sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature P92A to reconstruct the assayed variant sequence.

normalized_value_unit_evidence: km normalized_value=0.0256 normalized_unit=mM; unit conversion from 25.6 uM

raw_value_unit_evidence: Table 2. Kinetic parameters of the oxidoreductase activity of wild-type and mutant Ndi1 for ubiquinone-1; row P92A; column Km (uM); raw value 25.6 +/- 2.2 uM


organism: Saccharomyces cerevisiae

source_db: brenda

verification_status: verified

kcat_display: Not reported

ki_display: Not reported

kcat_over_km_display: Not reported

source_license: CC BY 4.0

km_unit: mM

sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature R479I to reconstruct the assayed variant sequence.

mutation_type: substitution

assay_conditions_summary: pH 6; cofactor NADH

normalized_value_unit_evidence: km normalized_value=0.0277 normalized_unit=mM; unit conversion from 27.7 uM

raw_value_unit_evidence: Table 2. Kinetic parameters of the oxidoreductase activity of wild-type and mutant Ndi1 for ubiquinone-1; row R479I; column Km (uM); raw value 27.7 +/- 1.2 uM

data_origin: experimental

status: verified

source: brenda


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature I459N to reconstruct the assayed variant sequence.

normalized_value_unit_evidence: km normalized_value=0.0303 normalized_unit=mM; unit conversion from 30.3 uM

raw_value_unit_evidence: Table 2. Kinetic parameters of the oxidoreductase activity of wild-type and mutant Ndi1 for ubiquinone-1; row I459N; column Km (uM); raw value 30.3 +/- 2.0 uM


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature Q394G to reconstruct the assayed variant sequence.

normalized_value_unit_evidence: km normalized_value=0.0308 normalized_unit=mM; unit conversion from 30.8 uM

raw_value_unit_evidence: Table 2. Kinetic parameters of the oxidoreductase activity of wild-type and mutant Ndi1 for ubiquinone-1; row Q394G; column Km (uM); raw value 30.8 +/- 3.4 uM


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature A393G to reconstruct the assayed variant sequence.

normalized_value_unit_evidence: km normalized_value=0.0325 normalized_unit=mM; unit conversion from 32.5 uM

raw_value_unit_evidence: Table 2. Kinetic parameters of the oxidoreductase activity of wild-type and mutant Ndi1 for ubiquinone-1; row A393G; column Km (uM); raw value 32.5 +/- 1.9 uM


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature H397A to reconstruct the assayed variant sequence.

normalized_value_unit_evidence: km normalized_value=0.035 normalized_unit=mM; unit conversion from 35.0 uM

raw_value_unit_evidence: Table 2. Kinetic parameters of the oxidoreductase activity of wild-type and mutant Ndi1 for ubiquinone-1; row H397A; column Km (uM); raw value 35.0 +/- 2.7 uM


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature L444D to reconstruct the assayed variant sequence.

normalized_value_unit_evidence: km normalized_value=0.0376 normalized_unit=mM; unit conversion from 37.6 uM

raw_value_unit_evidence: Table 2. Kinetic parameters of the oxidoreductase activity of wild-type and mutant Ndi1 for ubiquinone-1; row L444D; column Km (uM); raw value 37.6 +/- 3.9 uM


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature I459A to reconstruct the assayed variant sequence.

normalized_value_unit_evidence: km normalized_value=0.0402 normalized_unit=mM; unit conversion from 40.2 uM

raw_value_unit_evidence: Table 2. Kinetic parameters of the oxidoreductase activity of wild-type and mutant Ndi1 for ubiquinone-1; row I459A; column Km (uM); raw value 40.2 +/- 4.1 uM


enzyme_display_name: cytochrome-b5 reductase

organism: Homo sapiens

substrate_name: ferricytochrome b5

verification_status: unverified

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

enzyme: cytochrome-b5 reductase

substrate: ferricytochrome b5

status: unverified


substrate_name: ferricyanide

substrate: ferricyanide


mutation_signature: Unknown

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

proof_lines:
wild-type
wild-type, 25°C, pH 7.0
recombinant wild-type enzyme


proof_lines:
wild-type
wild-type, 25°C, pH 7.0


substrate_name: NADH

assay_conditions_summary: cofactor NADH

substrate: NADH


enzyme_display_name: azobenzene reductase

organism: Cereibacter sphaeroides

enzyme: azobenzene reductase


substrate_name: nitrofurazone

assay_conditions_summary: cofactor NADPH

substrate: nitrofurazone


substrate_name: Methyl red

substrate: Methyl red


proof_lines:
NADH-dependent, mutant K109H
mutant K109H


substrate_name: NADPH

proof_lines:
NADPH-dependent, mutant Y74W
mutant Y74W

substrate: NADPH


proof_lines:
NADH-dependent, wild-type enzyme
wild-type enzyme


proof_lines:
NADPH-dependent, wild-type enzyme
wild-type enzyme


proof_lines:
NADH-dependent, mutant K109A
mutant K109A


proof_lines:
NADPH-dependent, mutant K109H
mutant K109H


proof_lines:
mutant K109A
NADPH-dependent, mutant K109A


organism: Brevundimonas subvibrioides

substrate_name: acid Orange 7

substrate: acid Orange 7


substrate_name: acid Red 88

substrate: acid Red 88


substrate_name: Acid Red 151

substrate: Acid Red 151


substrate_name: Acid Orange 8

substrate: Acid Orange 8


enzyme_display_name: hydroxylamine reductase (NADH)

organism: Derxia gummosa

enzyme: hydroxylamine reductase (NADH)


substrate_name: hydroxylamine

smiles: NO

substrate: hydroxylamine


organism: Halomonas elongata

assay_conditions_summary: pH 7.6; 298.15 K; cofactor NADH

proof_lines: mutant enzyme L49C/D108C, at pH 7.6 and 25°C


organism: Neurospora crassa


organism: Shewanella oneidensis

assay_conditions_summary: pH 7; 313.15 K; cofactor NADPH

proof_lines: recombinant chimeric fusion protein AZR-GDH, pH 7.0, 40°C


assay_conditions_summary: pH 6.5; 298.15 K; cofactor NADH


assay_conditions_summary: pH 6.5; 298.15 K; cofactor NADPH


organism: Rattus norvegicus


organism: Shewanella xiamenensis

substrate_name: Methyl orange

substrate: Methyl orange


substrate_name: Congo red

substrate: Congo red


organism: Staphylococcus aureus

substrate_name: o-methyl red

assay_conditions_summary: 295.15 K; cofactor NADPH

proof_lines: pH not specified in the publication, 22°C

substrate: o-methyl red


substrate_name: flame orange

substrate: flame orange


organism: Rhodobacter capsulatus

substrate_name: methyl viologen

substrate: methyl viologen


proof_lines:
GST-fusion protein, pH 8.0, 40°C
native protein, pH 8.0, 40°C


assay_conditions_summary: pH 8; 313.15 K; cofactor NADH


substrate_name: Unknown

km_display: Not reported

ki_unit: mM

substrate: Unknown


organism: Enterococcus faecium

assay_conditions_summary: pH 7; 298.15 K; cofactor NADH


assay_conditions_summary: pH 7.1; 295.15 K; buffer 25 mM potassium phosphate; cofactor NADPH

proof_lines: in 25 mM potassium phosphate buffer (pH 7.1), 0.25 mM 2-[4-(dimethylamino)phenylazo]benzoic acid and 0.1 mM NADPH


assay_conditions_summary: pH 7.1; buffer 25 mM potassium phosphate; cofactor NADPH

proof_lines: in 25 mM potassium phosphate buffer (pH 7.1), 0.25 mM azo dye and 0.1 mM NADPH


proof_lines:
vs. NADPH
vs. methyl red


substrate_name: CO

smiles: [Co]

substrate: CO


enzyme_display_name: GMP reductase

organism: Artemia salina

enzyme: GMP reductase


assay_conditions_summary: pH 7; 298.15 K; cofactor NADPH

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


proof_lines: pH 7.0, wild type enzyme with cytochrome b5 as substrate

proof_lines: pH 7.0, wild type enzyme with ferricyanide as substrate


substrate_name: GMP

substrate: GMP


organism: Clostridium perfringens

substrate_name: FAD

assay_conditions_summary: cofactor FAD

proof_lines: pH not specified in the publication, temperature not specified in the publication

substrate: FAD


substrate_name: Direct blue 15

substrate: Direct blue 15


substrate_name: NADH

substrate: NADH


enzyme_display_name: nitroquinoline-N-oxide reductase

organism: Mus musculus

substrate_name: 4-Nitroquinoline N-oxide

enzyme: nitroquinoline-N-oxide reductase

substrate: 4-Nitroquinoline N-oxide


organism: Enterococcus faecalis

substrate_name: Methyl red

protein_accession_database: UniProt

kcat_over_km_origin: calculated

sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_mutation_ranked_match and applied mutation signature N106A with inferred residue-numbering offset -6 to reconstruct the assayed variant sequence.

assay_conditions_summary: pH 7

substrate: Methyl red


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_mutation_ranked_match and applied mutation signature L59G with inferred residue-numbering offset -6 to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_mutation_ranked_match and applied mutation signature E16G with inferred residue-numbering offset +10 to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_mutation_ranked_match and applied mutation signature R66A with inferred residue-numbering offset -5 to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_mutation_ranked_match and applied mutation signature N121A with inferred residue-numbering offset +11 to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_mutation_ranked_match and applied mutation signature A123F with inferred residue-numbering offset -2 to reconstruct the assayed variant sequence.


assay_conditions_summary: pH 7; cofactor NADH


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


organism: Escherichia coli

sequence_variant_note: Mutation signature Y120A 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 R59A with inferred residue-numbering offset +13 to reconstruct the assayed variant sequence.


substrate_name: p-methyl red

substrate: p-methyl red


sequence_variant_note: Mutation signature F162A references residue 162, which is outside the resolved sequence length.


organism: Pseudomonas aeruginosa

assay_conditions_summary: pH 8; cofactor NADH

proof_lines: wild type enzyme, using methyl red as cosubstrate, in 20 mM Tris?HCl pH 8.0, temperature not specified in the publication


assay_conditions_summary: pH 8; cofactor NADPH

proof_lines: wild type enzyme, using NADPH as cosubstrate, in 20 mM Tris?HCl pH 8.0, temperature not specified in the publication


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

proof_lines: mutant enzyme Y131F, using NADPH as cosubstrate, in 20 mM Tris?HCl pH 8.0, temperature not specified in the publication


substrate_name: balsalazide

substrate: balsalazide


proof_lines: mutant enzyme Y131F, using methyl red as cosubstrate, in 20 mM Tris?HCl pH 8.0, temperature not specified in the publication


substrate_name: NADPH

substrate: NADPH


substrate_name: sulfasalazine

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

assay_conditions_summary: pH 8

proof_lines: isoform AzoR3, pH 8.0, temperature not specified in the publication

proof_lines: isoform AzoR2, pH 8.0, temperature not specified in the publication

proof_lines: isoform AzoR1, pH 8.0, temperature not specified in the publication

substrate: sulfasalazine


organism: Leishmania major

source_db: uniprot

assay_conditions_summary: cofactor NADPH

proof_lines: kcat is 0.059 min(-1) with GMP as substrate and 0.043 min(-1) with NADPH as substrate.

source: uniprot


assay_conditions_summary: pH 7.4; cofactor FMN

proof_lines: cofactor roseoflavin mononucleotide, pH 7.4, 30°C


assay_conditions_summary: pH 7.4; cofactor NADH


substrate_name: ponceau SX

proof_lines:
azoreductase I
azoreductase II

substrate: ponceau SX


organism: Bacillus subtilis

substrate_name: Cr(VI)

assay_conditions_summary: pH 7.5; cofactor NADPH

proof_lines: pH 7.5, 30°C, recombinant His10-tagged enzyme

substrate: Cr(VI)


organism: Bacillus sp.

substrate_name: H+; Methyl red; NADPH

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

substrate: H+; Methyl red; NADPH


substrate_name: NADPH; Methyl red; H+

assay_conditions_summary: pH 7.5; buffer pH 7; cofactor NADPH

column_label: Table 5 columns rAZR-BSUa, Vmax (umole/min/mg), and kcat (S^-1); row Methyl Red.

normalized_value_unit_evidence: kcat 3.79 s^-1; Vmax 0.0002 mol s^-1 g^-1 from 12.0 umole/min/mg.

raw_value_unit_evidence: Table 5 row Methyl Red under rAZR-BSUa: Km 91.7 uM; Vmax 12.0 umole/min/mg; kcat 3.79 s^-1.

row_label: Methyl Red; rAZR-BSUa; Bacillus subtilis ATCC6633

table_label: Table 5, Apparent Kinetic Values of Recombinant Azoreductases against Methyl Red and Reactive Red 22

substrate: NADPH; Methyl red; H+


substrate_name: NADPH; H+; Reactive Red 22

column_label: Table 5 columns rAZR-BSUa, Km (uM) and Vmax (umole/min/mg); row Reactive Red 22.

normalized_value_unit_evidence: Km 0.0254 mM; Vmax 0.000095 mol s^-1 g^-1 from 5.7 umole/min/mg.

raw_value_unit_evidence: Table 5 row Reactive Red 22 under rAZR-BSUa: Km 25.4 uM; Vmax 5.7 umole/min/mg.

row_label: Reactive Red 22; rAZR-BSUa; Bacillus subtilis ATCC6633

substrate: NADPH; H+; Reactive Red 22


substrate_name: H+; NADH; Methyl red

assay_conditions_summary: cofactor NADH

substrate: H+; NADH; Methyl red


organism: Geobacillus stearothermophilus

substrate_name: H+; NADPH; Methyl red

column_label: Table 5 columns rAZR-GSTi, Vmax (umole/min/mg), and kcat (S^-1); row Methyl Red.

normalized_value_unit_evidence: kcat 4.13 s^-1; Vmax 0.000218333333333 mol s^-1 g^-1 from 13.1 umole/min/mg. Current sheet Vmax 0.0002183333 is rounded from the conversion.

raw_value_unit_evidence: Table 5 row Methyl Red under rAZR-GSTi: Km 134 uM; Vmax 13.1 umole/min/mg; kcat 4.13 s^-1.

