarium acuminatum


organism: Gloeophyllum sepiarium

substrate_name: D-glucose

substrate: D-glucose


organism: Lenzites styracinus


organism: Fusarium sambucinum

substrate_name: melibiose

proof_lines: recombinant His-tagged enzyme, pH 7.0, 30°C

substrate: melibiose


organism: Trametes ochracea

sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature V546C/T169G to reconstruct the assayed variant sequence.

proof_lines: V546C/T169G mutant, substrate 1,4-benzoquinone (constant D-glucose concentration, 100 mM), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-azinobis(

proof_lines: V546C/T169G mutant, substrate 1,4-benzoquinone (constant D-galactose concentration, 100 mM), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-azinobi


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature V546C/T169G/L537W to reconstruct the assayed variant sequence.

proof_lines: V546C/T169G/L537W mutant, substrate 1,4-benzoquinone (constant D-glucose concentration, 100 mM), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-azi

proof_lines: V546C/T169G/L537W mutant, substrate 1,4-benzoquinone (constant D-galactose concentration, 100 mM), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-a


substrate_name: ferricenium hexafluorophosphate

proof_lines: V546C/T169G/L537W mutant, substrate ferricenium hexafluorophosphate (constant D-glucose concentration, 100 mM), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay

proof_lines: mutant enzyme V546C/T169G/L537W, at pH 6.5 and 30°C

proof_lines: V546C/T169G/L537W mutant, substrate ferricenium hexafluorophosphate (constant D-galactose concentration, 100 mM), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled ass

substrate: ferricenium hexafluorophosphate


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature V546C/E542K to reconstruct the assayed variant sequence.

proof_lines: V546C/E542K mutant, substrate ferricenium hexafluorophosphate (constant D-galactose concentration, 100 mM), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay usi

proof_lines: mutant enzyme V546C/E542K, at pH 6.5 and 30°C

proof_lines: V546C/E542K mutant, substrate ferricenium hexafluorophosphate (constant D-glucose concentration, 100 mM), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using


proof_lines: V546C/T169G mutant, substrate ferricenium hexafluorophosphate (constant D-galactose concentration, 100 mM), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay usi

proof_lines: mutant enzyme V546C/T169G, at pH 6.5 and 30°C

proof_lines: V546C/T169G mutant, substrate ferricenium hexafluorophosphate (constant D-glucose concentration, 100 mM), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using


proof_lines: V546C/E542K mutant, substrate 1,4-benzoquinone (constant D-glucose concentration, 100 mM), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-azinobis(

proof_lines: V546C/E542K mutant, substrate 1,4-benzoquinone (constant D-galactose concentration, 100 mM), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-azinobi


proof_lines: V546C/T169G mutant, substrate D-galactose (constant O2 concentration), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-azinobis(3-ethylbenzthiazolin

proof_lines: V546C/T169G mutant, substrate D-glucose (constant O2 concentration), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-azinobis(3-ethylbenzthiazolines


proof_lines: V546C/T169G/L537W mutant, substrate D-glucose (constant O2 concentration), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-azinobis(3-ethylbenzthiaz

proof_lines: V546C/T169G/L537W mutant, substrate D-galactose (constant O2 concentration), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-azinobis(3-ethylbenzthi


proof_lines: V546C/E542K mutant, substrate D-glucose (constant O2 concentration), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-azinobis(3-ethylbenzthiazolines

proof_lines: V546C/E542K mutant, substrate D-galactose (constant O2 concentration), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-azinobis(3-ethylbenzthiazolin


proof_lines: substrate 2-methyl-1,4-benzoquinone (constant D-glucose concentration, 20 mM), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-azinobis(3-ethylbenzt


proof_lines: substrate 2-chloro-1,4-benzoquinone (constant D-glucose concentration, 20 mM), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-azinobis(3-ethylbenzt


substrate_name: ferricyanide

proof_lines: substrate ferricyanide (constant D-glucose concentration, 20 mM), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-azinobis(3-ethylbenzthiazolinesulf

substrate: ferricyanide


substrate_name: D-fructose

sequence_source: multiple

proof_lines: substrate D-fructose (constant O2 concentration), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) as th

proof_lines: substrate L-sorbose (constant O2 concentration), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) as the

source_databases_merged: brenda uniprot

substrate: D-fructose


source_license: CC BY 4.0; CC BY-NC-ND 4.0

proof_lines: substrate D-galactose (constant O2 concentration), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) as t

proof_lines: substrate D-glucose (constant O2 concentration), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) as the

source_databases_merged: brenda skid uniprot


proof_lines: substrate tetrafluoro-1,4-benzoquinone (constant D-glucose concentration, 20 mM), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-azinobis(3-ethylbe


proof_lines: substrate 2,6-dimethyl-1,4-benzoquinone (constant D-glucose concentration, 20 mM), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-azinobis(3-ethylb


proof_lines: substrate 2-methoxy-1,4-benzoquinone (constant D-glucose concentration, 20 mM), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-azinobis(3-ethylbenz


substrate_name: D-xylose

proof_lines: substrate D-xylose (constant O2 concentration), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) as the

substrate: D-xylose


proof_lines: substrate 2,6-dichloroindophenol (constant D-glucose concentration, 20 mM), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-azinobis(3-ethylbenzthia


proof_lines: substrate O2 (constant D-glucose concentration, 20 mM), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid)


proof_lines: substrate ferricenium hexafluorophosphate (constant D-glucose concentration, 20 mM), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-azinobis(3-ethy


proof_lines: substrate 1,4-benzoquinone (constant D-glucose concentration, 20 mM), activity determined spectrophotometrically at 420 nm by measuring formation of H2O2 with a horse-radish peroxidase-coupled assay using 2,2'-azinobis(3-ethylbenzthiazoline


sequence_variant_note: Mutation signature F454A/S455A/Y456A/H2P could not be applied safely because the resolved sequence does not match the expected reference residue(s).

assay_conditions_summary: pH 6.5; buffer 50 mM KH2PO4

proof_lines: mutant enzyme F454A/S455A/Y456A, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C


substrate_name: D-melibiose

substrate: D-melibiose


sequence_variant_note: Mutation signature F454P/H2P could not be applied safely because the resolved sequence does not match the expected reference residue(s).

proof_lines: mutant enzyme F454P, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C


sequence_variant_note: Mutation signature F454A/Y456A/H2P could not be applied safely because the resolved sequence does not match the expected reference residue(s).

proof_lines: mutant enzyme F454A/Y456A, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C


sequence_variant_note: Resolved sequence already matches the mutant residue(s) after applying an inferred residue-numbering offset +29, so it is treated as a source-provided variant sequence.

proof_lines: mutant enzyme F454N, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C


proof_lines: mutant enzyme F454A/S455A/Y456A, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C

proof_lines: mutant enzyme F454A/S455A/Y456A, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C


sequence_variant_note: Resolved sequence already matches the mutant residue(s) after applying an inferred residue-numbering offset +30, so it is treated as a source-provided variant sequence.

proof_lines: mutant enzyme Y456W, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C


sequence_variant_note: Mutation signature H450Q/H2P could not be applied safely because the resolved sequence does not match the expected reference residue(s).

proof_lines: mutant enzyme H450Q, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C


proof_lines: mutant enzyme F454P, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C

proof_lines: mutant enzyme F454P, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C


proof_lines: mutant enzyme F454N, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C

proof_lines: mutant enzyme F454N, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C


proof_lines: mutant enzyme F454A/Y456A, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C

proof_lines: mutant enzyme F454A/Y456A, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C


substrate_name: ferrocenium hexafluorophosphate

substrate: ferrocenium hexafluorophosphate


proof_lines: mutant enzyme H450Q, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C

proof_lines: mutant enzyme H450Q, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C


proof_lines: mutant enzyme Y456W, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C

proof_lines: mutant enzyme Y456W, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C


proof_lines: wild type enzyme, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C

proof_lines: wild-type, substrate 1,4-benzoquinone (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C

proof_lines: wild-type, substrate 1,4-benzoquinone (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C


proof_lines: mutant enzyme F454A/S455A/Y456A, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C


proof_lines: wild type enzyme, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature E542R to reconstruct the assayed variant sequence.

proof_lines: E542R mutant, substrate D-galactose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C

proof_lines: E542R mutant, substrate D-glucose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature L537G/E542K to reconstruct the assayed variant sequence.

proof_lines: L537G/E542K mutant, substrate D-galactose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C

proof_lines: L537G/E542K mutant, substrate D-glucose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature L537G/E542R to reconstruct the assayed variant sequence.

proof_lines: L537G/E542R mutant, substrate D-galactose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C

proof_lines: L537G/E542R mutant, substrate D-glucose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature L537W/E542R to reconstruct the assayed variant sequence.

proof_lines: L537W/E542R mutant, substrate D-galactose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C

proof_lines: L537W/E542R mutant, substrate D-glucose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature L537W/E542K to reconstruct the assayed variant sequence.

proof_lines: L537W/E542K mutant, substrate D-galactose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C

proof_lines: L537W/E542K mutant, substrate D-glucose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature L537G to reconstruct the assayed variant sequence.

proof_lines: L537G mutant, substrate D-galactose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C

proof_lines: L537G mutant, substrate D-glucose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature E542K to reconstruct the assayed variant sequence.

assay_conditions_summary: pH 6.5; buffer 50 mM phosphate

proof_lines: E542K mutant, substrate D-galactose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C

proof_lines: mutant enzyme E542K, in 50 mM phosphate buffer (pH 6.5) at 60°C

proof_lines: E542K mutant, substrate D-glucose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature L537W to reconstruct the assayed variant sequence.

