substrate_name: pyruvate

substrate: pyruvate


enzyme_display_name: Tauropine dehydrogenase

organism: Mazzaella japonica

substrate_name: taurine

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.

enzyme: Tauropine dehydrogenase

substrate: taurine

status: unverified

source: brenda


substrate_name: tauropine

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

substrate: tauropine


enzyme_display_name: beta-alanopine dehydrogenase

organism: Cellana grata

substrate_name: NADH

assay_conditions_summary: pH 7.1; 303.15 K; cofactor NADH

enzyme: beta-alanopine dehydrogenase

substrate: NADH


substrate_name: NAD+

assay_conditions_summary: pH 9; 303.15 K; cofactor NAD+

substrate: NAD+


substrate_name: beta-Alanine

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

substrate: beta-Alanine


substrate_name: beta-alanopine

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

substrate: beta-alanopine


organism: Haliotis discus hannai

assay_conditions_summary: cofactor NADH


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


organism: Patiria pectinifera


assay_conditions_summary: cofactor NAD+


enzyme_display_name: Strombine dehydrogenase

organism: Scolelepis fuliginosa

enzyme: Strombine dehydrogenase


substrate_name: Gly

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

substrate: Gly


organism: Mytilus edulis

substrate_name: L-Ala

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

substrate: L-Ala


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

protein_accession_database: UniProt

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

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

data_origin: experimental

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

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

substrate: L-Ser


substrate_name: L-Met

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

substrate: L-Met


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

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

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

sequence_source: multiple

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)

source_db: uniprot

verification_status: verified

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

status: verified

source: uniprot


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

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

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

verification_status: corrected

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

status: corrected


enzyme_display_name: Bifunctional protein STORR

substrate_name: (S)-reticuline

enzyme: Bifunctional protein STORR

substrate: (S)-reticuline


substrate_name: pyruvate

source_db: oed

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

source: oed


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

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


enzyme_display_name: glycine oxidase

organism: Bacillus subtilis

substrate_name: D-Pro

enzyme: glycine oxidase

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_db: skid

source_license: CC BY-NC-ND 4.0

enzyme: UniProt O76290

substrate: dihydrobiopterin

source: skid


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

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

mutation_type: substitution

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


km_display: Not reported

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


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


assay_conditions_summary: cofactor NADPH


organism: Cereibacter sphaeroides


organism: Rhodococcus erythropolis

assay_conditions_summary: cofactor NADH


substrate_name: Unknown

ki_unit: mM

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.


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

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

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


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

kcat_over_km_origin: calculated

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


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; 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


assay_conditions_summary: cofactor NADPH

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


organism: Streptomyces globisporus

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

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)

assay_conditions_summary: cofactor NADH

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

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

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


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

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)

assay_conditions_summary: cofactor NADH


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


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

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

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.


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

column_label: Table 1 has separate NADH and NADPH column groups; the export tuple mixes NADH KM with NADPH kcat for the row labeled NADPH.

normalized_value_unit_evidence: For NADPH, KM 7.3 uM = 0.0073 mM; kcat = 3.3 s^-1; kcat/Km = 450 s^-1*mM^-1. The export row uses KM 14.0 mM from the NADH block and kcat 3.3 s^-1 from the NADPH block.

raw_value_unit_evidence: Table 1 wild type row: NADH KM 14 +/- 4.2 mM, NADH kcat 2.2 +/- 0.4 s^-1, NADH kcat/Km 160 M^-1 s^-1; NADPH KM 7.3 +/- 1.0 uM, NADPH kcat 3.3 +/- 0.1 s^-1, NADPH kcat/Km 450 mM^-1 s^-1.

row_label: Wild type; NADPH substrate block

table_label: Table 1, Steady-state kinetic parameters for wild-type and mutant Tfu-FNOs using NADH and NADPH as substrate


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature R51A with inferred residue-numbering offset -18 to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature R51E/R55A with inferred residue-numbering offset -18 to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature R51E/R55N with inferred residue-numbering offset -18 to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature R51E/R55S with inferred residue-numbering offset -18 to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature R51V with inferred residue-numbering offset -18 to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature R51V/R55V with inferred residue-numbering offset -18 to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from accession_anchored:Q47RA9 and applied mutation signature R55A to reconstruct the assayed variant sequence.

column_label: PDF page 4, Table 1, R55A row under NADPH columns.

normalized_value_unit_evidence: Km 0.029 mM; kcat 8.8 s^-1; explicit kcat/Km 300 s^-1 mM^-1.

