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


enzyme_display_name: alanopine dehydrogenase

organism: Aphrodita aculeata

substrate_name: L-Ala

source_db: brenda

verification_status: unverified

kcat_display: Not reported

ki_display: Not reported

kcat_over_km_display: Not reported

mutation_signature: Unknown

source_license: CC BY 4.0

km_unit: mM

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

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

enzyme: alanopine dehydrogenase

substrate: L-Ala

status: unverified

source: brenda


substrate_name: NADH

assay_conditions_summary: cofactor NADH

substrate: NADH


substrate_name: L-Cys

smiles: C(C(C(=O)O)N)S

substrate: L-Cys


substrate_name: L-Thr

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

substrate: L-Thr


substrate_name: L-Ser

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

substrate: L-Ser


organism: Mercenaria mercenaria

substrate_name: pyruvate

substrate: pyruvate


substrate_name: glyoxylate

substrate: glyoxylate


substrate_name: L-Val

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

substrate: L-Val


substrate_name: Gly

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

substrate: Gly


enzyme_display_name: ephedrine dehydrogenase

organism: Arthrobacter sp. TS-15

enzyme: ephedrine dehydrogenase


substrate_name: Phenylglyoxal

substrate: Phenylglyoxal


substrate_name: methylbenzoyl formate

substrate: methylbenzoyl formate


organism: Escherichia coli

assay_conditions_summary: pH 7.2; 298.15 K; cofactor NADH

proof_lines: pH 7.2, 25°C, recombinant wild-type, CH3-H4folate-menadione oxidoreductase assay

proof_lines: pH 7.2, 25°C, recombinant wild-type, NADH-CH2-H4folate oxidoreductase assay


proof_lines: pH 7.2, 25°C, recombinant wild-type, NADH-menadione oxidoreductase assay


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

mutation_type: substitution

proof_lines: pH 7.2, 25°C, recombinant mutant Q183A, NADH-CH2-H4folate oxidoreductase assay

proof_lines: pH 7.2, 25°C, recombinant mutant Q183A, CH3-H4folate-menadione oxidoreductase assay


proof_lines: pH 7.2, 25°C, recombinant mutant Q183E, NADH-CH2-H4folate oxidoreductase assay

proof_lines: pH 7.2, 25°C, recombinant mutant Q183E, NADH-menadione oxidoreductase assay


proof_lines: pH 7.2, 25°C, recombinant mutant Q183E, CH3-H4folate-menadione oxidoreductase assay


proof_lines: pH 7.2, 25°C, recombinant mutant Q183A, NADH-menadione oxidoreductase assay


substrate_name: Unknown

km_display: Not reported

ki_unit: mM

substrate: Unknown


organism: Littorina littorea

proof_lines:
at pH 6.5
at pH 7.5


substrate_name: NAD+

assay_conditions_summary: cofactor NAD+

substrate: NAD+


substrate_name: meso-Alanopine

substrate: meso-Alanopine


organism: Homo sapiens

source_db: oed

verification_status: corrected

sequence_source: UniProt REST P13995

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

assay_conditions_summary: pH 8; 303.15 K; cofactor NAD+

normalized_value_unit_evidence: KM 133 uM = 0.133 mM; kcat 12.4 s^-1; kcat/KM 0.093 s^-1 uM^-1 = 93 s^-1*mM^-1; expected 93.23308270676692 is consistent with rounded kcat/KM from kcat/KM cells.

raw_value_unit_evidence: Table 1 row NAD+-dependent / CH2-H4PteGlu1: KM 133+/-20 uM, kcat 12.4+/-0.71 s^-1, kcat/KM 0.093 s^-1 uM^-1.

row_label: NAD+-dependent; CH2-H4PteGlu1

table_label: Table 1: Kinetic parameters for MTHFD2 5,10-CH2-THF dehydrogenase activity

data_origin: experimental

status: corrected

source: oed


normalized_value_unit_evidence: KM 359 uM = 0.359 mM; kcat 15.4 s^-1; kcat/KM 0.043 s^-1 uM^-1 = 43 s^-1*mM^-1; expected 42.89693593314764 is consistent with rounded kcat/KM from kcat/KM cells.

raw_value_unit_evidence: Table 1 row NAD+-dependent / CH2-H4PteGlu5: KM 359+/-32 uM, kcat 15.4+/-0.55 s^-1, kcat/KM 0.043 s^-1 uM^-1.

row_label: NAD+-dependent; CH2-H4PteGlu5


organism: Mycolicibacterium smegmatis


assay_conditions_summary: pH 7.6; 295.15 K; cofactor NADH


enzyme_display_name: (-)-ephedrine dehydrogenase

substrate_name: (R,S)-(-)-ephedrine

protein_accession_database: UniProt

assay_conditions_summary: pH 7.8; 298.15 K; cofactor NAD+

enzyme: (-)-ephedrine dehydrogenase

substrate: (R,S)-(-)-ephedrine


substrate_name: (R,R)-(-)-pseudoephedrine

substrate: (R,R)-(-)-pseudoephedrine


organism: Arabidopsis thaliana

assay_conditions_summary: pH 7.2; cofactor NADH


proof_lines: recombinant chimeric mutant enzyme, pH 7.2


substrate_name: NADPH

assay_conditions_summary: pH 7.2; cofactor NADPH

substrate: NADPH


proof_lines: recombinant chimeric mutant enzyme, with NADH, pH 7.2

proof_lines: recombinant chimeric mutant enzyme, with NADPH, pH 7.2


proof_lines: recombinant wild-type enzyme, with NADH, pH 7.2


organism: Acetobacterium woodii


organism: Concholepas concholepas


enzyme_display_name: D-lysopine dehydrogenase

organism: Mytilus edulis

substrate_name: L-arginine

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

enzyme: D-lysopine dehydrogenase

substrate: L-arginine


substrate_name: L-lysine

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

substrate: L-lysine


substrate_name: L-canavanine

smiles: C(CON=C(N)N)C(C(=O)O)N

substrate: L-canavanine


substrate_name: L-ornithine

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

substrate: L-ornithine


substrate_name: alanopine

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

substrate: alanopine


substrate_name: L-Ala

substrate: L-Ala


enzyme_display_name: D-nopaline dehydrogenase

organism: Helianthus annuus

assay_conditions_summary: cofactor NADPH

proof_lines:
nopaline synthesis
ornaline synthesis

enzyme: D-nopaline dehydrogenase


organism: Clostridium formicaceticum


substrate_name: NADH

substrate: NADH


organism: Spodoptera frugiperda


substrate_name: methenyltetrahydrofolate

substrate: methenyltetrahydrofolate


organism: Glycera alba

substrate_name: pyruvate

substrate: pyruvate


organism: Busycotypus canaliculatum

proof_lines:
enzyme from hepatopancreas
enzyme from ventricle
enzyme from gill


substrate_name: Gly

substrate: Gly


organism: Arthrobacter globiformis

substrate_name: (-)-ephedrine

substrate: (-)-ephedrine


organism: Scolelepis fuliginosa


organism: Rattus norvegicus


substrate_name: benzyl viologen

substrate: benzyl viologen


organism: Methanosarcina barkeri

substrate_name: NAD+

assay_conditions_summary: pH 6; 310.15 K; cofactor NAD+

substrate: NAD+


organism: Sus scrofa

substrate_name: methylenetetrahydrofolate

assay_conditions_summary: pH 6.7; 310.15 K; cofactor NADPH

proof_lines: methylenetetrahydrofolate reductase reaction, pH 6.7

substrate: methylenetetrahydrofolate


proof_lines: 5,10-methylenetetrahydrofolate reductase reaction, pH 7.2


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


organism: Thermus thermophilus


organism: Haemonchus contortus

proof_lines: at pH 6.5 temperature not specified in the publication


organism: Teladorsagia circumcincta

proof_lines: at pH 6.5, temperature not specified in the publication


enzyme_display_name: Bifunctional methylenetetrahydrofolate dehydrogenase/cyclohydrolase 2, mitochondrial

substrate_name: NAD(+)

source_db: uniprot

proof_lines: Has similar catalytic efficiencies, at physiological concentrations, with either NAD(+) or NADP(+) as substrates for the dehydrogenation of (6R)-5,10-methylene-5,6,7,8-tetrahydrofolate.

substrate: NAD(+)

source: uniprot


substrate_name: NADP(+)

enzyme: Bifunctional methylenetetrahydrofolate dehydrogenase/cyclohydrolase 2, mitochondrial

substrate: NADP(+)


substrate_name: 5,10-methylenetetrahydrofolate (in presence of NAD(+))


organism: Saccharomyces cerevisiae

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


substrate_name: NADPH

substrate: NADPH


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

substrate_name: Delta(1)-piperideine-2-carboxylate (at pH 7.0)

sequence_variant_note: Mutation signature H2C could not be applied safely because the resolved sequence does not match the expected reference residue(s).

assay_conditions_summary: pH 7

proof_lines: kcat is 2500 min(-1) for Pyr2C reduction. kcat is 2120 min(-1) for Pip2C reduction. kcat is 868 min(-1) for Pyr4SH2C reduction. kcat is 205 min(-1) for L-proline oxidation. kcat is 135 min(-1) for L-pipecolate oxidation. kcat is 272 min(-1)


substrate_name: Delta(1)-pyrroline-(4S)-hydroxy-2-carboxylate (at pH 7.0)


substrate_name: L-proline (at pH 10.0)

assay_conditions_summary: pH 10

substrate: L-proline (at pH 10.0)


substrate_name: L-pipecolate (at pH 10.0)

substrate: L-pipecolate (at pH 10.0)


substrate_name: trans-3-hydroxy-L-proline (at pH 10.0)

substrate: trans-3-hydroxy-L-proline (at pH 10.0)


organism: Arthrobacter sp. TS-15

substrate_name: (R,R)-(-)-pseudoephedrine

kcat_over_km_origin: calculated

substrate: (R,R)-(-)-pseudoephedrine


organism: Homo sapiens

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


organism: Arenicola marina

substrate_name: glycine

proof_lines: pH and temperature not specified in the publication

substrate: glycine


substrate_name: L-alanine

substrate: L-alanine


assay_conditions_summary: pH 6.6; cofactor NADPH

proof_lines: pH 6.6, 46°C, enzyme mutant HsMTHFR1-656

proof_lines: pH 6.6, 46°C, enzyme mutant HsMTHFR38-644


substrate_name: 5,10-Methylenetetrahydrofolate; NAD+

sequence_source: multiple

assay_conditions_summary: cofactor NADP+

substrate: 5,10-Methylenetetrahydrofolate; NAD+


organism: Mus musculus

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


substrate_name: NAD+; 5,10-Methylenetetrahydrofolate

substrate: NAD+; 5,10-Methylenetetrahydrofolate


organism: Arabidopsis thaliana


organism: Escherichia coli


substrate_name: H+; Menadione; NADPH

substrate: H+; Menadione; NADPH


substrate_name: tetrahydropteroylpentaglutamate/H4PteGlu5 (in presence of NAD(+))


substrate_name: Unknown

substrate: Unknown


substrate_name: Delta(1)-pyrroline-2-carboxylate (at pH 7.0)


organism: Leishmania major


organism: Pseudomonas aeruginosa

substrate_name: 1-Pyrroline-2-carboxylate; NADPH; H+

substrate: 1-Pyrroline-2-carboxylate; NADPH; H+


organism: Pseudomonas putida


verification_status: verified

assay_conditions_summary: pH 8; cofactor NADPH

column_label: Table IV reports Km(B) (microM) under column (B) Pip2C as 15; 15 microM = 0.015 mM.

raw_value_unit_evidence: Km raw_value=15 raw_unit=microM

row_label: Km(B)

table_label: Table IV - Kinetic parameters of DpkA in the forward direction

status: verified


substrate_name: L-proline

assay_conditions_summary: pH 10; cofactor NADP+

column_label: Table V reports Km(P) (microM) under column (P) L-Proline as 144,000; 144,000 microM = 144.0 mM.

raw_value_unit_evidence: Km raw_value=144,000 raw_unit=microM

row_label: Km(P)

table_label: Table V - Kinetic parameters of DpkA in the reverse direction

substrate: L-proline


substrate_name: L-pipecolate

column_label: Table V reports Km(P) (microM) under column (P) L-Pipecolate as 32,400; 32,400 microM = 32.4 mM.

raw_value_unit_evidence: Km raw_value=32,400 raw_unit=microM

substrate: L-pipecolate


substrate_name: NADPH (with Pyr2C as cosubstrate)

column_label: Table IV reports Km(NADPH) (microM) under column (B) Pyr2C as 34; 34 microM = 0.034 mM.

raw_value_unit_evidence: Km raw_value=34 raw_unit=microM

row_label: Km(NADPH)

substrate: NADPH (with Pyr2C as cosubstrate)


substrate_name: NADP(+) (with L-proline as cosubstrate)

substrate: NADP(+) (with L-proline as cosubstrate)


substrate_name: NADH

assay_conditions_summary: buffer phosphate buffer; cofactor NADPH

proof_lines:
dual-cofactor specificity
NADH
NADPH

substrate: NADH


substrate_name: NADPH

substrate: NADPH


proof_lines:
MTHFR
5-methyltetrahydrofolate
mutant folate phenotypes


assay_conditions_summary: pH 7.2; 298.15 K; cofactor NADH


proof_lines:
NADH
5-methyltetrahydrofolate
methylenetetrahydrofolate reductase


substrate_name: NAD+

assay_conditions_summary: cofactor NAD+

column_label: Abstract reports with the sequential reaction model, Km values for NAD+ and (6R,S)-5,10-methylene-THF are 1.6 and 0.06 mM, respectively.