proof_lines: L537W mutant, substrate D-galactose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C

proof_lines: L537W mutant, substrate D-glucose (O2 concentration constant), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C


proof_lines: E542R mutant, substrate 1,4-benzoquinone (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C

proof_lines: E542R mutant, substrate 1,4-benzoquinone (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C


proof_lines: L537G mutant, substrate 1,4-benzoquinone (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C

proof_lines: L537G mutant, substrate 1,4-benzoquinone (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C


proof_lines: L537G/E542K mutant, substrate 1,4-benzoquinone (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5

proof_lines: L537G/E542K mutant, substrate 1,4-benzoquinone (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5,


proof_lines: L537G/E542R mutant, substrate 1,4-benzoquinone (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5

proof_lines: L537G/E542R mutant, substrate 1,4-benzoquinone (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5,


proof_lines: L537W/E542R mutant, substrate 1,4-benzoquinone (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5

proof_lines: L537W/E542R mutant, substrate 1,4-benzoquinone (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5,


proof_lines: L537W/E542K mutant, substrate 1,4-benzoquinone (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5

proof_lines: L537W/E542K mutant, substrate 1,4-benzoquinone (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5,


proof_lines: L537W mutant, substrate 1,4-benzoquinone (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C

proof_lines: L537W mutant, substrate 1,4-benzoquinone (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C


proof_lines: E542K mutant, substrate 1,4-benzoquinone (D-galactose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C

proof_lines: E542K mutant, substrate 1,4-benzoquinone (D-glucose concentration constant, 100 mM), standard chromogenic ABTS assay (azino-bis-(3-ethylbenzthiazolin-6-sulfonic acid), horse-radish peroxidase, measuring absorbtion at 420 nm), pH 6.5, 30°C


sequence_variant_note: Sequence is treated as wild-type because the source explicitly marked the record as wild-type.

proof_lines: wild type enzyme, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C


proof_lines: mutant enzyme H450Q, using D-galactose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C

proof_lines: mutant enzyme H450Q, using D-glucose as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C


organism: Phlebiopsis gigantea

sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_mutation_ranked_match and applied mutation signature K312E to reconstruct the assayed variant sequence.

proof_lines: pH 7.3, 22°C, mutant enzyme K312E with a C-terminal His6-tag


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_mutation_ranked_match and applied mutation signature E540K to reconstruct the assayed variant sequence.

proof_lines: pH 7.3, 22°C, mutant enzyme E540K with a C-terminal His6-tag


substrate_name: L-sorbose

substrate: L-sorbose


smiles: C(C=O)C(C(C(CO)O)O)O


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature V546G/T169G to reconstruct the assayed variant sequence.

proof_lines: V546G/T169G, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay

proof_lines: V546G/T169G, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature V546P/T169G to reconstruct the assayed variant sequence.

proof_lines: V546P/T169G, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay

proof_lines: V546P/T169G, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature H548R with inferred residue-numbering offset +5 to reconstruct the assayed variant sequence.

proof_lines: H548R, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay

proof_lines: H548R, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature Q448N to reconstruct the assayed variant sequence.

proof_lines: Q448N, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay

proof_lines: Q448N, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature Q448C to reconstruct the assayed variant sequence.

proof_lines: Q448C, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay

proof_lines: Q448C, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature V546C/T169N to reconstruct the assayed variant sequence.

proof_lines: V546C/T169N, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay

proof_lines: V546C/T169N, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature Q448S to reconstruct the assayed variant sequence.

proof_lines: Q448S, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay

proof_lines: Q448S, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature N593R with inferred residue-numbering offset -8 to reconstruct the assayed variant sequence.

proof_lines: N593R, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay

proof_lines: N593R, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature V546P to reconstruct the assayed variant sequence.

proof_lines: V546P, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay

proof_lines: V546P, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature R472L to reconstruct the assayed variant sequence.

proof_lines: R472L, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay

proof_lines: R472L, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature R472G to reconstruct the assayed variant sequence.

proof_lines: R472G, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay

proof_lines: R472G, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature H548I with inferred residue-numbering offset +5 to reconstruct the assayed variant sequence.

proof_lines: H548I, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay

proof_lines: H548I, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature V546G to reconstruct the assayed variant sequence.

proof_lines: V546G, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay

proof_lines: V546G, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature V546C to reconstruct the assayed variant sequence.

proof_lines: V546C, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay

proof_lines: mutant enzyme V546C, in 50 mM phosphate buffer (pH 6.5) at 60°C

proof_lines: mutant enzyme V546C, pH and temperature not specified in the publication


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature M103S to reconstruct the assayed variant sequence.

smiles: CC(CO)O

column_label: Table 3 row (R)-1,2-Propanediol; column group M103S; metric columns KM, kcat, kcat/KM.

normalized_value_unit_evidence: kcat=0.4 s^-1; KM=110 mM; kcat_over_km=0.0036363636363636364 s^-1 mM^-1 from Table 3 kcat/KM; printed table cell 4 M^-1 s^-1 (= 0.004 s^-1 mM^-1) is rounded

raw_value_unit_evidence: Table 3 row (R)-1,2-Propanediol; M103S columns: KM=110 mM, kcat=0.4 s^-1, kcat/KM=4 M^-1 s^-1.

table_label: Table 3 - Apparent Steady-State Kinetic Parameters for PcAOX


substrate_name: methanol

column_label: Table 3 row methanol; column group M103S; metric columns KM, kcat, kcat/KM.

normalized_value_unit_evidence: kcat=1 s^-1; KM=2 mM; kcat_over_km=0.5 s^-1 mM^-1 from Table 3 kcat/KM; printed table cell 500 M^-1 s^-1 (= 0.5 s^-1 mM^-1) is rounded

raw_value_unit_evidence: Table 3 row methanol; M103S columns: KM=2 mM, kcat=1 s^-1, kcat/KM=500 M^-1 s^-1.

row_label: methanol

substrate: methanol


substrate_name: ethanol

column_label: Table 3 row ethanol; column group M103S; metric columns KM, kcat, kcat/KM.

normalized_value_unit_evidence: kcat=1 s^-1; KM=1 mM; kcat_over_km=1.0 s^-1 mM^-1 from Table 3 kcat/KM; printed table cell 1000 M^-1 s^-1 (= 1.0 s^-1 mM^-1) is rounded

raw_value_unit_evidence: Table 3 row ethanol; M103S columns: KM=1 mM, kcat=1 s^-1, kcat/KM=1000 M^-1 s^-1.

row_label: ethanol

substrate: ethanol


smiles: CCCO

column_label: Table 3 row 1-propanol; column group M103S; metric columns KM, kcat, kcat/KM.

raw_value_unit_evidence: Table 3 row 1-propanol; M103S columns: KM=2 mM, kcat=1 s^-1, kcat/KM=500 M^-1 s^-1.


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature F101S to reconstruct the assayed variant sequence.

column_label: Table 3 row methanol; column group F101S; metric columns KM, kcat, kcat/KM.

normalized_value_unit_evidence: kcat=4 s^-1; KM=2 mM; kcat_over_km=2.0 s^-1 mM^-1 from Table 3 kcat/KM; printed table cell 2000 M^-1 s^-1 (= 2.0 s^-1 mM^-1) is rounded

raw_value_unit_evidence: Table 3 row methanol; F101S columns: KM=2 mM, kcat=4 s^-1, kcat/KM=2000 M^-1 s^-1.


column_label: Table 3 row (R)-1,2-Propanediol; column group F101S; metric columns KM, kcat, kcat/KM.

normalized_value_unit_evidence: kcat=4 s^-1; KM=110 mM; kcat_over_km=0.03636363636363636 s^-1 mM^-1 from Table 3 kcat/KM; printed table cell 36 M^-1 s^-1 (= 0.036 s^-1 mM^-1) is rounded

raw_value_unit_evidence: Table 3 row (R)-1,2-Propanediol; F101S columns: KM=110 mM, kcat=4 s^-1, kcat/KM=36 M^-1 s^-1.


sequence_variant_note: Sequence is treated as wild-type because the source explicitly marked the record as wild-type.

column_label: PMID 30272958 Table 3; row (R)-1,2-Propanediol; column group wild-type; KM/kcat/kcat/KM cells: KM >1600 mM; kcat >5 s^-1; kcat/KM >3 M^-1 s^-1.

normalized_value_unit_evidence: KM >1600 mM; kcat >5 s^-1; kcat/KM >0.003 s^-1 mM^-1 (inequality/lower-bound cells)

row_label: (R)-1,2-Propanediol / wild-type

table_label: PMID 30272958 Table 3. Apparent Steady-State Kinetic Parameters for PcAOX


column_label: Table 3 row ethanol; column group F101S; metric columns KM, kcat, kcat/KM.

normalized_value_unit_evidence: kcat=5 s^-1; KM=1 mM; kcat_over_km=5.0 s^-1 mM^-1 from Table 3 kcat/KM; printed table cell 5000 M^-1 s^-1 (= 5.0 s^-1 mM^-1) is rounded

raw_value_unit_evidence: Table 3 row ethanol; F101S columns: KM=1 mM, kcat=5 s^-1, kcat/KM=5000 M^-1 s^-1.


column_label: Table 3 row 1-propanol; column group F101S; metric columns KM, kcat, kcat/KM.

normalized_value_unit_evidence: kcat=7 s^-1; KM=4 mM; kcat_over_km=1.75 s^-1 mM^-1 from Table 3 kcat/KM; printed table cell 1800 M^-1 s^-1 (= 1.8 s^-1 mM^-1) is rounded

raw_value_unit_evidence: Table 3 row 1-propanol; F101S columns: KM=4 mM, kcat=7 s^-1, kcat/KM=1800 M^-1 s^-1.


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature F101N to reconstruct the assayed variant sequence.

column_label: Table 3 row methanol; column group F101N; metric columns KM, kcat, kcat/KM.

normalized_value_unit_evidence: kcat=10 s^-1; KM=14 mM; kcat_over_km=0.7142857142857143 s^-1 mM^-1 from Table 3 kcat/KM; printed table cell 710 M^-1 s^-1 (= 0.71 s^-1 mM^-1) is rounded

raw_value_unit_evidence: Table 3 row methanol; F101N columns: KM=14 mM, kcat=10 s^-1, kcat/KM=710 M^-1 s^-1.


column_label: Table 3 row ethanol; column group F101N; metric columns KM, kcat, kcat/KM.

normalized_value_unit_evidence: kcat=10 s^-1; KM=11 mM; kcat_over_km=0.9090909090909091 s^-1 mM^-1 from Table 3 kcat/KM; printed table cell 910 M^-1 s^-1 (= 0.91 s^-1 mM^-1) is rounded

raw_value_unit_evidence: Table 3 row ethanol; F101N columns: KM=11 mM, kcat=10 s^-1, kcat/KM=910 M^-1 s^-1.


column_label: PMID 30272958 Table 3; row 1-propanol; column group wild-type; KM/kcat/kcat/KM cells: KM 45 mM; kcat 11 s^-1; kcat/KM 240 M^-1 s^-1.

row_label: 1-propanol / wild-type


column_label: Table 3 row 1-propanol; column group F101N; metric columns KM, kcat, kcat/KM.