row_label: Methyl Red; rAZR-GSTi; Geobacillus stearothermophilus IFO13737

substrate: H+; NADPH; Methyl red


column_label: Table 5 columns rAZR-GSTi, Km (uM) and Vmax (umole/min/mg); row Reactive Red 22.

normalized_value_unit_evidence: Km 0.0175 mM; Vmax 0.000055 mol s^-1 g^-1 from 3.3 umole/min/mg.

raw_value_unit_evidence: Table 5 row Reactive Red 22 under rAZR-GSTi: Km 17.5 uM; Vmax 3.3 umole/min/mg.

row_label: Reactive Red 22; rAZR-GSTi; Geobacillus stearothermophilus IFO13737


organism: Pantoea agglomerans


organism: Rhodobacter sphaeroides

substrate_name: H+; Methyl red; NADH

substrate: H+; Methyl red; NADH


organism: Pigmentiphaga kullae

substrate_name: Orange I

assay_conditions_summary: pH 6.8; cofactor NADPH

column_label: Table 2; row Orange I (NADPH); column Km (uM)

raw_value_unit_evidence: Table 2 row Orange I (NADPH) column Km (uM) raw value 3.0 uM

row_label: Orange I (NADPH)

table_label: Table 2 Michaelis constants (Km) and maximal velocities (Vmax) of recombinant AzoB of P. kullae K24 expressed in E. coli

substrate: Orange I


substrate_name: balsalzide

substrate: balsalzide


substrate_name: Unknown

substrate: Unknown


organism: Trypanosoma brucei brucei


source_db: oed

verification_status: corrected

sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:Q831B2 and applied mutation signature A123F to reconstruct the assayed variant sequence.

assay_conditions_summary: pH 7; 295.15 K; cofactor NADH

column_label: Table 3 (Kinetic parameters of wild-type and the mutants), printed line 'A123F'; column 'Km (uM, Methyl Red)' = 48.5; column 'kcat (s-1)' = 245; column 'kcat / KmMR (uM-1 s-1)' = 5.05. The same printed line also carries FMN* = 1.8, Km (uM, NADH) = 411, Vmax (U/ mg) = 350, kcat/KmNADH (uM-1 s-1) = 0.6 and Relative specific activity (%) = 36, none of which this row exports.

normalized_value_unit_evidence: kcat: printed Table 3 cell 245 in a column headed kcat (s-1); s-1 is already the canonical s^(-1), so no unit conversion applies and the value is 245.0 s^(-1). Km: printed Table 3 cell 48.5 in a column headed Km (uM, Methyl Red); 1 uM is 0.001 mM, so 48.5 / 1000 = 0.0485 mM. kcat/Km: printed Table 3 cell 5.05 in a column headed kcat / KmMR (uM-1 s-1); 1 uM^-1 s^-1 is 1000 s^-1 mM^-1, so 5.05 x 1000 = 5050.0 s^(-1)*mM^(-1). The catalog side of this record carries kcat 245.0 s^(-1), Km 0.0485 mM and kcat_over_km 5051.546391752578 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 3, printed line 'A123F': Km (uM, Methyl Red) = 48.5; kcat (s-1) = 245; printed kcat / KmMR = 5.05 in a column headed kcat / KmMR (uM-1 s-1).

row_label: Table 3, row A123F, Methyl Red columns

table_label: Table 3: Kinetic parameters of wild-type and the mutants (Arch Biochem Biophys 2012, 520(2):99-107). Printed column heads, in printed order: Enzyme; FMN*; Poly-merization; Km (uM, Methyl Red); Km (uM, NADH); Vmax (U/ mg); kcat (s-1); kcat / KmMR (uM-1 s-1); kcat/KmNADH (uM-1 s-1); Relative specific activity (%). Footnote * reads 'FMN per dimer of the purified protein' and footnote ** reads 'ND, not determined due to strong substrate Methyl Red inhibition'.

status: corrected

source: oed


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:Q831B2 and applied mutation signature L59G to reconstruct the assayed variant sequence.

column_label: Table 3 (Kinetic parameters of wild-type and the mutants), printed line 'L59G'; column 'Km (uM, Methyl Red)' = 28.8; column 'kcat (s-1)' = 6.61; column 'kcat / KmMR (uM-1 s-1)' = 0.23. The same printed line also carries FMN* = 1.8, Km (uM, NADH) = 2298, Vmax (U/ mg) = 9.4, kcat/KmNADH (uM-1 s-1) = 0.0029 and Relative specific activity (%) = 10, none of which this row exports.

normalized_value_unit_evidence: kcat: printed Table 3 cell 6.61 in a column headed kcat (s-1); s-1 is already the canonical s^(-1), so no unit conversion applies and the value is 6.61 s^(-1). Km: printed Table 3 cell 28.8 in a column headed Km (uM, Methyl Red); 1 uM is 0.001 mM, so 28.8 / 1000 = 0.0288 mM. kcat/Km: printed Table 3 cell 0.23 in a column headed kcat / KmMR (uM-1 s-1); 1 uM^-1 s^-1 is 1000 s^-1 mM^-1, so 0.23 x 1000 = 230.0 s^(-1)*mM^(-1). The catalog side of this record carries kcat 6.61 s^(-1), Km 0.0288 mM and kcat_over_km 229.5138888888889 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 3, printed line 'L59G': Km (uM, Methyl Red) = 28.8; kcat (s-1) = 6.61; printed kcat / KmMR = 0.23 in a column headed kcat / KmMR (uM-1 s-1).

row_label: Table 3, row L59G, Methyl Red columns


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:Q831B2 and applied mutation signature N106A to reconstruct the assayed variant sequence.

column_label: Table 3 (Kinetic parameters of wild-type and the mutants), printed line 'N106A'; column 'Km (uM, Methyl Red)' = 11.2; column 'kcat (s-1)' = 1.2; column 'kcat / KmMR (uM-1 s-1)' = 0.11. The same printed line also carries FMN* = 1.6, Km (uM, NADH) = 793, Vmax (U/ mg) = 1.7, kcat/KmNADH (uM-1 s-1) = 0.0015 and Relative specific activity (%) = 3, none of which this row exports.

normalized_value_unit_evidence: kcat: printed Table 3 cell 1.2 in a column headed kcat (s-1); s-1 is already the canonical s^(-1), so no unit conversion applies and the value is 1.2 s^(-1). Km: printed Table 3 cell 11.2 in a column headed Km (uM, Methyl Red); 1 uM is 0.001 mM, so 11.2 / 1000 = 0.0112 mM. kcat/Km: printed Table 3 cell 0.11 in a column headed kcat / KmMR (uM-1 s-1); 1 uM^-1 s^-1 is 1000 s^-1 mM^-1, so 0.11 x 1000 = 110.0 s^(-1)*mM^(-1). The catalog side of this record carries kcat 1.2 s^(-1), Km 0.0112 mM and kcat_over_km 107.14285714285714 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 3, printed line 'N106A': Km (uM, Methyl Red) = 11.2; kcat (s-1) = 1.2; printed kcat / KmMR = 0.11 in a column headed kcat / KmMR (uM-1 s-1).

row_label: Table 3, row N106A, Methyl Red columns


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:Q831B2 and applied mutation signature N121A to reconstruct the assayed variant sequence.

column_label: Table 3 (Kinetic parameters of wild-type and the mutants), printed line 'N121A'; column 'Km (uM, Methyl Red)' = 8.2; column 'kcat (s-1)' = 111; column 'kcat / KmMR (uM-1 s-1)' = 13.54. The same printed line also carries FMN* = 1.9, Km (uM, NADH) = 292, Vmax (U/ mg) = 158, kcat/KmNADH (uM-1 s-1) = 0.38 and Relative specific activity (%) = 170, none of which this row exports.

normalized_value_unit_evidence: kcat: printed Table 3 cell 111 in a column headed kcat (s-1); s-1 is already the canonical s^(-1), so no unit conversion applies and the value is 111.0 s^(-1). Km: printed Table 3 cell 8.2 in a column headed Km (uM, Methyl Red); 1 uM is 0.001 mM, so 8.2 / 1000 = 0.0082 mM. kcat/Km: printed Table 3 cell 13.54 in a column headed kcat / KmMR (uM-1 s-1); 1 uM^-1 s^-1 is 1000 s^-1 mM^-1, so 13.54 x 1000 = 13540.0 s^(-1)*mM^(-1). The catalog side of this record carries kcat 111.0 s^(-1), Km 0.0082 mM and kcat_over_km 13536.585365853658 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 3, printed line 'N121A': Km (uM, Methyl Red) = 8.2; kcat (s-1) = 111; printed kcat / KmMR = 13.54 in a column headed kcat / KmMR (uM-1 s-1).

row_label: Table 3, row N121A, Methyl Red columns


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:Q831B2 and applied mutation signature R18G to reconstruct the assayed variant sequence.

column_label: Table 3 (Kinetic parameters of wild-type and the mutants), printed line 'R18G'; column 'Km (uM, Methyl Red)' = 16.7; column 'kcat (s-1)' = 3.17; column 'kcat / KmMR (uM-1 s-1)' = 0.19. The same printed line also carries FMN* = 1.7, Km (uM, NADH) = 624, Vmax (U/ mg) = 4.5, kcat/KmNADH (uM-1 s-1) = 0.005 and Relative specific activity (%) = 9, none of which this row exports.

normalized_value_unit_evidence: kcat: printed Table 3 cell 3.17 in a column headed kcat (s-1); s-1 is already the canonical s^(-1), so no unit conversion applies and the value is 3.17 s^(-1). Km: printed Table 3 cell 16.7 in a column headed Km (uM, Methyl Red); 1 uM is 0.001 mM, so 16.7 / 1000 = 0.0167 mM. kcat/Km: printed Table 3 cell 0.19 in a column headed kcat / KmMR (uM-1 s-1); 1 uM^-1 s^-1 is 1000 s^-1 mM^-1, so 0.19 x 1000 = 190.0 s^(-1)*mM^(-1). The catalog side of this record carries kcat 3.17 s^(-1), Km 0.0167 mM and kcat_over_km 189.82035928143713 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 3, printed line 'R18G': Km (uM, Methyl Red) = 16.7; kcat (s-1) = 3.17; printed kcat / KmMR = 0.19 in a column headed kcat / KmMR (uM-1 s-1).

row_label: Table 3, row R18G, Methyl Red columns


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:Q831B2 and applied mutation signature R21G to reconstruct the assayed variant sequence.

column_label: Table 3 (Kinetic parameters of wild-type and the mutants), printed line 'R21G'; column 'Km (uM, Methyl Red)' = 12.6; column 'kcat (s-1)' = 140; column 'kcat / KmMR (uM-1 s-1)' = 11.11. The same printed line also carries FMN* = 1.9, Km (uM, NADH) = 380, Vmax (U/ mg) = 200, kcat/KmNADH (uM-1 s-1) = 0.37 and Relative specific activity (%) = 382, none of which this row exports.

normalized_value_unit_evidence: kcat: printed Table 3 cell 140 in a column headed kcat (s-1); s-1 is already the canonical s^(-1), so no unit conversion applies and the value is 140.0 s^(-1). Km: printed Table 3 cell 12.6 in a column headed Km (uM, Methyl Red); 1 uM is 0.001 mM, so 12.6 / 1000 = 0.0126 mM. kcat/Km: printed Table 3 cell 11.11 in a column headed kcat / KmMR (uM-1 s-1); 1 uM^-1 s^-1 is 1000 s^-1 mM^-1, so 11.11 x 1000 = 11110.0 s^(-1)*mM^(-1). The catalog side of this record carries kcat 140.0 s^(-1), Km 0.0126 mM and kcat_over_km 11111.111111111111 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 3, printed line 'R21G': Km (uM, Methyl Red) = 12.6; kcat (s-1) = 140; printed kcat / KmMR = 11.11 in a column headed kcat / KmMR (uM-1 s-1).

row_label: Table 3, row R21G, Methyl Red columns


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:Q831B2 and applied mutation signature R66A to reconstruct the assayed variant sequence.

column_label: Table 3 (Kinetic parameters of wild-type and the mutants), printed line 'R66A'; column 'Km (uM, Methyl Red)' = 5.3; column 'kcat (s-1)' = 77.3; column 'kcat / KmMR (uM-1 s-1)' = 14.58. The same printed line also carries FMN* = 1.9, Km (uM, NADH) = 581, Vmax (U/ mg) = 110, kcat/KmNADH (uM-1 s-1) = 0.13 and Relative specific activity (%) = 47, none of which this row exports.

normalized_value_unit_evidence: kcat: printed Table 3 cell 77.3 in a column headed kcat (s-1); s-1 is already the canonical s^(-1), so no unit conversion applies and the value is 77.3 s^(-1). Km: printed Table 3 cell 5.3 in a column headed Km (uM, Methyl Red); 1 uM is 0.001 mM, so 5.3 / 1000 = 0.0053 mM. kcat/Km: printed Table 3 cell 14.58 in a column headed kcat / KmMR (uM-1 s-1); 1 uM^-1 s^-1 is 1000 s^-1 mM^-1, so 14.58 x 1000 = 14580.0 s^(-1)*mM^(-1). The catalog side of this record carries kcat 77.3 s^(-1), Km 0.00053 mM and kcat_over_km 145849.05660377358 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 3, printed line 'R66A': Km (uM, Methyl Red) = 5.3; kcat (s-1) = 77.3; printed kcat / KmMR = 14.58 in a column headed kcat / KmMR (uM-1 s-1).

row_label: Table 3, row R66A, Methyl Red columns


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:Q831B2 and applied mutation signature V122Y to reconstruct the assayed variant sequence.

column_label: Table 3 (Kinetic parameters of wild-type and the mutants), printed line 'V122Y'; column 'Km (uM, Methyl Red)' = 109.2; column 'kcat (s-1)' = 17.5; column 'kcat / KmMR (uM-1 s-1)' = 0.16. The same printed line also carries FMN* = 1.7, Km (uM, NADH) = 405, Vmax (U/ mg) = 24.8, kcat/KmNADH (uM-1 s-1) = 0.044 and Relative specific activity (%) = 14, none of which this row exports.