raw_value_unit_evidence: Km raw_value=1.6 mM raw_unit=mM

row_label: Saccharomyces cerevisiae NAD+-dependent 5,10-methylene-THF dehydrogenase / NAD+

table_label: PubMed XML abstract PMID 2223762

substrate: NAD+


enzyme_display_name: UniProt P13995

source_db: skid

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

assay_conditions_summary: pH 7.3; cofactor NAD+

source_databases_merged: skid uniprot

enzyme: UniProt P13995

source: skid


proof_lines:
trans-3-hydroxy-L-proline
Pyr2C reductase
Azospirillum brasilense


substrate_name: L-Pipecolate

assay_conditions_summary: pH 10; 303.15 K; cofactor NADP+

column_label: Table 2. Kinetic parameters of AbLhpI, CpLhpI and PaLhpD in the reverse direction; row PaLhpD / L-Pipecolate / NADP+; columns Km (mM); kcat (min^-1); kcat/Km (min^-1*mM^-1); raw table cells: Km 34.8 ± 4.0 mM | kcat 135 ± 14 min^-1 | kcat/Km 3.88 ± 0.05 min^-1*mM^-1

normalized_value_unit_evidence: kcat normalized_value=2.25 normalized_unit=s^-1; Km normalized_value=34.8 normalized_unit=mM; kcat/Km normalized_value=0.06465517241379311 normalized_unit=s^-1*mM^-1

raw_value_unit_evidence: kcat raw_value=135 ± 14 raw_unit=min^-1; Km raw_value=34.8 ± 4.0 raw_unit=mM; kcat/Km raw_value=3.88 ± 0.05 raw_unit=min^-1*mM^-1

row_label: PaLhpD / L-Pipecolate / NADP+

substrate: L-Pipecolate


assay_conditions_summary: pH 7; 303.15 K; cofactor NADP+

column_label: Table 2. Kinetic parameters of AbLhpI, CpLhpI and PaLhpD in the reverse direction; row PaLhpD / L-Proline / NADP+; columns Km (mM); kcat (min^-1); kcat/Km (min^-1*mM^-1); raw table cells: Km 18.5 ± 2.1 mM | kcat 205 ± 18 min^-1 | kcat/Km 11.1 ± 0.3 min^-1*mM^-1

normalized_value_unit_evidence: kcat normalized_value=3.42 normalized_unit=s^-1; Km normalized_value=18.5 normalized_unit=mM; kcat/Km normalized_value=0.18486486486486486 normalized_unit=s^-1*mM^-1

raw_value_unit_evidence: kcat raw_value=205 ± 18 raw_unit=min^-1; Km raw_value=18.5 ± 2.1 raw_unit=mM; kcat/Km raw_value=11.1 ± 0.3 raw_unit=min^-1*mM^-1

row_label: PaLhpD / L-Proline / NADP+


column_label: Table 2. Kinetic parameters of AbLhpI, CpLhpI and PaLhpD in the reverse direction; row PaLhpD / T3LHyp / NADP+; columns Km (mM); kcat (min^-1); kcat/Km (min^-1*mM^-1); raw table cells: Km 132 ± 26 mM | kcat 272 ± 53 min^-1 | kcat/Km 2.07 ± 0.01 min^-1*mM^-1

normalized_value_unit_evidence: kcat normalized_value=4.53 normalized_unit=s^-1; Km normalized_value=132.0 normalized_unit=mM; kcat/Km normalized_value=0.03431818181818182 normalized_unit=s^-1*mM^-1

raw_value_unit_evidence: kcat raw_value=272 ± 53 raw_unit=min^-1; Km raw_value=132 ± 26 raw_unit=mM; kcat/Km raw_value=2.07 ± 0.01 raw_unit=min^-1*mM^-1

row_label: PaLhpD / T3LHyp / NADP+


assay_conditions_summary: pH 7; 303.15 K; cofactor NADPH

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


substrate_name: diglutamate

assay_conditions_summary: cofactor NADH

substrate: diglutamate


organism: Papaver somniferum


enzyme_display_name: glycine oxidase

organism: Bacillus cereus

substrate_name: D-alanine

enzyme: glycine oxidase

substrate: D-alanine


substrate_name: D-proline

substrate: D-proline


substrate_name: glycine

substrate: glycine


enzyme_display_name: alanine dehydrogenase

proof_lines:
soluble enzyme
Sepharose-bound enzyme

enzyme: alanine dehydrogenase


substrate_name: pyruvate

substrate: pyruvate


enzyme_display_name: Strombine dehydrogenase

organism: Aphrodita aculeata

enzyme: Strombine dehydrogenase


substrate_name: oxaloacetate

substrate: oxaloacetate


substrate_name: Hydroxypyruvate

substrate: Hydroxypyruvate


substrate_name: L-Cys

substrate: L-Cys


substrate_name: glyoxylate

substrate: glyoxylate


substrate_name: Gly

substrate: Gly


substrate_name: L-Thr

substrate: L-Thr


substrate_name: L-Ala

substrate: L-Ala


substrate_name: L-Val

substrate: L-Val


substrate_name: L-Ser

substrate: L-Ser


organism: Marphysa sanguinea

assay_conditions_summary: pH 7.2; 303.15 K; cofactor NAD+

proof_lines: apparent value, isozyme St/AlDH1, with 20 mM D-strombine, in 0.1 M PIPES-NaOH (pH 7.2), at 30°C

proof_lines: apparent value, isozyme St/AlDH1, with 20 mM meso-alanopine, in 0.1 M PIPES-NaOH (pH 7.2), at 30°C


substrate_name: D-strombine

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

assay_conditions_summary: pH 9.1; 303.15 K; cofactor NAD+

proof_lines: apparent value, isozyme St/AlDH1, with 1 mM NAD+, in 0.1 M PIPES-NaOH (pH 9.1), at 30°C

substrate: D-strombine


substrate_name: meso-Alanopine

substrate: meso-Alanopine


assay_conditions_summary: pH 7.2; 303.15 K; cofactor NADH

proof_lines: apparent value, isozyme St/AlDH1, with 0.3 mM NADH and 100 mM Gly, in 0.1 M PIPES-NaOH (pH 7.2), at 30°C

proof_lines: apparent value, isozyme St/AlDH1, with 0.3 mM NADH and 100 mM L-Ala, in 0.1 M PIPES-NaOH (pH 7.2), at 30°C


proof_lines: apparent value, isozyme St/AlDH1, with 0.3 mM NADH and 5 mM pyruvate, in 0.1 M PIPES-NaOH (pH 7.2), at 30°C


substrate_name: NADH

proof_lines: apparent value, isozyme St/AlDH1, with 5 mM pyruvate and 100 mM Gly, in 0.1 M PIPES-NaOH (pH 7.2), at 30°C

proof_lines: apparent value, isozyme St/AlDH1, with 5 mM pyruvate and 100 mM L-Ala, in 0.1 M PIPES-NaOH (pH 7.2), at 30°C

substrate: NADH


organism: Meretrix lusoria


substrate_name: beta-Ala

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

substrate: beta-Ala


substrate_name: Unknown

substrate: Unknown


organism: Mercenaria mercenaria


enzyme_display_name: Tauropine dehydrogenase

organism: Halichondria japonica

enzyme: Tauropine dehydrogenase


substrate_name: NAD+

substrate: NAD+


substrate_name: tauropine

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

substrate: tauropine


substrate_name: taurine

substrate: taurine


organism: Haliotis tuberculata lamellosa


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


substrate_name: Ala

substrate: Ala


organism: Mazzaella japonica


substrate_name: pyruvate

substrate: pyruvate


enzyme_display_name: beta-alanopine dehydrogenase

organism: Cellana grata

assay_conditions_summary: pH 7.1; 303.15 K; cofactor NADH

enzyme: beta-alanopine dehydrogenase


assay_conditions_summary: pH 9; 303.15 K; cofactor NAD+


substrate_name: beta-Alanine

substrate: beta-Alanine


substrate_name: beta-alanopine

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

substrate: beta-alanopine


organism: Haliotis discus hannai


organism: Patiria pectinifera


organism: Scolelepis fuliginosa


substrate_name: Gly

substrate: Gly


organism: Mytilus edulis


organism: Glycera alba


organism: Suberites domuncula

assay_conditions_summary: pH 7; 303.15 K; cofactor NADH

proof_lines: determined using constant cosubstrate concentrations (100 mM glycine and 0.3 mM NADH, pH 7.0)


enzyme_display_name: Opine dehydrogenase

organism: Arthrobacter sp.

substrate_name: L-Cys

enzyme: Opine dehydrogenase

substrate: L-Cys


substrate_name: L-Leu

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

substrate: L-Leu


substrate_name: oxaloacetate

substrate: oxaloacetate


substrate_name: L-Ser

substrate: L-Ser


substrate_name: L-Met

smiles: CSCCC(C(=O)O)N

substrate: L-Met


substrate_name: L-Ala

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

substrate: L-Ala


substrate_name: L-Ile

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

substrate: L-Ile


substrate_name: L-Thr

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

substrate: L-Thr


substrate_name: L-Phe

substrate: L-Phe


substrate_name: glyoxylate

substrate: glyoxylate


smiles: CCCCC(=O)C(=O)[O-]


substrate_name: beta-chloro-(S)-alanine

substrate: beta-chloro-(S)-alanine


substrate_name: (S)-norvaline

smiles: CCCC(C(=O)O)N

substrate: (S)-norvaline


substrate_name: L-Val

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

substrate: L-Val


substrate_name: (S)-norleucine

smiles: CCCCC(C(=O)O)N

substrate: (S)-norleucine


substrate_name: (R,S)-2,3-diaminopropionic acid

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

substrate: (R,S)-2,3-diaminopropionic acid


substrate_name: L-phenylglycine

substrate: L-phenylglycine


enzyme_display_name: N5-(carboxyethyl)ornithine synthase

organism: Clostridium botulinum

substrate_name: L-Ornithine; Pyruvate; H+; NADPH

assay_conditions_summary: cofactor NADPH

enzyme: N5-(carboxyethyl)ornithine synthase

substrate: L-Ornithine; Pyruvate; H+; NADPH


organism: Lactococcus lactis subsp. lactis

substrate_name: NADPH; Pyruvate; L-Ornithine

substrate: NADPH; Pyruvate; L-Ornithine


enzyme_display_name: Ketimine reductase mu-crystallin

organism: Bos taurus

substrate_name: 3,4-dehydro-thiomorpholine-3-carboxylate (at pH 5.0 and 37 degrees Celsius)

assay_conditions_summary: pH 5; cofactor NADH

column_label: Table V (Apparent kinetic constants), row S-Aminoethylcysteine ketimine, column Km (mM), value 0.24; Table V note says NADH for AECK and pH 5.0; Materials and Methods says spectrophotometry at 340 nm and 37 C

raw_value_unit_evidence: Table V row S-Aminoethylcysteine ketimine, column Km (mM) = 0.24

row_label: S-Aminoethylcysteine ketimine

table_label: Table V

enzyme: Ketimine reductase mu-crystallin


organism: Homo sapiens

assay_conditions_summary: pH 5

substrate: 3,4-dehydro-thiomorpholine-3-carboxylate (at pH 5.0 and 37 degrees Celsius)


enzyme_display_name: N(5)-(carboxyethyl)ornithine synthase

substrate_name: L-ornithine

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

column_label: PubMed abstract substrate Km sentence reports ornithine K_M 3.3 mM.

normalized_value_unit_evidence: K_M 3.3 mM

raw_value_unit_evidence: Abstract sentence: Substrate Km values were determined to be NADPH, 6.6 microM; pyruvate, 150 microM; ornithine, 3.3 mM; and lysine, 18.2 mM.

row_label: Abstract substrate Km sentence / ornithine

table_label: PubMed abstract

enzyme: N(5)-(carboxyethyl)ornithine synthase

substrate: L-ornithine


substrate_name: L-lysine

column_label: PubMed abstract substrate Km sentence reports lysine K_M 18.2 mM.

row_label: Abstract substrate Km sentence / lysine

substrate: L-lysine


substrate_name: pyruvate

column_label: PubMed abstract substrate Km sentence reports pyruvate K_M 150 microM, normalized to 0.15 mM.

row_label: Abstract substrate Km sentence / pyruvate

substrate: pyruvate


substrate_name: NADPH

column_label: PubMed abstract substrate Km sentence reports NADPH K_M 6.6 microM, normalized to 0.0066 mM.

row_label: Abstract substrate Km sentence / NADPH

substrate: NADPH


organism: Arabella iricolor

substrate_name: NADH

assay_conditions_summary: pH 7; cofactor NADH

column_label: Apparent Michaelis Constant section; Km metric; NADH; raw Km value 0.036 +/- 0.002 mM.

raw_value_unit_evidence: Apparent Km for NADH = 0.036 +/- 0.002 mM.

row_label: Apparent Km prose; NADH row/value

table_label: PMID 8840516, Apparent Michaelis Constant prose section

source_databases_merged: brenda uniprot

substrate: NADH


substrate_name: NAD+

assay_conditions_summary: pH 8.5; cofactor NAD+

column_label: Apparent Michaelis Constant section; Km metric; NAD+; raw Km value 0.051 +/- 0.002 mM.

raw_value_unit_evidence: Apparent Km for NAD+ = 0.051 +/- 0.002 mM.

row_label: Apparent Km prose; NAD+ row/value

substrate: NAD+


column_label: Apparent Michaelis Constant section; Km metric; pyruvate; raw Km value 0.34 +/- 0.02 mM.

raw_value_unit_evidence: Apparent Km for pyruvate = 0.34 +/- 0.02 mM.

row_label: Apparent Km prose; pyruvate row/value


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

column_label: Apparent Michaelis Constant section; Km metric; 2-ketobutyrate / 2-oxobutanoate; raw Km value 3.3 +/- 0.211 mM.

raw_value_unit_evidence: Apparent Km for 2-ketobutyrate / 2-oxobutanoate = 3.3 +/- 0.211 mM.

row_label: Apparent Km prose; 2-ketobutyrate / 2-oxobutanoate row/value


substrate_name: tauropine

column_label: Apparent Michaelis Constant section; Km metric; tauropine; raw Km value 4.8 +/- 0.31 mM.

raw_value_unit_evidence: Apparent Km for tauropine = 4.8 +/- 0.31 mM.

row_label: Apparent Km prose; tauropine row/value

substrate: tauropine


substrate_name: oxaloacetate

column_label: Apparent Michaelis Constant section; Km metric; oxalacetate / oxaloacetate; raw Km value 4.9 +/- 0.24 mM.

raw_value_unit_evidence: Apparent Km for oxalacetate / oxaloacetate = 4.9 +/- 0.24 mM.

row_label: Apparent Km prose; oxalacetate / oxaloacetate row/value

substrate: oxaloacetate


substrate_name: taurine

column_label: Apparent Michaelis Constant section; Km metric; taurine; raw Km value 35.7 +/- 3.1 mM.

raw_value_unit_evidence: Apparent Km for taurine = 35.7 +/- 3.1 mM.