row_label: R55A; NADPH columns


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature R55N with inferred residue-numbering offset -18 to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from accession_anchored:Q47RA9 and applied mutation signature R55S to reconstruct the assayed variant sequence.

column_label: PDF page 4, Table 1, R55S row under NADPH Km/kcat/kcat-over-Km columns.

normalized_value_unit_evidence: Km 0.170 mM; kcat 6.9 s^-1; explicit kcat/Km 41 s^-1 mM^-1.

row_label: R55S; NADPH columns


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature R55V with inferred residue-numbering offset -18 to reconstruct the assayed variant sequence.


organism: Bacillus subtilis (strain 168)

substrate_name: 4-nitrophenol (NP)(at pH 8 and 25 degrees Celsius)

column_label: Table 1; row NP; column Km; raw value 0.2 uM; table abbreviation NP is defined as 4-nitrophenol.

raw_value_unit_evidence: Table 1 NP row, Km column: 0.2 uM.

row_label: NP (4-nitrophenol)

table_label: Table 1: Kinetic Parameters for Nitro-organic Compounds and Chromate


substrate_name: 5-nitro-2-furaldehyde semicarbazone (NF)(at pH 8 and 25 degrees Celsius)

assay_conditions_summary: pH 8; 298.15 K; cofactor NADH

column_label: Table 1; row NF; column Km; raw value 0.7 uM. Figure 3A independently states Km = 0.7 uM.

raw_value_unit_evidence: Table 1 NF row, Km column: 0.7 uM.

row_label: NF (nitrofurazone; 5-nitro-2-furaldehyde semicarbazone)


substrate_name: FMN (at pH 8 and 25 degrees Celsius)

assay_conditions_summary: pH 8; 298.15 K; cofactor FMN

column_label: Figure 2A-associated prose; segment external FMN with NADPH; metric Km; raw value 4.2 uM.

raw_value_unit_evidence: Primary prose: reciprocal-FMN replot yields Km = 4.2 uM for FMN.

row_label: External FMN as electron-accepting substrate with NADPH

table_label: Figure 2A-associated primary prose

substrate: FMN (at pH 8 and 25 degrees Celsius)


substrate_name: NADPH (at pH 8 and 25 degrees Celsius)

assay_conditions_summary: pH 8; cofactor NADPH

column_label: The abstract reports Km values for NADPH and NADH of 4.4 microM and 6.4 microM, respectively.

raw_value_unit_evidence: Km raw_value=4.4 microM raw_unit=microM

row_label: Prose segment: NADPH Km

table_label: PubMed PMID 16229462 abstract

substrate: NADPH (at pH 8 and 25 degrees Celsius)


substrate_name: chromate (at pH 8 and 25 degrees Celsius)

column_label: Table 1; row chromate (CrVI); column Km; raw value 5 uM.

raw_value_unit_evidence: Table 1 chromate (CrVI) row, Km column: 5 uM.

row_label: chromate (CrVI)


substrate_name: NADH (at pH 8 and 25 degrees Celsius)

raw_value_unit_evidence: Km raw_value=6.4 microM raw_unit=microM

row_label: Prose segment: NADH Km

substrate: NADH (at pH 8 and 25 degrees Celsius)


substrate_name: methyl-4-nitrobenzenesulfonate (NBS)(at pH 8 and 25 degrees Celsius)

column_label: Table 1; row NBS; column Km; raw value 96 uM. Figure 3B independently states Km = 96 uM.

raw_value_unit_evidence: Table 1 NBS row, Km column: 96 uM.

row_label: NBS


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:O29370 and applied mutation signature I135A to reconstruct the assayed variant sequence.

column_label: Table 3: NADPH 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 I135A under the 'Phase 2' block; column 'kcat(s-1)' = 1.24; column 'KmNADPH(uM)' = 2.9; column 'kcat/Km(M-1s-1)' = 4.2 x 10^5.

normalized_value_unit_evidence: kcat: printed cell 1.24 s^-1; already in s^-1, no conversion; the exported value is 1.24 s^(-1). Km: printed cell 2.9 uM; 1 uM is 0.001 mM, so 2.9 / 1000 = 0.0029 mM; the exported value is 0.0029 mM. kcat/Km: printed cell 4.2 x 10^5 M^-1 s^-1; 1 M^-1 s^-1 is 0.001 s^-1 mM^-1, so 4.2 x 10^5 / 1000 = 420 s^-1 mM^-1, while the export stores 427.58620689655174, the full-precision quotient 1.24 / 0.0029 of the two printed component cells.