normalized_value_unit_evidence: kcat: printed Table 3 cell 17.5 in a column headed kcat (s-1); s-1 is already the canonical s^(-1), so no unit conversion applies and the value is 17.5 s^(-1). Km: printed Table 3 cell 109.2 in a column headed Km (uM, Methyl Red); 1 uM is 0.001 mM, so 109.2 / 1000 = 0.1092 mM. kcat/Km: printed Table 3 cell 0.16 in a column headed kcat / KmMR (uM-1 s-1); 1 uM^-1 s^-1 is 1000 s^-1 mM^-1, so 0.16 x 1000 = 160.0 s^(-1)*mM^(-1). The catalog side of this record carries kcat 17.5 s^(-1), Km 0.109 mM and kcat_over_km 160.55045871559633 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 3, printed line 'V122Y': Km (uM, Methyl Red) = 109.2; kcat (s-1) = 17.5; printed kcat / KmMR = 0.16 in a column headed kcat / KmMR (uM-1 s-1).

row_label: Table 3, row V122Y, Methyl Red columns


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:Q831B2 and applied mutation signature W62A to reconstruct the assayed variant sequence.

column_label: Table 3 (Kinetic parameters of wild-type and the mutants), printed line 'W62A'; column 'Km (uM, Methyl Red)' = 33.5; column 'kcat (s-1)' = 21.1; column 'kcat / KmMR (uM-1 s-1)' = 0.63. The same printed line also carries FMN* = 1.7, Km (uM, NADH) = 3, Vmax (U/ mg) = 30, kcat/KmNADH (uM-1 s-1) = 7.03 and Relative specific activity (%) = 35, none of which this row exports.

normalized_value_unit_evidence: kcat: printed Table 3 cell 21.1 in a column headed kcat (s-1); s-1 is already the canonical s^(-1), so no unit conversion applies and the value is 21.1 s^(-1). Km: printed Table 3 cell 33.5 in a column headed Km (uM, Methyl Red); 1 uM is 0.001 mM, so 33.5 / 1000 = 0.0335 mM. kcat/Km: printed Table 3 cell 0.63 in a column headed kcat / KmMR (uM-1 s-1); 1 uM^-1 s^-1 is 1000 s^-1 mM^-1, so 0.63 x 1000 = 630.0 s^(-1)*mM^(-1). The catalog side of this record carries kcat 21.1 s^(-1), Km 0.0335 mM and kcat_over_km 629.8507462686567 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 3, printed line 'W62A': Km (uM, Methyl Red) = 33.5; kcat (s-1) = 21.1; printed kcat / KmMR = 0.63 in a column headed kcat / KmMR (uM-1 s-1).

row_label: Table 3, row W62A, Methyl Red columns


assay_conditions_summary: pH 7; 295.15 K; buffer pH 7; cofactor NADH

column_label: Table 3. Kinetic parameters of wild-type and the mutants; row Wild-type; columns Km (uM, Methyl Red), kcat (s^-1), kcat/Km MR (uM^-1 s^-1).

normalized_value_unit_evidence: kcat paper=59.1 s^-1; Km paper=0.024 mM; kcat/Km paper=2460 s^-1 mM^-1

raw_value_unit_evidence: Table 3 row Wild-type: Km (Methyl Red) 24 uM; kcat 59.1 s^-1; kcat/Km MR 2.46 uM^-1 s^-1.

row_label: Wild-type


organism: Staphylococcus aureus

sequence_variant_note: Mutation signature C2C could not be applied safely because the resolved sequence does not match the expected reference residue(s).


organism: Leishmania donovani

substrate_name: GMP; NADPH; H+

substrate: GMP; NADPH; H+


substrate_name: Methyl red

substrate: Methyl red


substrate_name: Reactive Red 22

assay_conditions_summary: pH 7.5; 298.15 K; buffer pH 7; cofactor NADPH

column_label: Table 5; row Reactive Red 22; column group rAZR-BSUa; column Km (uM) cell 25.4; column Vmax (umole/min/mg) cell 5.7; column kcat (S^-1) cell 1.8; Table 5 prints no kcat/Km column.

normalized_value_unit_evidence: kcat 1.8 S^-1 = 1.8 s^-1. Km 25.4 uM = 0.0254 mM (25.4 / 1000). kcat/Km is not printed in Table 5; the paper-cell quotient is paper_kcat / paper_km = 1.8 / 0.0254 = 70.86614173228347 s^-1 mM^-1, while the catalog holds 72.0 s^-1 mM^-1 computed from the catalog's rounded Km 0.025 mM.

raw_value_unit_evidence: Table 5, row Reactive Red 22, columns under rAZR-BSUa: Km 25.4 uM; Vmax 5.7 umole/min/mg; kcat 1.8 S^-1.

row_label: Reactive Red 22; rAZR-BSUa (Bacillus subtilis ATCC6633)

table_label: Table 5: Apparent Kinetic Values of Recombinant Azoreductases against Methyl Red and Reactive Red 22

substrate: Reactive Red 22


column_label: Table 5; row Methyl Red; column group rAZR-BSPo; column Km (uM) cell 395; column Vmax (umole/min/mg) cell 10.4; column kcat (S^-1) cell 3.3; Table 5 prints no kcat/Km column.

normalized_value_unit_evidence: kcat 3.3 S^-1 = 3.3 s^-1. Km 395 uM = 0.395 mM (395 / 1000). kcat/Km is not printed in Table 5; the paper-cell quotient is paper_kcat / paper_km = 3.3 / 0.395 = 8.354430379746834 s^-1 mM^-1, which equals the catalog value 8.354430379746834 s^-1 mM^-1.

raw_value_unit_evidence: Table 5, row Methyl Red, columns under rAZR-BSPo: Km 395 uM; Vmax 10.4 umole/min/mg; kcat 3.3 S^-1.

row_label: Methyl Red; rAZR-BSPo (Bacillus sp. OY1-2)


column_label: Table 5; row Reactive Red 22; column group rAZR-BSPo; column Km (uM) cell 24.3; column Vmax (umole/min/mg) cell 22.1; column kcat (S^-1) cell 7.11; Table 5 prints no kcat/Km column.

normalized_value_unit_evidence: kcat 7.11 S^-1 = 7.11 s^-1. Km 24.3 uM = 0.024300000000000002 mM (24.3 / 1000). kcat/Km is not printed in Table 5; the paper-cell quotient is paper_kcat / paper_km = 7.11 / 0.024300000000000002 = 292.59259259259255 s^-1 mM^-1, while the catalog holds 296.25 s^-1 mM^-1 computed from the catalog's rounded Km 0.024 mM.

raw_value_unit_evidence: Table 5, row Reactive Red 22, columns under rAZR-BSPo: Km 24.3 uM; Vmax 22.1 umole/min/mg; kcat 7.11 S^-1.

row_label: Reactive Red 22; rAZR-BSPo (Bacillus sp. OY1-2)


enzyme_display_name: UniProt P41407

source_db: skid

source_license: CC BY-NC-ND 4.0

enzyme: UniProt P41407

source: skid


enzyme_display_name: UniProt P24474

substrate_name: nitrite

proof_lines:
Raw SKiD Env: Temp=25°C, pH=6.2
Raw SKiD Env: Temp=-----, pH=-----

enzyme: UniProt P24474

substrate: nitrite


enzyme_display_name: UniProt O68601

organism: Achromobacter xylosoxidans

enzyme: UniProt O68601


enzyme_display_name: UniProt P38501

organism: Alcaligenes faecalis

enzyme: UniProt P38501


enzyme_display_name: UniProt Q8VNU2

organism: Desulfovibrio desulfuricans

substrate_name: reduced methyl viologen

enzyme: UniProt Q8VNU2

substrate: reduced methyl viologen


enzyme_display_name: UniProt Q9S1E5

organism: Wolinella succinogenes

enzyme: UniProt Q9S1E5


substrate_name: hydroxylamine

substrate: hydroxylamine


enzyme_display_name: UniProt B3E641

organism: Trichlorobacter lovleyi

enzyme: UniProt B3E641


enzyme_display_name: trimethylamine-N-oxide reductase

organism: Shewanella sp.

substrate_name: Trimethylamine N-oxide

enzyme: trimethylamine-N-oxide reductase

substrate: Trimethylamine N-oxide


organism: Rhodospirillum rubrum

substrate_name: Dimethylsulfoxide

smiles: CS(=O)C

substrate: Dimethylsulfoxide


substrate_name: N,N-dimethyldodecylamine N-oxide

smiles: CCCCCCCCCCCC[N+](C)(C)[O-]

substrate: N,N-dimethyldodecylamine N-oxide


substrate_name: gamma-picoline N-oxide

substrate: gamma-picoline N-oxide


substrate_name: Trimethylamine-N-oxide

substrate: Trimethylamine-N-oxide


organism: Enterococcus faecium


substrate_name: NADH

assay_conditions_summary: pH 7; 298.15 K; cofactor NADPH

substrate: NADH


substrate_name: NADPH

column_label: Table 2; row NADPH; column Km (uM)

raw_value_unit_evidence: Table 2 row NADPH column Km (uM) raw value 1.0 uM

row_label: NADPH

substrate: NADPH


column_label: Table 2; row NADH; column Km (uM)

raw_value_unit_evidence: Table 2 row NADH column Km (uM) raw value 170 uM

row_label: NADH


organism: Escherichia coli

column_label: PMC Table 2 row Anthaquinone-2-sulfonate reports Km 8.1 uM; kcat 1.1 s^-1; kcat/Km 135.8 s^-1 mM^-1.

normalized_value_unit_evidence: Km normalized_value=0.0081 mM; kcat normalized_value=1.1 s^-1; kcat/Km normalized_value=135.8 s^-1 mM^-1

raw_value_unit_evidence: Km raw_value=8.1 raw_unit=uM; kcat raw_value=1.1 raw_unit=s^-1; kcat/Km raw_value=135.8 raw_unit=s^-1 mM^-1

table_label: Table 2


column_label: PMC Table 2 row Antraquinone-2,6-disulfonate reports Km 14.6 uM; kcat 2.6 s^-1; kcat/Km 178.1 s^-1 mM^-1.

normalized_value_unit_evidence: Km normalized_value=0.0146 mM; kcat normalized_value=2.6 s^-1; kcat/Km normalized_value=178.1 s^-1 mM^-1

raw_value_unit_evidence: Km raw_value=14.6 raw_unit=uM; kcat raw_value=2.6 raw_unit=s^-1; kcat/Km raw_value=178.1 raw_unit=s^-1 mM^-1


column_label: PMC Table 2 row 2-Hydroxy-1,4-naphthoquinone reports Km 87.3 uM; kcat 5.8 s^-1; kcat/Km 66.4 s^-1 mM^-1.

normalized_value_unit_evidence: Km normalized_value=0.0873 mM; kcat normalized_value=5.8 s^-1; kcat/Km normalized_value=66.4 s^-1 mM^-1

raw_value_unit_evidence: Km raw_value=87.3 raw_unit=uM; kcat raw_value=5.8 raw_unit=s^-1; kcat/Km raw_value=66.4 raw_unit=s^-1 mM^-1


column_label: PMC Table 2 row 2-Methyl-1,4-benzoquinone reports Km 1704.5 uM; kcat 72.0 s^-1; kcat/Km 42.2 s^-1 mM^-1.

normalized_value_unit_evidence: Km normalized_value=1.7045 mM; kcat normalized_value=72.0 s^-1; kcat/Km normalized_value=42.2 s^-1 mM^-1

raw_value_unit_evidence: Km raw_value=1704.5 raw_unit=uM; kcat raw_value=72.0 raw_unit=s^-1; kcat/Km raw_value=42.2 raw_unit=s^-1 mM^-1


column_label: Table 5; row Methyl Red; column group rAZR-GSTi; column Km (uM) cell 134; column Vmax (umole/min/mg) cell 13.1; column kcat (S^-1) cell 4.13; Table 5 prints no kcat/Km column.

normalized_value_unit_evidence: kcat 4.13 S^-1 = 4.13 s^-1. Km 134 uM = 0.134 mM (134 / 1000). kcat/Km is not printed in Table 5; the paper-cell quotient is paper_kcat / paper_km = 4.13 / 0.134 = 30.820895522388057 s^-1 mM^-1, which equals the catalog value 30.820895522388057 s^-1 mM^-1.

raw_value_unit_evidence: Table 5, row Methyl Red, columns under rAZR-GSTi: Km 134 uM; Vmax 13.1 umole/min/mg; kcat 4.13 S^-1.

row_label: Methyl Red; rAZR-GSTi (Geobacillus stearothermophilus IFO13737)


enzyme_display_name: nitrobenzene nitroreductase

organism: Psychrobacter sp. ANT206

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

enzyme: nitrobenzene nitroreductase


substrate_name: nitrobenzene

substrate: nitrobenzene


enzyme_display_name: nitrite reductase (cytochrome; ammonia-forming)

organism: Campylobacter curvus

proof_lines: oxidation reaction, recombinant MBP-tagged wild-type enzyme, pH 7.0, 37°C

proof_lines: reduction reaction, recombinant MBP-tagged wild-type enzyme, pH 7.0, 37°C


proof_lines: recombinant MBP-tagged wild-type enzyme, pH 7.0, 37°C

enzyme: nitrite reductase (cytochrome; ammonia-forming)


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

proof_lines: recombinant MBP-tagged mutant W434Y, pH 7.0, 37°C


proof_lines: oxidation reaction, recombinant MBP-tagged mutant W434Y, pH 7.0, 37°C

proof_lines: reduction reaction, recombinant MBP-tagged mutant W434Y, pH 7.0, 37°C


enzyme_display_name: nitric oxide reductase [NAD(P)+, nitrous oxide-forming]

organism: Aspergillus oryzae

assay_conditions_summary: pH 7.2; 310.15 K; cofactor NADPH

proof_lines: in 50 mM sodium phosphate (pH 7.2), at 37°C


enzyme: nitric oxide reductase [NAD(P)+, nitrous oxide-forming]


enzyme_display_name: nitrite reductase (NADH)

organism: Rhodobacter capsulatus

assay_conditions_summary: pH 9; 303.15 K; cofactor NADH

enzyme: nitrite reductase (NADH)


organism: Gluconobacter oxydans

assay_conditions_summary: pH 8; 310.15 K; cofactor NADPH


organism: Caminibacter mediatlanticus


organism: Trypanosoma brucei brucei

substrate_name: GMP

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

sequence_source: multiple

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

assay_conditions_summary: pH 8; 308.15 K; cofactor NADPH

proof_lines:
pH 8.0, 35°C
Raw SKiD Env: Temp=35°C, pH=8

source_databases_merged: brenda skid

substrate: GMP


assay_conditions_summary: pH 8; 308.15 K; buffer 50 mM sodium phosphate; cofactor NADPH

column_label: PMC4701174 Table 1 row TbGMPR column K_m NADPH reports 12.3 +/- 0.8 uM.