row_label: Apparent Km prose; taurine row/value

substrate: taurine


substrate_name: 2-ketobutyrate for the reverse reaction of the tauropine dehydrogenase activity (at pH 7.0)


organism: Clostridium botulinum (strain Hall / ATCC 3502 / NCTC 13319 / Type A)


substrate_name: delta(1)-piperideine-2-carboxylate (at pH 7.2 and 37 degrees Celsius)


substrate_name: 1-pyrroline-2-carboxylate (at pH 7.2 and 37 degrees Celsius)


substrate_name: delta(2)-thiazoline-2-carboxylate (at pH 7.2 and 37 degrees Celsius)


substrate_name: NADPH (at pH 5.5 and 37 degrees Celsius)

assay_conditions_summary: pH 5.5; buffer pH 5; cofactor NADPH

column_label: Table V, Apparent kinetic constants, columns Substrate, Km (mM), Vmax (umol/min per mg); row NADPH a has Km 0.015 mM

raw_value_unit_evidence: Table V row NADPH a reports Km 0.015 mM and Vmax 20.8 umol/min per mg; footnote a gives pH 5.5 NAD(P)H assay system

row_label: NADPH a


substrate_name: NADH (at pH 5.5 and 37 degrees Celsius)

assay_conditions_summary: pH 5.5; buffer pH 5; cofactor NADH

column_label: Table V, Apparent kinetic constants, columns Substrate, Km (mM), Vmax (umol/min per mg); row NADH a has Km 0.26 mM

normalized_value_unit_evidence: Km 0.26 mM

raw_value_unit_evidence: Table V row NADH a reports Km 0.26 mM and Vmax 149 umol/min per mg; footnote a gives pH 5.5 NAD(P)H assay system

row_label: NADH a

substrate: NADH (at pH 5.5 and 37 degrees Celsius)


substrate_name: delta(1)-piperideine-2-carboxylate (at pH 5.0 and 37 degrees Celsius)

assay_conditions_summary: pH 5; cofactor NADPH

column_label: Table V (Apparent kinetic constants), row delta1-Piperidine 2-carboxylate, column Km (mM), value 0.33; Table V note says pH 5.0; Materials and Methods says spectrophotometry at 340 nm and 37 C

raw_value_unit_evidence: Table V row delta1-Piperidine 2-carboxylate, column Km (mM) = 0.33

row_label: delta1-Piperidine 2-carboxylate


substrate_name: (R)-lanthionine ketimine (at pH 5.0 and 37 degrees Celsius)

assay_conditions_summary: pH 5; buffer pH 5; cofactor NADPH

column_label: Table V, Apparent kinetic constants, columns Substrate, Km (mM), Vmax (umol/min per mg); row Lanthionine ketimine has Km 1.17 mM

normalized_value_unit_evidence: Km 1.17 mM

raw_value_unit_evidence: Table V row Lanthionine ketimine reports Km 1.17 mM and Vmax 2.2 umol/min per mg; standard Table V note gives pH 5.0

row_label: Lanthionine ketimine


substrate_name: (S)-cystathionine ketimine (at pH 5.0 and 37 degrees Celsius)

column_label: Table V, Apparent kinetic constants, columns Substrate, Km (mM), Vmax (umol/min per mg); row Cystathionine ketimine has Km 10 mM

normalized_value_unit_evidence: Km 10.0 mM

raw_value_unit_evidence: Table V row Cystathionine ketimine reports Km 10 mM and Vmax 52 umol/min per mg; standard Table V note gives pH 5.0

row_label: Cystathionine ketimine


organism: Papaver somniferum

assay_conditions_summary: pH 8; cofactor NADPH

column_label: Supplementary PDF page 15 (printed page 14), Fig. S3B, blue substrate-inhibition legend: Vmax = 45 +/- 11, Km = 14 +/- 5.7; x-axis 1,2-Dehydroreticuline (uM).

raw_value_unit_evidence: Blue fit legend reports Km = 14 +/- 5.7 uM.

row_label: Panel B: full-length STORR (CYP82Y2-Oxired), substrate 1,2-dehydroreticuline, substrate-inhibition model

table_label: Supplementary Fig. S3B


enzyme_display_name: Bifunctional protein STORR

substrate_name: (S)-reticuline

enzyme: Bifunctional protein STORR

substrate: (S)-reticuline


substrate_name: pyruvate

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


substrate_name: NADH (at pH 5.0 and 37 degrees Celsius)

proof_lines:
mu-crystallin
ketimine reductase
human mu-crystallin

substrate: NADH (at pH 5.0 and 37 degrees Celsius)


organism: Lactococcus lactis

substrate_name: NADPH


substrate_name: ornithine

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

substrate: ornithine


substrate_name: lysine

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

substrate: lysine


organism: Sus scrofa

substrate_name: NADH

assay_conditions_summary: cofactor NADH

proof_lines: reaction with S-aminoethylcysteine ketimine


substrate_name: S-Aminoethylcysteine ketimine

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

substrate: S-Aminoethylcysteine ketimine


substrate_name: Lanthionine ketimine

substrate: Lanthionine ketimine


substrate_name: Cystathionine ketimine

substrate: Cystathionine ketimine


organism: Bacillus subtilis

substrate_name: D-Pro

substrate: D-Pro


substrate_name: N-ethylglycine

substrate: N-ethylglycine


substrate_name: sarcosine

substrate: sarcosine


enzyme_display_name: diaminopimelate dehydrogenase

organism: Bacteroides fragilis

enzyme: diaminopimelate dehydrogenase


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


enzyme_display_name: pyrroline-5-carboxylate reductase

enzyme: pyrroline-5-carboxylate reductase

substrate: NADPH


substrate: NADH


enzyme_display_name: UniProt O76290

organism: Trypanosoma brucei brucei

substrate_name: dihydrobiopterin

source_license: CC BY-NC-ND 4.0

enzyme: UniProt O76290

substrate: dihydrobiopterin


proof_lines: pH 7.5, temperature not specified in the publication, Vmax: 0.9 micromol/min/mg protein

substrate: pyruvate


assay_conditions_summary: pH 7.5; cofactor NADPH

proof_lines: pH 7.5, temperature not specified in the publication


substrate_name: L-ornithine

substrate: L-ornithine


enzyme_display_name: flavin reductase (NADPH)

organism: Vibrio harveyi

assay_conditions_summary: pH 7; 295.15 K; buffer 20 mM N-tris[hydroxymethyl]methyl-3-aminopropanesulfonic acid; cofactor NADPH

proof_lines: mutant enzyme N134A, in 20 mM N-tris[hydroxymethyl]methyl-3-aminopropanesulfonic acid buffer, pH 7.0, at 22°C

enzyme: flavin reductase (NADPH)


proof_lines: mutant enzyme R133A, in 20 mM N-tris[hydroxymethyl]methyl-3-aminopropanesulfonic acid buffer, pH 7.0, at 22°C


proof_lines: mutant enzyme R225A, in 20 mM N-tris[hydroxymethyl]methyl-3-aminopropanesulfonic acid buffer, pH 7.0, at 22°C


proof_lines: wild type enzyme, in 20 mM N-tris[hydroxymethyl]methyl-3-aminopropanesulfonic acid buffer, pH 7.0, at 22°C


proof_lines: mutant enzyme R15A, in 20 mM N-tris[hydroxymethyl]methyl-3-aminopropanesulfonic acid buffer, pH 7.0, at 22°C


proof_lines: mutant enzyme K167A, in 20 mM N-tris[hydroxymethyl]methyl-3-aminopropanesulfonic acid buffer, pH 7.0, at 22°C


enzyme_display_name: 6,7-dihydropteridine reductase

substrate_name: quinoid 6-methyl-7,8-dihydropterin

enzyme: 6,7-dihydropteridine reductase

substrate: quinoid 6-methyl-7,8-dihydropterin


proof_lines: recombinant Y150E glutathione-S-transferase fusion mutant enzyme


proof_lines: recombinant Y150K glutathione-S-transferase fusion mutant enzyme


proof_lines: recombinant Y150F glutathione-S-transferase fusion mutant enzyme


proof_lines: recombinant wild-type glutathione-S-transferase fusion enzyme


proof_lines: recombinant Y150S glutathione-S-transferase fusion mutant enzyme


substrate_name: quinoid 7,8-dihydrobiopterin

proof_lines: recombinant Y150H glutathione-S-transferase fusion mutant enzyme

substrate: quinoid 7,8-dihydrobiopterin


substrate_name: quinoid 7,8-dihydropterin

substrate: quinoid 7,8-dihydropterin


enzyme_display_name: pteridine reductase

organism: Agrobacterium fabrum

enzyme: pteridine reductase


proof_lines: recombinant mutant Y161F/Y163F, pH 6.0, 25°C


proof_lines: recombinant wild-type enzyme, pH 6.0, 25°C


substrate_name: Cibacron Marine

assay_conditions_summary: pH 7.5; 310.15 K; cofactor FMN

proof_lines:
pH 7.5, 37°C
substrate FMN, pH 7.5, 37°C
substrate NADPH, pH 7.5, 37°C

substrate: Cibacron Marine


substrate_name: FMN

assay_conditions_summary: pH 7.5; 310.15 K; cofactor NADPH

proof_lines:
pH 7.5, 37°C
pH 7.0, 23°C, recombinant enzyme

substrate: FMN


substrate_name: NADPH

proof_lines: pH 7.0, 23°C, recombinant enzyme, with FMN


assay_conditions_summary: pH 7.5; 310.15 K; cofactor NADH


organism: Entamoeba histolytica

assay_conditions_summary: cofactor FAD


substrate_name: FAD

substrate: FAD


substrate_name: riboflavin

substrate: riboflavin


substrate_name: galactoflavin

substrate: galactoflavin


assay_conditions_summary: cofactor NADPH


enzyme_display_name: FAD reductase (NADH)

organism: Pseudomonas protegens Pf-5

substrate_name: NADH

assay_conditions_summary: pH 7; 303.15 K; cofactor NADH

enzyme: FAD reductase (NADH)


substrate_name: Lumiflavin

substrate: Lumiflavin


organism: Streptomyces viridifaciens


assay_conditions_summary: cofactor FMN


organism: Cereibacter sphaeroides


organism: Rhodococcus erythropolis


substrate_name: Unknown

substrate: Unknown


assay_conditions_summary: pH 7; 296.15 K; buffer 50 mM phosphate; cofactor NADPH

proof_lines: mutant enzyme E99K, in 50 mM phosphate buffer, at pH 7.0, at 23°C


assay_conditions_summary: pH 7; 296.15 K; buffer 50 mM phosphate; cofactor FMN


proof_lines: enzyme reconstituted from apoenzyme and 2-thioFMN


organism: Crithidia fasciculata


sequence_variant_note: Resolved canonical sequence from source_record and applied mutation signature R203A to reconstruct the assayed variant sequence.


assay_conditions_summary: pH 7; buffer 50 mM phosphate; cofactor NADPH


enzyme_display_name: flavin reductase (NADH)

organism: Mycolicibacterium goodii

assay_conditions_summary: pH 7; 328.15 K; cofactor FMN

enzyme: flavin reductase (NADH)


assay_conditions_summary: pH 7; 328.15 K; cofactor NADH


organism: Corynebacterium glutamicum

assay_conditions_summary: pH 7.5; 303.15 K; cofactor NADPH

proof_lines: with FMN, pH 7.5, 30°C, recombinant enzyme

proof_lines: with FAD, pH 7.5, 30°C, recombinant enzyme


proof_lines: enzyme fused to luciferase, at 23°C in 50 mM phosphate buffer, pH 7.0

proof_lines: native enzyme, at 23°C in 50 mM phosphate buffer, pH 7.0


organism: Trypanosoma cruzi


organism: Photobacterium leiognathi

assay_conditions_summary: pH 8; buffer 50 mM Tris-Cl; cofactor FMN


assay_conditions_summary: pH 8; buffer 50 mM Tris-Cl; cofactor NADH

column_label: PMID 20836540 Table 1 row KMb reports 15 uM for NADH; PMID 18156264 text reports Km for NADH calculated from steady-state kinetics at 4 deg C pH 8.0.

normalized_value_unit_evidence: Km normalized_value=0.015 normalized_unit=mM; current_export_km=0.015 mM

raw_value_unit_evidence: Km raw_value=15 raw_unit=uM for NADH in PMID 20836540 Table 1; previous report text gives Km for NADH = 15.1 uM

row_label: LuxG reaction / NADH steady-state parameter

table_label: PMID 20836540 Table 1; PMID 18156264 steady-state kinetics prose


assay_conditions_summary: pH 7.5; 303.15 K; cofactor NADH


organism: Rattus norvegicus

assay_conditions_summary: cofactor NADH


substrate_name: quinonoid dihydrobiopterin

substrate: quinonoid dihydrobiopterin


organism: Bombyx mori

assay_conditions_summary: pH 7.4; 295.15 K; cofactor NADPH


organism: Leishmania tarentolae

substrate_name: folate

proof_lines: dihydrobiopterin, recombinant enzyme, pH 4.7

substrate: folate


substrate_name: dihydrofolate

assay_conditions_summary: pH 6

substrate: dihydrofolate


assay_conditions_summary: pH 4.7; cofactor NADPH

proof_lines: with dihydrobiopterin as cosubstrate, recombinant enzyme, pH 4.7

proof_lines: with dihydrofolate as cosubstrate, recombinant enzyme, pH 6.0

proof_lines: with biopterin as cosubstrate, recombinant enzyme, pH 4.7

substrate: NADPH


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_mutation_ranked_match and applied mutation signature W104F with inferred residue-numbering offset +5 to reconstruct the assayed variant sequence.

substrate: NADH


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_mutation_ranked_match and applied mutation signature A6V to reconstruct the assayed variant sequence.


enzyme_display_name: Dihydropteridine reductase

enzyme: Dihydropteridine reductase


organism: Paracoccus denitrificans

assay_conditions_summary: pH 7.4; 303.15 K; cofactor NADH

proof_lines:
versus FMN, pH 7.4, 30°C
versus NADH, pH 7.4, 30°C


organism: Escherichia coli

assay_conditions_summary: pH 7; buffer 100 mM Na+/K+ phosphate; cofactor NADPH

proof_lines: 25°C, enzyme in complex with monooxygenase SsuD, presence of octanesulfonate

proof_lines: coupled oxidoreductase-luciferase assay, kcat/Km = 73/(M*min), in 100 mM Na+/K+ phosphate buffer with 100 mM NaCl, pH 7.0, 1 microM Fre oxidoreductase, 10 microM decanal, 10 microM NADPH, 5 microM luciferase


assay_conditions_summary: cofactor NADPH


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_mutation_ranked_match and applied mutation signature A133S with inferred residue-numbering offset +8 to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_mutation_ranked_match and applied mutation signature K150E with inferred residue-numbering offset -9 to reconstruct the assayed variant sequence.