normalized_value_unit_evidence: kcat=12 s^-1; KM=11 mM; kcat_over_km=1.0909090909090908 s^-1 mM^-1 from Table 3 kcat/KM; printed table cell 1100 M^-1 s^-1 (= 1.1 s^-1 mM^-1) is rounded

raw_value_unit_evidence: Table 3 row 1-propanol; F101N columns: KM=11 mM, kcat=12 s^-1, kcat/KM=1100 M^-1 s^-1.


column_label: Table 3 row (R)-1,2-Propanediol; column group F101N; metric columns KM, kcat, kcat/KM.

normalized_value_unit_evidence: kcat=13 s^-1; KM=1200 mM; kcat_over_km=0.010833333333333334 s^-1 mM^-1 from Table 3 kcat/KM; printed table cell 11 M^-1 s^-1 (= 0.011 s^-1 mM^-1) is rounded

raw_value_unit_evidence: Table 3 row (R)-1,2-Propanediol; F101N columns: KM=1200 mM, kcat=13 s^-1, kcat/KM=11 M^-1 s^-1.


column_label: PMID 30272958 Table 3; row methanol; column group wild-type; KM/kcat/kcat/KM cells: KM 2 mM; kcat 18 s^-1; kcat/KM 9000 M^-1 s^-1.

normalized_value_unit_evidence: KM 2 mM; kcat 18 s^-1; kcat/KM 9 s^-1 mM^-1

row_label: methanol / wild-type


column_label: PMID 30272958 Table 3; row ethanol; column group wild-type; KM/kcat/kcat/KM cells: KM 15 mM; kcat 19 s^-1; kcat/KM 1300 M^-1 s^-1.

row_label: ethanol / wild-type


substrate_name: D-cellobiose

substrate: D-cellobiose


substrate_name: D-maltoheptaose

substrate: D-maltoheptaose


substrate_name: D-maltopentaose

substrate: D-maltopentaose


substrate_name: D-fucose

smiles: CC(C(C(C(C=O)O)O)O)O

substrate: D-fucose


substrate_name: D-maltotriose

substrate: D-maltotriose


substrate_name: D-trehalose

substrate: D-trehalose


sequence_variant_note: Sequence identity was resolved, but no explicit wild-type or mutant call was captured.

smiles: CCCCCCO

column_label: Table 2 columns Substrate/KM/KI/kcat/kcat/KM, row Compound 9; table footnote [a] says HRP-coupled assay in 50 mM potassium phosphate pH 7.5 and units KM=mM, kcat=s^-1, kcat/KM=M^-1 s^-1.

raw_value_unit_evidence: Table 2 Compound 9: Km=3.0 mM; kcat=0.56 s^-1; kcat/KM=187 M^-1 s^-1.

row_label: Compound 9

table_label: Table 2: Apparent steady-state kinetic parameters for AOX*


substrate_name: diethylene glycol

smiles: C(COCCO)O

column_label: Table 2 columns Substrate/KM/KI/kcat/kcat/KM, row Compound 4; table footnote [a] says HRP-coupled assay in 50 mM potassium phosphate pH 7.5 and units KM=mM, kcat=s^-1, kcat/KM=M^-1 s^-1.

row_label: Compound 4

substrate: diethylene glycol


smiles: CC(CCCO)O

column_label: Table 2 columns Substrate/KM/KI/kcat/kcat/KM, row Compound 2; table footnote [a] says HRP-coupled assay in 50 mM potassium phosphate pH 7.5 and units KM=mM, kcat=s^-1, kcat/KM=M^-1 s^-1.

row_label: Compound 2


smiles: C(CCO)CCO

column_label: Table 2 columns Substrate/KM/KI/kcat/kcat/KM, row Compound 3; table footnote [a] says HRP-coupled assay in 50 mM potassium phosphate pH 7.5 and units KM=mM, kcat=s^-1, kcat/KM=M^-1 s^-1.

row_label: Compound 3


smiles: C(CSCCO)O

column_label: Table 2 columns Substrate/KM/KI/kcat/kcat/KM, row Compound 5; table footnote [a] says HRP-coupled assay in 50 mM potassium phosphate pH 7.5 and units KM=mM, kcat=s^-1, kcat/KM=M^-1 s^-1.

raw_value_unit_evidence: Table 2 Compound 5: Km=119 mM; kcat=3.56 s^-1; kcat/KM=29.9 M^-1 s^-1.

row_label: Compound 5


smiles: C(CCCO)CCO

column_label: Table 2 columns Substrate/KM/KI/kcat/kcat/KM, row Compound 6; table footnote [a] says HRP-coupled assay in 50 mM potassium phosphate pH 7.5 and units KM=mM, kcat=s^-1, kcat/KM=M^-1 s^-1.

raw_value_unit_evidence: Table 2 Compound 6: Km=12.9 mM; kcat=4.07 s^-1; kcat/KM=316 M^-1 s^-1.

row_label: Compound 6


smiles: C(CCO)CO

column_label: Table 2 columns Substrate/KM/KI/kcat/kcat/KM, row Compound 1; table footnote [a] says HRP-coupled assay in 50 mM potassium phosphate pH 7.5 and units KM=mM, kcat=s^-1, kcat/KM=M^-1 s^-1.

raw_value_unit_evidence: Table 2 Compound 1: Km=198 mM; kcat=6.74 s^-1; kcat/KM=34.1 M^-1 s^-1.

row_label: Compound 1


proof_lines:
pH 6.5, substrate: D-glucose
at pH 6.5 and 30°C
pH 4.5, substrate: D-glucose


proof_lines:
pH 6.5, substrate: D-glucose
at pH 6.5 and 30°C


proof_lines:
pH 4.5, substrate: D-glucose
at pH 4.5 and 30°C
pH 6.5, substrate: D-glucose


proof_lines:
pH 4.5, substrate: D-glucose
pH 6.5, substrate: D-glucose


substrate_name: L-arabinose

assay_conditions_summary: pH 10.5; buffer the publication 100 mM ethanolamine

proof_lines: pH and temperature not specified in the publication

substrate: L-arabinose


organism: Fusarium sp.

substrate_name: guar

sequence_variant_note: Source notes indicate a mutant or variant enzyme, but no machine-applicable substitution signature was available.

substrate: guar


proof_lines:
mutant Y436H/v494A, pH 7.0, 25°C
mutant Y436H, pH 7.0, 25°C


substrate_name: D-galactose

proof_lines:
mutant Y436H, pH 7.0, 25°C
mutant Y436H/v494A, pH 7.0, 25°C

substrate: D-galactose


organism: Streptomyces sp. SA-COO

substrate_name: cholesterol

sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:P12676 and applied mutation signature A32C/S129C/T371C/A423C with inferred residue-numbering offset +37 to reconstruct the assayed variant sequence.

assay_conditions_summary: pH 7; 310.15 K; buffer 50 mM sodium phosphate

column_label: Table 1: Michaelis-Menten Constants and Tryptophan Fluorescence Emission Maxima for Wild-Type ChoA and ChoA Mutants (Biochemistry 2018, 57(36):5370-5378); Table 1, row 'mutant 3, A32C/S129C/T371C/A423C'; column 'kcat (s^-1)' prints 23; column 'Km (uM)' prints 11; column 'kcat/Km (M^-1 s^-1)' prints 2.0 x 10^6; column 'lambda-max Trp (nm)' prints 332.

normalized_value_unit_evidence: kcat: printed Table 1 cell 23 s^-1; already in s^-1, no conversion; the exported value is 23.0 s^(-1) and the printed cell reproduces it. Km: printed Table 1 cell 11 uM; 11 / 1000 = 0.011 mM; the exported value is 0.011 mM and the printed cell reproduces it. kcat/Km: printed Table 1 cell 2.0 x 10^6 M^-1 s^-1; 1 M^-1 s^-1 is 0.001 s^-1 mM^-1, so 2.0 x 10^6 / 1000 = 2000.0 s^-1 mM^-1. The exported ratio is 2090.909090909091 s^(-1)*mM^(-1), which is the quotient 23.0 / 0.011 of the row's own kcat and Km rather than the printed efficiency cell, so the ratio is corrected to the printed value 2000.0 and kcat and Km are unchanged.

substrate: cholesterol


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:P12676 and applied mutation signature A32C/T168C/S312C/A465C with inferred residue-numbering offset +37 to reconstruct the assayed variant sequence.

column_label: Table 1: Michaelis-Menten Constants and Tryptophan Fluorescence Emission Maxima for Wild-Type ChoA and ChoA Mutants (Biochemistry 2018, 57(36):5370-5378); Table 1, row 'mutant 7, A32C/T168C/S312C/A465C'; column 'kcat (s^-1)' prints 30; column 'Km (uM)' prints 12; column 'kcat/Km (M^-1 s^-1)' prints 2.4 x 10^6; column 'lambda-max Trp (nm)' prints 328.

normalized_value_unit_evidence: kcat: printed Table 1 cell 30 s^-1; already in s^-1, no conversion; the exported value is 30.0 s^(-1) and the printed cell reproduces it. Km: printed Table 1 cell 12 uM; 12 / 1000 = 0.012 mM; the exported value is 0.012 mM and the printed cell reproduces it. kcat/Km: printed Table 1 cell 2.4 x 10^6 M^-1 s^-1; 1 M^-1 s^-1 is 0.001 s^-1 mM^-1, so 2.4 x 10^6 / 1000 = 2400.0 s^-1 mM^-1. The exported ratio is 2500.0 s^(-1)*mM^(-1), which is the quotient 30.0 / 0.012 of the row's own kcat and Km rather than the printed efficiency cell, so the ratio is corrected to the printed value 2400.0 and kcat and Km are unchanged.


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:P12676 and applied mutation signature A184C/A301C/T394C with inferred residue-numbering offset +37 to reconstruct the assayed variant sequence.

column_label: Table 1: Michaelis-Menten Constants and Tryptophan Fluorescence Emission Maxima for Wild-Type ChoA and ChoA Mutants (Biochemistry 2018, 57(36):5370-5378); Table 1, row 'mutant 8, A184C/A301C/T394C'; column 'kcat (s^-1)' prints 57; column 'Km (uM)' prints 11; column 'kcat/Km (M^-1 s^-1)' prints 5.0 x 10^6; column 'lambda-max Trp (nm)' prints 329.

normalized_value_unit_evidence: kcat: printed Table 1 cell 57 s^-1; already in s^-1, no conversion; the exported value is 57.0 s^(-1) and the printed cell reproduces it. Km: printed Table 1 cell 11 uM; 11 / 1000 = 0.011 mM; the exported value is 0.011 mM and the printed cell reproduces it. kcat/Km: printed Table 1 cell 5.0 x 10^6 M^-1 s^-1; 1 M^-1 s^-1 is 0.001 s^-1 mM^-1, so 5.0 x 10^6 / 1000 = 5000.0 s^-1 mM^-1. The exported ratio is 5181.818181818182 s^(-1)*mM^(-1), which is the quotient 57.0 / 0.011 of the row's own kcat and Km rather than the printed efficiency cell, so the ratio is corrected to the printed value 5000.0 and kcat and Km are unchanged.