raw_value_unit_evidence: Table 3, Phase 2 block, body row I135A: kcat = 1.24 s^-1; KmNADPH = 2.9 uM; printed kcat/Km = 4.2 x 10^5 M^-1 s^-1.

row_label: Table 3 Phase 2 body row whose Fno cell reads I135A, the NADPH steady-state second-phase row for that enzyme form

substrate: NADPH


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:O29370 and applied mutation signature I135G to reconstruct the assayed variant sequence.

column_label: Table 3: NADPH 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 I135G under the 'Phase 2' block; column 'kcat(s-1)' = 0.33; column 'KmNADPH(uM)' = 654; column 'kcat/Km(M-1s-1)' = 5.0 x 10^2.

normalized_value_unit_evidence: kcat: printed cell 0.33 s^-1; already in s^-1, no conversion; the exported value is 0.33 s^(-1). Km: printed cell 654 uM; 1 uM is 0.001 mM, so 654 / 1000 = 0.654 mM; the exported value is 0.654 mM. kcat/Km: printed cell 5.0 x 10^2 M^-1 s^-1; 1 M^-1 s^-1 is 0.001 s^-1 mM^-1, so 5.0 x 10^2 / 1000 = 0.5 s^-1 mM^-1, while the export stores 0.5045871559633027, the full-precision quotient 0.33 / 0.654 of the two printed component cells.

raw_value_unit_evidence: Table 3, Phase 2 block, body row I135G: kcat = 0.33 s^-1; KmNADPH = 654 uM; printed kcat/Km = 5.0 x 10^2 M^-1 s^-1.

row_label: Table 3 Phase 2 body row whose Fno cell reads I135G, the NADPH steady-state second-phase row for that enzyme form


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:O29370 and applied mutation signature I135V to reconstruct the assayed variant sequence.

column_label: Table 3: NADPH 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 I135V under the 'Phase 2' block; column 'kcat(s-1)' = 2.16; column 'KmNADPH(uM)' = 51; column 'kcat/Km(M-1s-1)' = 4.2 x 10^4.

normalized_value_unit_evidence: kcat: printed cell 2.16 s^-1; already in s^-1, no conversion; the exported value is 2.16 s^(-1). Km: printed cell 51 uM; 1 uM is 0.001 mM, so 51 / 1000 = 0.051 mM; the exported value is 0.051 mM. kcat/Km: printed cell 4.2 x 10^4 M^-1 s^-1; 1 M^-1 s^-1 is 0.001 s^-1 mM^-1, so 4.2 x 10^4 / 1000 = 42 s^-1 mM^-1, while the export stores 42.352941176470594, the full-precision quotient 2.16 / 0.051 of the two printed component cells.

raw_value_unit_evidence: Table 3, Phase 2 block, body row I135V: kcat = 2.16 s^-1; KmNADPH = 51 uM; printed kcat/Km = 4.2 x 10^4 M^-1 s^-1.

row_label: Table 3 Phase 2 body row whose Fno cell reads I135V, the NADPH steady-state second-phase row for that enzyme form


substrate_name: oxidized coenzyme F420

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 I135A; column 'kcat(s-1)' = 1.6; column 'KmFO(uM)' = 3.6; column 'kcat/Km(M-1s-1)' = 4.5 x 10^5.

normalized_value_unit_evidence: kcat: printed cell 1.6 s^-1; already in s^-1, no conversion; the exported value is 1.6 s^(-1). Km: printed cell 3.6 uM; 1 uM is 0.001 mM, so 3.6 / 1000 = 0.0036 mM; the exported value is 0.0036 mM. kcat/Km: printed cell 4.5 x 10^5 M^-1 s^-1; 1 M^-1 s^-1 is 0.001 s^-1 mM^-1, so 4.5 x 10^5 / 1000 = 450 s^-1 mM^-1, while the export stores 444.44444444444446, the full-precision quotient 1.6 / 0.0036 of the two printed component cells.

raw_value_unit_evidence: Table 2, body row I135A: kcat = 1.6 s^-1; KmFO = 3.6 uM; printed kcat/Km = 4.5 x 10^5 M^-1 s^-1.

row_label: Table 2 body row whose Fno cell reads I135A, the FO steady-state row for that enzyme form