organism: Pseudomonas putida

assay_conditions_summary: pH 7.5; buffer Primary measurement evidence: evidence location type: prose; cofactor NADH

column_label: Journal page 3287, Results prose subsection Substrate specificity and kinetic properties of Fred (there is no kinetic table for Fred in this paper). The sentence reports two flavin measurements: for FMN the printed values are Km 3.6 uM, kcat 283 s^-1 and kcat/Km 7.9 x 10^7; for FAD they are Km 19 uM, kcat 128 s^-1 and kcat/Km 6.7 x 10^6. The measurement read for this record is the FAD one: printed Km 19 uM, printed kcat 128 s^-1 and printed kcat/Km 6.7 x 10^6.

normalized_value_unit_evidence: kcat: printed 128 s^-1; the value is already in s^-1, so no unit conversion applies; the catalog value is 128.0 s^(-1). Km: printed 19 uM; 19 / 1000 = 0.019 mM; the catalog value is 0.019 mM. kcat/Km: the catalog value 6736.842105263158 s^(-1)*mM^(-1) is the full-precision quotient 128 / 0.019 of those same two printed cells. The paper additionally prints its own kcat/Km numeral 6.7 x 10^6 M^-1 s^-1; 1 M^-1 s^-1 is 0.001 s^-1 mM^-1, so that numeral converts to 6700.0 s^-1 mM^-1, and the full-precision quotient rounds to the same two significant digits the paper prints, so the printed numeral corroborates the catalog quotient and no metric changes.

raw_value_unit_evidence: Reviewed tuple for FAD. The reviewed primary measurement is stated in the Results subsection Substrate specificity and kinetic properties of Fred, journal page 3287. The matched measurement is FAD reduction by purified Fred. The measured construct is purified Fred flavin reductase of Pseudomonas putida ATCC 17453. The measured substrate is FAD, the flavin whose reduction by Fred was assayed. The assay condition is pH 7.5, 25 C, 50 mM Tris-HCl buffer. Fred with FAD as the varied substrate: kcat 128 s^-1. Fred with FAD as the varied substrate: Km 19 uM. Fred with FAD as the varied substrate: printed kcat/Km 6.7 x 10^6 M^-1 s^-1, where the numeral is printed in the paper and the unit is resolved from the printed components.

row_label: FAD reduction by purified Fred

table_label: No table: the values are printed in the Results prose on journal page 3287


assay_conditions_summary: pH 7.5; cofactor NADH

proof_lines: kcat is 283 sec(-1) for the reduction of FMN. kcat is 128 sec(-1) for the reduction of FAD.


organism: Leishmania major

proof_lines:
recombinant enzyme, pH 6.0
recombinant enzyme, pH 4.7


substrate_name: biopterin

proof_lines: NADPH, with folate as cosubstrate, recombinant enzyme, pH 6.0

substrate: biopterin


proof_lines:
recombinant enzyme, pH 4.7
recombinant enzyme, pH 6.0
native enzyme, pH 6.0


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_consensus_sequence and applied mutation signature Y175F to reconstruct the assayed variant sequence.


organism: Ovis aries

proof_lines: reduction of quinonoid 2-amino-4-hydroxy-6,7-dimetyldihydropteridine


proof_lines: reduction of quinonoid 2-amino-4-hydroxy-6,7-dimethyldihydropteridine


organism: Streptomyces globisporus

assay_conditions_summary: pH 7.5; 295.15 K; cofactor NADH

proof_lines: kcat is 3.1 sec(-1) with NADH as substrate. kcat is 4.5 sec(-1) with FAD as substrate.

source_databases_merged: brenda skid uniprot


organism: Homo sapiens

assay_conditions_summary: pH 7.5; cofactor FMN

proof_lines: pH and temperature not specified in the publication


proof_lines: coupled oxidoreductase-luciferase assay, kcat/Km = 1257/(M*min), in 100 mM Na+/K+ phosphate buffer with 100 mM NaCl, pH 7.0, 1 microM Fre oxidoreductase, 10 microM decanal, 10 microM NADPH, 5 microM luciferase, riboflavin is favoured by Fre

proof_lines: coupled oxidoreductase-luciferase assay, in 100 mM Na+/K+ phosphate buffer with 100 mM NaCl, pH 7.0, 1 microM Fre oxidoreductase, 10 microM decanal, 10 microM NADPH, 5 microM luciferase, riboflavin is favoured by Fre oxidoreductase but a po


organism: Microcebus murinus

assay_conditions_summary: cofactor FAD


enzyme_display_name: Vomilenine reductase

organism: Rauvolfia serpentina

substrate_name: vomilenine

proof_lines: kcat is 1.7 sec(-1) with vomilenine as substrate.

enzyme: Vomilenine reductase

substrate: vomilenine


enzyme_display_name: Flavin reductase (NADPH)

organism: Lentzea aerocolonigenes

substrate_name: NADH (with FAD)

enzyme: Flavin reductase (NADPH)

substrate: NADH (with FAD)


assay_conditions_summary: pH 7.4; buffer 50 mM HEPES; cofactor NADPH

column_label: PMC HTML Table 1 and Europe PMC PDF text lines 241-256 report LmPTR1 qH2B columns Kappm, kcat, and kcat/Kappm.

raw_value_unit_evidence: Table 1 LmPTR1 qH2B column: Kappm=10.3 +/- 1.5 uM; kcat=1.84 s^-1; kcat/Kappm=1.8 x 10^5 M^-1 s^-1.

row_label: LmPTR1 / qH2B

table_label: Table 1. Summary and comparison of the kinetic characterization of TbPTR1, LmPTR1, and LmQDPR with various substrates


substrate_name: dihydrobiopterin

column_label: PMC HTML Table 1 and Europe PMC PDF text lines 241-256 report LmPTR1 H2B columns Kappm, kcat, and kcat/Kappm.

normalized_value_unit_evidence: paper kcat=0.57 s^-1; paper Km-like Kappm=3.32 uM=0.00332 mM; paper kcat_over_km=170 s^-1 mM^-1; expected Km is 0.0032 mM.

raw_value_unit_evidence: Table 1 LmPTR1 H2B column: Kappm=3.32 +/- 0.48 uM; kcat=0.57 s^-1; kcat/Kappm=1.7 x 10^5 M^-1 s^-1.

row_label: LmPTR1 / H2B

substrate: dihydrobiopterin


enzyme_display_name: p-hydroxyphenylacetate 3-hydroxylase, reductase component

organism: Acinetobacter baumannii

substrate_name: NADH (with FMN as cosubstrate)

proof_lines: Values measured using the C1-C2 complex. kcat is 343 min(-1) for the reduction of FMN with NADH.

substrate: NADH (with FMN as cosubstrate)


organism: Trypanosoma brucei

assay_conditions_summary: pH 7.4; buffer 50 mM HEPES

proof_lines:
in 50 mM HEPES buffer, pH 7.4, at 25°C
pH 5.0, 25°C, recombinant enzyme
pH 3.7


enzyme_display_name: Flavin-dependent trigonelline monooxygenase, reductase component

organism: Acinetobacter baylyi (strain ATCC 33305 / BD413 / ADP1)

proof_lines: kcat is 72 sec(-1) for NADH as substrate.


organism: Dictyostelium discoideum

assay_conditions_summary: pH 7.5; 298.15 K; cofactor NADH

proof_lines: apparent value, in 50 mM Tris-HCl, pH 7.5, at 25°C


organism: Bacillus subtilis

substrate_name: Riboflavin-5-phosphate; H+; NADPH

substrate: Riboflavin-5-phosphate; H+; NADPH


assay_conditions_summary: pH 7.5; 298.15 K; cofactor FAD


organism: Mus musculus

substrate_name: INSR substrate

substrate: INSR substrate


organism: Leishmania tarentolae

substrate_name: H+; Biopterin; NADPH

substrate: H+; Biopterin; NADPH


substrate_name: H+; 7,8-Dihydrobiopterin; NADPH

substrate: H+; 7,8-Dihydrobiopterin; NADPH


substrate_name: NADPH; H+; Quinonoid 6,7-dimethyl dihydropterin


organism: Pseudomonas sp.


substrate: NADPH; H+; Quinonoid 6,7-dimethyl dihydropterin


organism: Acinetobacter baylyi

substrate_name: NADH; H+; Riboflavin-5-phosphate

substrate: NADH; H+; Riboflavin-5-phosphate


organism: Burkholderia cepacia

substrate_name: FAD; NADH; H+

substrate: FAD; NADH; H+


organism: Cupriavidus necator

assay_conditions_summary: cofactor NAD+


organism: Escherichia coli

substrate_name: H+; FAD; NADH

substrate: H+; FAD; NADH


organism: Saccharomyces cerevisiae

substrate_name: NADH; H+; FAD

substrate: NADH; H+; FAD


substrate_name: Unknown

substrate: Unknown


organism: Leishmania major

substrate_name: NADPH; H+; Biopterin

assay_conditions_summary: pH 7.4; 298.15 K; buffer 50 mM HEPES; cofactor NADPH

proof_lines: pH 7.5, temperature not specified in the publication

substrate: NADPH; H+; Biopterin


assay_conditions_summary: pH 7.4; 298.15 K; buffer preserved note previews about HEPES


organism: Ovis aries

proof_lines:
cofactor NADPH
enzyme from brain
cofactor NADH


organism: Rattus norvegicus

substrate_name: H+; Quinonoid 6,7-dimethyl dihydropteridine; NADPH


substrate_name: quinoid dihydrobiopterin

sequence_variant_note: Resolved canonical sequence from identity matching and applied mutation signature K150Q with inferred residue-numbering offset -9 to reconstruct the assayed variant sequence.

substrate: quinoid dihydrobiopterin


organism: Pseudomonas putida


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_mutation_ranked_match and applied mutation signature K150I with inferred residue-numbering offset -9 to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_mutation_ranked_match and applied mutation signature K150M with inferred residue-numbering offset -9 to reconstruct the assayed variant sequence.


organism: Acinetobacter baumannii

substrate_name: apo-C1_hpah; Riboflavin-5-phosphate

substrate: apo-C1_hpah; Riboflavin-5-phosphate


enzyme_display_name: NADH-dependent flavin reductase

organism: Rhodococcus sp

enzyme: NADH-dependent flavin reductase


substrate_name: FAD

substrate: FAD


substrate_name: FMN

proof_lines:
SMOB-ADP1
Acinetobacter baylyi
NADH

substrate: FMN


substrate_name: NADH

substrate: NADH


substrate_name: riboflavin

substrate: riboflavin


organism: Agrobacterium fabrum

assay_conditions_summary: pH 6; 298.15 K; cofactor NADPH

column_label: Table 1. Summary of kinetic parameters and substrate specificities for PruA from A. tumefaciens compared to homologs from B. abortus and P. aeruginosa; row A. tumefaciens PruA / H2BPt; columns kcat s^-1 raw 4.7, Km uM raw 34.7 +/- 3.8, kcat/Km M^-1*s^-1 raw 1.3 x 10^5; normalized paper kcat 4.7 s^-1, Km 0.0347 mM, same-row kcat/Km 135.44668587896254 s^-1*mM^-1; export values kcat 4.7 s^-1, Km 0.0347 mM, kcat/Km 135.44668587896254 s^-1*mM^-1

normalized_value_unit_evidence: kcat normalized_value=4.7 normalized_unit=s^-1; Km normalized_value=0.0347 normalized_unit=mM from 34.7 +/- 3.8 uM / 1000; kcat/Km normalized_value=135.44668587896254 normalized_unit=s^-1*mM^-1 from same-row kcat and Km; printed efficiency=1.3 x 10^5 M^-1*s^-1

raw_value_unit_evidence: kcat raw_value=4.7 raw_unit=s^-1; Km raw_value=34.7 +/- 3.8 raw_unit=uM; kcat/Km raw_value=1.3 x 10^5 raw_unit=M^-1*s^-1

row_label: A. tumefaciens PruA / H2BPt


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature R24A to reconstruct the assayed variant sequence.

column_label: Table 2. Kinetic parameters for PruA variants with H2MPt; row PruA R24A / H2MPt; columns kcat s^-1 raw 0.8, Km uM raw 169.4 +/- 50.2, kcat/Km M^-1*s^-1 raw 4.8 x 10^3; normalized paper kcat 0.8 s^-1, Km 0.1694 mM, same-row kcat/Km 4.722550177095632 s^-1*mM^-1; export values kcat 0.8 s^-1, Km 0.1694 mM, kcat/Km 4.722550177095632 s^-1*mM^-1

normalized_value_unit_evidence: kcat normalized_value=0.8 normalized_unit=s^-1; Km normalized_value=0.1694 normalized_unit=mM from 169.4 +/- 50.2 uM / 1000; kcat/Km normalized_value=4.722550177095632 normalized_unit=s^-1*mM^-1 from same-row kcat and Km; printed efficiency=4.8 x 10^3 M^-1*s^-1

raw_value_unit_evidence: kcat raw_value=0.8 raw_unit=s^-1; Km raw_value=169.4 +/- 50.2 raw_unit=uM; kcat/Km raw_value=4.8 x 10^3 raw_unit=M^-1*s^-1

row_label: PruA R24A / H2MPt


column_label: Table 1. Summary of kinetic parameters and substrate specificities for PruA from A. tumefaciens compared to homologs from B. abortus and P. aeruginosa; row A. tumefaciens PruA / H2MPt; columns kcat s^-1 raw 2.3, Km uM raw 71.3 +/- 11.1, kcat/Km M^-1*s^-1 raw 3.2 x 10^4; normalized paper kcat 2.3 s^-1, Km 0.0713 mM, same-row kcat/Km 32.25806451612903 s^-1*mM^-1; export values kcat 2.3 s^-1, Km 0.0713 mM, kcat/Km 32.25806451612903 s^-1*mM^-1

normalized_value_unit_evidence: kcat normalized_value=2.3 normalized_unit=s^-1; Km normalized_value=0.0713 normalized_unit=mM from 71.3 +/- 11.1 uM / 1000; kcat/Km normalized_value=32.25806451612903 normalized_unit=s^-1*mM^-1 from same-row kcat and Km; printed efficiency=3.2 x 10^4 M^-1*s^-1

raw_value_unit_evidence: kcat raw_value=2.3 raw_unit=s^-1; Km raw_value=71.3 +/- 11.1 raw_unit=uM; kcat/Km raw_value=3.2 x 10^4 raw_unit=M^-1*s^-1

row_label: A. tumefaciens PruA / H2MPt


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature Y161A with inferred residue-numbering offset +2 to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature Y161F/Y163F to reconstruct the assayed variant sequence.