raw_value_unit_evidence: Table 1 reports TbGMPR K_m NADPH raw_value=12.3 raw_unit=uM.

row_label: TbGMPR

table_label: PMC4701174 Table 1


organism: Arabidopsis thaliana

proof_lines: N-terminal plus C-terminal fragment, pH 7, 22°C


organism: Rattus norvegicus

substrate_name: Amaranth

substrate: Amaranth


organism: Priestia megaterium


enzyme_display_name: nitrite reductase (NO-forming)

organism: Paramagnetospirillum magnetotacticum

proof_lines: electron donor: reduced tetramethyl-4-phenylenediamine

enzyme: nitrite reductase (NO-forming)


organism: Proteus vulgaris


organism: Homo sapiens

proof_lines: pH 7.4, temperature not specified in the publication


organism: Histoplasma capsulatum

substrate_name: NO

smiles: [No]

proof_lines: in 100 mM potassium phosphate (pH 7.5), at 25°C

substrate: NO


organism: Mycobacterium tuberculosis


organism: Paracoccus pantotrophus

assay_conditions_summary: pH 7

proof_lines: 25°C, pH 7.0, wild-type, without pre-reduction with dithionite

proof_lines: 25°C, pH 7.0, wild-type, pre-reduction with dithionite

proof_lines: 25°C, pH 7.0, activated wild-type enzyme


smiles: O=O


sequence_variant_note: Mutation signature Y25S could not be applied safely because the resolved sequence does not match the expected reference residue(s).


organism: Ectopseudomonas oleovorans

assay_conditions_summary: pH 8

proof_lines: imobilized enzyme, 25°C, pH not specified in the publication

proof_lines: temperature not specified in the publication, pH 8.0

source_databases_merged: brenda uniprot


proof_lines: 25°C, pH not specified in the publication


substrate_name: nitrofurazone

substrate: nitrofurazone


enzyme_display_name: FMN-dependent NAD(P)H:quinone oxidoreductase 1

organism: Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1)

substrate_name: methyl red (in the presence of NADPH)

assay_conditions_summary: cofactor NADPH

column_label: PMC Table 2 row PaAzoR1 / Km / MRE reports Km 92.7 +/- 7.6 uM.

normalized_value_unit_evidence: Km normalized_value=0.0927 mM

raw_value_unit_evidence: Km raw_value=92.7 +/- 7.6 raw_unit=uM

row_label: PaAzoR1 / Km / MRE

substrate: methyl red (in the presence of NADPH)


substrate_name: sulfasalazine (in the presence of NADPH)

column_label: JMB 2007 PMID 17904577 Table 2 row paAzoR1 / Km / Sulfasalazine reports Km 69 +/- 2 uM.

normalized_value_unit_evidence: Km normalized_value=0.069 mM

raw_value_unit_evidence: Km raw_value=69 +/- 2 raw_unit=uM

row_label: paAzoR1 / Km / Sulfasalazine

enzyme: FMN-dependent NAD(P)H:quinone oxidoreductase 1


substrate_name: balsalazide (in the presence of NADPH)

column_label: PMC Table 2 row PaAzoR1 / Km / Balsalazide reports Km 98.6 +/- 4.2 uM.

normalized_value_unit_evidence: Km normalized_value=0.0986 mM

raw_value_unit_evidence: Km raw_value=98.6 +/- 4.2 raw_unit=uM

row_label: PaAzoR1 / Km / Balsalazide

substrate: balsalazide (in the presence of NADPH)


substrate_name: olsalazine (in the presence of NADPH)

column_label: JMB 2007 PMID 17904577 Table 2 row paAzoR1 / Km / Olsalazine reports Km 104 +/- 3 uM.

normalized_value_unit_evidence: Km normalized_value=0.104 mM

raw_value_unit_evidence: Km raw_value=104 +/- 3 raw_unit=uM

row_label: paAzoR1 / Km / Olsalazine

substrate: olsalazine (in the presence of NADPH)


substrate_name: NADH (in the presence of methyl red)

column_label: PMC Table 2 row PaAzoR1 / Km / NADH reports Km 538 +/- 21 uM.

normalized_value_unit_evidence: Km normalized_value=0.538 mM

raw_value_unit_evidence: Km raw_value=538 +/- 21 raw_unit=uM

row_label: PaAzoR1 / Km / NADH

substrate: NADH (in the presence of methyl red)


substrate_name: NADPH (in the presence of methyl red)

column_label: PMC Table 2 row PaAzoR1 / Km / NADPH reports Km 1197 +/- 33 uM.

normalized_value_unit_evidence: Km normalized_value=1.197 mM

raw_value_unit_evidence: Km raw_value=1197 +/- 33 raw_unit=uM

row_label: PaAzoR1 / Km / NADPH

substrate: NADPH (in the presence of methyl red)


organism: Alcaligenes faecalis

substrate_name: nitrite

sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_mutation_ranked_match and applied mutation signature M150H with inferred residue-numbering offset +5 to reconstruct the assayed variant sequence.

substrate: nitrite


sequence_variant_note: Resolved sequence already matches the mutant residue(s) after applying an inferred residue-numbering offset +3, so it is treated as a source-provided variant sequence.


sequence_variant_note: Mutation signature M150G could not be applied safely because the resolved sequence does not match the expected reference residue(s).


enzyme_display_name: FMN-dependent NADH-azoreductase

organism: Shewanella oneidensis

substrate_name: Methyl red

sequence_variant_note: Resolved canonical sequence from ncbi_refseq_accession:NP_719919.1 and applied mutation signature N96A to reconstruct the assayed variant sequence.

assay_conditions_summary: pH 7.4; cofactor NADH

column_label: Table 2. Kinetic parameters of wild-type and mutant soAzoR; row Methyl red / N96A; columns Km (uM), Kcat (s^-1), and Kcat/Km (uM^-1 s^-1); Methyl red / N96A: Km 93.15 uM; kcat 2.90 s^-1; kcat/Km 0.03 uM^-1*s^-1. Direct unit conversion divides Km by 1000 and multiplies printed Kcat/Km by 1000 to give Km=0.09315 mM; kcat=2.9 s^-1; kcat/Km=30 s^-1*mM^-1.

row_label: Methyl red / N96A

enzyme: FMN-dependent NADH-azoreductase

substrate: Methyl red


sequence_variant_note: Resolved canonical sequence from ncbi_refseq_accession:NP_719919.1 and applied mutation signature H143A to reconstruct the assayed variant sequence.

column_label: Table 2. Kinetic parameters of wild-type and mutant soAzoR; row Methyl red / H143A; columns Km (uM), Kcat (s^-1), and Kcat/Km (uM^-1 s^-1); Methyl red / H143A: Km 68.72 uM; kcat 6.35 s^-1; kcat/Km 0.09 uM^-1*s^-1. Direct unit conversion divides Km by 1000 and multiplies printed Kcat/Km by 1000 to give Km=0.06872 mM; kcat=6.35 s^-1; kcat/Km=90 s^-1*mM^-1.

row_label: Methyl red / H143A


sequence_variant_note: Resolved canonical sequence from ncbi_refseq_accession:NP_719919.1 and applied mutation signature S16A to reconstruct the assayed variant sequence.

column_label: Table 2. Kinetic parameters of wild-type and mutant soAzoR; row Methyl red / S16A; columns Km (uM), Kcat (s^-1), and Kcat/Km (uM^-1 s^-1); Methyl red / S16A: Km 76.76 uM; kcat 9.80 s^-1; kcat/Km 0.13 uM^-1*s^-1. Direct unit conversion divides Km by 1000 and multiplies printed Kcat/Km by 1000 to give Km=0.07676 mM; kcat=9.8 s^-1; kcat/Km=130 s^-1*mM^-1.

row_label: Methyl red / S16A


organism: Campylobacter fetus subsp. fetus


substrate_name: hydroxylamine

proof_lines: oxidation reaction, recombinant MBP-tagged wild-type enzyme, pH 7.0, 37°C

proof_lines: reduction reaction, recombinant MBP-tagged wild-type enzyme, pH 7.0, 37°C

substrate: hydroxylamine


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

proof_lines: recombinant MBP-tagged mutant W434Y, pH 7.0, 37°C


organism: Shewanella massilia

substrate_name: Dimethylsulfoxide

substrate: Dimethylsulfoxide


substrate_name: DL-methionine sulfoxide

smiles: CS(=O)CCC(C(=O)O)N

substrate: DL-methionine sulfoxide


substrate_name: diphenylsulfoxide

substrate: diphenylsulfoxide


substrate_name: dibutylsulfoxide

smiles: CCCCS(=O)CCCC

substrate: dibutylsulfoxide


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

proof_lines: initially oxidized enzyme, electron donor cytochrome c550

proof_lines: 25°C, pH 7.0, electron donor cytochrome c550

proof_lines: initially oxidized enzyme, electron donor horse heart cytochrome c


proof_lines: 25°C, pH 7.0, electron donor horse heart cytochrome c


organism: Wolinella succinogenes

sequence_variant_note: Resolved sequence already matches the mutant residue(s) after applying an inferred residue-numbering offset -5, so it is treated as a source-provided variant sequence.

proof_lines: mutant H264N, pH 7.0, temperature not specified in the publication


proof_lines: wild-type, pH 7.0, temperature not specified in the publication


substrate_name: menadione

substrate: menadione


substrate_name: NADPH

assay_conditions_summary: pH 8; cofactor NADPH

substrate: NADPH


organism: Bacillus subtilis (strain 168)

substrate_name: 7-cyano-7-deazaguanine (at pH 7.5 and 30 degrees Celsius)

assay_conditions_summary: pH 7.5; cofactor NADPH

proof_lines: kcat is 0.66 min(-1) (at pH 7.5 and 30 degrees Celsius).


substrate_name: NADPH (at pH 7.5 and 30 degrees Celsius)


organism: Thioalkalivibrio nitratireducens

substrate_name: sulfite

assay_conditions_summary: pH 7; buffer 0.1 M K+-phosphate

proof_lines: modified enzyme that contains an additional covalent bond between residues Tyr303 and Gln360, pH 7.0, 20°C

proof_lines: in 0.1 M K+-phosphate buffer, at pH 7.0 and 30°C

substrate: sulfite


organism: Geobacillus kaustophilus (strain HTA426)

substrate_name: 7-cyano-7-deazaguanine (at 55 degrees Celsius)

proof_lines: kcat is 3.9 min(-1) for the reduction of 7-cyano-7-deazaguanine, and 9.3 min(-1) for that of 5-cyanopyrrolo[2,3-d]pyrimidin-4-one (at 55 degrees Celsius).


substrate_name: NADPH (at 55 degrees Celsius)

substrate: NADPH (at 55 degrees Celsius)


organism: Escherichia coli (strain K12)

proof_lines: KM for preQ0 appears to be inferior to 1.5 uM, and kcat is 0.1268 sec(-1) (PubMed:23410922). kcat is 6.5 min(-1) with preQ0 as substrate and 3.6 min(-1) with 5-cyanopyrrolo[2,3-d]pyrimidin-4-one as substrate (PubMed:23595998).


organism: Paracoccus denitrificans


organism: Fusarium oxysporum

substrate_name: NO

proof_lines: in 0.1 mM sodium phosphate, pH 7.2, at 10°C

substrate: NO


organism: Escherichia coli

substrate_name: NADH

assay_conditions_summary: pH 7.4; 303.15 K; cofactor NADH

proof_lines: kcat is 1.1 sec(-1) with anthaquinone-2-sulfonate as substrate. kcat is 2.6 sec(-1) with antraquinone-2,6-disulfonate as substrate. kcat is 5.8 sec(-1) with 2-hydroxy-1,4-naphthoquinone as substrate. kcat is 72.0 sec(-1) with 2-methyl-1,4-b

source_databases_merged: skid uniprot

substrate: NADH


column_label: PMC Table 2 row 2-Hydroxy-1,4-naphthoquinone reports Km 87.3 uM.

normalized_value_unit_evidence: Km normalized_value=0.0873 mM

raw_value_unit_evidence: Km raw_value=87.3 raw_unit=uM


column_label: PMC Table 2 row 2-Methyl-1,4-benzoquinone reports Km 1704.5 uM.

normalized_value_unit_evidence: Km normalized_value=1.7045 mM

raw_value_unit_evidence: Km raw_value=1704.5 raw_unit=uM


substrate_name: methyl red

substrate: methyl red


organism: Rhodococcus opacus

assay_conditions_summary: pH 6; 298.15 K; cofactor NADH

proof_lines:
recombinant enzyme, pH 6.0, 25°C
Raw SKiD Env: Temp=25°C, pH=6


organism: Achromobacter xylosoxidans

substrate_name: nitrite

sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_mutation_ranked_match and applied mutation signature N90S with inferred residue-numbering offset +15 to reconstruct the assayed variant sequence.


substrate_name: p-methyl red

substrate: p-methyl red


proof_lines: kcat is 824 sec(-1) with nitrite as substrate. kcat is 2380 sec(-1) with hydroxylamine as substrate.

source_databases_merged: brenda skid uniprot


substrate_name: nitrofurazone

proof_lines: kcat is 13 sec(-1) with methyl red as substrate. kcat is 13 sec(-1) with sulfasalazine as substrate. kcat is 37 sec(-1) with balsalazide as substrate. kcat is 2.5 sec(-1) with olsalazine as substrate. kcat is 10 sec(-1) with NADH as substra

substrate: nitrofurazone


enzyme_display_name: Cytochrome c-552

organism: Shewanella oneidensis (strain ATCC 700550 / JCM 31522 / CIP 106686 / LMG 19005 / NCIMB 14063 / MR-1)

substrate_name: hydroxylamine

enzyme: Cytochrome c-552


enzyme_display_name: preQ1 synthase

organism: Bacillus subtilis

substrate_name: 7-Cyano-7-deazaguanine; NADPH

assay_conditions_summary: cofactor NADP+

enzyme: preQ1 synthase

substrate: 7-Cyano-7-deazaguanine; NADPH


enzyme_display_name: nitric oxide reductase [NAD(P), nitrous oxide-forming]

organism: Cupriavidus necator

substrate_name: NADH; Nitric oxide; H+

assay_conditions_summary: cofactor NAD+

substrate: NADH; Nitric oxide; H+


substrate_name: H+; Nitric oxide; NADH

enzyme: nitric oxide reductase [NAD(P), nitrous oxide-forming]

substrate: H+; Nitric oxide; NADH


organism: Saccharomyces cerevisiae

substrate_name: NADH; H+; Nitric oxide

substrate: NADH; H+; Nitric oxide


enzyme_display_name: FMN-dependent NADH:quinone oxidoreductase 2

substrate_name: 2,6-dichloroondophenol (DCIP) (at pH 7.5 and at 25 degrees Celsius)


enzyme_display_name: UniProt Q831B2

organism: Enterococcus faecalis

assay_conditions_summary: pH 7.1; 296.15 K; cofactor NADH

column_label: Table 3, Methyl red row, Km column, 0.024 mM.

