organism: Klebsiella sp. 601

sequence_source: Reconstructed from direct UniProt B0FMT2 canonical sequence plus PMID 21624144 alpha351-380M substitutions; not accepted because the selected batch row aggregates multiple substrate source rows.

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

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant alpha351-380M


enzyme_display_name: laccase

substrate_name: 2,6-Dimethoxyphenol; O2

ki_display: Not reported

kcat_over_km_display: Not reported

mutation_signature: Unknown

protein_accession_database: UniProt

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

km_unit: mM

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

enzyme: laccase

substrate: 2,6-Dimethoxyphenol; O2


substrate_name: O2; 2,6-Dimethoxyphenol

sequence_source: Reconstructed from direct UniProt B0FMT2 canonical sequence with E106F substitution confirmed by PMID 21624144; accepted status still blocked by exact kinetic tuple verification.

mutation_type: substitution

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant E106F

substrate: O2; 2,6-Dimethoxyphenol


organism: Haloferax volcanii

substrate_name: bilirubin

source_db: oed

verification_status: corrected

source_license: CC BY 4.0

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

data_origin: experimental

substrate: bilirubin

status: corrected

source: oed


substrate_name: syringaldazine

kcat_over_km_origin: calculated

substrate: syringaldazine


organism: Marinomonas mediterranea

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

proof_lines: recombinant His-tagged enzyme MmPPOA-695, pH 5.0, 25°C


organism: Bacillus subtilis

sequence_variant_note: Resolved canonical sequence from UniProtKB reviewed entry P07788 (COTA_BACSU) plus direct single-substitution reconstruction from the selected mutation signature. and applied mutation signature R476K to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant R476K


sequence_variant_note: Resolved canonical sequence from UniProtKB reviewed entry P07788 (COTA_BACSU) plus direct single-substitution reconstruction from the selected mutation signature. and applied mutation signature T480A to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant T480A


sequence_variant_note: Resolved canonical sequence from UniProtKB reviewed entry P07788 (COTA_BACSU) plus direct single-substitution reconstruction from the selected mutation signature. and applied mutation signature T480F to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant T480F


organism: Paenibacillus glucanolyticus

sequence_source: UniProtKB:A0A165SC98 direct REST lookup on 2026-05-30


organism: Ochrobactrum sp. 531


organism: Pleurotus ostreatus

substrate_name: 2,2'-Azino-bis(3-ethylbenzthiazoline-6-sulfonic acid); O2; 2,2'-Azino-bis(3-ethylbenzthiazoline-6-sulfonic acid); O2; H2O; ...

sequence_source: UniProtKB:Q12739 FASTA retrieved 2026-05-30


substrate_name: 2,2'-azino-bis(3-ethyl-benzothiazoline-6-sulfonic acid); syringaldazine; 2,6-dimethoxyphenol

source_db: brenda

kcat_display: Not reported

mutation_signature: Δ4

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

mutation_type: deletion

proof_lines: pH 3.0, 22°C, recombinant truncated mutant POXA1bDELTA4

proof_lines: pH 6.0, 22°C, recombinant truncated mutant POXA1bDELTA4

proof_lines: pH 5.3, 22°C, recombinant truncated mutant POXA1bDELTA4

source: brenda


organism: Pseudomonas fluorescens


organism: Geobacillus sp. JS12


organism: Pyrobaculum aerophilum

sequence_variant_note: Resolved canonical sequence from UniProtKB:Q8ZWA8 direct REST lookup 2026-05-30; mutation reconstructed from selected SABIO note and applied mutation signature M470F to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant M470F N-terminal His-tagged


organism: Rheinheimera sp.