column_label: Table 2. Kinetic parameters for PruA variants with H2MPt; row PruA Y161F Y163F / H2MPt; columns kcat s^-1 raw 2.0, Km uM raw 333.2 +/- 122.6, kcat/Km M^-1*s^-1 raw 6.0 x 10^3; normalized paper kcat 2.0 s^-1, Km 0.3332 mM, same-row kcat/Km 6.002400960384154 s^-1*mM^-1; export values kcat 2.0 s^-1, Km 0.3332 mM, kcat/Km 6.002400960384154 s^-1*mM^-1

normalized_value_unit_evidence: kcat normalized_value=2.0 normalized_unit=s^-1; Km normalized_value=0.3332 normalized_unit=mM from 333.2 +/- 122.6 uM / 1000; kcat/Km normalized_value=6.002400960384154 normalized_unit=s^-1*mM^-1 from same-row kcat and Km; printed efficiency=6.0 x 10^3 M^-1*s^-1

raw_value_unit_evidence: kcat raw_value=2.0 raw_unit=s^-1; Km raw_value=333.2 +/- 122.6 raw_unit=uM; kcat/Km raw_value=6.0 x 10^3 raw_unit=M^-1*s^-1

row_label: PruA Y161F Y163F / H2MPt


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature Y163F with inferred residue-numbering offset -2 to reconstruct the assayed variant sequence.

column_label: Table 2. Kinetic parameters for PruA variants with H2MPt; row PruA Y163F / H2MPt; columns kcat s^-1 raw 4.2, Km uM raw 212.7 +/- 36.6, kcat/Km M^-1*s^-1 raw 2.0 x 10^4; normalized paper kcat 4.2 s^-1, Km 0.2127 mM, same-row kcat/Km 19.746121297602258 s^-1*mM^-1; export values kcat 4.2 s^-1, Km 0.2127 mM, kcat/Km 19.746121297602258 s^-1*mM^-1

normalized_value_unit_evidence: kcat normalized_value=4.2 normalized_unit=s^-1; Km normalized_value=0.2127 normalized_unit=mM from 212.7 +/- 36.6 uM / 1000; kcat/Km normalized_value=19.746121297602258 normalized_unit=s^-1*mM^-1 from same-row kcat and Km; printed efficiency=2.0 x 10^4 M^-1*s^-1

raw_value_unit_evidence: kcat raw_value=4.2 raw_unit=s^-1; Km raw_value=212.7 +/- 36.6 raw_unit=uM; kcat/Km raw_value=2.0 x 10^4 raw_unit=M^-1*s^-1

row_label: PruA Y163F / H2MPt


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

column_label: Table 1. Summary of kinetic parameters and substrate specificities for PruA from A. tumefaciens compared to homologs from B. abortus and P. aeruginosa; row A. tumefaciens PruA / H2NPt; columns kcat s^-1 raw 2.4, Km uM raw 95.1 +/- 11.9, kcat/Km M^-1*s^-1 raw 2.5 x 10^4; normalized paper kcat 2.4 s^-1, Km 0.0951 mM, same-row kcat/Km 25.236593059936908 s^-1*mM^-1; export values kcat 2.4 s^-1, Km 0.0951 mM, kcat/Km 25.236593059936908 s^-1*mM^-1

normalized_value_unit_evidence: kcat normalized_value=2.4 normalized_unit=s^-1; Km normalized_value=0.0951 normalized_unit=mM from 95.1 +/- 11.9 uM / 1000; kcat/Km normalized_value=25.236593059936908 normalized_unit=s^-1*mM^-1 from same-row kcat and Km; printed efficiency=2.5 x 10^4 M^-1*s^-1

raw_value_unit_evidence: kcat raw_value=2.4 raw_unit=s^-1; Km raw_value=95.1 +/- 11.9 raw_unit=uM; kcat/Km raw_value=2.5 x 10^4 raw_unit=M^-1*s^-1

row_label: A. tumefaciens PruA / H2NPt


organism: Homo sapiens

assay_conditions_summary: pH 7.5; cofactor FAD


assay_conditions_summary: pH 7.5; 298.15 K; buffer phosphate; cofactor FAD

column_label: Table 1; row FAD; column Km (uM) raw value 125.3+/-31.4; column kcat (s^-1) raw value 0.088; there is no kcat/Km column in Table 1.

normalized_value_unit_evidence: kcat 0.088 s^-1 = 0.088 s^-1. Km 125.3 uM = 0.1253 mM (125.3 / 1000 = 0.1253). kcat/Km is not printed by the paper, so paper kcat_over_km = paper_kcat / paper_km = 0.088 / 0.1253 = 0.7023144453312051 s^-1 mM^-1. The catalog's 2.4 is 0.3 / 0.125, built on the catalog kcat 0.3 that the paper contradicts.

raw_value_unit_evidence: Table 1 (Kinetic constants for FR at pH 7.5), row FAD: Km = 125.3+/-31.4 uM; Vmax = 0.24+/-0.06 umol min^-1 mg^-1; kcat = 0.088 s^-1. The table prints no kcat/Km column.

row_label: FAD

table_label: Table 1: Kinetic constants for FR at pH 7.5


substrate_name: NADPH

assay_conditions_summary: pH 5; 298.15 K; cofactor NADPH

substrate: NADPH


assay_conditions_summary: pH 7.5; 298.15 K; cofactor NADPH


substrate_name: oxidized riboflavin

column_label: Table 1; row Riboflavin; column Km (uM) raw value 53.2+/-12.6; column kcat (s^-1) raw value 0.107; there is no kcat/Km column in Table 1.

normalized_value_unit_evidence: kcat 0.107 s^-1 = 0.107 s^-1. Km 53.2 uM = 0.0532 mM (53.2 / 1000 = 0.0532). kcat/Km is not printed by the paper, so paper kcat_over_km = paper_kcat / paper_km = 0.107 / 0.0532 = 2.011278195488722 s^-1 mM^-1. The catalog's 5.660377358490566 is 0.3 / 0.053, built on the catalog kcat 0.3 that the paper contradicts.

raw_value_unit_evidence: Table 1 (Kinetic constants for FR at pH 7.5), row Riboflavin: Km = 53.2+/-12.6 uM; Vmax = 0.29+/-0.07 umol min^-1 mg^-1; kcat = 0.107 s^-1. The table prints no kcat/Km column.

row_label: Riboflavin

substrate: oxidized riboflavin


assay_conditions_summary: pH 5; 298.15 K; cofactor FAD


normalized_value_unit_evidence: kcat 0.107 s^-1 = 0.107 s^-1. Km 53.2 uM = 0.0532 mM (53.2 / 1000 = 0.0532). kcat/Km is not printed by the paper, so paper kcat_over_km = paper_kcat / paper_km = 0.107 / 0.0532 = 2.011278195488722 s^-1 mM^-1. The catalog value 2.018867924528302 is 0.107 / 0.053, i.e. the ratio recomputed from the rounded Km rather than the paper's 0.0532 mM.


substrate_name: NADP

assay_conditions_summary: cofactor NADPH

proof_lines:
NADPH
NAD(P)H
flavin reductase

substrate: NADP


assay_conditions_summary: cofactor NADH

proof_lines:
NAD(P)H
FMN
pyridine nucleotide


substrate_name: biliverdin IX-beta

proof_lines:
biliverdin-IXbeta reductase
FMN
ferric reductase

substrate: biliverdin IX-beta


column_label: Abstract reports the enzyme catalyses reduction of FMN, FAD and riboflavin, with K(m) values of 52 microM, 125 microM and 53 microM respectively.

raw_value_unit_evidence: Km raw_value=53 microM raw_unit=microM

row_label: Human biliverdin-IXbeta reductase flavin reductase reaction / riboflavin

table_label: PubMed XML abstract PMID 10620517


organism: Bacillus subtilis

assay_conditions_summary: pH 7.5; 310.15 K; cofactor NADPH


proof_lines: biopterin NADPH


substrate_name: folate

substrate: folate


substrate_name: quinonoid dihydrobiopterin

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

proof_lines:
dihydropteridine reductase
quinonoid dihydropteridine
L-threo-neopterin

substrate: quinonoid dihydrobiopterin


proof_lines:
dihydropteridine reductase
tetrahydropteridine
phenylalanine hydroxylase


substrate_name: L-threo-neopterin

proof_lines:
dihydropteridine reductase
L-threo-neopterin
quinonoid dihydropteridine

substrate: L-threo-neopterin


enzyme_display_name: NADH:FAD oxidoreductase

substrate_name: FMN (with fixed NADH concentration at 300 uM)

enzyme: NADH:FAD oxidoreductase


organism: Trypanosoma brucei

assay_conditions_summary: pH 7.4; 298.15 K; cofactor NADPH

column_label: PMC HTML Table 1 and Europe PMC PDF text lines 241-256 report TbPTR1 H2B columns Kappm, kcat, and kcat/Kappm.

normalized_value_unit_evidence: kcat=0.05 s^-1; Km-like Kappm=0.0294 uM=0.0000294 mM; kcat_over_km table value=1700 s^-1 mM^-1, consistent with expected 1666.6666666666667 from rounded Km/kcat cells.

row_label: TbPTR1 / H2B


organism: Leishmania tarentolae

substrate_name: biopterin

substrate: biopterin


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature Y175F to reconstruct the assayed variant sequence.


organism: Escherichia coli (strain K12)


substrate_name: nitrofurazone

substrate: nitrofurazone


substrate_name: Tetrahydrobiopterin

substrate: Tetrahydrobiopterin


substrate_name: NADH (with 0.5 mM of FAD at 22 degrees Celsius and at pH 7.5)


substrate_name: FAD (with 0.3 mM of NADH at 22 degrees Celsius and at pH 7.5)


proof_lines:
cofactor NADH
cofactor NADPH


enzyme_display_name: 4-hydroxyphenylacetate 3-monooxygenase, reductase component

organism: Thermus thermophilus (strain ATCC 27634 / DSM 579 / HB8)

substrate_name: FAD (at 30 degrees Celsius and pH 7.0)

assay_conditions_summary: cofactor FAD

substrate: FAD (at 30 degrees Celsius and pH 7.0)


substrate_name: FAD

assay_conditions_summary: pH 7; cofactor FAD

enzyme: 4-hydroxyphenylacetate 3-monooxygenase, reductase component


assay_conditions_summary: pH 7.5; 298.15 K; buffer Primary measurement evidence: evidence location type: prose; cofactor FAD


substrate_name: FMN

assay_conditions_summary: pH 7.5; 298.15 K; cofactor FMN


substrate_name: dihydrobiopterin

substrate: dihydrobiopterin


normalized_value_unit_evidence: paper kcat=0.57 s^-1; paper Km-like Kappm=0.00332 mM; paper kcat_over_km=170 s^-1 mM^-1; expected tuple has Km=0.003 mM and kcat_over_km=190 s^-1 mM^-1.


substrate_name: S-nitroso-CoA

assay_conditions_summary: cofactor CoA

substrate: S-nitroso-CoA


organism: Trypanosoma brucei brucei


substrate_name: quinonoid 7,8-dihydropterin

proof_lines:
Leishmania major
QDPR
quinonoid pterins

substrate: quinonoid 7,8-dihydropterin


substrate_name: NADPH

sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature D37I with inferred residue-numbering offset +1 to reconstruct the assayed variant sequence.

assay_conditions_summary: pH 6.8; 295.15 K; buffer pH 6; cofactor NADPH

normalized_value_unit_evidence: kcat 26 s^-1; Km/K_NADPH 0.44 mM; kcat/Km 60 s^-1 mM^-1.

raw_value_unit_evidence: Table I row NADPH D-37-I: kcat 26 s^-1; kcat/K_NADPH 0.06 uM^-1 s^-1; K_NADPH 440 uM.