normalized_value_unit_evidence: Methyl red Km=0.024 mM; the target chemical name is the same compound.

raw_value_unit_evidence: Table 3 Methyl red row reports Km 0.024 mM.

row_label: Methyl red

table_label: Table 3. Michaelis constants and maximal velocities of recombinant AzoA

enzyme: UniProt Q831B2


organism: Achromobacter cycloclastes

substrate_name: Reduced pseudoazurin; H+; Nitrite

substrate: Reduced pseudoazurin; H+; Nitrite


substrate_name: H+; Nitrite; Reduced pseudoazurin

substrate: H+; Nitrite; Reduced pseudoazurin


organism: Alcaligenes faecalis

substrate_name: Nitrite; Reduced pseudoazurin; H+

substrate: Nitrite; Reduced pseudoazurin; H+


organism: Candidatus Jettenia caeni

substrate_name: ferricytochrome c B0428

substrate: ferricytochrome c B0428


substrate_name: ferricytochrome c

substrate: ferricytochrome c


organism: Desulfovibrio desulfuricans

substrate_name: reduced phenosafranine

substrate: reduced phenosafranine


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_mutation_ranked_match and applied mutation signature G198E with inferred residue-numbering offset +14 to reconstruct the assayed variant sequence.

substrate: nitrite


substrate_name: 5-cyanopyrrolo[2,3-d]pyrimidin-4-one (at 55 degrees Celsius)


enzyme_display_name: Nitrobenzene nitroreductase

organism: Ectopseudomonas oleovorans

substrate_name: Unknown

enzyme: Nitrobenzene nitroreductase

substrate: Unknown


enzyme_display_name: UniProt P41407

enzyme: UniProt P41407


organism: Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1)


organism: Shewanella oneidensis

substrate_name: NADH

sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:Q8E990 and applied mutation signature R59A to reconstruct the assayed variant sequence.

column_label: Table 2 (Kinetic parameters of wild-type and mutant soAzoR), substrate block 'NADH', enzyme row 'R59A'; column 'Vmax (U mg-1)' = 175.27; column 'Km (uM)' = 343.3; column 'Kcat (s-1)' = 32.13; column 'Kcat/Km (uM-1 s-1)' = 0.09; column 'Relative activity (%)' = 83.

normalized_value_unit_evidence: Km: printed Table 2 cell 343.3 uM; 343.3 / 1000 = 0.3433 mM is the canonical paper value. kcat: printed Table 2 cell 32.13 s^(-1); the printed column head is Kcat (s-1), so no unit conversion applies and 32.13 s^(-1) is the canonical paper value. kcat/Km: printed Table 2 cell 0.09 uM^-1 s^-1; 1 uM^-1 s^-1 is 1000 s^-1 mM^-1, so 0.09 x 1000 = 90 s^-1 mM^-1 is the canonical paper value.

raw_value_unit_evidence: Table 2 (Kinetic parameters of wild-type and mutant soAzoR), substrate block 'NADH', enzyme row 'R59A': Km = 343.3 uM; Kcat = 32.13 s^(-1); printed Kcat/Km = 0.09 uM^-1 s^-1; printed Vmax = 175.27 U mg^-1; printed relative activity = 83 %.

row_label: Table 2, substrate block 'NADH', enzyme row 'R59A'

table_label: Table 2: Kinetic parameters of wild-type and mutant soAzoR (Journal of Molecular Graphics and Modelling 78 (2017) 213-220)


enzyme_display_name: Trimethylamine-N-oxide reductase 1

organism: Escherichia coli (strain K12)

substrate_name: TMAO with menadiol as the electron donor (at pH 6.5 and 37 degrees Celsius)

proof_lines: kcat is 95.24 sec(-1) with TMAO as substrate (at pH 6.5) (PubMed:30945846). kcat is 0.145 sec(-1) with TMAO as substrate and menadiol as the electron donor (at pH 6.5 and 37 degrees Celsius) (PubMed:39769096). kcat is 227.65 sec(-1) with TM

enzyme: Trimethylamine-N-oxide reductase 1


sequence_variant_note: Resolved canonical sequence from ncbi_refseq_accession:NP_719919.1 and applied mutation signature R59A to reconstruct the assayed variant sequence.

column_label: Table 2. Kinetic parameters of wild-type and mutant soAzoR; row NADH / R59A; columns Km (uM), Kcat (s^-1), and Kcat/Km (uM^-1 s^-1); NADH / R59A: Km 343.3 uM; kcat 32.13 s^-1; kcat/Km 0.09 uM^-1*s^-1. Direct unit conversion divides Km by 1000 and multiplies printed Kcat/Km by 1000 to give Km=0.3433 mM; kcat=32.13 s^-1; kcat/Km=90 s^-1*mM^-1.

row_label: NADH / R59A


column_label: Table 2. Kinetic parameters of wild-type and mutant soAzoR; row NADH / H143A; columns Km (uM), Kcat (s^-1), and Kcat/Km (uM^-1 s^-1); NADH / H143A: Km 433.80 uM; kcat 29.39 s^-1; kcat/Km 0.07 uM^-1*s^-1. Direct unit conversion divides Km by 1000 and multiplies printed Kcat/Km by 1000 to give Km=0.4338 mM; kcat=29.39 s^-1; kcat/Km=70 s^-1*mM^-1.

row_label: NADH / H143A


substrate_name: Methyl red

sequence_variant_note: Resolved canonical sequence from ncbi_refseq_accession:NP_719919.1 and applied mutation signature R139A to reconstruct the assayed variant sequence.

column_label: Table 2. Kinetic parameters of wild-type and mutant soAzoR; row Methyl red / R139A; columns Km (uM), Kcat (s^-1), and Kcat/Km (uM^-1 s^-1); Methyl red / R139A: Km 48.09 uM; kcat 21.48 s^-1; kcat/Km 0.45 uM^-1*s^-1. Direct unit conversion divides Km by 1000 and multiplies printed Kcat/Km by 1000 to give Km=0.04809 mM; kcat=21.48 s^-1; kcat/Km=450 s^-1*mM^-1.

row_label: Methyl red / R139A

substrate: Methyl red


sequence_variant_note: Resolved canonical sequence from ncbi_refseq_accession:NP_719919.1 and applied mutation signature Y119A to reconstruct the assayed variant sequence.

column_label: Table 2. Kinetic parameters of wild-type and mutant soAzoR; row Methyl red / Y119A; columns Km (uM), Kcat (s^-1), and Kcat/Km (uM^-1 s^-1); Methyl red / Y119A: Km 19.28 uM; kcat 20.02 s^-1; kcat/Km 1.04 uM^-1*s^-1. Direct unit conversion divides Km by 1000 and multiplies printed Kcat/Km by 1000 to give Km=0.01928 mM; kcat=20.02 s^-1; kcat/Km=1040 s^-1*mM^-1.

row_label: Methyl red / Y119A


column_label: Table 2. Kinetic parameters of wild-type and mutant soAzoR; row NADH / S16A; columns Km (uM), Kcat (s^-1), and Kcat/Km (uM^-1 s^-1); NADH / S16A: Km 530.7 uM; kcat 19.18 s^-1; kcat/Km 0.04 uM^-1*s^-1. Direct unit conversion divides Km by 1000 and multiplies printed Kcat/Km by 1000 to give Km=0.5307 mM; kcat=19.18 s^-1; kcat/Km=40 s^-1*mM^-1.

row_label: NADH / S16A

substrate: NADH


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

column_label: Table 2 columns substrate, enzyme, Km (uM), Kcat (s^-1), and Kcat/Km (uM^-1 s^-1); row matched by substrate MR/Methyl red and enzyme WT.

normalized_value_unit_evidence: kcat 33.86 s^-1; Km 0.01155 mM; printed kcat/Km 2930 s^-1 mM^-1; current export kcat and kcat/Km appear scaled 1000x above the paper values.

raw_value_unit_evidence: Table 2 MR WT row: Vmax 184.70 U mg^-1; Km 11.55 uM; Kcat 33.86 s^-1; Kcat/Km 2.93 uM^-1 s^-1; relative activity 100%.

row_label: MR; WT

table_label: Table 2, Kinetic parameters of wild-type and mutant soAzoR


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:Q8E990 and applied mutation signature F117A to reconstruct the assayed variant sequence.

column_label: Table 2 (Kinetic parameters of wild-type and mutant soAzoR), substrate block 'MR', enzyme row 'F117A'; column 'Vmax (U mg-1)' = 383.76; column 'Km (uM)' = 16.03; column 'Kcat (s-1)' = 70.36; column 'Kcat/Km (uM-1 s-1)' = 4.39; column 'Relative activity (%)' = 208.

normalized_value_unit_evidence: Km: printed Table 2 cell 16.03 uM; 16.03 / 1000 = 0.01603 mM is the canonical paper value. kcat: printed Table 2 cell 70.36 s^(-1); the printed column head is Kcat (s-1), so no unit conversion applies and 70.36 s^(-1) is the canonical paper value. kcat/Km: printed Table 2 cell 4.39 uM^-1 s^-1; 1 uM^-1 s^-1 is 1000 s^-1 mM^-1, so 4.39 x 1000 = 4390 s^-1 mM^-1 is the canonical paper value.

raw_value_unit_evidence: Table 2 (Kinetic parameters of wild-type and mutant soAzoR), substrate block 'MR', enzyme row 'F117A': Km = 16.03 uM; Kcat = 70.36 s^(-1); printed Kcat/Km = 4.39 uM^-1 s^-1; printed Vmax = 383.76 U mg^-1; printed relative activity = 208 %.

row_label: Table 2, substrate block 'MR', enzyme row 'F117A'


column_label: Table 2 (Kinetic parameters of wild-type and mutant soAzoR), substrate block 'MR', enzyme row 'R59A'; column 'Vmax (U mg-1)' = 129.15; column 'Km (uM)' = 29.60; column 'Kcat (s-1)' = 23.68; column 'Kcat/Km (uM-1 s-1)' = 0.80; column 'Relative activity (%)' = 70.

normalized_value_unit_evidence: Km: printed Table 2 cell 29.60 uM; 29.60 / 1000 = 0.0296 mM is the canonical paper value. kcat: printed Table 2 cell 23.68 s^(-1); the printed column head is Kcat (s-1), so no unit conversion applies and 23.68 s^(-1) is the canonical paper value. kcat/Km: printed Table 2 cell 0.80 uM^-1 s^-1; 1 uM^-1 s^-1 is 1000 s^-1 mM^-1, so 0.80 x 1000 = 800 s^-1 mM^-1 is the canonical paper value.

raw_value_unit_evidence: Table 2 (Kinetic parameters of wild-type and mutant soAzoR), substrate block 'MR', enzyme row 'R59A': Km = 29.60 uM; Kcat = 23.68 s^(-1); printed Kcat/Km = 0.80 uM^-1 s^-1; printed Vmax = 129.15 U mg^-1; printed relative activity = 70 %.

row_label: Table 2, substrate block 'MR', enzyme row 'R59A'


column_label: Table 2 (Kinetic parameters of wild-type and mutant soAzoR), substrate block 'NADH', enzyme row 'F117A'; column 'Vmax (U mg-1)' = 372.18; column 'Km (uM)' = 27.14; column 'Kcat (s-1)' = 68.23; column 'Kcat/Km (uM-1 s-1)' = 2.51; column 'Relative activity (%)' = 176.

normalized_value_unit_evidence: Km: printed Table 2 cell 27.14 uM; 27.14 / 1000 = 0.02714 mM is the canonical paper value. kcat: printed Table 2 cell 68.23 s^(-1); the printed column head is Kcat (s-1), so no unit conversion applies and 68.23 s^(-1) is the canonical paper value. kcat/Km: printed Table 2 cell 2.51 uM^-1 s^-1; 1 uM^-1 s^-1 is 1000 s^-1 mM^-1, so 2.51 x 1000 = 2510 s^-1 mM^-1 is the canonical paper value.

raw_value_unit_evidence: Table 2 (Kinetic parameters of wild-type and mutant soAzoR), substrate block 'NADH', enzyme row 'F117A': Km = 27.14 uM; Kcat = 68.23 s^(-1); printed Kcat/Km = 2.51 uM^-1 s^-1; printed Vmax = 372.18 U mg^-1; printed relative activity = 176 %.

row_label: Table 2, substrate block 'NADH', enzyme row 'F117A'


column_label: Table 2 (Kinetic parameters of wild-type and mutant soAzoR), substrate block 'MR', enzyme row 'WT'; column 'Vmax (U mg-1)' = 184.70; column 'Km (uM)' = 11.55; column 'Kcat (s-1)' = 33.86; column 'Kcat/Km (uM-1 s-1)' = 2.93; column 'Relative activity (%)' = 100.

normalized_value_unit_evidence: Km: printed Table 2 cell 11.55 uM; 11.55 / 1000 = 0.01155 mM is the canonical paper value. kcat: printed Table 2 cell 33.86 s^(-1); the printed column head is Kcat (s-1), so no unit conversion applies and 33.86 s^(-1) is the canonical paper value. kcat/Km: printed Table 2 cell 2.93 uM^-1 s^-1; 1 uM^-1 s^-1 is 1000 s^-1 mM^-1, so 2.93 x 1000 = 2930 s^-1 mM^-1 is the canonical paper value.