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant deltaRhLacc (367 amino acids)


organism: Streptomyces californicus

substrate_name: Potassium ferrocyanide; O2

substrate: Potassium ferrocyanide; O2


organism: Streptomyces coelicolor

km_display: Not reported

sequence_source: UniProt Q9XAL8 canonical sequence with Y108F applied during 2026-05-30 worker review

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant Y108F


sequence_source: UniProt Q9XAL8 canonical sequence with Y108A applied during 2026-05-30 worker review

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant Y108A


proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant M168G


organism: Rhodococcus erythropolis

mutation_signature: laccase( ) deletion -terminal residues

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant deletion 21 C-terminal residues, N-terminal His6-tag


sequence_source: UniProtKB:Q8ZWA8 canonical context only; source row is M470L N-terminal His-tagged and exact construct sequence is unresolved.

sequence_variant_note: Canonical sequence is preserved, but mutation context is present; variant sequence was not attached without safely applying the mutation.

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant M470L N-terminal His-tagged


sequence_variant_note: Resolved canonical sequence from UniProtKB Q9XAL8 REST FASTA checked 2026-05-30; M266W reconstructed with shared +30 residue-numbering offset inferred from selected batch signatures. and applied mutation signature M266W with inferred residue-numbering offset +30 to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant M266W


sequence_variant_note: Resolved canonical sequence from UniProtKB Q9XAL8 REST FASTA checked 2026-05-30; M266A reconstructed with shared +30 residue-numbering offset inferred from selected batch signatures. and applied mutation signature M266A with inferred residue-numbering offset +30 to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant M266A


sequence_variant_note: Resolved canonical sequence from UniProtKB Q9XAL8 REST FASTA checked 2026-05-30; M168G/M266A reconstructed with shared +30 residue-numbering offset inferred from selected batch signatures. and applied mutation signature M168G/M266A with inferred residue-numbering offset +30 to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant M168G/M266A


sequence_variant_note: Resolved canonical sequence from UniProtKB Q9XAL8 REST FASTA checked 2026-05-30; M168A reconstructed with shared +30 residue-numbering offset inferred from selected batch signatures. and applied mutation signature M168A with inferred residue-numbering offset +30 to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant M168A


proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant Y199W


proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant M168G/M266W


sequence_source: reconstructed_from_uniprot_accession Q9XAL8, checked 2026-05-30

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant SLAC V290A


organism: Streptomyces cyaneus

sequence_source: UniProtKB REST direct lookup F6L7B5, checked 2026-05-30

proof_lines: pH and temperature not specified in the publication


sequence_source: UniProtKB Q9XAL8 FASTA direct check; reconstructed S292A after confirming canonical residue S at position 292.

sequence_variant_note: Mutation signature S292A matches the mutant residue at offset +0 and the reference residue at offset +2; variant reconstruction was not applied because multiple plausible residue-numbering offsets are present.

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant S292A


sequence_source: UniProtKB Q9XAL8 FASTA direct check; reconstructed M298F after confirming canonical residue M at position 298.

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant SLAC M298F


sequence_source: UniProtKB Q9XAL8 FASTA direct check; reconstructed V290N after confirming canonical residue V at position 290.

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant SLAC V290N


organism: Streptomyces lividans

verification_status: unverified

status: unverified


organism: Streptomyces sviceus

sequence_variant_note: Mutation signature M220L matches the mutant residue at offset +0 and the reference residue at offset +12; variant reconstruction was not applied because multiple plausible residue-numbering offsets are present.

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant M220L, N-terminal His6-tag


substrate_name: Syringaldazine; O2

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant M295F Ssl1

substrate: Syringaldazine; O2


proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant M293G Ssl1


proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant M295A Ssl1


substrate_name: O2; Syringaldazine

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant M293G/M295L Ssl1

substrate: O2; Syringaldazine


sequence_source: UniProtKB B5HSR1 canonical sequence checked 2026-05-30; variant reconstructed from M295Y after matching the canonical residue(s).

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant M295Y Ssl1


sequence_source: UniProtKB B5HSR1 canonical sequence checked 2026-05-30; variant reconstructed from M195A after matching the canonical residue(s).

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant M195A Ssl1


sequence_source: UniProtKB B5HSR1 canonical sequence checked 2026-05-30; variant reconstructed from M295I after matching the canonical residue(s).