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 I135G; column 'kcat(s-1)' = 0.7; column 'KmFO(uM)' = 3.6; column 'kcat/Km(M-1s-1)' = 2.0 x 10^5.

normalized_value_unit_evidence: kcat: printed cell 0.7 s^-1; already in s^-1, no conversion; the exported value is 0.7 s^(-1). Km: printed cell 3.6 uM; 1 uM is 0.001 mM, so 3.6 / 1000 = 0.0036 mM; the exported value is 0.0036 mM. kcat/Km: printed cell 2.0 x 10^5 M^-1 s^-1; 1 M^-1 s^-1 is 0.001 s^-1 mM^-1, so 2.0 x 10^5 / 1000 = 200 s^-1 mM^-1, while the export stores 194.44444444444443, the full-precision quotient 0.7 / 0.0036 of the two printed component cells.

raw_value_unit_evidence: Table 2, body row I135G: kcat = 0.7 s^-1; KmFO = 3.6 uM; printed kcat/Km = 2.0 x 10^5 M^-1 s^-1.

row_label: Table 2 body row whose Fno cell reads I135G, the FO steady-state row for that enzyme form


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 I135V; column 'kcat(s-1)' = 1.8; column 'KmFO(uM)' = 3.7; column 'kcat/Km(M-1s-1)' = 4.9 x 10^5.

normalized_value_unit_evidence: kcat: printed cell 1.8 s^-1; already in s^-1, no conversion; the exported value is 1.8 s^(-1). Km: printed cell 3.7 uM; 1 uM is 0.001 mM, so 3.7 / 1000 = 0.0037 mM; the exported value is 0.0037 mM. kcat/Km: printed cell 4.9 x 10^5 M^-1 s^-1; 1 M^-1 s^-1 is 0.001 s^-1 mM^-1, so 4.9 x 10^5 / 1000 = 490 s^-1 mM^-1, while the export stores 486.48648648648646, the full-precision quotient 1.8 / 0.0037 of the two printed component cells.

raw_value_unit_evidence: Table 2, body row I135V: kcat = 1.8 s^-1; KmFO = 3.7 uM; printed kcat/Km = 4.9 x 10^5 M^-1 s^-1.

row_label: Table 2 body row whose Fno cell reads I135V, the FO steady-state row for that enzyme form


substrate_name: NADPH (with nitrofurazone at pH 7 and at 23 degrees Celsius)

proof_lines: NfrA1 FMN nitrofurazone


substrate_name: FMN (with NADPH at pH 7 and at 23 degrees Celsius)

assay_conditions_summary: pH 7; cofactor FMN


substrate_name: nitrofurazone (with NADPH at pH 7 and at 23 degrees Celsius)


substrate_name: reduced coenzyme F420

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


substrate_name: NADPH


substrate_name: NADP+


substrate_name: NADP(+) (at pH 8.0)

substrate: NADP(+) (at pH 8.0)


organism: Escherichia coli

substrate_name: FMN

assay_conditions_summary: pH 7.5; 298.15 K; cofactor FMN

column_label: Table 1 row wild-type SsuE; columns kcat (min^-1), Km (uM), and kcat/Km (uM^-1 min^-1).

normalized_value_unit_evidence: kcat normalized_value=4.25 normalized_unit=s^-1 | km normalized_value=0.00011 normalized_unit=mM | kcat_over_km normalized_value=38333.333333333336 normalized_unit=s^-1 mM^-1

row_label: wild-type SsuE

table_label: Table 1. Steady-State Kinetic Parameters for the Y118 SsuE Variants Measuring NADPH-Dependent FMN Reductase Activity

substrate: FMN


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature Y118F with inferred residue-numbering offset +1 to reconstruct the assayed variant sequence.


assay_conditions_summary: pH 7.5; cofactor NADP+

column_label: Primary FEBS article, PDF page 2 / journal page 125, Results and discussion paragraph beginning with the purified-enzyme properties; the Km sentence explicitly pairs F420, 0.3 mM, and pH 6.0.

raw_value_unit_evidence: The Km values were determined to be 0.3 mM for F420 (pH 6.0).

table_label: Results and discussion kinetic prose, PDF page 2 / journal page 125


substrate_name: NADPH (at pH 6.0)

column_label: Primary FEBS article, PDF page 2 / journal page 125, Results and discussion purified-enzyme paragraph; the Km sentence explicitly pairs NADPH, 50 uM, and pH 6.0.

normalized_value_unit_evidence: Km=0.05 mM after exact conversion of 50 uM.