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_mutation_ranked_match and applied mutation signature A184C/T239C/A407C/A465C with inferred residue-numbering offset +37 to reconstruct the assayed variant sequence.

proof_lines: pH 7.0, 37°C, mutant enzyme A184C/T239C/A407C/A465C, cholesterol solubilized in detergent micelles as a substrate


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:P12676 and applied mutation signature S153C/A205C/S312C/T435C with inferred residue-numbering offset +37 to reconstruct the assayed variant sequence.

column_label: Table 1: Michaelis-Menten Constants and Tryptophan Fluorescence Emission Maxima for Wild-Type ChoA and ChoA Mutants (Biochemistry 2018, 57(36):5370-5378); Table 1, row 'mutant 4, S153C/A205C/S312C/T435C'; column 'kcat (s^-1)' prints 68; column 'Km (uM)' prints 10; column 'kcat/Km (M^-1 s^-1)' prints 7.0 x 10^6; column 'lambda-max Trp (nm)' prints 328.

normalized_value_unit_evidence: kcat: printed Table 1 cell 68 s^-1; already in s^-1, no conversion; the exported value is 68.0 s^(-1) and the printed cell reproduces it. Km: printed Table 1 cell 10 uM; 10 / 1000 = 0.01 mM; the exported value is 0.01 mM and the printed cell reproduces it. kcat/Km: printed Table 1 cell 7.0 x 10^6 M^-1 s^-1; 1 M^-1 s^-1 is 0.001 s^-1 mM^-1, so 7.0 x 10^6 / 1000 = 7000.0 s^-1 mM^-1. The exported ratio is 6800.0 s^(-1)*mM^(-1), which is the quotient 68.0 / 0.01 of the row's own kcat and Km rather than the printed efficiency cell, so the ratio is corrected to the printed value 7000.0 and kcat and Km are unchanged.


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:P12676 and applied mutation signature L274C with inferred residue-numbering offset +37 to reconstruct the assayed variant sequence.

column_label: Table 1: Michaelis-Menten Constants and Tryptophan Fluorescence Emission Maxima for Wild-Type ChoA and ChoA Mutants (Biochemistry 2018, 57(36):5370-5378); Table 1, row 'mutant 2, L274C'; column 'kcat (s^-1)' prints 75; column 'Km (uM)' prints 17; column 'kcat/Km (M^-1 s^-1)' prints 4.5 x 10^6; column 'lambda-max Trp (nm)' prints 328.

normalized_value_unit_evidence: kcat: printed Table 1 cell 75 s^-1; already in s^-1, no conversion; the exported value is 75.0 s^(-1) and the printed cell reproduces it. Km: printed Table 1 cell 17 uM; 17 / 1000 = 0.017 mM; the exported value is 0.017 mM and the printed cell reproduces it. kcat/Km: printed Table 1 cell 4.5 x 10^6 M^-1 s^-1; 1 M^-1 s^-1 is 0.001 s^-1 mM^-1, so 4.5 x 10^6 / 1000 = 4500.0 s^-1 mM^-1. The exported ratio is 4411.764705882352 s^(-1)*mM^(-1), which is the quotient 75.0 / 0.017 of the row's own kcat and Km rather than the printed efficiency cell, so the ratio is corrected to the printed value 4500.0 and kcat and Km are unchanged.

row_label: Table 1, row 'mutant 2, L274C'


organism: Trametes sp.

substrate_name: D-glucose

sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature T166A/E539K to reconstruct the assayed variant sequence.

substrate: D-glucose


sequence_variant_note: Mutation signature E359K could not be applied safely because the resolved sequence does not match the expected reference residue(s).


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature K312E/E539K to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature K312E to reconstruct the assayed variant sequence.


organism: Brevibacterium sterolicum

assay_conditions_summary: pH 7.5; buffer product formation followed

proof_lines: assay relies on spectroscopic detection of 4-cholesten-3-one formation, decreasing length of C17 chain affects turnover negatively

proof_lines: substrate cholesterol, polarographic determination of the rate of oxygen consumption. 1% thesit (polyoxyethylene(9)-lauryl-ether), 10% propan-2-ol and 50 mM phosphate, 25°C and pH 7.5

source_databases_merged: brenda skid


assay_conditions_summary: pH 7.5; buffer H2O2 production assayed

proof_lines: assay relies on detection of H2O2 formation

proof_lines: product formation followed at 240 nm in the presence of 0.02 ml H2O2, substrate dissolved in Triton X-100, 100 mM phosphate buffer

proof_lines: H2O2 production assayed, 100 mM phosphate buffer


substrate_name: pregnenolone

assay_conditions_summary: pH 7.5; buffer substrate pregnenolone

proof_lines: assay relies on spectroscopic detection of 4-cholesten-3-one formation

proof_lines: substrate pregnenolone, detection of product formation at 240 nm. 0.1 M potassium phosphate, 1% Triton X-100, 1.25% propan-2-ol, 25°C and pH 7.5

substrate: pregnenolone


substrate_name: cholestanol

assay_conditions_summary: pH 7.5; buffer substrate cholestanol

proof_lines: substrate cholestanol, rate of H2O2 formation detected with o-dianisidine and horseradish peroxidase. 0.1 M potassium phosphate, 1% Triton X-100, 1.25% propan-2-ol, 25°C and pH 7.5

substrate: cholestanol


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature N593C with inferred residue-numbering offset -8 to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme N593C, with O2 as electron acceptor, at pH 6.5 and 30°C

proof_lines: mutant enzyme N593C, with 2,6-dichlorophenolindophenol as electron acceptor, at pH 6.5 and 30°C

proof_lines: mutant enzyme N593C, with 1,4-benzoquinone as electron acceptor, at pH 6.5 and 30°C


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature Q448H to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme Q448H, with O2 as electron acceptor, at pH 6.5 and 30°C

proof_lines: mutant enzyme Q448H, with 2,6-dichlorophenolindophenol as electron acceptor, at pH 6.5 and 30°C

proof_lines: mutant enzyme Q448H, with 1,4-benzoquinone as electron acceptor, at pH 6.5 and 30°C


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature L547R to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme L547R, with O2 as electron acceptor, at pH 6.5 and 30°C

proof_lines: mutant enzyme L547R, with 1,4-benzoquinone as electron acceptor, at pH 6.5 and 30°C

proof_lines: mutant enzyme L547R, with 2,6-dichlorophenolindophenol as electron acceptor, at pH 6.5 and 30°C


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature L545C to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme L545C, with O2 as electron acceptor, at pH 6.5 and 30°C

proof_lines: mutant enzyme L545C, with 2,6-dichlorophenolindophenol as electron acceptor, at pH 6.5 and 30°C

proof_lines: mutant enzyme L545C, with 1,4-benzoquinone as electron acceptor, at pH 6.5 and 30°C


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature T166R to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme T166R, with O2 as electron acceptor, at pH 6.5 and 30°C

proof_lines: mutant enzyme T166R, with 1,4-benzoquinone as electron acceptor, at pH 6.5 and 30°C

proof_lines: mutant enzyme T166R, with 2,6-dichlorophenolindophenol as electron acceptor, at pH 6.5 and 30°C


organism: Streptomyces sp.

proof_lines: mutant enzyme N485D, determined by H2O2 detection

proof_lines: mutant enzyme N485D, determined by cholest-4-en-3-one detection

proof_lines: apparent value, mutant enzyme N485D, pH 7.0, temperature not specified in the publication


proof_lines: mutant enzyme G347N, determined by H2O2 detection

proof_lines: mutant enzyme G347N, determined by cholest-4-en-3-one detection


proof_lines: mutant enzyme F359W, determined by cholest-4-en-3-one detection

proof_lines: mutant enzyme F359W, determined by H2O2 detection


organism: Aspergillus terreus

substrate_name: benzyl alcohol

sequence_variant_note: Sequence is treated as wild-type because the source explicitly marked the record as wild-type.

proof_lines:
recombinant enzyme, at pH 6.0 and 30°C
pH 8.0, 30°C

substrate: benzyl alcohol


substrate_name: 3,4-dimethoxybenzyl alcohol

assay_conditions_summary: pH 6; buffer 0.1 M sodium phosphate

column_label: Table 2 row Veratryl alcohol columns Km (mM), kcat (min^-1), and kcat/Km (min^-1 mM^-1); explicit required kcat/Km cell does not match expected value.

raw_value_unit_evidence: Expected substrate 3,4-dimethoxybenzyl alcohol maps to paper row Veratryl alcohol: Km 4.91 mM, kcat 6925.42 min^-1, kcat/Km 1410.47 min^-1 mM^-1. The expected values instead match paper row m-anisyl alcohol / 3-methoxybenzyl alcohol.

row_label: Veratryl alcohol

table_label: Table 2. Steady-state kinetic parameters of in-vitro refolded recombinant alcohol oxidase from E. coli.

substrate: 3,4-dimethoxybenzyl alcohol


substrate_name: 3-Methoxybenzyl alcohol

column_label: Table 2 row m-anisyl alcohol reports Km 2.88 mM, kcat 6341.70 min^-1, and kcat/Km 2201.98 min^-1 mM^-1; the expected values instead align with the Veratryl alcohol row.

raw_value_unit_evidence: m-anisyl alcohol row: Km raw_value=2.88 raw_unit=mM; kcat raw_value=6341.70 raw_unit=min^-1; kcat/Km raw_value=2201.98 raw_unit=min^-1 mM^-1. Veratryl alcohol row: Km raw_value=4.91 raw_unit=mM; kcat raw_value=6925.42 raw_unit=min^-1; kcat/Km raw_value=1410.47 raw_unit=min^-1 mM^-1.