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant M295I Ssl1


sequence_variant_note: Resolved canonical sequence from UniProtKB B5HSR1 canonical sequence reconstructed with selected mutation_signature and applied mutation signature M295T to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant M295T Ssl1


sequence_variant_note: Resolved canonical sequence from UniProtKB B5HSR1 canonical sequence reconstructed with selected mutation_signature and applied mutation signature V287N to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant V287N Ssl1


sequence_variant_note: Resolved canonical sequence from UniProtKB B5HSR1 canonical sequence reconstructed with selected mutation_signature and applied mutation signature M295V to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant M295V Ssl1


sequence_variant_note: Resolved canonical sequence from UniProtKB B5HSR1 canonical sequence reconstructed with selected mutation_signature and applied mutation signature M195A/V287N/M293G/M295L to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant M195A/V287N/M293G/M295L Ssl1


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


substrate_name: Syringaldazine

substrate: Syringaldazine


sequence_source: UniProtKB:B5HSR1 canonical sequence only

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant M293L, N-terminal His6-tag


sequence_source: UniProtKB B5HSR1 direct lookup resolved the canonical protein, but the selected SABIO-RK note includes an N-terminal His6-tagged M220L/T222L construct and the exact tagged construct sequence was not provided.

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant M220L/T222L, N-terminal His6-tag


sequence_source: UniProtKB B5HSR1 direct lookup resolved the canonical protein, but the selected SABIO-RK note includes an N-terminal His6-tagged M195L construct and the exact tagged construct sequence was not provided.

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant M195L, N-terminal His6-tag


sequence_source: UniProtKB B5HSR1 direct lookup resolved the canonical protein and residue 295=M, but the selected SABIO-RK context includes an N-terminal His6-tagged M295L note and a separate M295L Ssl1 note across two PMID/substrate contexts.

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant M295L , N-terminal His6-tag

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant M295L Ssl1


sequence_source: UniProtKB B5HSR1 direct lookup resolved a 325-aa canonical sequence, but the selected SABIO-RK note says deletion of C-terminal residues 312-328 with an N-terminal His6 tag, leaving the coordinate system and exact construct sequence unresolved.

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant del C-terminal residues 312-328, N-terminal His6-tag


sequence_source: UniProtKB B5HSR1 FASTA fetched 2026-05-30; M195A/M293G/M295L reconstructed from canonical residues M195, M293, and M295.

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

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant M195A/M293G/M295L Ssl1


organism: Streptomyces viridochromogenes

sequence_source: UniProtKB J9S5G3 FASTA fetched 2026-05-30.

assay_conditions_summary: pH 4

proof_lines: pH 4.0, temperature not specified in the publication

proof_lines: pH 8.5, temperature not specified in the publication


organism: Streptomyces viridosporus

sequence_source: UniProtKB J9PBR2 FASTA fetched 2026-05-30.

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


organism: Thermus thermophilus

substrate_name: O2; Guaiacol

verification_status: verified

sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:F6DF14 and applied mutation signature D106L to reconstruct the assayed variant sequence.

column_label: Table 1: Apparent steady-state kinetic constants measured at the saturating concentrations of O2 for guaiacol by the lacTT proteins (Appl Microbiol Biotechnol 2018, 102:4049-4061); printed column heads LacTT | Km (uM) | kcat (s^-1) | kcat/Km (s^-1 mM^-1); enzyme row D106L; column 'Km (uM)' = 488; column 'kcat (s^-1)' = 2.1; column 'kcat/Km (s^-1 mM^-1)' = 4.3. captured primary segment: Table 1 row D106L | construct lacTT D106L | substrate guaiacol | condition pH 6.0 | Km 488 uM | kcat 2.1 s^-1 | kcat/Km 4.3 s^-1 mM^-1

normalized_value_unit_evidence: Km: printed Table 1 cell 488 uM; 488 / 1000 = 0.488 mM. kcat: printed Table 1 cell 2.1 s^-1; the printed turnover column is per second, so no unit conversion applies and the value is 2.1 s^(-1). kcat/Km: printed Table 1 cell 4.3 s^-1 mM^-1; the printed efficiency column is already per second per millimolar, so no unit conversion applies and the value is 4.3 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 1, enzyme row D106L: Km = 488 uM; kcat = 2.1 s^-1; kcat/Km = 4.3 s^-1 mM^-1.