row_label: NADPH / D-37-I

table_label: Table I. Comparative kinetic properties of the mutant D-37-I and wild-type rat DHPR


substrate_name: tetrahydrobiopterin

proof_lines:
cofactor NADPH
cofactor NADH

substrate: tetrahydrobiopterin


substrate_name: quinoid dihydrobiopterin

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

substrate: quinoid dihydrobiopterin


substrate_name: quinonoid dihydrobiopterin


substrate_name: NADH

substrate: NADH


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


proof_lines: Y146F/K150E mutant enzyme, approx. value


substrate: NADPH


organism: Escherichia coli W

assay_conditions_summary: pH 7.8; 295.15 K; cofactor FMN

proof_lines: pH 7.8, 22°C, the value is obtained with NADH as the electron donor

substrate: FMN


substrate_name: riboflavin

assay_conditions_summary: pH 7.8; 295.15 K; cofactor NADH

substrate: riboflavin


assay_conditions_summary: pH 7.8; 295.15 K; cofactor FAD

substrate: FAD


proof_lines: pH 7.8, 22°C, the value is obtained with FMN as electron acceptor


organism: Archaeoglobus fulgidus

substrate_name: reduced coenzyme F420

substrate: reduced coenzyme F420


substrate_name: NADP+

substrate: NADP+


enzyme_display_name: F420-dependent NADP reductase

organism: Archaeoglobus fulgidus (strain ATCC 49558 / DSM 4304 / JCM 9628 / NBRC 100126 / VC-16)

substrate_name: NADP(+) (at pH 8.0 and 65 degrees Celsius)

enzyme: F420-dependent NADP reductase


organism: Methanothermobacter thermautotrophicus (strain ATCC 29096 / DSM 1053 / JCM 10044 / NBRC 100330 / Delta H)

substrate_name: NADP(+)

substrate: NADP(+)


enzyme_display_name: glutamate dehydrogenase (NADP+)

organism: Escherichia coli

assay_conditions_summary: pH 8; 298.15 K; cofactor NADPH

proof_lines: wild-type EcGDH, Vmax: 503 micromol/min/mg, pH 8.0, 25°C

enzyme: glutamate dehydrogenase (NADP+)


proof_lines: wild-type EcGDH, Vmax: 464 micromol/min/mg, pH 8.0, 25°C


proof_lines: wild-type EcGDH, Vmax: 298 micromol/min/mg, pH 8.0, 25°C


enzyme_display_name: dihydrofolate reductase

proof_lines: pH 7, 20°C, dimeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli prepared by introducing residues located at the Thermotoga maritima dihydrofolate reductase dimer interface

proof_lines: pH 7, 20°C, monomeric variant (Xet-3) of dihydrofolate reductase from Escherichia coli prepared by introducing residues located at the Thermotoga maritima dihydrofolate reductase dimer interface

enzyme: dihydrofolate reductase


assay_conditions_summary: pH 7; 293.15 K; cofactor NADPH


proof_lines:
pH 7, 20°C, wild-type enzyme
wild-type enzyme from plasmid


organism: Enterococcus faecium

substrate_name: folate

substrate: folate


enzyme_display_name: FMN reductase (NADH)

organism: Aminobacter aminovorans

substrate_name: Nitrilotriacetic acid; O2; H+; NADH

enzyme: FMN reductase (NADH)

substrate: Nitrilotriacetic acid; O2; H+; NADH


organism: Nocardioides sp. PD653

assay_conditions_summary: pH 7.5; 293.15 K; cofactor FMN


assay_conditions_summary: pH 7.5; 293.15 K; cofactor NADH


organism: Thermobifida fusca


assay_conditions_summary: pH 6; 298.15 K; cofactor NADPH

proof_lines: above, pH 6.0, 25°C, recombinant mutant R51V/R55V


proof_lines: above, pH 6.0, 25°C, recombinant mutant T28A/R51V/R55V


proof_lines: above, pH 6.0, 25°C, recombinant mutant S50Q


proof_lines: above, pH 6.0, 25°C, recombinant mutant R51E/R55S


substrate_name: oxidized coenzyme F420

substrate: oxidized coenzyme F420


assay_conditions_summary: pH 6; 298.15 K; cofactor NADP+

proof_lines: above, pH 6.0, 25°C, recombinant mutant R51V


proof_lines: above, pH 6.0, 25°C, recombinant mutant R51E/R55A


proof_lines: above, pH 6.0, 25°C, recombinant mutant R51A


proof_lines: above, pH 6.0, 25°C, recombinant mutant S50E/R55V


proof_lines: pH 6.0, 25°C, recombinant wild-type enzyme


proof_lines: above, pH 6.0, 25°C, recombinant mutant S50E/R55A


proof_lines: pH 6.0, 25°C, recombinant mutant T28A/R55A


proof_lines: above, pH 6.0, 25°C, recombinant mutant S50E


proof_lines: above, pH 6.0, 25°C, recombinant mutant R51E/R55N


proof_lines: above, pH 6.0, 25°C, recombinant mutant T28A/R51V


proof_lines: above, pH 6.0, 25°C, recombinant mutant R55N


organism: Streptomyces griseus


substrate_name: coenzyme F420

substrate: coenzyme F420


assay_conditions_summary: pH 6; 295.15 K; cofactor NADP+


assay_conditions_summary: pH 6; 295.15 K; cofactor NADPH


enzyme_display_name: FMN reductase [NAD(P)H]

organism: Lacticaseibacillus rhamnosus

assay_conditions_summary: pH 5.5; 308.15 K; cofactor NADH

proof_lines: recombinant wild-type enzyme, pH 5.5, 35°C

enzyme: FMN reductase [NAD(P)H]


organism: Methanococcus vannielii


assay_conditions_summary: pH 7; 293.15 K; cofactor NADP+


organism: Methanosphaera stadtmanae

proof_lines: pH and temperature not specified in the publication


assay_conditions_summary: cofactor NADPH


assay_conditions_summary: cofactor NADP+


organism: Vibrio harveyi

assay_conditions_summary: pH 7; 296.15 K; cofactor NADPH


substrate_name: FMN

assay_conditions_summary: pH 8.5; 296.15 K; cofactor FMN

proof_lines:
pH 8.5, 23°C, spectrometric assay
pH 5.6, 23°C, spectrometric assay
pH 6.8, 23°C


assay_conditions_summary: pH 8.5; 296.15 K; cofactor NADPH

proof_lines:
pH 8.5, 23°C, spectrometric assay
pH 5.6, 23°C, spectrometric assay
pH 7.0, 23°C


assay_conditions_summary: pH 7.5; 298.15 K; cofactor NADH

proof_lines: pH 7.5, 25°C, recombinant wild-type enzyme


assay_conditions_summary: pH 7.5; 298.15 K; cofactor NADPH


organism: Paracoccus denitrificans

assay_conditions_summary: pH 7; 303.15 K; cofactor NADPH


assay_conditions_summary: pH 7; 303.15 K; cofactor NADH


substrate_name: Unknown

assay_conditions_summary: pH 7; 303.15 K; cofactor FMN

proof_lines: versus NADH, pH 7.0, 30°C, recombinant wild-type, competitive

proof_lines: versus FMN, pH 7.0, 30°C, recombinant wild-type, uncompetitive

proof_lines: versus NADH, pH 7.0, 30°C, recombinant wild-type, mixed noncompetitive

substrate: Unknown


assay_conditions_summary: pH 7.5; 293.15 K; cofactor NADH


assay_conditions_summary: pH 7.5; 293.15 K; cofactor FMN


enzyme_display_name: riboflavin reductase [NAD(P)H]

organism: Corynebacterium glutamicum

assay_conditions_summary: pH 7.5; 303.15 K; cofactor NADPH

proof_lines: with FAD, pH 7.5, 30°C, recombinant enzyme

proof_lines: with FMN, pH 7.5, 30°C, recombinant enzyme

enzyme: riboflavin reductase [NAD(P)H]


proof_lines: with riboflavin, pH 7.5, 30°C, recombinant enzyme


organism: Methanobrevibacter smithii


enzyme_display_name: FMN reductase (NADPH)

sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature N134A with inferred residue-numbering offset +14 to reconstruct the assayed variant sequence.

assay_conditions_summary: pH 7; buffer 50 mM phosphate; cofactor NADPH

proof_lines: mutant enzyme N134A, in 50 mM phosphate buffer, pH 7.0, at 22°C

enzyme: FMN reductase (NADPH)


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature R133A to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme R133A, in 50 mM phosphate buffer, pH 7.0, at 22°C


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature R225A to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme R225A, in 50 mM phosphate buffer, pH 7.0, at 22°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 50 mM phosphate buffer, pH 7.0, at 22°C


assay_conditions_summary: pH 7.5; cofactor NADPH

proof_lines:
second substrate: NADH, pH 7.5, 23°C
second substrate: NADPH, pH 7.5, 23°C


substrate_name: FAD


assay_conditions_summary: pH 7.5; cofactor NADH

proof_lines: second substrate riboflavin, pH 7.5, 23°C


substrate_name: NADPH


enzyme_display_name: NADPH-flavin oxidoreductase

organism: Streptomyces viridifaciens

assay_conditions_summary: cofactor FAD

column_label: Table III columns Substrate, Second substrate, kcat, Km, kcat/Km; row FAD

raw_value_unit_evidence: FAD row: second substrate 100 mM NADPH; kcat 51 s^-1; Km 3.6 +/- 0.4 uM; kcat/Km 14.2 s^-1 uM^-1

row_label: FAD row

table_label: JBC Table III - Apparent kinetic data for substrates

enzyme: NADPH-flavin oxidoreductase


assay_conditions_summary: cofactor FMN

column_label: Table III columns Substrate, Second substrate, kcat, Km, kcat/Km; row FMN

row_label: FMN row


assay_conditions_summary: cofactor NADPH

column_label: Table III columns Substrate, Second substrate, kcat, Km, kcat/Km; row Riboflavin

raw_value_unit_evidence: Riboflavin row: kcat 6.7 s^-1; Km 11.1 +/- 2 uM; kcat/Km 0.6 s^-1 uM^-1

row_label: Riboflavin row


substrate_name: NADPH (with 40 mM FAD)

column_label: Table III columns Substrate, Second substrate, kcat, Km, kcat/Km; row NADPH

raw_value_unit_evidence: NADPH row: second substrate 40 mM FAD; kcat 62 s^-1; Km 32 +/- 2 uM; kcat/Km 1.9 s^-1 uM^-1

row_label: NADPH row

substrate: NADPH (with 40 mM FAD)


organism: Methanothermobacter thermautotrophicus

assay_conditions_summary: pH 6; cofactor NADPH

proof_lines: pH 6.0, temperature not specified in the publication


assay_conditions_summary: pH 8; cofactor NADP+

proof_lines: pH 8.0, temperature not specified in the publication


assay_conditions_summary: pH 8


organism: Archaeoglobus fulgidus

sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature I135G to reconstruct the assayed variant sequence.

assay_conditions_summary: pH 6.5; cofactor NADPH

proof_lines: phase I, pH 6.5, 22°C, recombinant mutant I135G

proof_lines: phase II, pH 6.5, 22°C, recombinant mutant I135G


sequence_variant_note: Mutation signature I135A could not be applied safely because the resolved sequence does not match the expected reference residue(s).

proof_lines: phase I, pH 6.5, 22°C, recombinant mutant I135A

proof_lines: phase II, pH 6.5, 22°C, recombinant mutant I135A


sequence_variant_note: Mutation signature I135V could not be applied safely because the resolved sequence does not match the expected reference residue(s).

proof_lines: phase I, pH 6.5, 22°C, recombinant mutant I135V

proof_lines: phase II, pH 6.5, 22°C, recombinant mutant I135V


proof_lines: phase I, pH 6.5, 22°C, recombinant wild-type enzyme

proof_lines: phase II, pH 6.5, 22°C, recombinant wild-type enzyme


enzyme_display_name: Flavin reductase

organism: Pseudomonas putida

proof_lines: kcat is 283 sec(-1) for the reduction of FMN. kcat is 128 sec(-1) for the reduction of FAD.

enzyme: Flavin reductase


assay_conditions_summary: cofactor NADH


assay_conditions_summary: pH 7; cofactor NADPH


enzyme_display_name: Chorismate synthase aro-2

organism: Neurospora crassa (strain ATCC 24698 / 74-OR23-1A / CBS 708.71 / DSM 1257 / FGSC 987)

proof_lines: kcat is 0.87 sec(-1) for chorismate formation.

enzyme: Chorismate synthase aro-2


enzyme_display_name: NADH-dependent FMN reductase SfnF

organism: Pseudomonas fluorescens (strain Pf0-1)

proof_lines: kcat is 14 sec(-1) for FMNH(2) as substrate. kcat is 10 sec(-1) for NADH as substrate.

enzyme: NADH-dependent FMN reductase SfnF


enzyme_display_name: NADH-dependent FMN reductase AsuE2

organism: Streptomyces nodosus subsp. asukaensis

proof_lines: kcat is 2.34 sec(-1) with NADH as substrate. kcat is 5.31 sec(-1) with NADPH as substrate.

enzyme: NADH-dependent FMN reductase AsuE2


assay_conditions_summary: pH 6.8; cofactor NADPH

proof_lines: pH 7.0, 23°C, single-enzyme spectrophotometric assay monitoring the NADPH oxidation


proof_lines:
pH 6.5, 22°C, recombinant enzyme
pH 5.5, 65°C


proof_lines: pH 7.0, 23°C, enzyme derivative reconstituted from apoenzyme and 2-thioFMN


enzyme_display_name: FMN reductase (NADH) SmoA

organism: Agrobacterium fabrum (strain C58 / ATCC 33970)

proof_lines: kcat is 14.5 sec(-1) with NADH as substrate.

enzyme: FMN reductase (NADH) SmoA


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature R15A to reconstruct the assayed variant sequence.

assay_conditions_summary: pH 7; 295.15 K; buffer 50 mM phosphate; cofactor NADPH

proof_lines: mutant enzyme R15A, in 50 mM phosphate buffer, pH 7.0, at 22°C


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature K167A to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme K167A, in 50 mM phosphate buffer, pH 7.0, at 22°C


substrate_name: H+; Riboflavin-5-phosphate; NADPH

assay_conditions_summary: pH 7; 296.15 K; cofactor NADPH

substrate: H+; Riboflavin-5-phosphate; NADPH


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature I135V to reconstruct the assayed variant sequence.

proof_lines: with F420 precursor, FO, pH 6.5, 22°C, recombinant mutant I135V


assay_conditions_summary: pH 6.5; 295.15 K; cofactor NADPH

column_label: Table 2: FO steady-state kinetics parameters for wtFno and I135 variants (Biochemistry and Biophysics Reports 2017); the transcribed row is the body row whose 'Fno' cell reads wtFno; column 'kcat(s-1)' = 5.3; column 'KmFO(uM)' = 4.0; column 'kcat/Km(M-1s-1)' = 1.3 x 10^6.

normalized_value_unit_evidence: kcat: printed cell 5.3 s^-1; already in s^-1, no conversion; the exported value is 5.3 s^(-1). Km: printed cell 4.0 uM; 1 uM is 0.001 mM, so 4.0 / 1000 = 0.004 mM; the exported value is 0.004 mM. kcat/Km: printed cell 1.3 x 10^6 M^-1 s^-1; 1 M^-1 s^-1 is 0.001 s^-1 mM^-1, so 1.3 x 10^6 / 1000 = 1300 s^-1 mM^-1; the export carries no kcat_over_km field for this row, so the printed cell is recorded as evidence only and no ratio is written.

raw_value_unit_evidence: Table 2, body row wtFno: kcat = 5.3 s^-1; KmFO = 4.0 uM; printed kcat/Km = 1.3 x 10^6 M^-1 s^-1.

row_label: Table 2 body row whose Fno cell reads wtFno, the FO steady-state row for that enzyme form


organism: Bacillus subtilis

substrate_name: H+; NADPH; Nitrofurazone

substrate: H+; NADPH; Nitrofurazone


unit: Mswedgeone


substrate_name: NADH; H+; Riboflavin-5-phosphate

sequence: SRLPPATRDKLADEITFFPA

wild_type_sequence: SRLPPATRDKLADEITFFPA

substrate: NADH; H+; Riboflavin-5-phosphate


organism: Methanogenium organophilum

sequence: MKVGIMGGTGNIGEGLARRIXIGGKYDVMIKGRDKA

wild_type_sequence: MKVGIMGGTGNIGEGLARRIXIGGKYDVMIKGRDKA

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


organism: Methanothermobacter marburgensis (strain ATCC BAA-927 / DSM 2133 / JCM 14651 / NBRC 100331 / OCM 82 / Marburg)


enzyme_display_name: carboxynorspermidine synthase

organism: Vibrio alginolyticus

substrate_name: 1,3-Diaminopropane; L-Aspartate 4-semialdehyde; H+; NADPH

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

enzyme: carboxynorspermidine synthase


organism: Thermobifida fusca

sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature P89Y to reconstruct the assayed variant sequence.