row_label: m-anisyl alcohol / 3-methoxybenzyl alcohol

table_label: PMC3994049 Table 2

substrate: 3-Methoxybenzyl alcohol


substrate_name: 4-methoxybenzyl alcohol

column_label: Table 2 row p-anisyl alcohol, columns Km (mM), kcat (min^-1), and kcat/Km (min^-1 mM^-1), raw cells 1.08, 8455.86, and 7829.50, normalized to 1.08 mM, 140.931 s^(-1), and 130.4916666667 s^(-1)*mM^(-1)

normalized_value_unit_evidence: Km normalized_value=1.08 normalized_unit=mM; kcat normalized_value=140.931 normalized_unit=s^(-1); kcat/Km normalized_value=130.4916666667 normalized_unit=s^(-1)*mM^(-1)

raw_value_unit_evidence: Km raw_value=1.08 mM raw_unit=mM raw_cell=1.08; kcat raw_value=8455.86 min^-1 raw_unit=min^-1 raw_cell=8455.86; kcat/Km raw_value=7829.50 min^-1 mM^-1 raw_unit=min^-1 mM^-1 raw_cell=7829.50

row_label: p-anisyl alcohol

substrate: 4-methoxybenzyl alcohol


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature H167A/H548R to reconstruct the assayed variant sequence.

proof_lines:
pH 6.0, 25°C, mutant H167A/H548R
pH 7.0, 25°C, mutant H167A/H548R
pH 8.0, 25°C, mutant H167A/H548R


sequence_variant_note: Resolved sequence already matches the mutant residue(s), so it is treated as a source-provided variant sequence.

assay_conditions_summary: pH 7; buffer 50 mM KH2PO4

proof_lines: pH 7.0, 25°C, reductive half-reaction, wild-type enzyme

proof_lines: wild type enzyme, in 50 mM sodium phosphate (pH 7.0), at 25°C

proof_lines: wild type enzyme, using O2 as cosubstrate, in 50 mM KH2PO4 buffer (pH 6.5), at 30°C


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature T169S to reconstruct the assayed variant sequence.

proof_lines: pH 7.0, 25°C, reductive half-reaction, mutant T169S

proof_lines: T169S, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay

proof_lines: mutant enzyme T169S, in 50 mM sodium phosphate (pH 7.0), at 25°C


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature T169N to reconstruct the assayed variant sequence.

proof_lines: pH 7.0, 25°C, reductive half-reaction, mutant T169N

proof_lines: T169N, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay

proof_lines: T169N, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay


sequence_variant_note: Source notes indicate a mutant or variant enzyme, but no machine-applicable substitution signature was available.

proof_lines: mutant enzyme T169G, pH and temperature not specified in the publication

proof_lines: T169G, mutant, substrate D-glucose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay

proof_lines: T169G, mutant, substrate D-galactose, 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)-peroxidase assay


proof_lines: mutant enzyme T196G/V546C, pH and temperature not specified in the publication


proof_lines: mutant enzyme H167A, pH and temperature not specified in the publication


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_mutation_ranked_match and applied mutation signature Q286R to reconstruct the assayed variant sequence.


enzyme_display_name: Cholesterol oxidase

organism: Streptomyces sp. (strain SA-COO)

source_db: uniprot

proof_lines: kcat is 44 sec(-1) for the oxidation of cholesterol. kcat is 64 sec(-1) for the isomerization of cholest-5-en-3-one.

enzyme: Cholesterol oxidase

source: uniprot


proof_lines: mutant enzyme H450G/E542K/V546C, in 50 mM phosphate buffer (pH 6.5) at 60°C


proof_lines: mutant enzyme H450G, in 50 mM phosphate buffer (pH 6.5) at 60°C

proof_lines: mutant enzyme H450G, pH and temperature not specified in the publication


proof_lines: mutant enzyme H450G/V546C, in 50 mM phosphate buffer (pH 6.5) at 60°C

proof_lines: mutant enzyme H450G/V546C, pH and temperature not specified in the publication


proof_lines: mutant enzyme T169A, in 50 mM sodium phosphate (pH 7.0), at 4°C

proof_lines: mutant enzyme T169A, in 50 mM sodium phosphate (pH 7.0), at 25°C


proof_lines: wild type enzyme, using D-galactose as cosubstrate, in 50 mM sodium phosphate (pH 7.0), at 25°C

proof_lines: wild type enzyme, using D-glucose as cosubstrate, in 50 mM sodium phosphate (pH 7.0), at 4°C

source_databases_merged: brenda oed

source_db: oed


proof_lines: mutant enzyme T169S, using D-galactose as cosubstrate, in 50 mM sodium phosphate (pH 7.0), at 25°C

proof_lines: mutant enzyme T169S, using D-glucose as cosubstrate ,in 50 mM sodium phosphate (pH 7.0), at 4°C

proof_lines: mutant enzyme T169S, using D-glucose as cosubstrate, in 50 mM sodium phosphate (pH 7.0), at 4°C


proof_lines: mutant enzyme T169A, using D-glucose as cosubstrate, in 50 mM sodium phosphate (pH 7.0), at 4°C

proof_lines: mutant enzyme T169A, using D-galactose as cosubstrate,in 50 mM sodium phosphate (pH 7.0), at 25°C


proof_lines: mutant enzyme T169N, using D-galactose as cosubstrate,in 50 mM sodium phosphate (pH 7.0), at 25°C

proof_lines: mutant enzyme T169N, using D-glucose as cosubstrate, in 50 mM sodium phosphate (pH 7.0), at 4°C


proof_lines: mutant enzyme T169G, using D-galactose as cosubstrate,in 50 mM sodium phosphate (pH 7.0), at 25°C

proof_lines: mutant enzyme T169G, using D-glucose as cosubstrate, in 50 mM sodium phosphate (pH 7.0), at 4°C


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_mutation_ranked_match and applied mutation signature H447E/E361Q with inferred residue-numbering offset +37 to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_mutation_ranked_match and applied mutation signature H447Q/E361Q with inferred residue-numbering offset +37 to reconstruct the assayed variant sequence.


organism: Lyophyllum shimeji

column_label: LsP2Ox row DCIP; columns Km [mM], kcat [s^-1], and kcat/Km [mM^-1 s^-1] report Km 0.187 mM; kcat 67.3 s^-1; kcat/Km 361 mM^-1 s^-1.

row_label: LsP2Ox row DCIP

table_label: Table 4. Apparent kinetic constants for electron acceptor substrates


column_label: LsP2Ox row BQ; columns Km [mM], kcat [s^-1], and kcat/Km [mM^-1 s^-1] report Km 0.033 mM; kcat 92.3 s^-1; kcat/Km 2760 mM^-1 s^-1.

row_label: LsP2Ox row BQ


assay_conditions_summary: pH 7.5; buffer 50 mM phosphate

proof_lines: H69A mutant enzyme, steady state, 50 mM phosphate buffer, 1% thesit, 1% 2-propanol

proof_lines: H69A mutant enzyme, cholesterol oxidation, 50 mM phosphate buffer, 1% thesit, 1% 2-propanol

proof_lines: H69A mutant enzyme, 50 mM phosphate buffer, pH 7.5, 1% thesit, 1% 2-propanol


assay_conditions_summary: buffer 50 mM phosphate

proof_lines: H69A mutant enzyme, 5-cholesten-3-one isomerization, 50 mM phosphate buffer, 1% thesit, 1% 2-propanol


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_mutation_ranked_match and applied mutation signature V191A with inferred residue-numbering offset +3 to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_mutation_ranked_match and applied mutation signature N485L with inferred residue-numbering offset -12 to reconstruct the assayed variant sequence.

proof_lines: apparent value, mutant enzyme N485L, pH 5.1, temperature not specified in the publication

proof_lines: apparent value, mutant enzyme N485L, pH 7.0, temperature not specified in the publication


organism: Phanerodontia chrysosporium

column_label: Table 5, row 2-Deoxy-D-glucose, Km (mM) column: 55.0 +/- 1.4.

raw_value_unit_evidence: Table 5 row 2-Deoxy-D-glucose, Km=55.0 +/- 1.4 mM.

table_label: Table 5, PDF page 5 / printed page 512


substrate_name: maltose

column_label: Table 5, row Maltose, Km (mM) column: 257.0 +/- 10.0.

raw_value_unit_evidence: Table 5 row Maltose, Km=257.0 +/- 10.0 mM. Kinetic-properties prose qualification: the maltose result must be interpreted with care because the substrate contained trace glucose as measured by HPLC; TLC detection of the corresponding oxidation product was negative; the authors state that the trace glucose could not account for the high H2O2-production rate.

row_label: Maltose

substrate: maltose


organism: Mycolicibacterium neoaurum

sequence_variant_note: Sequence identity was resolved, but no explicit wild-type or mutant call was captured.


mutation_signature: stopped-flow measurement / stopped-flow measurement

sequence_variant_note: Mutation context was captured, but the signature could not be translated into an exact residue substitution safely.

mutation_type: truncation

proof_lines: isomerization to 4-cholesten-3-one, rapid reaction, stopped-flow measurement, 500 mM phosphate buffer, pH 7.5, in the presence of 1% thesit and 1.25% 2-propanol


organism: Bacillus sp. (in: firmicutes)

proof_lines:
pH 7, 30°C, enzyme CO2
pH 7, 30°C, enzyme CO1


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature F454A with inferred residue-numbering offset -15 to reconstruct the assayed variant sequence.


organism: Brevibacterium sp.

proof_lines: non-tagged recombinant enzyme, pH 7.2, 37°C

proof_lines: N-terminally His-tagged recombinant enzyme , pH 7.2, 37°C


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature T169G/E542K/V546C to reconstruct the assayed variant sequence.

proof_lines: T169G/E542K/V546C, mutant, 30°C, substrate D-glucose (O2 saturated), pH 6.5, standard 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) assay

proof_lines: T169G/E542K/V546C, mutant, 30°C, substrate D-galactose (O2 saturated), pH 6.5, standard 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) assay

proof_lines: T169G/E542K/V546C, mutant, 50°C, substrate D-glucose (O2 saturated), pH 6.5, standard 2,2'-azinobis(3-ethylbenzthiazolinesulfonic acid) assay


assay_conditions_summary: pH 5

proof_lines: soluble enzyme, 500 mM glucose used as electron acceptor, pH 5.0


proof_lines: pH 5.5, substrate D-glucose, measuring oxygen consumption (computer-interfaced Oxy-32 oxygen-monitoring system), 30°C

proof_lines: pH 7.5, substrate D-glucose, measuring oxygen consumption (computer-interfaced Oxy-32 oxygen-monitoring system), 30°C

proof_lines: pH 6.5, substrate D-glucose, measuring oxygen consumption (computer-interfaced Oxy-32 oxygen-monitoring system), 30°C


proof_lines: substrate 1,4-benzoquinone (100 mM D-glucose), pH 4.5

proof_lines: substrate D-glucose (air saturated), pH 6.5


substrate_name: trans-androsterone

assay_conditions_summary: pH 7.5; buffer o-dianisidine/horseradish peroxidase under phosphate

proof_lines: substrate trans-androsterone, rate of H2O2 formation detected with o-dianisidine and horseradish peroxidase. 0.1 M potassium phosphate, 1% Triton X-100, 1.25% propan-2-ol, 25°C and pH 7.5

substrate: trans-androsterone


substrate_name: ferrocenium hexafluorophosphate

sequence_variant_note: Mutation signature F454A/Y456A/H2P could not be applied safely because the resolved sequence does not match the expected reference residue(s).