raw_value_unit_evidence: Table 2 (Kinetic parameters of wild-type and mutant soAzoR), substrate block 'MR', enzyme row 'WT': Km = 11.55 uM; Kcat = 33.86 s^(-1); printed Kcat/Km = 2.93 uM^-1 s^-1; printed Vmax = 184.70 U mg^-1; printed relative activity = 100 %.

row_label: Table 2, substrate block 'MR', enzyme row 'WT'


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:Q8E990 and applied mutation signature Y119A to reconstruct the assayed variant sequence.

column_label: Table 2 (Kinetic parameters of wild-type and mutant soAzoR), substrate block 'MR', enzyme row 'Y119A'; column 'Vmax (U mg-1)' = 109.19; column 'Km (uM)' = 19.28; column 'Kcat (s-1)' = 20.02; column 'Kcat/Km (uM-1 s-1)' = 1.04; column 'Relative activity (%)' = 59.

normalized_value_unit_evidence: Km: printed Table 2 cell 19.28 uM; 19.28 / 1000 = 0.01928 mM is the canonical paper value. kcat: printed Table 2 cell 20.02 s^(-1); the printed column head is Kcat (s-1), so no unit conversion applies and 20.02 s^(-1) is the canonical paper value. kcat/Km: printed Table 2 cell 1.04 uM^-1 s^-1; 1 uM^-1 s^-1 is 1000 s^-1 mM^-1, so 1.04 x 1000 = 1040 s^-1 mM^-1 is the canonical paper value.

raw_value_unit_evidence: Table 2 (Kinetic parameters of wild-type and mutant soAzoR), substrate block 'MR', enzyme row 'Y119A': Km = 19.28 uM; Kcat = 20.02 s^(-1); printed Kcat/Km = 1.04 uM^-1 s^-1; printed Vmax = 109.19 U mg^-1; printed relative activity = 59 %.

row_label: Table 2, substrate block 'MR', enzyme row 'Y119A'


enzyme_display_name: FMN-dependent NADPH-azoreductase

organism: Staphylococcus aureus

enzyme: FMN-dependent NADPH-azoreductase


enzyme_display_name: Cytochrome c nitrite reductase subunit NrfA

organism: Nitratidesulfovibrio vulgaris (strain ATCC 29579 / DSM 644 / CCUG 34227 / NCIMB 8303 / VKM B-1760 / Hildenborough)


organism: Thioalkalivibrio nitratireducens


column_label: Table 2. Kinetic parameters of wild-type and mutant soAzoR; row NADH / R139A; columns Km (uM), Kcat (s^-1), and Kcat/Km (uM^-1 s^-1); NADH / R139A: Km 224.10 uM; kcat 30.2 s^-1; kcat/Km 0.14 uM^-1*s^-1. Direct unit conversion divides Km by 1000 and multiplies printed Kcat/Km by 1000 to give Km=0.2241 mM; kcat=30.2 s^-1; kcat/Km=140 s^-1*mM^-1.

row_label: NADH / R139A


column_label: Table 2. Kinetic parameters of wild-type and mutant soAzoR; row NADH / N96A; columns Km (uM), Kcat (s^-1), and Kcat/Km (uM^-1 s^-1); NADH / N96A: Km 315.80 uM; kcat 14.42 s^-1; kcat/Km 0.05 uM^-1*s^-1. Direct unit conversion divides Km by 1000 and multiplies printed Kcat/Km by 1000 to give Km=0.3158 mM; kcat=14.42 s^-1; kcat/Km=50 s^-1*mM^-1.

row_label: NADH / N96A


column_label: Table 2. Kinetic parameters of wild-type and mutant soAzoR; row NADH / Y119A; columns Km (uM), Kcat (s^-1), and Kcat/Km (uM^-1 s^-1); NADH / Y119A: Km 655.10 uM; kcat 17.17 s^-1; kcat/Km 0.03 uM^-1*s^-1. Direct unit conversion divides Km by 1000 and multiplies printed Kcat/Km by 1000 to give Km=0.6551 mM; kcat=17.17 s^-1; kcat/Km=30 s^-1*mM^-1.

row_label: NADH / Y119A


table_label: Table 2. Kinetic parameters of wild-type and mutant soAzoR


sequence_variant_note: Resolved canonical sequence from ncbi_refseq_accession:NP_719919.1 and applied mutation signature F117A to reconstruct the assayed variant sequence.

column_label: Table 2. Kinetic parameters of wild-type and mutant soAzoR; row Methyl red / F117A; columns Km (uM), Kcat (s^-1), and Kcat/Km (uM^-1 s^-1); Methyl red / F117A: Km 16.03 uM; kcat 70.36 s^-1; kcat/Km 4.39 uM^-1*s^-1. Direct unit conversion divides Km by 1000 and multiplies printed Kcat/Km by 1000 to give Km=0.01603 mM; kcat=70.36 s^-1; kcat/Km=4390 s^-1*mM^-1.

row_label: Methyl red / F117A


enzyme_display_name: nitrobenzene nitroreductase

organism: Gluconobacter oxydans


assay_conditions_summary: pH 8; 310.15 K; buffer 50 mM phosphate; cofactor NADPH

column_label: Table 3 columns are Substrate/cofactor, Km (uM), Vmax (U/mg), kcat (S^-1), kcat/Km (S^-1/uM); CB1954 row cells 54 +/- 11, 26.24 +/- 2.74, 1.06, 0.020.

normalized_value_unit_evidence: Km 54 uM = 0.054 mM; kcat 1.06 s^-1; kcat/Km 0.020 s^-1/uM = 20 s^-1 mM^-1, matching expected 19.62962962962963 within table precision/rounding.

table_label: Springer official article Table 3, Kinetic parameters of Gox0834 from G. oxydans


substrate_name: NADPH

column_label: Table 3 columns are Substrate/cofactor, Km (uM), Vmax (U/mg), kcat (S^-1), kcat/Km (S^-1/uM); NADPH row cells 153 +/- 66, 26.20 +/- 2.70, 1.05, 0.0069.

normalized_value_unit_evidence: Km 153 uM = 0.153 mM; kcat 1.05 s^-1; kcat/Km 0.0069 s^-1/uM = 6.9 s^-1 mM^-1, matching expected 6.862745098039216 within table precision/rounding.

row_label: NADPH

substrate: NADPH


substrate_name: nitrobenzene

column_label: Table 3 columns are Substrate/cofactor, Km (uM), Vmax (U/mg), kcat (S^-1), kcat/Km (S^-1/uM); nitrobenzene row cells 545 +/- 70, 4.59 +/- 0.33, 0.18, 0.00033.

normalized_value_unit_evidence: Km 545 uM = 0.545 mM; kcat 0.18 s^-1; kcat/Km 0.00033 s^-1/uM = 0.33 s^-1 mM^-1, matching expected 0.33027522935779813 within table precision/rounding.

raw_value_unit_evidence: Table 3 nitrobenzene row: Km 545 +/- 70 uM, Vmax 4.59 +/- 0.33 U/mg, kcat 0.18 s^-1, kcat/Km 0.00033 s^-1/uM.

row_label: nitrobenzene

substrate: nitrobenzene


enzyme_display_name: nitrite reductase (cytochrome; ammonia-forming)

organism: Escherichia coli

substrate_name: Reduced methyl viologen; Nitrite; H+

substrate: Reduced methyl viologen; Nitrite; H+


substrate_name: Nitrite; H+; Reduced methyl viologen

substrate: Nitrite; H+; Reduced methyl viologen


column_label: Table 2. Kinetic parameters of wild-type and mutant soAzoR; row Methyl red / H143A; columns Km (uM), Kcat (s^-1), and Kcat/Km (uM^-1 s^-1); Methyl red / H143A: Km 68.72 uM; kcat 6.35 s^-1; kcat/Km 0.09 uM^-1*s^-1. Direct unit conversion divides Km by 1000 and multiplies printed Kcat/Km by 1000 to give Km=0.06872 mM; kcat=6.35 s^-1; kcat/Km=90 s^-1*mM^-1.

row_label: Methyl red / H143A


column_label: Table 2. Kinetic parameters of wild-type and mutant soAzoR; row Methyl red / N96A; columns Km (uM), Kcat (s^-1), and Kcat/Km (uM^-1 s^-1); Methyl red / N96A: Km 93.15 uM; kcat 2.90 s^-1; kcat/Km 0.03 uM^-1*s^-1. Direct unit conversion divides Km by 1000 and multiplies printed Kcat/Km by 1000 to give Km=0.09315 mM; kcat=2.9 s^-1; kcat/Km=30 s^-1*mM^-1.

row_label: Methyl red / N96A


column_label: Table 2. Kinetic parameters of wild-type and mutant soAzoR; row Methyl red / S16A; columns Km (uM), Kcat (s^-1), and Kcat/Km (uM^-1 s^-1); Methyl red / S16A: Km 76.76 uM; kcat 9.80 s^-1; kcat/Km 0.13 uM^-1*s^-1. Direct unit conversion divides Km by 1000 and multiplies printed Kcat/Km by 1000 to give Km=0.07676 mM; kcat=9.8 s^-1; kcat/Km=130 s^-1*mM^-1.

row_label: Methyl red / S16A


organism: Pseudomonas denitrificans (nom. rej.)


enzyme_display_name: trimethylamine-N-oxide reductase

organism: Shewanella massilia

substrate_name: alpha-picoline N-oxide

substrate: alpha-picoline N-oxide


substrate_name: 4-methylmorpholine N-oxide

substrate: 4-methylmorpholine N-oxide


substrate_name: Trimethylamine N-oxide

substrate: Trimethylamine N-oxide


substrate_name: N,N-dimethyldodecylamine N-oxide

substrate: N,N-dimethyldodecylamine N-oxide


substrate_name: methyl red

column_label: Abstract reports apparent Km values for NADPH and Methyl Red substrates were 0.074 and 0.057 mM, respectively.

raw_value_unit_evidence: Km raw_value=0.057 mM raw_unit=mM

row_label: S. aureus Azo1 / Methyl Red

table_label: PubMed XML abstract PMID 15870453

substrate: methyl red


enzyme_display_name: UniProt P0ABK9

substrate_name: nitrite

column_label: Official ACS article visible text, Experimental Procedures/Results context for purified periplasmic cytochrome c nitrite reductase

normalized_value_unit_evidence: paper kcat 770 s^-1 and Km 0.028 mM; sheet kcat 1350 s^-1 and Km 0.046 mM

raw_value_unit_evidence: NrfA turnover with nitrite 770 NO2- s-1; KM 28 uM (NO2-)

row_label: Purified E. coli NrfA; nitrite substrate

table_label: Official ACS article visible prose

enzyme: UniProt P0ABK9


normalized_value_unit_evidence: paper Km 28 uM = 0.028 mM; sheet Km 0.022 mM


substrate_name: hydroxylamine

substrate: hydroxylamine


substrate_name: Hydroxylamine

column_label: Results and Discussion purified-NrfA activity paragraph: hydroxylamine turnover 2380 NH2OH s^-1 and KM 30 mM (NH2OH); no separate kcat/KM cell.

normalized_value_unit_evidence: kcat 2380 s^-1; Km 30 mM; same-measurement derived kcat/Km 79.33333333333333 s^-1*mM^-1.

raw_value_unit_evidence: Hydroxylamine measurement: turnover 2380 NH2OH s^-1; KM 30 mM (NH2OH); no independently printed kcat/KM cell.

row_label: hydroxylamine measurement

table_label: Results and Discussion activity paragraph

substrate: Hydroxylamine


organism: Bacillus subtilis

assay_conditions_summary: pH 7.5; 303.15 K; cofactor NADPH


substrate_name: reduced diquat

substrate: reduced diquat


substrate_name: reduced methyl viologen

substrate: reduced methyl viologen


substrate_name: reduced phenosafranine

substrate: reduced phenosafranine


substrate_name: NADH


substrate_name: Methyl red


column_label: Table 2 (Kinetic parameters of wild-type and mutant soAzoR), substrate block 'NADH', enzyme row 'WT'; column 'Vmax (U mg-1)' = 211.90; column 'Km (uM)' = 187.77; column 'Kcat (s-1)' = 33.90; column 'Kcat/Km (uM-1 s-1)' = 0.18; column 'Relative activity (%)' = 100.

normalized_value_unit_evidence: Km: printed Table 2 cell 187.77 uM; 187.77 / 1000 = 0.18777 mM is the canonical paper value. kcat: printed Table 2 cell 33.90 s^(-1); the printed column head is Kcat (s-1), so no unit conversion applies and 33.90 s^(-1) is the canonical paper value. kcat/Km: printed Table 2 cell 0.18 uM^-1 s^-1; 1 uM^-1 s^-1 is 1000 s^-1 mM^-1, so 0.18 x 1000 = 180 s^-1 mM^-1 is the canonical paper value.

raw_value_unit_evidence: Table 2 (Kinetic parameters of wild-type and mutant soAzoR), substrate block 'NADH', enzyme row 'WT': Km = 187.77 uM; Kcat = 33.90 s^(-1); printed Kcat/Km = 0.18 uM^-1 s^-1; printed Vmax = 211.90 U mg^-1; printed relative activity = 100 %.

row_label: Table 2, substrate block 'NADH', enzyme row 'WT'


organism: Bacillus subtilis (strain 168)

substrate_name: NADH (at pH 7.5 and at 25 degrees Celsius)

assay_conditions_summary: pH 7.5; cofactor NADH

enzyme: FMN-dependent NADH:quinone oxidoreductase 2


organism: Enterococcus faecalis (strain ATCC 700802 / V583)

assay_conditions_summary: cofactor NADH

column_label: The abstract reports apparent Km values for NADH and Methyl red of 0.14 and 0.024 mM, respectively.

raw_value_unit_evidence: Km raw_value=0.024 mM raw_unit=mM

row_label: Prose segment: Methyl red Km

table_label: PubMed PMID 15003265 abstract


substrate: nitrite


organism: Enterococcus faecalis

column_label: Table 3; row NADH; Km (mM) column; cell 0.14.

raw_value_unit_evidence: Table 3 NADH row, Km column: 0.14 mM.

row_label: NADH

table_label: Table 3. Michaelis constants (Km) and maximal velocities (Vmax) of recombinant AzoA of E. faecalis expressed in E. coli


enzyme_display_name: GMP reductase

organism: Leishmania donovani

substrate_name: GMP

column_label: Abstract states the K'm for GMP was 21 microM at pH 6.9.

raw_value_unit_evidence: Km raw_value=21 raw_unit=microM

row_label: Leishmania donovani GMP reductase with GMP

table_label: PubMed abstract PMID 7159467

enzyme: GMP reductase

substrate: GMP


enzyme_display_name: azobenzene reductase

organism: Pantoea agglomerans

assay_conditions_summary: pH 7; cofactor NADH

enzyme: azobenzene reductase

substrate: NADH


organism: Mycolicibacterium smegmatis (strain ATCC 700084 / mc(2)155)

substrate_name: NADPH (at pH 7.8)

assay_conditions_summary: pH 6.6; cofactor NADPH

substrate: NADPH (at pH 7.8)


organism: Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)

assay_conditions_summary: pH 7.8; cofactor NADPH

column_label: Table 1 lists Mtb pH 7.8 NADPH Kmapp 63 +/- 7 microM; Results state apparent Km values for NADPH were 30 +/- 4 microM and 63 +/- 7 microM for Msm and Mtb GMPR respectively.

raw_value_unit_evidence: Km raw_value=63 +/- 7 raw_unit=microM

row_label: Mtb GMPR at pH 7.8 with NADPH varied

table_label: PMC full-text PMID 35338694 / Table 1 and kinetic-parameters Results paragraph


enzyme_display_name: nitrite reductase (NO-forming)

organism: Ralstonia pickettii


organism: Salmonella enterica subsp. enterica serovar Typhimurium

proof_lines:
GMP reductase
GMP
TPNH


enzyme_display_name: Cytochrome c-552

organism: Escherichia coli (strain K12)

substrate_name: sulfite

assay_conditions_summary: pH 7

column_label: Abstract reports the Km (~70 microM) for sulfite reduction at pH 7.

raw_value_unit_evidence: Km raw_value=~70 microM raw_unit=microM

row_label: Escherichia coli NrfA / sulfite reduction

table_label: PubMed XML abstract PMID 20629638

substrate: sulfite


organism: Alcaligenes faecalis


organism: Pseudomonas sp.