column_label: table label: Table 1 (Steady-state kinetic parameters for wild-type Tfu-FNO and selected Tfu-FNO G29X and P89X variants toward selected NCBs); row label: ProNA+ / P89Y; column label: kcat (min^-1); Km (mM); derived kcat/Km from normalized kcat and Km cells; printed kcat/Km column (s^-1 M^-1) kept as table context; raw value/unit and normalized value/unit preserved per metric; assay condition: 25 C spectrophotometric assay in 50 mM Tris-HCl, 100 mM NaCl, 3% (v/v) DMSO, pH 8.0, with 40 microM F420H2 and varied NCB concentration; rates followed at 400 nm.; captured paper segment: Table 1 (Steady-state kinetic parameters for wild-type Tfu-FNO and selected Tfu-FNO G29X and P89X variants toward selected NCBs); ProNA+ / P89Y; kcat 0.0098 s^-1, kcat_over_km 0.00028 s^-1 mM^-1, km 35 mM.

normalized_value_unit_evidence: kcat normalized value: 0.0098; normalized unit: s^-1; kcat_over_km normalized value: 0.00028; normalized unit: s^-1 mM^-1; km normalized value: 35; normalized unit: mM

raw_value_unit_evidence: kcat raw value: 0.59 ± 0.11; raw unit: min^-1; km raw value: 35 ± 14; raw unit: mM; kcat_over_km raw value: 0.0098 / 35; raw unit: s^-1 per mM; printed kcat/Km column: 0.28 ± 0.12 s^-1 M^-1

row_label: ProNA+ / P89Y


column_label: table label: Table 1 (Steady-state kinetic parameters for wild-type Tfu-FNO and selected Tfu-FNO G29X and P89X variants toward selected NCBs); row label: ProNA+ / wt; column label: kcat (min^-1); Km (mM); derived kcat/Km from normalized kcat and Km cells; printed kcat/Km column (s^-1 M^-1) kept as table context; raw value/unit and normalized value/unit preserved per metric; assay condition: 25 C spectrophotometric assay in 50 mM Tris-HCl, 100 mM NaCl, 3% (v/v) DMSO, pH 8.0, with 40 microM F420H2 and varied NCB concentration; rates followed at 400 nm.; captured paper segment: Table 1 (Steady-state kinetic parameters for wild-type Tfu-FNO and selected Tfu-FNO G29X and P89X variants toward selected NCBs); ProNA+ / wt; kcat 0.0015 s^-1, kcat_over_km 0.00015000000000000001 s^-1 mM^-1, km 10 mM.

normalized_value_unit_evidence: kcat normalized value: 0.0015; normalized unit: s^-1; kcat_over_km normalized value: 0.00015000000000000001; normalized unit: s^-1 mM^-1; km normalized value: 10; normalized unit: mM

raw_value_unit_evidence: kcat raw value: 0.09 ± 0.007; raw unit: min^-1; km raw value: 10 ± 2.8; raw unit: mM; kcat_over_km raw value: 0.0015 / 10; raw unit: s^-1 per mM; printed kcat/Km column: 0.15 ± 0.044 s^-1 M^-1

row_label: ProNA+ / wt


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature G29S to reconstruct the assayed variant sequence.

column_label: table label: Table 1 (Steady-state kinetic parameters for wild-type Tfu-FNO and selected Tfu-FNO G29X and P89X variants toward selected NCBs); row label: AmNA+ / G29S; column label: kcat (min^-1); Km (mM); derived kcat/Km from normalized kcat and Km cells; printed kcat/Km column (s^-1 M^-1) kept as table context; raw value/unit and normalized value/unit preserved per metric; assay condition: 25 C spectrophotometric assay in 50 mM Tris-HCl, 100 mM NaCl, 3% (v/v) DMSO, pH 8.0, with 40 microM F420H2 and varied NCB concentration; rates followed at 400 nm.; captured paper segment: Table 1 (Steady-state kinetic parameters for wild-type Tfu-FNO and selected Tfu-FNO G29X and P89X variants toward selected NCBs); AmNA+ / G29S; kcat 0.16 s^-1, kcat_over_km 0.008888888888888889 s^-1 mM^-1, km 18 mM.

normalized_value_unit_evidence: kcat normalized value: 0.16; normalized unit: s^-1; kcat_over_km normalized value: 0.008888888888888889; normalized unit: s^-1 mM^-1; km normalized value: 18; normalized unit: mM

raw_value_unit_evidence: kcat raw value: 9.6 ± 0.56; raw unit: min^-1; km raw value: 18 ± 2.5; raw unit: mM; kcat_over_km raw value: 0.16 / 18; raw unit: s^-1 per mM; printed kcat/Km column: 8.8 ± 1.3 s^-1 M^-1

row_label: AmNA+ / G29S


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature G29Y to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:Q47RA9 and applied mutation signature P89H to reconstruct the assayed variant sequence.

column_label: table label: Table 1 (Steady-state kinetic parameters for wild-type Tfu-FNO and selected Tfu-FNO G29X and P89X variants toward selected NCBs); row label: AmNA+ / P89H; column label: kcat (min^-1); Km (mM); derived kcat/Km from normalized kcat and Km cells; printed kcat/Km column (s^-1 M^-1) kept as table context; raw value/unit and normalized value/unit preserved per metric; assay condition: 25 C spectrophotometric assay in 50 mM Tris-HCl, 100 mM NaCl, 3% (v/v) DMSO, pH 8.0, with 40 microM F420H2 and varied NCB concentration; rates followed at 400 nm.; captured paper segment: Table 1 (Steady-state kinetic parameters for wild-type Tfu-FNO and selected Tfu-FNO G29X and P89X variants toward selected NCBs); AmNA+ / P89H; kcat 0.042 s^-1, kcat_over_km 0.003 s^-1 mM^-1, km 14 mM.

normalized_value_unit_evidence: kcat normalized value: 0.042; normalized unit: s^-1; kcat_over_km normalized value: 0.003; normalized unit: s^-1 mM^-1; km normalized value: 14; normalized unit: mM

raw_value_unit_evidence: kcat raw value: 2.5 ± 0.21; raw unit: min^-1; km raw value: 14 ± 3.0; raw unit: mM; kcat_over_km raw value: 0.042 / 14; raw unit: s^-1 per mM; printed kcat/Km column: 3.0 ± 0.69 s^-1 M^-1

row_label: AmNA+ / P89H


column_label: table label: Table 1 (Steady-state kinetic parameters for wild-type Tfu-FNO and selected Tfu-FNO G29X and P89X variants toward selected NCBs); row label: AmNA+ / wt; column label: kcat (min^-1); Km (mM); derived kcat/Km from normalized kcat and Km cells; printed kcat/Km column (s^-1 M^-1) kept as table context; raw value/unit and normalized value/unit preserved per metric; assay condition: 25 C spectrophotometric assay in 50 mM Tris-HCl, 100 mM NaCl, 3% (v/v) DMSO, pH 8.0, with 40 microM F420H2 and varied NCB concentration; rates followed at 400 nm.; captured paper segment: Table 1 (Steady-state kinetic parameters for wild-type Tfu-FNO and selected Tfu-FNO G29X and P89X variants toward selected NCBs); AmNA+ / wt; kcat 0.0233 s^-1, kcat_over_km 0.0019416666666666668 s^-1 mM^-1, km 12 mM.

normalized_value_unit_evidence: kcat normalized value: 0.0233; normalized unit: s^-1; kcat_over_km normalized value: 0.0019416666666666668; normalized unit: s^-1 mM^-1; km normalized value: 12; normalized unit: mM

raw_value_unit_evidence: kcat raw value: 1.4 ± 0.07; raw unit: min^-1; km raw value: 12 ± 1.7; raw unit: mM; kcat_over_km raw value: 0.0233 / 12; raw unit: s^-1 per mM; printed kcat/Km column: 1.9 ± 0.29 s^-1 M^-1

row_label: AmNA+ / wt


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature G29W to reconstruct the assayed variant sequence.


column_label: table label: Table 1 (Steady-state kinetic parameters for wild-type Tfu-FNO and selected Tfu-FNO G29X and P89X variants toward selected NCBs); row label: BNA+ / G29W; column label: kcat (min^-1); Km (mM); derived kcat/Km from normalized kcat and Km cells; printed kcat/Km column (s^-1 M^-1) kept as table context; raw value/unit and normalized value/unit preserved per metric; assay condition: 25 C spectrophotometric assay in 50 mM Tris-HCl, 100 mM NaCl, 3% (v/v) DMSO, pH 8.0, with 40 microM F420H2 and varied NCB concentration; rates followed at 400 nm.; captured paper segment: Table 1 (Steady-state kinetic parameters for wild-type Tfu-FNO and selected Tfu-FNO G29X and P89X variants toward selected NCBs); BNA+ / G29W; kcat 0.417 s^-1, kcat_over_km 0.09697674418604652 s^-1 mM^-1, km 4.3 mM.

normalized_value_unit_evidence: kcat normalized value: 0.417; normalized unit: s^-1; kcat_over_km normalized value: 0.09697674418604652; normalized unit: s^-1 mM^-1; km normalized value: 4.3; normalized unit: mM

raw_value_unit_evidence: kcat raw value: 25 ± 1.1; raw unit: min^-1; km raw value: 4.3 ± 0.68; raw unit: mM; kcat_over_km raw value: 0.417 / 4.3; raw unit: s^-1 per mM; printed kcat/Km column: 97 ± 15 s^-1 M^-1

row_label: BNA+ / G29W


column_label: table label: Table 1 (Steady-state kinetic parameters for wild-type Tfu-FNO and selected Tfu-FNO G29X and P89X variants toward selected NCBs); row label: BNA+ / G29Y; column label: kcat (min^-1); Km (mM); derived kcat/Km from normalized kcat and Km cells; printed kcat/Km column (s^-1 M^-1) kept as table context; raw value/unit and normalized value/unit preserved per metric; assay condition: 25 C spectrophotometric assay in 50 mM Tris-HCl, 100 mM NaCl, 3% (v/v) DMSO, pH 8.0, with 40 microM F420H2 and varied NCB concentration; rates followed at 400 nm.; captured paper segment: Table 1 (Steady-state kinetic parameters for wild-type Tfu-FNO and selected Tfu-FNO G29X and P89X variants toward selected NCBs); BNA+ / G29Y; kcat 0.145 s^-1, kcat_over_km 0.015263157894736841 s^-1 mM^-1, km 9.5 mM.

normalized_value_unit_evidence: kcat normalized value: 0.145; normalized unit: s^-1; kcat_over_km normalized value: 0.015263157894736841; normalized unit: s^-1 mM^-1; km normalized value: 9.5; normalized unit: mM

raw_value_unit_evidence: kcat raw value: 8.7 ± 0.64; raw unit: min^-1; km raw value: 9.5 ± 2.0; raw unit: mM; kcat_over_km raw value: 0.145 / 9.5; raw unit: s^-1 per mM; printed kcat/Km column: 15 ± 3.4 s^-1 M^-1

row_label: BNA+ / G29Y


column_label: table label: Table 1 (Steady-state kinetic parameters for wild-type Tfu-FNO and selected Tfu-FNO G29X and P89X variants toward selected NCBs); row label: BNA+ / wt; column label: kcat (min^-1); Km (mM); derived kcat/Km from normalized kcat and Km cells; printed kcat/Km column (s^-1 M^-1) kept as table context; raw value/unit and normalized value/unit preserved per metric; assay condition: 25 C spectrophotometric assay in 50 mM Tris-HCl, 100 mM NaCl, 3% (v/v) DMSO, pH 8.0, with 40 microM F420H2 and varied NCB concentration; rates followed at 400 nm.; captured paper segment: Table 1 (Steady-state kinetic parameters for wild-type Tfu-FNO and selected Tfu-FNO G29X and P89X variants toward selected NCBs); BNA+ / wt; kcat 0.0045 s^-1, kcat_over_km 0.0007031249999999999 s^-1 mM^-1, km 6.4 mM.

normalized_value_unit_evidence: kcat normalized value: 0.0045; normalized unit: s^-1; kcat_over_km normalized value: 0.0007031249999999999; normalized unit: s^-1 mM^-1; km normalized value: 6.4; normalized unit: mM

raw_value_unit_evidence: kcat raw value: 0.27 ± 0.01; raw unit: min^-1; km raw value: 6.4 ± 0.60; raw unit: mM; kcat_over_km raw value: 0.0045 / 6.4; raw unit: s^-1 per mM; printed kcat/Km column: 0.70 ± 0.071 s^-1 M^-1

row_label: BNA+ / wt


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature I135A to reconstruct the assayed variant sequence.

proof_lines: with F420 precursor, FO, pH 6.5, 22°C, recombinant mutant I135A


proof_lines: with F420 precursor, FO, pH 6.5, 22°C, recombinant mutant I135G


assay_conditions_summary: pH 6

proof_lines: pH 6.0, temperature not specified in the publication


enzyme_display_name: NAD(P)H-dependent FAD/FMN reductase GTNG_3158

organism: Geobacillus thermodenitrificans (strain NG80-2)

proof_lines: With FAD as substrate NADH can be used as an electron donor, with 95% activity relative to NADPH.


column_label: table label: Table 1 (Steady-state kinetic parameters for wild-type Tfu-FNO and selected Tfu-FNO G29X and P89X variants toward selected NCBs); row label: EtNA+ / wt; column label: kcat (min^-1); Km (mM); derived kcat/Km from normalized kcat and Km cells; printed kcat/Km column (s^-1 M^-1) kept as table context; raw value/unit and normalized value/unit preserved per metric; assay condition: 25 C spectrophotometric assay in 50 mM Tris-HCl, 100 mM NaCl, 3% (v/v) DMSO, pH 8.0, with 40 microM F420H2 and varied NCB concentration; rates followed at 400 nm.; captured paper segment: Table 1 (Steady-state kinetic parameters for wild-type Tfu-FNO and selected Tfu-FNO G29X and P89X variants toward selected NCBs); EtNA+ / wt; kcat 0.0073 s^-1, kcat_over_km 0.000292 s^-1 mM^-1, km 25 mM.