substrate: ferrocenium hexafluorophosphate

source: oed


sequence_variant_note: Resolved sequence already matches the mutant residue(s) after applying an inferred residue-numbering offset +29, so it is treated as a source-provided variant sequence.


sequence_variant_note: Mutation signature F454P/H2P could not be applied safely because the resolved sequence does not match the expected reference residue(s).


sequence_variant_note: Mutation signature H450Q/H2P could not be applied safely because the resolved sequence does not match the expected reference residue(s).


sequence_variant_note: Sequence is treated as wild-type because the source explicitly marked the record as wild-type.

column_label: Table 4; row label Wild-type | Fc+/Glc; column label relevant Km, kcat, and kcat/Km columns; raw value cells K m[Fc+] 400 ± 110 μM, kcat[Fc+/Glc] 210 ± 30 s^-1, kcat[Fc+/Glc]/Km[Fc+] 0.51 × 10^6 M^-1 s^-1

normalized_value_unit_evidence: Paper central values normalize to kcat 210.0 s^-1, Km 0.4 mM, and explicit kcat/Km 510.0 s^-1 mM^-1. Explicit paper kcat/Km is 510 s^-1 mM^-1; export carries 525.0 from central-cell division, so not strict exact.

raw_value_unit_evidence: Table 4, row Wild-type | Fc+/Glc; columns: K m[Fc+] 400 ± 110 μM; kcat[Fc+/Glc] 210 ± 30 s^-1; kcat[Fc+/Glc]/Km[Fc+] 0.51 × 10^6 M^-1 s^-1.

row_label: Wild-type | Fc+/Glc

table_label: Table 4


sequence_variant_note: Resolved sequence already matches the mutant residue(s) after applying an inferred residue-numbering offset +30, so it is treated as a source-provided variant sequence.


sequence_variant_note: Mutation signature F454A/S455A/Y456A/H2P could not be applied safely because the resolved sequence does not match the expected reference residue(s).


organism: Coprinopsis cinerea

substrate_name: 4-anisyl alcohol

sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:A0A2Z5WP81 and applied mutation signature L416W to reconstruct the assayed variant sequence.

assay_conditions_summary: pH 7; 298.15 K; buffer 50 mM potassium phosphate

column_label: Table 2 (Steady-state kinetic constants of rCcAAO and the mutants), body row 'p-Anisyl alcohol' with enzyme-form cell 'L416W'; column 'Km (uM)' = 7.56; column 'kcat (s-1)' = 3.49; column 'kcat/Km (s-1 uM-1)' = 0.461. The value printed after each ± in the same cell is the fit error and is not an exported metric.

normalized_value_unit_evidence: kcat: printed Table 2 cell 3.49 in a column headed kcat (s-1); s-1 is the canonical s^(-1), so no unit conversion applies and the value is 3.49 s^(-1). Km: printed Table 2 cell 7.56 in a column headed Km (uM); 7.56 / 1000 = 0.00756 mM. kcat/Km: printed Table 2 cell 0.461 in a column headed kcat/Km (s-1 uM-1); 1 s^-1 uM^-1 is 1000 s^-1 mM^-1, so 0.461 x 1000 = 461 s^-1 mM^-1. The exported values are kcat 3.49 s^(-1), Km 0.0076 mM and kcat_over_km 459.2105263157895 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 2, body row 'p-Anisyl alcohol', enzyme-form cell 'L416W': Km = 7.56 uM; kcat = 3.49 s-1; printed kcat/Km = 0.461 in a column headed (s-1 uM-1), that is 0.461 s^(-1)*uM^(-1).

row_label: Table 2, row p-Anisyl alcohol / L416W

table_label: Table 2: Steady-state kinetic constants of rCcAAO and the mutants (Biotechnology Letters 2018, 40(7):1077-1086). Printed column heads: Substrate; enzyme form; Km (uM); kcat (s-1); kcat/Km (s-1 uM-1)

substrate: 4-anisyl alcohol


column_label: Official Springer Table 2 row p-Anisyl alcohol / rCcAAO: Km column 11.6 +/- 1.0 uM, kcat column 12.5 +/- 0.3 s^-1, kcat/Km column 1.08 +/- 0.09 s^-1 uM^-1.

normalized_value_unit_evidence: Km normalized_value=0.0116 normalized_unit=mM; kcat normalized_value=12.5 normalized_unit=s^-1; kcat/Km normalized_value=1080 normalized_unit=s^-1 mM^-1

raw_value_unit_evidence: Km raw_value=11.6 +/- 1.0 raw_unit=uM; kcat raw_value=12.5 +/- 0.3 raw_unit=s^-1; kcat/Km raw_value=1.08 +/- 0.09 raw_unit=s^-1 uM^-1

row_label: p-Anisyl alcohol / rCcAAO

table_label: Table 2


substrate_name: 4-Hydroxybenzyl alcohol

sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:A0A2Z5WP81 and applied mutation signature L416F to reconstruct the assayed variant sequence.

column_label: Table 2 (Steady-state kinetic constants of rCcAAO and the mutants), body row '4-Hydroxybenzyl alcohol' with enzyme-form cell 'L416F'; column 'Km (uM)' = 163; column 'kcat (s-1)' = 8.77; column 'kcat/Km (s-1 uM-1)' = 0.0539. The value printed after each ± in the same cell is the fit error and is not an exported metric.

normalized_value_unit_evidence: kcat: printed Table 2 cell 8.77 in a column headed kcat (s-1); s-1 is the canonical s^(-1), so no unit conversion applies and the value is 8.77 s^(-1). Km: printed Table 2 cell 163 in a column headed Km (uM); 163 / 1000 = 0.163 mM. kcat/Km: printed Table 2 cell 0.0539 in a column headed kcat/Km (s-1 uM-1); 1 s^-1 uM^-1 is 1000 s^-1 mM^-1, so 0.0539 x 1000 = 53.9 s^-1 mM^-1. The exported values are kcat 8.77 s^(-1), Km 0.163 mM and kcat_over_km 53.803680981595086 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 2, body row '4-Hydroxybenzyl alcohol', enzyme-form cell 'L416F': Km = 163 uM; kcat = 8.77 s-1; printed kcat/Km = 0.0539 in a column headed (s-1 uM-1), that is 0.0539 s^(-1)*uM^(-1).

row_label: Table 2, row 4-Hydroxybenzyl alcohol / L416F

substrate: 4-Hydroxybenzyl alcohol


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature L416W to reconstruct the assayed variant sequence.

column_label: Official Springer Table 2 row 4-Hydroxybenzyl alcohol / L416W: Km column 466 +/- 74 uM, kcat column 0.539 +/- 0.051 s^-1, kcat/Km column 0.00119 +/- 0.00007 s^-1 uM^-1.

normalized_value_unit_evidence: Km normalized_value=0.466 normalized_unit=mM; kcat normalized_value=0.539 normalized_unit=s^-1; kcat/Km normalized_value=1.19 normalized_unit=s^-1 mM^-1

raw_value_unit_evidence: Km raw_value=466 +/- 74 raw_unit=uM; kcat raw_value=0.539 +/- 0.051 raw_unit=s^-1; kcat/Km raw_value=0.00119 +/- 0.00007 raw_unit=s^-1 uM^-1

row_label: 4-Hydroxybenzyl alcohol / L416W


column_label: Official Springer Table 2 row 4-Hydroxybenzyl alcohol / rCcAAO: Km column 56.3 +/- 7.6 uM, kcat column 4.81 +/- 0.06 s^-1, kcat/Km column 0.0855 +/- 0.0120 s^-1 uM^-1.

normalized_value_unit_evidence: Km normalized_value=0.0563 normalized_unit=mM; kcat normalized_value=4.81 normalized_unit=s^-1; kcat/Km normalized_value=85.5 normalized_unit=s^-1 mM^-1

raw_value_unit_evidence: Km raw_value=56.3 +/- 7.6 raw_unit=uM; kcat raw_value=4.81 +/- 0.06 raw_unit=s^-1; kcat/Km raw_value=0.0855 +/- 0.0120 raw_unit=s^-1 uM^-1

row_label: 4-Hydroxybenzyl alcohol / rCcAAO


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature L416F to reconstruct the assayed variant sequence.


column_label: Table 2 (Steady-state kinetic constants of rCcAAO and the mutants), body row 'Benzyl alcohol' with enzyme-form cell 'L416W'; column 'Km (uM)' = 5.87; column 'kcat (s-1)' = 3.84; column 'kcat/Km (s-1 uM-1)' = 0.665. The value printed after each ± in the same cell is the fit error and is not an exported metric.

normalized_value_unit_evidence: kcat: printed Table 2 cell 3.84 in a column headed kcat (s-1); s-1 is the canonical s^(-1), so no unit conversion applies and the value is 3.84 s^(-1). Km: printed Table 2 cell 5.87 in a column headed Km (uM); 5.87 / 1000 = 0.00587 mM. kcat/Km: printed Table 2 cell 0.665 in a column headed kcat/Km (s-1 uM-1); 1 s^-1 uM^-1 is 1000 s^-1 mM^-1, so 0.665 x 1000 = 665 s^-1 mM^-1. The exported values are kcat 3.84 s^(-1), Km 0.0059 mM and kcat_over_km 650.8474576271186 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 2, body row 'Benzyl alcohol', enzyme-form cell 'L416W': Km = 5.87 uM; kcat = 3.84 s-1; printed kcat/Km = 0.665 in a column headed (s-1 uM-1), that is 0.665 s^(-1)*uM^(-1).

row_label: Table 2, row Benzyl alcohol / L416W


column_label: Official Springer Table 2 row Benzyl alcohol / rCcAAO: Km column 1.21 +/- 0.27 uM, kcat column 6.13 +/- 0.12 s^-1, kcat/Km column 5.06 +/- 1.08 s^-1 uM^-1.