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


organism: Thioalkalivibrio nitratireducens

enzyme: nitrite reductase (cytochrome; ammonia-forming)


enzyme_display_name: FMN-dependent NADH-azoreductase

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

proof_lines: mutant R21G/N121A, pH 7.1, temperature not specified in the publication


proof_lines: mutant R21G/N121Q, pH 7.1, temperature not specified in the publication


proof_lines: mutant R21K, pH 7.1, temperature not specified in the publication


proof_lines: wild-type, pH 7.1, temperature not specified in the publication


proof_lines: mutant N121Q, pH 7.1, temperature not specified in the publication


proof_lines: mutant R21K/N121A, pH 7.1, temperature not specified in the publication


proof_lines: mutant R21K/N121Q, pH 7.1, temperature not specified in the publication


assay_conditions_summary: pH 7.1; cofactor NADH


organism: Roseobacter denitrificans

enzyme: trimethylamine-N-oxide reductase


enzyme_display_name: cytochrome-b5 reductase

organism: Sus scrofa

substrate_name: ferricytochrome b5

assay_conditions_summary: pH 7; 298.15 K; cofactor NADH

enzyme: cytochrome-b5 reductase

substrate: ferricytochrome b5


substrate_name: ferricyanide

substrate: ferricyanide


organism: Achromobacter xylosoxidans

enzyme: nitrite reductase (NO-forming)


proof_lines:
25°C, wild-type
wild-type, pH 7.1


organism: Escherichia coli

substrate_name: N,N-dimethylhexylamine N-oxide

proof_lines:
tungsten-restored enzyme
molybdenum-restored enzyme
with trimethylamine N-oxide, pH 6.9

substrate: N,N-dimethylhexylamine N-oxide


substrate_name: N-cyclohexyldimethylamine N-oxide

substrate: N-cyclohexyldimethylamine N-oxide


substrate_name: 3-hydroxypyridine N-oxide

substrate: 3-hydroxypyridine N-oxide


substrate_name: gamma-picoline N-oxide

proof_lines:
molybdenum-restored enzyme
tungsten-restored enzyme

substrate: gamma-picoline N-oxide


substrate_name: 4-chloropyridine N-oxide

substrate: 4-chloropyridine N-oxide


substrate_name: 2-chloropyridine N-oxide

substrate: 2-chloropyridine N-oxide


substrate_name: 3-picoline N-oxide

substrate: 3-picoline N-oxide


substrate_name: N,N-dimethyldodecylamine N-oxide

substrate: N,N-dimethyldodecylamine N-oxide


enzyme: FMN-dependent NADH-azoreductase


substrate_name: diphenylsulfoxide

substrate: diphenylsulfoxide


substrate_name: Dimethylsulfoxide

substrate: Dimethylsulfoxide


substrate_name: Pyridine N-oxide

proof_lines:
tungsten-restored enzyme
molybdenum-restored enzyme

substrate: Pyridine N-oxide


substrate_name: 4-methylmorpholine N-oxide

substrate: 4-methylmorpholine N-oxide


substrate_name: alpha-picoline N-oxide

substrate: alpha-picoline N-oxide


substrate_name: tetramethylene sulfoxide

substrate: tetramethylene sulfoxide


substrate_name: Trimethylamine N-oxide

proof_lines:
tungsten-restored enzyme
molybdenum-restored enzyme
with methyl viologen, pH 6.9

substrate: Trimethylamine N-oxide


organism: Homo sapiens

substrate_name: GMP

substrate: GMP


substrate_name: dGMP

substrate: dGMP


substrate_name: NADPH

assay_conditions_summary: pH 7.8; 298.15 K; cofactor NADPH

proof_lines: wild type enzyme, with GMP as cosubstrate, at pH 7.8 and 25°C

substrate: NADPH


substrate_name: IMP

substrate: IMP


substrate_name: NADP+

assay_conditions_summary: pH 7.8; 298.15 K; cofactor NADP+

proof_lines: wild type enzyme, with IMP and NH3 as cosubstrates, at pH 7.8 and 25°C

substrate: NADP+


proof_lines: mutant enzyme D219A, with GMP as cosubstrate, at pH 7.8 and 25°C


substrate_name: dIMP

substrate: dIMP


proof_lines: mutant enzyme D219A, with IMP and NH3 as cosubstrates, at pH 7.8 and 25°C


substrate_name: NADH

assay_conditions_summary: pH 7.2; 298.15 K; cofactor NADH

substrate: NADH


assay_conditions_summary: pH 7.2; 298.15 K; cofactor NADPH


enzyme_display_name: UniProt Q8X1D8

organism: Fusarium oxysporum

smiles: CCCCCC[N+](=O)[O-]

enzyme: UniProt Q8X1D8


smiles: CCCCCCCC[N+](=O)[O-]


enzyme_display_name: UniProt Q9FDD4

organism: Streptomyces ansochromogenes

enzyme: UniProt Q9FDD4


substrate_name: nitroethane

substrate: nitroethane


substrate_name: ferricytochrome b5

substrate: ferricytochrome b5


enzyme_display_name: nitroalkane oxidase

organism: Pseudomonas aeruginosa

proof_lines: mutant enzyme H183C, at 30°C, pH not specified in the publication

enzyme: nitroalkane oxidase


proof_lines: mutant enzyme H183S, at 30°C, pH not specified in the publication


proof_lines: wild type enzyme, at 30°C, pH not specified in the publication


substrate_name: Ferrocytochrome c-551

proof_lines: recombinant wild-type enzyme, at pH 6.2 and 27°C

proof_lines: recombinant wild-type enzyme, O2 reduction

substrate: Ferrocytochrome c-551


proof_lines: recombinant H369A mutant enzyme, O2 reduction


proof_lines: recombinant H327A mutant enzyme, O2 reduction


proof_lines: recombinant H327A and H369a mutant enzymes


substrate_name: nitrite

substrate: nitrite


proof_lines:
recombinant wild-type enzyme
pH 6.2, 25°C, wild-type enzyme


enzyme_display_name: hydroxylamine dehydrogenase

organism: Campylobacter curvus

enzyme: hydroxylamine dehydrogenase


substrate_name: hydroxylamine

substrate: hydroxylamine


enzyme_display_name: NADPH-hemoprotein reductase

assay_conditions_summary: pH 7.5; 298.15 K; cofactor NADPH

enzyme: NADPH-hemoprotein reductase


substrate_name: ferricyanide

substrate: ferricyanide


substrate_name: Unknown

substrate: Unknown


enzyme_display_name: nitric oxide reductase (cytochrome c)

organism: Aspergillus oryzae

substrate_name: NO

enzyme: nitric oxide reductase (cytochrome c)

substrate: NO


assay_conditions_summary: pH 7.2; 310.15 K; cofactor NADPH


organism: Fusarium lichenicola

assay_conditions_summary: cofactor NADPH

proof_lines:
NADH as electron acceptor, cNor2
NADH as electron acceptor, cNor1
NADPH as electron acceptor, cNor2


organism: Campylobacter fetus


enzyme_display_name: nitrous-oxide reductase

organism: Wolinella succinogenes

substrate_name: benzyl viologen

enzyme: nitrous-oxide reductase

substrate: benzyl viologen


enzyme_display_name: hydrazine dehydrogenase

organism: Candidatus Brocadia anammoxidans

substrate_name: hydrazine

smiles: NN

proof_lines: pH 8.0, 35°C, reduction of methylthiazolyltetrazolium bromide via phenazine methosulfate

enzyme: hydrazine dehydrogenase

substrate: hydrazine


proof_lines: pH 8.0, 65°C, reduction of methylthiazolyltetrazolium bromide via phenazine methosulfate


organism: Pyrobaculum aerophilum


enzyme_display_name: hydroxylamine oxidase

organism: Candidatus Kuenenia stuttgartensis

proof_lines: 37°C, pH not specified in the publication

enzyme: hydroxylamine oxidase


proof_lines:
native enzyme
NADH as electron acceptor


organism: Nitratidesulfovibrio vulgaris

substrate_name: nitric oxide

substrate: nitric oxide


proof_lines: with bovine cytochrome c as electron acceptor, at pH 7.5 and 60°C


organism: Neomoorella thermoacetica


organism: Nitrososphaera viennensis


organism: Caminibacter mediatlanticus


organism: Candidatus Jettenia caeni


organism: Candidatus Brocadia fulgida


organism: Fusarium oxysporum


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:Q8X1D8 and applied mutation signature S276A to reconstruct the assayed variant sequence.

assay_conditions_summary: pH 8; 303.15 K; cofactor FAD

column_label: Table 2, substrate block '1-nitrooctane', enzyme-form column 'S276A NAO'; column 'kcat(nitrooctane) (s-1)' = 0.2; column 'K nitrooctane (mM)' = 0.3; column 'kcat/K nitrooctane (mM-1 s-1)' = 1.3

normalized_value_unit_evidence: kcat: printed Table 2 cell 0.2 s^-1; already in s^-1, so no unit conversion applies; the exported value is 0.2 s^(-1). Km: printed Table 2 cell 0.3 mM; already in mM, so no unit conversion applies; the exported value is 0.3 mM. kcat/Km: printed Table 2 cell 1.3 mM^-1 s^-1; 1 mM^-1 s^-1 is 1 s^-1 mM^-1, so no unit conversion applies and the printed cell normalizes to 1.3 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 2, 1-nitrooctane block, S276A NAO column: kcat = 0.2 s^-1; Km = 0.3 mM; printed kcat/Km = 1.3 mM^-1 s^-1.

table_label: Table 2: Kinetic parameters for S276A nitroalkane oxidase, S276A NAO column (Biochemistry 2009, 48(15):3407-3416)


column_label: Table 2, substrate block '1-nitrohexane', enzyme-form column 'S276A NAO'; column 'kcat(nitrohexane) (s-1)' = 0.30; column 'Knitrohexane (mM)' = 0.20; column 'kcat/Knitrohexane (mM-1 s-1)' = 1.5

normalized_value_unit_evidence: kcat: printed Table 2 cell 0.30 s^-1; already in s^-1, so no unit conversion applies; the exported value is 0.3 s^(-1). Km: printed Table 2 cell 0.20 mM; already in mM, so no unit conversion applies; the exported value is 0.2 mM. kcat/Km: printed Table 2 cell 1.5 mM^-1 s^-1; 1 mM^-1 s^-1 is 1 s^-1 mM^-1, so no unit conversion applies and the printed cell normalizes to 1.5 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 2, 1-nitrohexane block, S276A NAO column: kcat = 0.30 s^-1; Km = 0.20 mM; printed kcat/Km = 1.5 mM^-1 s^-1.


column_label: Table 2, substrate block 'nitroethane', enzyme-form column 'S276A NAO'; column 'kcat(nitroethane) (s-1)' = 1.0; column 'K(nitroethane) (mM)' = 3.3; column 'kcat/K(nitroethane) (mM-1 s-1)' = 0.30

normalized_value_unit_evidence: kcat: printed Table 2 cell 1.0 s^-1; already in s^-1, so no unit conversion applies; the exported value is 1.0 s^(-1). Km: printed Table 2 cell 3.3 mM; already in mM, so no unit conversion applies; the exported value is 3.3 mM. kcat/Km: printed Table 2 cell 0.30 mM^-1 s^-1; 1 mM^-1 s^-1 is 1 s^-1 mM^-1, so no unit conversion applies and the printed cell normalizes to 0.3 s^(-1)*mM^(-1). The exact quotient of the same two printed cells is 1.0 / 3.3 = 0.30303030303030304 s^-1 mM^-1, which the paper prints rounded to its own two-decimal precision as 0.30; the exported ratio keeps the exact quotient and is corroborated by that printed cell.

raw_value_unit_evidence: Table 2, nitroethane block, S276A NAO column: kcat = 1.0 s^-1; Km = 3.3 mM; printed kcat/Km = 0.30 mM^-1 s^-1.

source_databases_merged: brenda oed


column_label: Table 2, substrate block '1-nitrohexane', enzyme-form column 'wild-type NAO'; column 'kcat(nitrohexane) (s-1)' = 2.0; column 'Knitrohexane (mM)' = 0.04; column 'kcat/Knitrohexane (mM-1 s-1)' = 47

normalized_value_unit_evidence: kcat: printed Table 2 cell 2.0 s^-1; already in s^-1, so no unit conversion applies; the exported value is 2.0 s^(-1). Km: printed Table 2 cell 0.04 mM; already in mM, so no unit conversion applies; the exported value is 0.04 mM. kcat/Km: printed Table 2 cell 47 mM^-1 s^-1; 1 mM^-1 s^-1 is 1 s^-1 mM^-1, so no unit conversion applies and the printed cell normalizes to 47.0 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 2, 1-nitrohexane block, wild-type NAO column: kcat = 2.0 s^-1; Km = 0.04 mM; printed kcat/Km = 47 mM^-1 s^-1.