normalized_value_unit_evidence: kcat normalized value: 0.0073; normalized unit: s^-1; kcat_over_km normalized value: 0.000292; normalized unit: s^-1 mM^-1; km normalized value: 25; normalized unit: mM

raw_value_unit_evidence: kcat raw value: 0.44 ± 0.01; raw unit: min^-1; km raw value: 25 ± 1.4; raw unit: mM; kcat_over_km raw value: 0.0073 / 25; raw unit: s^-1 per mM; printed kcat/Km column: 0.29 ± 0.018 s^-1 M^-1

row_label: EtNA+ / wt


column_label: table label: Table 1 (Steady-state kinetic parameters for wild-type Tfu-FNO and selected Tfu-FNO G29X and P89X variants toward selected NCBs); row label: EtOHNA+ / P89H; column label: kcat (min^-1); Km (mM); derived kcat/Km from normalized kcat and Km cells; printed kcat/Km column (s^-1 M^-1) kept as table context; raw value/unit and normalized value/unit preserved per metric; assay condition: 25 C spectrophotometric assay in 50 mM Tris-HCl, 100 mM NaCl, 3% (v/v) DMSO, pH 8.0, with 40 microM F420H2 and varied NCB concentration; rates followed at 400 nm.; captured paper segment: Table 1 (Steady-state kinetic parameters for wild-type Tfu-FNO and selected Tfu-FNO G29X and P89X variants toward selected NCBs); EtOHNA+ / P89H; kcat 0.048 s^-1, kcat_over_km 0.0024000000000000002 s^-1 mM^-1, km 20 mM.

normalized_value_unit_evidence: kcat normalized value: 0.048; normalized unit: s^-1; kcat_over_km normalized value: 0.0024000000000000002; normalized unit: s^-1 mM^-1; km normalized value: 20; normalized unit: mM

raw_value_unit_evidence: kcat raw value: 2.9 ± 0.22; raw unit: min^-1; km raw value: 20 ± 3.3; raw unit: mM; kcat_over_km raw value: 0.048 / 20; raw unit: s^-1 per mM; printed kcat/Km column: 2.4 ± 0.44 s^-1 M^-1

row_label: EtOHNA+ / P89H


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:Q47RA9 and applied mutation signature P89Y to reconstruct the assayed variant sequence.

column_label: table label: Table 1 (Steady-state kinetic parameters for wild-type Tfu-FNO and selected Tfu-FNO G29X and P89X variants toward selected NCBs); row label: EtOHNA+ / P89Y; column label: kcat (min^-1); Km (mM); derived kcat/Km from normalized kcat and Km cells; printed kcat/Km column (s^-1 M^-1) kept as table context; raw value/unit and normalized value/unit preserved per metric; assay condition: 25 C spectrophotometric assay in 50 mM Tris-HCl, 100 mM NaCl, 3% (v/v) DMSO, pH 8.0, with 40 microM F420H2 and varied NCB concentration; rates followed at 400 nm.; captured paper segment: Table 1 (Steady-state kinetic parameters for wild-type Tfu-FNO and selected Tfu-FNO G29X and P89X variants toward selected NCBs); EtOHNA+ / P89Y; kcat 0.083 s^-1, kcat_over_km 0.0025151515151515154 s^-1 mM^-1, km 33 mM.

normalized_value_unit_evidence: kcat normalized value: 0.083; normalized unit: s^-1; kcat_over_km normalized value: 0.0025151515151515154; normalized unit: s^-1 mM^-1; km normalized value: 33; normalized unit: mM

raw_value_unit_evidence: kcat raw value: 5.0 ± 0.63; raw unit: min^-1; km raw value: 33 ± 9.2; raw unit: mM; kcat_over_km raw value: 0.083 / 33; raw unit: s^-1 per mM; printed kcat/Km column: 2.5 ± 0.77 s^-1 M^-1

row_label: EtOHNA+ / P89Y


column_label: table label: Table 1 (Steady-state kinetic parameters for wild-type Tfu-FNO and selected Tfu-FNO G29X and P89X variants toward selected NCBs); row label: EtOHNA+ / wt; column label: kcat (min^-1); Km (mM); derived kcat/Km from normalized kcat and Km cells; printed kcat/Km column (s^-1 M^-1) kept as table context; raw value/unit and normalized value/unit preserved per metric; assay condition: 25 C spectrophotometric assay in 50 mM Tris-HCl, 100 mM NaCl, 3% (v/v) DMSO, pH 8.0, with 40 microM F420H2 and varied NCB concentration; rates followed at 400 nm.; captured paper segment: Table 1 (Steady-state kinetic parameters for wild-type Tfu-FNO and selected Tfu-FNO G29X and P89X variants toward selected NCBs); EtOHNA+ / wt; kcat 0.0233 s^-1, kcat_over_km 0.001165 s^-1 mM^-1, km 20 mM.

normalized_value_unit_evidence: kcat normalized value: 0.0233; normalized unit: s^-1; kcat_over_km normalized value: 0.001165; normalized unit: s^-1 mM^-1; km normalized value: 20; normalized unit: mM

raw_value_unit_evidence: kcat raw value: 1.4 ± 0.06; raw unit: min^-1; km raw value: 20 ± 2.5; raw unit: mM; kcat_over_km raw value: 0.0233 / 20; raw unit: s^-1 per mM; printed kcat/Km column: 1.2 ± 0.15 s^-1 M^-1

row_label: EtOHNA+ / wt


column_label: table label: Table 1 (Steady-state kinetic parameters for wild-type Tfu-FNO and selected Tfu-FNO G29X and P89X variants toward selected NCBs); row label: ProOHNA+ / P89Y; column label: kcat (min^-1); Km (mM); derived kcat/Km from normalized kcat and Km cells; printed kcat/Km column (s^-1 M^-1) kept as table context; raw value/unit and normalized value/unit preserved per metric; assay condition: 25 C spectrophotometric assay in 50 mM Tris-HCl, 100 mM NaCl, 3% (v/v) DMSO, pH 8.0, with 40 microM F420H2 and varied NCB concentration; rates followed at 400 nm.; captured paper segment: Table 1 (Steady-state kinetic parameters for wild-type Tfu-FNO and selected Tfu-FNO G29X and P89X variants toward selected NCBs); ProOHNA+ / P89Y; kcat 0.0118 s^-1, kcat_over_km 0.00033714285714285714 s^-1 mM^-1, km 35 mM.

normalized_value_unit_evidence: kcat normalized value: 0.0118; normalized unit: s^-1; kcat_over_km normalized value: 0.00033714285714285714; normalized unit: s^-1 mM^-1; km normalized value: 35; normalized unit: mM

raw_value_unit_evidence: kcat raw value: 0.71 ± 0.096; raw unit: min^-1; km raw value: 35 ± 10; raw unit: mM; kcat_over_km raw value: 0.0118 / 35; raw unit: s^-1 per mM; printed kcat/Km column: 0.34 ± 0.11 s^-1 M^-1

row_label: ProOHNA+ / P89Y


column_label: table label: Table 1 (Steady-state kinetic parameters for wild-type Tfu-FNO and selected Tfu-FNO G29X and P89X variants toward selected NCBs); row label: ProOHNA+ / wt; column label: kcat (min^-1); Km (mM); derived kcat/Km from normalized kcat and Km cells; printed kcat/Km column (s^-1 M^-1) kept as table context; raw value/unit and normalized value/unit preserved per metric; assay condition: 25 C spectrophotometric assay in 50 mM Tris-HCl, 100 mM NaCl, 3% (v/v) DMSO, pH 8.0, with 40 microM F420H2 and varied NCB concentration; rates followed at 400 nm.; captured paper segment: Table 1 (Steady-state kinetic parameters for wild-type Tfu-FNO and selected Tfu-FNO G29X and P89X variants toward selected NCBs); ProOHNA+ / wt; kcat 0.0042 s^-1, kcat_over_km 0.000168 s^-1 mM^-1, km 25 mM.

normalized_value_unit_evidence: kcat normalized value: 0.0042; normalized unit: s^-1; kcat_over_km normalized value: 0.000168; normalized unit: s^-1 mM^-1; km normalized value: 25; normalized unit: mM

raw_value_unit_evidence: kcat raw value: 0.25 ± 0.013; raw unit: min^-1; km raw value: 25 ± 3.0; raw unit: mM; kcat_over_km raw value: 0.0042 / 25; raw unit: s^-1 per mM; printed kcat/Km column: 0.17 ± 0.022 s^-1 M^-1

row_label: ProOHNA+ / wt


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature G29L to reconstruct the assayed variant sequence.

column_label: table label: Table 1 (Steady-state kinetic parameters for wild-type Tfu-FNO and selected Tfu-FNO G29X and P89X variants toward selected NCBs); row label: ProNA+ / G29L; column label: kcat (min^-1); Km (mM); derived kcat/Km from normalized kcat and Km cells; printed kcat/Km column (s^-1 M^-1) kept as table context; raw value/unit and normalized value/unit preserved per metric; assay condition: 25 C spectrophotometric assay in 50 mM Tris-HCl, 100 mM NaCl, 3% (v/v) DMSO, pH 8.0, with 40 microM F420H2 and varied NCB concentration; rates followed at 400 nm.; captured paper segment: Table 1 (Steady-state kinetic parameters for wild-type Tfu-FNO and selected Tfu-FNO G29X and P89X variants toward selected NCBs); ProNA+ / G29L; kcat 0.006 s^-1, kcat_over_km 0.0003333333333333333 s^-1 mM^-1, km 18 mM.

normalized_value_unit_evidence: kcat normalized value: 0.006; normalized unit: s^-1; kcat_over_km normalized value: 0.0003333333333333333; normalized unit: s^-1 mM^-1; km normalized value: 18; normalized unit: mM

raw_value_unit_evidence: kcat raw value: 0.36 ± 0.057; raw unit: min^-1; km raw value: 18 ± 7.9; raw unit: mM; kcat_over_km raw value: 0.006 / 18; raw unit: s^-1 per mM; printed kcat/Km column: 0.33 ± 0.16 s^-1 M^-1

row_label: ProNA+ / G29L


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature P89H to reconstruct the assayed variant sequence.

column_label: table label: Table 1 (Steady-state kinetic parameters for wild-type Tfu-FNO and selected Tfu-FNO G29X and P89X variants toward selected NCBs); row label: ProNA+ / P89H; column label: kcat (min^-1); Km (mM); derived kcat/Km from normalized kcat and Km cells; printed kcat/Km column (s^-1 M^-1) kept as table context; raw value/unit and normalized value/unit preserved per metric; assay condition: 25 C spectrophotometric assay in 50 mM Tris-HCl, 100 mM NaCl, 3% (v/v) DMSO, pH 8.0, with 40 microM F420H2 and varied NCB concentration; rates followed at 400 nm.; captured paper segment: Table 1 (Steady-state kinetic parameters for wild-type Tfu-FNO and selected Tfu-FNO G29X and P89X variants toward selected NCBs); ProNA+ / P89H; kcat 0.0045 s^-1, kcat_over_km 0.0007894736842105262 s^-1 mM^-1, km 5.7 mM.

normalized_value_unit_evidence: kcat normalized value: 0.0045; normalized unit: s^-1; kcat_over_km normalized value: 0.0007894736842105262; normalized unit: s^-1 mM^-1; km normalized value: 5.7; normalized unit: mM

raw_value_unit_evidence: kcat raw value: 0.27 ± 0.026; raw unit: min^-1; km raw value: 5.7 ± 1.9; raw unit: mM; kcat_over_km raw value: 0.0045 / 5.7; raw unit: s^-1 per mM; printed kcat/Km column: 0.79 ± 0.27 s^-1 M^-1

row_label: ProNA+ / P89H


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature P89L to reconstruct the assayed variant sequence.

column_label: table label: Table 1 (Steady-state kinetic parameters for wild-type Tfu-FNO and selected Tfu-FNO G29X and P89X variants toward selected NCBs); row label: ProNA+ / P89L; column label: kcat (min^-1); Km (mM); derived kcat/Km from normalized kcat and Km cells; printed kcat/Km column (s^-1 M^-1) kept as table context; raw value/unit and normalized value/unit preserved per metric; assay condition: 25 C spectrophotometric assay in 50 mM Tris-HCl, 100 mM NaCl, 3% (v/v) DMSO, pH 8.0, with 40 microM F420H2 and varied NCB concentration; rates followed at 400 nm.; captured paper segment: Table 1 (Steady-state kinetic parameters for wild-type Tfu-FNO and selected Tfu-FNO G29X and P89X variants toward selected NCBs); ProNA+ / P89L; kcat 0.0073 s^-1, kcat_over_km 0.00027037037037037036 s^-1 mM^-1, km 27 mM.

normalized_value_unit_evidence: kcat normalized value: 0.0073; normalized unit: s^-1; kcat_over_km normalized value: 0.00027037037037037036; normalized unit: s^-1 mM^-1; km normalized value: 27; normalized unit: mM

raw_value_unit_evidence: kcat raw value: 0.44 ± 0.048; raw unit: min^-1; km raw value: 27 ± 6.9; raw unit: mM; kcat_over_km raw value: 0.0073 / 27; raw unit: s^-1 per mM; printed kcat/Km column: 0.27 ± 0.075 s^-1 M^-1

row_label: ProNA+ / P89L


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature S50E with inferred residue-numbering offset +2 to reconstruct the assayed variant sequence.

assay_conditions_summary: pH 6; 298.15 K; cofactor NADPH


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature S50E/R55A to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature S50E/R55V to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature S50Q with inferred residue-numbering offset +2 to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from accession_anchored:Q47RA9 and applied mutation signature T28A to reconstruct the assayed variant sequence.

column_label: PDF page 4, Table 1, T28A row under NADPH columns.

normalized_value_unit_evidence: Km 0.019 mM; kcat 14 s^-1; explicit kcat/Km 720 s^-1 mM^-1.

row_label: T28A; NADPH columns


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature T28A/R51V to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature T28A/R51V/R55V to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature T28A/R55A to reconstruct the assayed variant sequence.