normalized_value_unit_evidence: Km normalized_value=0.00121 normalized_unit=mM; kcat normalized_value=6.13 normalized_unit=s^-1; kcat/Km normalized_value=5060 normalized_unit=s^-1 mM^-1

raw_value_unit_evidence: Km raw_value=1.21 +/- 0.27 raw_unit=uM; kcat raw_value=6.13 +/- 0.12 raw_unit=s^-1; kcat/Km raw_value=5.06 +/- 1.08 raw_unit=s^-1 uM^-1

row_label: Benzyl alcohol / rCcAAO


substrate_name: isovanillyl alcohol

column_label: Official Springer Table 2 row Isovanillyl alcohol / L416F: Km column 318 +/- 19 uM, kcat column 8.67 +/- 0.55 s^-1, kcat/Km column 0.0273 +/- 0.0001 s^-1 uM^-1.

normalized_value_unit_evidence: Km normalized_value=0.318 normalized_unit=mM; kcat normalized_value=8.67 normalized_unit=s^-1; kcat/Km normalized_value=27.3 normalized_unit=s^-1 mM^-1

raw_value_unit_evidence: Km raw_value=318 +/- 19 raw_unit=uM; kcat raw_value=8.67 +/- 0.55 raw_unit=s^-1; kcat/Km raw_value=0.0273 +/- 0.0001 raw_unit=s^-1 uM^-1

row_label: Isovanillyl alcohol / L416F

substrate: isovanillyl alcohol


column_label: Official Springer Table 2 row Isovanillyl alcohol / L416W: Km column 154 +/- 2 uM, kcat column 2.05 +/- 0.05 s^-1, kcat/Km column 0.0133 +/- 0.0005 s^-1 uM^-1.

normalized_value_unit_evidence: Km normalized_value=0.154 normalized_unit=mM; kcat normalized_value=2.05 normalized_unit=s^-1; kcat/Km normalized_value=13.3 normalized_unit=s^-1 mM^-1

raw_value_unit_evidence: Km raw_value=154 +/- 2 raw_unit=uM; kcat raw_value=2.05 +/- 0.05 raw_unit=s^-1; kcat/Km raw_value=0.0133 +/- 0.0005 raw_unit=s^-1 uM^-1

row_label: Isovanillyl alcohol / L416W


organism: Gibberella zeae

substrate_name: D-Galactose

sequence_variant_note: Mutation signature C383A could not be applied safely because the resolved sequence does not match the expected reference residue(s).

substrate: D-Galactose


sequence_variant_note: Mutation signature C383S could not be applied safely because the resolved sequence does not match the expected reference residue(s).


sequence_variant_note: Resolved sequence already matches the mutant residue(s) after applying an inferred residue-numbering offset -39, so it is treated as a source-provided variant sequence.


sequence_variant_note: Mutation signature C383S/Y436A could not be applied safely because the resolved sequence does not match the expected reference residue(s).


sequence_variant_note: Resolved sequence already matches the mutant residue(s) after applying an inferred residue-numbering offset +20, so it is treated as a source-provided variant sequence.


sequence_variant_note: Mutation signature C383S/Y436H/V494A could not be applied safely because the resolved sequence does not match the expected reference residue(s).


sequence_variant_note: Resolved sequence already matches the mutant residue(s) after applying an inferred residue-numbering offset -15, so it is treated as a source-provided variant sequence.


sequence_variant_note: Mutation signature Y436A could not be applied safely because the resolved sequence does not match the expected reference residue(s).


sequence_variant_note: Mutation signature Y436A/V494A could not be applied safely because the resolved sequence does not match the expected reference residue(s).


sequence_variant_note: Mutation signature Y436H could not be applied safely because the resolved sequence does not match the expected reference residue(s).


sequence_variant_note: Sequence is treated as wild-type because the source explicitly marked the record as wild-type.

proof_lines:
rCcAAO
aryl-alcohol oxidase
aromatic alcohols


substrate_name: Vanillyl alcohol

column_label: Table 2 (Steady-state kinetic constants of rCcAAO and the mutants), body row 'Vanillyl alcohol' with enzyme-form cell 'L416F'; column 'Km (uM)' = 980; column 'kcat (s-1)' = 21.3; column 'kcat/Km (s-1 uM-1)' = 0.0218. The value printed after each ± in the same cell is the fit error and is not an exported metric.

normalized_value_unit_evidence: kcat: printed Table 2 cell 21.3 in a column headed kcat (s-1); s-1 is the canonical s^(-1), so no unit conversion applies and the value is 21.3 s^(-1). Km: printed Table 2 cell 980 in a column headed Km (uM); 980 / 1000 = 0.98 mM. kcat/Km: printed Table 2 cell 0.0218 in a column headed kcat/Km (s-1 uM-1); 1 s^-1 uM^-1 is 1000 s^-1 mM^-1, so 0.0218 x 1000 = 21.8 s^-1 mM^-1. The exported values are kcat 21.3 s^(-1), Km 0.98 mM and kcat_over_km 21.73469387755102 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 2, body row 'Vanillyl alcohol', enzyme-form cell 'L416F': Km = 980 uM; kcat = 21.3 s-1; printed kcat/Km = 0.0218 in a column headed (s-1 uM-1), that is 0.0218 s^(-1)*uM^(-1).

row_label: Table 2, row Vanillyl alcohol / L416F

substrate: Vanillyl alcohol


column_label: Table 2 (Steady-state kinetic constants of rCcAAO and the mutants), body row 'Vanillyl alcohol' with enzyme-form cell 'L416W'; column 'Km (uM)' = 419; column 'kcat (s-1)' = 0.811; column 'kcat/Km (s-1 uM-1)' = 0.00194. The value printed after each ± in the same cell is the fit error and is not an exported metric.

normalized_value_unit_evidence: kcat: printed Table 2 cell 0.811 in a column headed kcat (s-1); s-1 is the canonical s^(-1), so no unit conversion applies and the value is 0.811 s^(-1). Km: printed Table 2 cell 419 in a column headed Km (uM); 419 / 1000 = 0.419 mM. kcat/Km: printed Table 2 cell 0.00194 in a column headed kcat/Km (s-1 uM-1); 1 s^-1 uM^-1 is 1000 s^-1 mM^-1, so 0.00194 x 1000 = 1.94 s^-1 mM^-1. The exported values are kcat 0.81 s^(-1), Km 0.419 mM and kcat_over_km 1.9331742243436756 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 2, body row 'Vanillyl alcohol', enzyme-form cell 'L416W': Km = 419 uM; kcat = 0.811 s-1; printed kcat/Km = 0.00194 in a column headed (s-1 uM-1), that is 0.00194 s^(-1)*uM^(-1).

row_label: Table 2, row Vanillyl alcohol / L416W


column_label: Table 2 (Steady-state kinetic constants of rCcAAO and the mutants), body row 'Vanillyl alcohol' with enzyme-form cell 'rCcAAO'; column 'Km (uM)' = 6.27; column 'kcat (s-1)' = 14.7; column 'kcat/Km (s-1 uM-1)' = 2.35. The value printed after each ± in the same cell is the fit error and is not an exported metric.

normalized_value_unit_evidence: kcat: printed Table 2 cell 14.7 in a column headed kcat (s-1); s-1 is the canonical s^(-1), so no unit conversion applies and the value is 14.7 s^(-1). Km: printed Table 2 cell 6.27 in a column headed Km (uM); 6.27 / 1000 = 0.00627 mM. kcat/Km: printed Table 2 cell 2.35 in a column headed kcat/Km (s-1 uM-1); 1 s^-1 uM^-1 is 1000 s^-1 mM^-1, so 2.35 x 1000 = 2350 s^-1 mM^-1. The exported values are kcat 14.7 s^(-1), Km 0.0063 mM and kcat_over_km 2333.333333333333 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 2, body row 'Vanillyl alcohol', enzyme-form cell 'rCcAAO': Km = 6.27 uM; kcat = 14.7 s-1; printed kcat/Km = 2.35 in a column headed (s-1 uM-1), that is 2.35 s^(-1)*uM^(-1).

row_label: Table 2, row Vanillyl alcohol / rCcAAO


substrate_name: veratryl alcohol

column_label: Table 2 (Steady-state kinetic constants of rCcAAO and the mutants), body row 'Veratryl alcohol' with enzyme-form cell 'L416F'; column 'Km (uM)' = 424; column 'kcat (s-1)' = 12.4; column 'kcat/Km (s-1 uM-1)' = 0.0293. The value printed after each ± in the same cell is the fit error and is not an exported metric.

normalized_value_unit_evidence: kcat: printed Table 2 cell 12.4 in a column headed kcat (s-1); s-1 is the canonical s^(-1), so no unit conversion applies and the value is 12.4 s^(-1). Km: printed Table 2 cell 424 in a column headed Km (uM); 424 / 1000 = 0.424 mM. kcat/Km: printed Table 2 cell 0.0293 in a column headed kcat/Km (s-1 uM-1); 1 s^-1 uM^-1 is 1000 s^-1 mM^-1, so 0.0293 x 1000 = 29.3 s^-1 mM^-1. The exported values are kcat 12.4 s^(-1), Km 0.424 mM and kcat_over_km 29.245283018867926 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 2, body row 'Veratryl alcohol', enzyme-form cell 'L416F': Km = 424 uM; kcat = 12.4 s-1; printed kcat/Km = 0.0293 in a column headed (s-1 uM-1), that is 0.0293 s^(-1)*uM^(-1).

row_label: Table 2, row Veratryl alcohol / L416F

substrate: veratryl alcohol


column_label: Table 2 (Steady-state kinetic constants of rCcAAO and the mutants), body row 'Veratryl alcohol' with enzyme-form cell 'L416W'; column 'Km (uM)' = 686; column 'kcat (s-1)' = 2.96; column 'kcat/Km (s-1 uM-1)' = 0.00432. The value printed after each ± in the same cell is the fit error and is not an exported metric.

normalized_value_unit_evidence: kcat: printed Table 2 cell 2.96 in a column headed kcat (s-1); s-1 is the canonical s^(-1), so no unit conversion applies and the value is 2.96 s^(-1). Km: printed Table 2 cell 686 in a column headed Km (uM); 686 / 1000 = 0.686 mM. kcat/Km: printed Table 2 cell 0.00432 in a column headed kcat/Km (s-1 uM-1); 1 s^-1 uM^-1 is 1000 s^-1 mM^-1, so 0.00432 x 1000 = 4.32 s^-1 mM^-1. The exported values are kcat 2.96 s^(-1), Km 0.686 mM and kcat_over_km 4.314868804664723 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 2, body row 'Veratryl alcohol', enzyme-form cell 'L416W': Km = 686 uM; kcat = 2.96 s-1; printed kcat/Km = 0.00432 in a column headed (s-1 uM-1), that is 0.00432 s^(-1)*uM^(-1).