table_label: Table 2: Kinetic parameters for wild-type nitroalkane oxidase, wild-type NAO column (Biochemistry 2009, 48(15):3407-3416)


column_label: Table 2, substrate block '1-nitrooctane', enzyme-form column 'wild-type NAO'; column 'kcat(nitrooctane) (s-1)' = 4.4; column 'K nitrooctane (mM)' = 0.03; column 'kcat/K nitrooctane (mM-1 s-1)' = 150

normalized_value_unit_evidence: kcat: printed Table 2 cell 4.4 s^-1; already in s^-1, so no unit conversion applies; the exported value is 4.4 s^(-1). Km: printed Table 2 cell 0.03 mM; already in mM, so no unit conversion applies; the exported value is 0.03 mM. kcat/Km: printed Table 2 cell 150 mM^-1 s^-1; 1 mM^-1 s^-1 is 1 s^-1 mM^-1, so no unit conversion applies and the printed cell normalizes to 150.0 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 2, 1-nitrooctane block, wild-type NAO column: kcat = 4.4 s^-1; Km = 0.03 mM; printed kcat/Km = 150 mM^-1 s^-1.


column_label: Table 2, substrate block 'nitroethane', enzyme-form column 'wild-type NAO'; column 'kcat(nitroethane) (s-1)' = 15; column 'K(nitroethane) (mM)' = 2.3; column 'kcat/K(nitroethane) (mM-1 s-1)' = 6.3

normalized_value_unit_evidence: kcat: printed Table 2 cell 15 s^-1; already in s^-1, so no unit conversion applies; the exported value is 15.0 s^(-1). Km: printed Table 2 cell 2.3 mM; already in mM, so no unit conversion applies; the exported value is 2.3 mM. kcat/Km: printed Table 2 cell 6.3 mM^-1 s^-1; 1 mM^-1 s^-1 is 1 s^-1 mM^-1, so no unit conversion applies and the printed cell normalizes to 6.3 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 2, nitroethane block, wild-type NAO column: kcat = 15 s^-1; Km = 2.3 mM; printed kcat/Km = 6.3 mM^-1 s^-1.


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


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature D403E with inferred residue-numbering offset -1 to reconstruct the assayed variant sequence.

assay_conditions_summary: pH 8


sequence_variant_note: Resolved canonical sequence from multiple and applied mutation signature R409K to reconstruct the assayed variant sequence.

proof_lines: pH 8.0, 30°C, mutant enzyme R409K, substrate inhibition constant


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature R409K to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature D402E to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from multiple and applied mutation signature S171A to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme S171A, in 200 mM HEPES, 0.1 mM FAD, pH 8.0, 30°C


sequence_variant_note: Mutation signature S171V could not be applied safely because the resolved sequence does not match the expected reference residue(s).

assay_conditions_summary: pH 8; cofactor FAD

proof_lines: mutant enzyme S171V, in 200 mM HEPES, 0.1 mM FAD, pH 8.0, 30°C


sequence_variant_note: Mutation signature S171T could not be applied safely because the resolved sequence does not match the expected reference residue(s).

proof_lines: mutant enzyme S171T, in 200 mM HEPES, 0.1 mM FAD, pH 8.0, 30°C


sequence_variant_note: Resolved canonical sequence from multiple and applied mutation signature Y398F to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme Y398F, in 200 mM HEPES, 0.1 mM FAD, pH 8.0, 30°C


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature Y398F to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature S171A to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature C397S to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme C397S, in 200 mM HEPES, 0.1 mM FAD, pH 8.0, 30°C


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

proof_lines: wild type enzyme, in 200 mM HEPES, 0.1 mM FAD, pH 8.0, 30°C


organism: Podospora anserina

sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:B2AM55 and applied mutation signature D399N to reconstruct the assayed variant sequence.

column_label: Table 1 (Steady-state kinetic parameters for wild-type and mutant PODANSg2158 with nitroethane as substrate), D399N column: parameter row 'kcat (s-1)' = 0.037; parameter row 'KNE (mM)' = 10; parameter row 'kcat/KNE (mM-1s-1)' = 0.004.

normalized_value_unit_evidence: kcat: printed Table 1 cell 0.037 s^-1; the cell is already in s^-1, so no unit conversion applies and the normalized value is 0.037 s^(-1). Km: printed Table 1 cell 10 mM; the cell is already in mM, so no unit conversion applies and the normalized value is 10.0 mM. kcat/Km: printed Table 1 cell 0.004 mM^-1 s^-1; 1 mM^-1 s^-1 is 1 s^-1 mM^-1, so the printed cell converts to 0.004 s^(-1)*mM^(-1) with no numeric change.

raw_value_unit_evidence: Table 1, D399N column, nitroethane parameter rows: kcat (s-1) = 0.037; KNE (mM) = 10; kcat/KNE (mM-1s-1) = 0.004.

table_label: Table 1: Steady-state kinetic parameters for wild-type and mutant PODANSg2158 with nitroethane as substrate (Biochemistry 2010, 49(24):5035-5041)


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:B2AM55 and applied mutation signature S273A to reconstruct the assayed variant sequence.

column_label: Table 1 (Steady-state kinetic parameters for wild-type and mutant PODANSg2158 with nitroethane as substrate), S273A column: parameter row 'kcat (s-1)' = 5.0; parameter row 'KNE (mM)' = 24; parameter row 'kcat/KNE (mM-1s-1)' = 0.21.

normalized_value_unit_evidence: kcat: printed Table 1 cell 5.0 s^-1; the cell is already in s^-1, so no unit conversion applies and the normalized value is 5.0 s^(-1). Km: printed Table 1 cell 24 mM; the cell is already in mM, so no unit conversion applies and the normalized value is 24.0 mM. kcat/Km: printed Table 1 cell 0.21 mM^-1 s^-1; 1 mM^-1 s^-1 is 1 s^-1 mM^-1, so the printed cell converts to 0.21 s^(-1)*mM^(-1) with no numeric change.

raw_value_unit_evidence: Table 1, S273A column, nitroethane parameter rows: kcat (s-1) = 5.0; KNE (mM) = 24; kcat/KNE (mM-1s-1) = 0.21.


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:B2AM55 and applied mutation signature R406K to reconstruct the assayed variant sequence.

column_label: Table 1 (Steady-state kinetic parameters for wild-type and mutant PODANSg2158 with nitroethane as substrate), R406K column: parameter row 'kcat (s-1)' = 9.0; parameter row 'KNE (mM)' = 25; parameter row 'kcat/KNE (mM-1s-1)' = 0.37.

normalized_value_unit_evidence: kcat: printed Table 1 cell 9.0 s^-1; the cell is already in s^-1, so no unit conversion applies and the normalized value is 9.0 s^(-1). Km: printed Table 1 cell 25 mM; the cell is already in mM, so no unit conversion applies and the normalized value is 25.0 mM. kcat/Km: printed Table 1 cell 0.37 mM^-1 s^-1; 1 mM^-1 s^-1 is 1 s^-1 mM^-1, so the printed cell converts to 0.37 s^(-1)*mM^(-1) with no numeric change.

raw_value_unit_evidence: Table 1, R406K column, nitroethane parameter rows: kcat (s-1) = 9.0; KNE (mM) = 25; kcat/KNE (mM-1s-1) = 0.37.


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

proof_lines: wild type enzyme, in 50 mM HEPES, pH 8.0, 30°C


column_label: Table 1 (Steady-state kinetic parameters for wild-type and mutant PODANSg2158 with nitroethane as substrate), Wild-Type column: parameter row 'kcat (s-1)' = 15; parameter row 'KO2 (mM)' = 0.39; parameter row 'kcat/KO2 (mM-1s-1)' = 38.

normalized_value_unit_evidence: kcat: printed Table 1 cell 15 s^-1; the cell is already in s^-1, so no unit conversion applies and the normalized value is 15.0 s^(-1). Km: printed Table 1 cell 0.39 mM; the cell is already in mM, so no unit conversion applies and the normalized value is 0.39 mM. kcat/Km: printed Table 1 cell 38 mM^-1 s^-1; 1 mM^-1 s^-1 is 1 s^-1 mM^-1, so the printed cell converts to 38.0 s^(-1)*mM^(-1) with no numeric change.

raw_value_unit_evidence: Table 1, Wild-Type column, oxygen parameter rows: kcat (s-1) = 15; KO2 (mM) = 0.39; kcat/KO2 (mM-1s-1) = 38.


organism: Achromobacter cycloclastes

proof_lines: pH 7.1, temperature not specified in the publication


organism: Nitrosococcus oceani

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

proof_lines: pH 7.8, temperature not specified in the publication


organism: Nitrosomonas europaea

proof_lines: pH 7.5, 25°C, with enzyme form HAO-A, that forms only the monocyano derivative of heme P460

proof_lines: pH 7.5, 25°C, with enzyme form HAO-B, that forms a mono- and dicyano complex


enzyme_display_name: Hydroxylamine oxidoreductase

organism: Kuenenia stuttgartiensis

substrate_name: hydroxylamine (at 37 degrees Celsius)

proof_lines: kcat is 15 sec(-1) with hydroxylamine as substrate (at 37 degrees Celsius) (PubMed:24302732). kcat is 4.9 sec(-1) with hydroxylamine as substrate (at 37 degrees Celsius) (PubMed:24302732).

enzyme: Hydroxylamine oxidoreductase

substrate: hydroxylamine (at 37 degrees Celsius)


substrate_name: hydrazine (at 37 degrees Celsius)

substrate: hydrazine (at 37 degrees Celsius)


enzyme_display_name: factor-independent urate hydroxylase

organism: Vicia faba

substrate_name: uric acid

enzyme: factor-independent urate hydroxylase

substrate: uric acid


enzyme_display_name: UniProt D0VWQ1

organism: Arthrobacter globiformis

substrate_name: Urate

enzyme: UniProt D0VWQ1

substrate: Urate


enzyme_display_name: UniProt C5HDG5

organism: Metabacillus fastidiosus

proof_lines:
Raw SKiD Env: Temp=40°C, pH=9.2
Raw SKiD Env: Temp=25°C, pH=9.2
Raw SKiD Env: Temp=25°C, pH=7.4

enzyme: UniProt C5HDG5


enzyme_display_name: UniProt Q5FZI9

organism: Canis lupus familiaris

enzyme: UniProt Q5FZI9


enzyme_display_name: ferredoxin-nitrite reductase

organism: Pyropia yezoensis

enzyme: ferredoxin-nitrite reductase


organism: Cucurbita pepo


enzyme_display_name: UniProt Q76KA9

organism: Nicotiana tabacum

enzyme: UniProt Q76KA9


enzyme_display_name: UniProt Q76KB0

enzyme: UniProt Q76KB0


substrate_name: NADH; Nitric oxide; H+

substrate: NADH; Nitric oxide; H+


organism: Sus scrofa domestica

substrate_name: 2-Nitropropane; H2O; O2

substrate: 2-Nitropropane; H2O; O2


substrate_name: nitroethane

proof_lines:
cosubstrate D2O, 30°C, pH 8.5
Raw SKiD Env: Temp=°C, pH=8
Raw SKiD Env: Temp=30°C, pH=8

substrate: nitroethane


substrate_name: reduced benzyl viologen

substrate: reduced benzyl viologen


organism: Paracoccus pantotrophus


substrate_name: ferricytochrome c-554

proof_lines: source of substrate Nitrosococcus oceani, pH 7.8, temperature not specified in the publication

substrate: ferricytochrome c-554


substrate_name: ferricytochrome c

proof_lines: from Rhodospirillum rubrum, Saccharomyces oviformis and cow

substrate: ferricytochrome c


substrate_name: H2O2; Acetaldehyde; Nitrite

source_license: CC BY 4.0; SABIO-RK terms (free for academic use; see sabiork.h-its.org)

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

substrate: H2O2; Acetaldehyde; Nitrite


substrate_name: benzoate

substrate: benzoate


substrate_name: phenylacetic acid

substrate: phenylacetic acid


enzyme_display_name: Nitroalkane oxidase

assay_conditions_summary: cofactor FAD

column_label: PubMed XML abstract primary prose reports the Michaelis constant for 1-nitropropane as 1.54 mM.

raw_value_unit_evidence: Km raw_value=1.54 raw_unit=mM for 1-nitropropane.

table_label: Primary article abstract prose: Michaelis constants for nitroalkane oxidase substrates/cofactor

enzyme: Nitroalkane oxidase


column_label: PubMed XML abstract primary prose reports the Michaelis constant for 2-nitropropane as 7.40 mM.

raw_value_unit_evidence: Km raw_value=7.40 raw_unit=mM for 2-nitropropane.


column_label: PubMed XML abstract primary prose reports the Michaelis constant for 3-nitro-2-pentanol as 3.08 mM.

raw_value_unit_evidence: Km raw_value=3.08 raw_unit=mM for 3-nitro-2-pentanol.


substrate_name: nitrocyclohexane

column_label: PubMed XML abstract primary prose reports the Michaelis constant for nitrocyclohexane as 0.90 mM.

raw_value_unit_evidence: Km raw_value=0.90 raw_unit=mM for nitrocyclohexane.

row_label: nitrocyclohexane

substrate: nitrocyclohexane


substrate_name: FAD

column_label: PubMed XML abstract primary prose reports the Michaelis constant for FAD cofactor as 1.33 microM.

raw_value_unit_evidence: Km raw_value=1.33 raw_unit=microM for FAD cofactor.

row_label: FAD cofactor

substrate: FAD


substrate_name: nitrite

proof_lines:
hydroxylamine oxidoreductase
Nitrosomonas europaea

substrate: nitrite


enzyme_display_name: UniProt B2AM55

row_label: Table 1, Wild-Type column, oxygen parameter rows

enzyme: UniProt B2AM55


enzyme_display_name: nitroalkane oxidase

sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature D402E to reconstruct the assayed variant sequence.

enzyme: nitroalkane oxidase