raw_value_unit_evidence: The Km values were determined to be 50 uM for NADPH (pH 6.0).

row_label: NADPH at pH 6.0

substrate: NADPH (at pH 6.0)


column_label: Primary FEBS article, PDF page 2 / journal page 125, Results and discussion purified-enzyme paragraph; the Km sentence explicitly pairs NADP+, 70 uM, and pH 8.0.

normalized_value_unit_evidence: Km=0.07 mM after exact conversion of 70 uM.

raw_value_unit_evidence: The Km values were determined to be 70 uM for NADP+ (pH 8.0).

row_label: NADP+ at pH 8.0


substrate_name: NADH

assay_conditions_summary: pH 7.5; 298.15 K; cofactor NADPH

substrate: NADH


substrate_name: FAD

assay_conditions_summary: pH 7.5; 298.15 K; cofactor FAD

substrate: FAD


assay_conditions_summary: pH 7.5; cofactor NADPH

column_label: Abstract reports the Km value for NADPH was 1.51 mM.

raw_value_unit_evidence: Km raw_value=1.51 mM raw_unit=mM

row_label: Vibrio alginolyticus carboxynorspermidine synthase / NADPH

table_label: PubMed XML abstract PMID 1955861


organism: Sulfitobacter sp.

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


organism: Bacillus cereus

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

assay_conditions_summary: pH 7.2; 310.15 K; cofactor NADPH


organism: Vibrio harveyi


enzyme_display_name: 2-methyl-1-pyrroline reductase

organism: Streptomyces sp.

assay_conditions_summary: pH 7; 303.15 K; cofactor NADPH

enzyme: 2-methyl-1-pyrroline reductase


organism: Hyphomicrobium nitrativorans

substrate_name: dihydrosarcinapterin

substrate: dihydrosarcinapterin


organism: Paracoccus denitrificans

assay_conditions_summary: pH 7.4; 303.15 K; buffer 25 mM Tris/HCl; cofactor NADH

column_label: Official Wiley PDF text extract, printed page 560, Table 3: columns Substrate (variable concentration), Second substrate (fixed concentrations), Third substrate, and Km (uM); accompanying caption gives kcat values for FerA Fe(III) reductase and FMN reductase activities.

normalized_value_unit_evidence: For the queued NADH substrate, the printed NADH Km normalizes to 0.011 mM, not 0.02 mM. The exported 0.02 mM Km corresponds to the FMN row, and 28 / 0.020 = 1400 s^-1 mM^-1 is a derived FMN ratio rather than a primary NADH kcat/Km cell.

raw_value_unit_evidence: Table 3 reports FerA FMN reductase kinetic parameters: variable NADH with FMN as second substrate has Km 11 +/- 5 uM; variable FMN with NADH as second substrate has Km 20 +/- 8 uM. The text states kcat is 28 +/- 6 s^-1 for FerA FMN reductase activity. No kcat/Km column is printed.

row_label: FerA, FMN reductase activity

table_label: Table 3


organism: Methylobacterium nodulans


organism: Paenibacillus lactis


organism: Streptomyces sp. GF3587

assay_conditions_summary: cofactor NADPH

column_label: Table 3. Kinetics data for cyclic imine substrates 1-7; row 5e (R = cyclohexyl); columns kcat, Km, kcat/Km.

raw_value_unit_evidence: Table 3 row 5e (R = cyclohexyl) reports Vmax 0.0388; kcat 0.0215 s^-1; Km 1.55 mM; kcat/Km 0.0139 s^-1 mM^-1.

row_label: Table 3 row 5e (R = cyclohexyl); 6-cyclohexyl-2,3,4,5-tetrahydropyridine


column_label: Table 3. Kinetics data for cyclic imine substrates 1-7; row 5j (R = isopropenyl); columns kcat.

raw_value_unit_evidence: Table 3 row 5j (R = isopropenyl) reports Vmax 4.24; kcat 2.35 s^-1.

row_label: Table 3 row 5j (R = isopropenyl); 6-(prop-1-en-2-yl)-2,3,4,5-tetrahydropyridine


column_label: Table 3. Kinetics data for cyclic imine substrates 1-7; row 5l (R = 2-thienyl); columns Km.

raw_value_unit_evidence: Table 3 row 5l (R = 2-thienyl) reports Vmax 1.16; Km 0.244 mM.

row_label: Table 3 row 5l (R = 2-thienyl); 6-(thiophen-2-yl)-2,3,4,5-tetrahydropyridine