row_label: Table 2, row Veratryl alcohol / L416W


sequence_variant_note: Mutation signature H450Q/H2P could not be applied safely because the resolved sequence does not match the expected reference residue(s).


sequence_variant_note: Resolved sequence already matches the mutant residue(s) after applying an inferred residue-numbering offset +30, so it is treated as a source-provided variant sequence.


substrate_name: Methyl alpha-D-galactopyranoside

sequence_variant_note: Resolved canonical sequence from UniProtKB:P0CS93 and applied mutation signature Y436A/V494A with inferred residue-numbering offset +41 to reconstruct the assayed variant sequence.

column_label: Official OUP publisher PDF, Table II, row Y436A/V494A under 1-methyl-alpha-D-galactopyranoside, columns KM (mM), kcat (s^-1), and kcat/KM (M^-1 s^-1): 46 +/- 3; 1548 +/- 95; 33600 +/- 3000.

normalized_value_unit_evidence: KM 46 mM; kcat 1548 s^-1; printed kcat/KM 33.6 s^-1 mM^-1 after exact /1000 unit normalization.

raw_value_unit_evidence: Table II Y436A/V494A, 1-methyl-alpha-D-galactopyranoside: KM 46 +/- 3 mM; kcat 1548 +/- 95 s^-1; kcat/KM 33600 +/- 3000 M^-1 s^-1.

table_label: Table II

substrate: Methyl alpha-D-galactopyranoside


sequence_variant_note: Resolved canonical sequence from UniProtKB:P0CS93 and applied mutation signature Y436H with inferred residue-numbering offset +41 to reconstruct the assayed variant sequence.

column_label: Official OUP publisher PDF, Table II, row Y436H under 1-methyl-alpha-D-galactopyranoside, columns KM (mM), kcat (s^-1), and kcat/KM (M^-1 s^-1): 55 +/- 5; 1267 +/- 97; 23000 +/- 2700.

normalized_value_unit_evidence: KM 55 mM; kcat 1267 s^-1; printed kcat/KM 23.0 s^-1 mM^-1 after exact /1000 unit normalization.

raw_value_unit_evidence: Table II Y436H, 1-methyl-alpha-D-galactopyranoside: KM 55 +/- 5 mM; kcat 1267 +/- 97 s^-1; kcat/KM 23000 +/- 2700 M^-1 s^-1.


sequence_variant_note: Resolved canonical sequence from UniProtKB:P0CS93 and applied mutation signature Y436H/V494A with inferred residue-numbering offset +41 to reconstruct the assayed variant sequence.

column_label: Official OUP publisher PDF, Table II, row Y436H/V494A under 1-methyl-alpha-D-galactopyranoside, columns KM (mM), kcat (s^-1), and kcat/KM (M^-1 s^-1): 45 +/- 7; 851 +/- 91; 18900 +/- 3600.

normalized_value_unit_evidence: KM 45 mM; kcat 851 s^-1; printed kcat/KM 18.9 s^-1 mM^-1 after exact /1000 unit normalization.

raw_value_unit_evidence: Table II Y436H/V494A, 1-methyl-alpha-D-galactopyranoside: KM 45 +/- 7 mM; kcat 851 +/- 91 s^-1; kcat/KM 18900 +/- 3600 M^-1 s^-1.


enzyme_display_name: Galactose oxidase

substrate_name: D-fructose

enzyme: Galactose oxidase

substrate: D-fructose


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:A0A2Z5WP81 and applied mutation signature L416F to reconstruct the assayed variant sequence.

smiles: C/C=C/C=C/CO

column_label: Table 2 (Steady-state kinetic constants of rCcAAO and the mutants), body row 'trans,trans-2,4-Hexadien-1-ol' with enzyme-form cell 'L416F'; column 'Km (uM)' = 16.4; column 'kcat (s-1)' = 68.4; column 'kcat/Km (s-1 uM-1)' = 4.18. The value printed after each ± in the same cell is the fit error and is not an exported metric.

normalized_value_unit_evidence: kcat: printed Table 2 cell 68.4 in a column headed kcat (s-1); s-1 is the canonical s^(-1), so no unit conversion applies and the value is 68.4 s^(-1). Km: printed Table 2 cell 16.4 in a column headed Km (uM); 16.4 / 1000 = 0.0164 mM. kcat/Km: printed Table 2 cell 4.18 in a column headed kcat/Km (s-1 uM-1); 1 s^-1 uM^-1 is 1000 s^-1 mM^-1, so 4.18 x 1000 = 4180 s^-1 mM^-1. The exported values are kcat 68.4 s^(-1), Km 0.0164 mM and kcat_over_km 4170.731707317073 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 2, body row 'trans,trans-2,4-Hexadien-1-ol', enzyme-form cell 'L416F': Km = 16.4 uM; kcat = 68.4 s-1; printed kcat/Km = 4.18 in a column headed (s-1 uM-1), that is 4.18 s^(-1)*uM^(-1).

row_label: Table 2, row trans,trans-2,4-Hexadien-1-ol / L416F


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:A0A2Z5WP81 and applied mutation signature L416W to reconstruct the assayed variant sequence.

column_label: Table 2 (Steady-state kinetic constants of rCcAAO and the mutants), body row 'trans,trans-2,4-Hexadien-1-ol' with enzyme-form cell 'L416W'; column 'Km (uM)' = 7.04; column 'kcat (s-1)' = 17.3; column 'kcat/Km (s-1 uM-1)' = 2.46. The value printed after each ± in the same cell is the fit error and is not an exported metric.

normalized_value_unit_evidence: kcat: printed Table 2 cell 17.3 in a column headed kcat (s-1); s-1 is the canonical s^(-1), so no unit conversion applies and the value is 17.3 s^(-1). Km: printed Table 2 cell 7.04 in a column headed Km (uM); 7.04 / 1000 = 0.00704 mM. kcat/Km: printed Table 2 cell 2.46 in a column headed kcat/Km (s-1 uM-1); 1 s^-1 uM^-1 is 1000 s^-1 mM^-1, so 2.46 x 1000 = 2460 s^-1 mM^-1. The exported values are kcat 17.3 s^(-1), Km 0.007 mM and kcat_over_km 2471.4285714285716 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 2, body row 'trans,trans-2,4-Hexadien-1-ol', enzyme-form cell 'L416W': Km = 7.04 uM; kcat = 17.3 s-1; printed kcat/Km = 2.46 in a column headed (s-1 uM-1), that is 2.46 s^(-1)*uM^(-1).

row_label: Table 2, row trans,trans-2,4-Hexadien-1-ol / L416W


column_label: Official Springer Table 2 row trans,trans-2,4-Hexadien-1-ol / rCcAAO: Km column 15.6 +/- 0.8 uM, kcat column 48.3 +/- 1.4 s^-1, kcat/Km column 3.10 +/- 0.23 s^-1 uM^-1.

normalized_value_unit_evidence: Km normalized_value=0.0156 normalized_unit=mM; kcat normalized_value=48.3 normalized_unit=s^-1; kcat/Km normalized_value=3100 normalized_unit=s^-1 mM^-1

raw_value_unit_evidence: Km raw_value=15.6 +/- 0.8 raw_unit=uM; kcat raw_value=48.3 +/- 1.4 raw_unit=s^-1; kcat/Km raw_value=3.10 +/- 0.23 raw_unit=s^-1 uM^-1

row_label: trans,trans-2,4-Hexadien-1-ol / rCcAAO


substrate_name: 3-anisyl alcohol

column_label: Table 2 (Steady-state kinetic constants of rCcAAO and the mutants), body row 'm-Anisyl alcohol' with enzyme-form cell 'L416F'; column 'Km (uM)' = 34.2; column 'kcat (s-1)' = 12.4; column 'kcat/Km (s-1 uM-1)' = 0.361. The value printed after each ± in the same cell is the fit error and is not an exported metric.

normalized_value_unit_evidence: kcat: printed Table 2 cell 12.4 in a column headed kcat (s-1); s-1 is the canonical s^(-1), so no unit conversion applies and the value is 12.4 s^(-1). Km: printed Table 2 cell 34.2 in a column headed Km (uM); 34.2 / 1000 = 0.0342 mM. kcat/Km: printed Table 2 cell 0.361 in a column headed kcat/Km (s-1 uM-1); 1 s^-1 uM^-1 is 1000 s^-1 mM^-1, so 0.361 x 1000 = 361 s^-1 mM^-1. The exported values are kcat 12.4 s^(-1), Km 0.034 mM and kcat_over_km 364.70588235294116 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 2, body row 'm-Anisyl alcohol', enzyme-form cell 'L416F': Km = 34.2 uM; kcat = 12.4 s-1; printed kcat/Km = 0.361 in a column headed (s-1 uM-1), that is 0.361 s^(-1)*uM^(-1).

row_label: Table 2, row m-Anisyl alcohol / L416F

substrate: 3-anisyl alcohol


column_label: Table 2 (Steady-state kinetic constants of rCcAAO and the mutants), body row 'm-Anisyl alcohol' with enzyme-form cell 'L416W'; column 'Km (uM)' = 30.8; column 'kcat (s-1)' = 11.3; column 'kcat/Km (s-1 uM-1)' = 0.367. The value printed after each ± in the same cell is the fit error and is not an exported metric.

normalized_value_unit_evidence: kcat: printed Table 2 cell 11.3 in a column headed kcat (s-1); s-1 is the canonical s^(-1), so no unit conversion applies and the value is 11.3 s^(-1). Km: printed Table 2 cell 30.8 in a column headed Km (uM); 30.8 / 1000 = 0.0308 mM. kcat/Km: printed Table 2 cell 0.367 in a column headed kcat/Km (s-1 uM-1); 1 s^-1 uM^-1 is 1000 s^-1 mM^-1, so 0.367 x 1000 = 367 s^-1 mM^-1. The exported values are kcat 11.3 s^(-1), Km 0.031 mM and kcat_over_km 364.5161290322581 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 2, body row 'm-Anisyl alcohol', enzyme-form cell 'L416W': Km = 30.8 uM; kcat = 11.3 s-1; printed kcat/Km = 0.367 in a column headed (s-1 uM-1), that is 0.367 s^(-1)*uM^(-1).

row_label: Table 2, row m-Anisyl alcohol / L416W