row_label: Table 1, enzyme row D106L

substrate: O2; Guaiacol

status: verified


substrate_name: Guaiacol; O2

sequence_variant_note: Resolved canonical sequence from UniProtKB:F6DF14 FASTA; direct REST lookup on 2026-05-30 and applied mutation signature E456L to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant E456L

substrate: Guaiacol; O2


sequence_variant_note: Resolved canonical sequence from UniProtKB:F6DF14 FASTA; direct REST lookup on 2026-05-30 and applied mutation signature D423A to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant D423A


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


sequence_source: UniProtKB:Q72HW2 direct lookup 2026-05-30

proof_lines: pH and temperature not specified in the publication


organism: Streptomyces griseoflavus

sequence_source: UniProt A0A0M4FJ81 canonical sequence fetched via UniProt REST on 2026-05-30; row mutation reconstructed during review.

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant H165F


organism: Thermobaculum terrenum


organism: Thermobifida fusca

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

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant R49K, N-terminal His-tag


organism: Thermus sp.


sequence_source: UniProtKB REST accession F6DF14, checked 2026-05-30; row-specific variant reconstructed from canonical sequence

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant E356A


sequence_variant_note: Mutation signature D106R matches the mutant residue at offset +0 and the reference residue at offset +1; variant reconstruction was not applied because multiple plausible residue-numbering offsets are present.

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant D106R


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:F6DF14 and applied mutation signature E456D to reconstruct the assayed variant sequence.

column_label: Table 1: Apparent steady-state kinetic constants measured at the saturating concentrations of O2 for guaiacol by the lacTT proteins (Appl Microbiol Biotechnol 2018, 102:4049-4061); printed column heads LacTT | Km (uM) | kcat (s^-1) | kcat/Km (s^-1 mM^-1); enzyme row E456D; column 'Km (uM)' = 380; column 'kcat (s^-1)' = 0.78; column 'kcat/Km (s^-1 mM^-1)' = 2.1. captured primary segment: Table 1 row E456D | construct lacTT E456D | substrate guaiacol | condition pH 6.0 | Km 380 uM | kcat 0.78 s^-1 | kcat/Km 2.1 s^-1 mM^-1

normalized_value_unit_evidence: Km: printed Table 1 cell 380 uM; 380 / 1000 = 0.38 mM. kcat: printed Table 1 cell 0.78 s^-1; the printed turnover column is per second, so no unit conversion applies and the value is 0.78 s^(-1). kcat/Km: printed Table 1 cell 2.1 s^-1 mM^-1; the printed efficiency column is already per second per millimolar, so no unit conversion applies and the value is 2.1 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 1, enzyme row E456D: Km = 380 uM; kcat = 0.78 s^-1; kcat/Km = 2.1 s^-1 mM^-1.

row_label: Table 1, enzyme row E456D


sequence_variant_note: Mutation signature D106E matches the mutant residue at offset +0 and the reference residue at offset +1; variant reconstruction was not applied because multiple plausible residue-numbering offsets are present.

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant D106E


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:F6DF14 and applied mutation signature E456R to reconstruct the assayed variant sequence.

column_label: Table 1: Apparent steady-state kinetic constants measured at the saturating concentrations of O2 for guaiacol by the lacTT proteins (Appl Microbiol Biotechnol 2018, 102:4049-4061); printed column heads LacTT | Km (uM) | kcat (s^-1) | kcat/Km (s^-1 mM^-1); enzyme row E456R; column 'Km (uM)' = 512; column 'kcat (s^-1)' = 0.32; column 'kcat/Km (s^-1 mM^-1)' = 0.62. captured primary segment: Table 1 row E456R | construct lacTT E456R | substrate guaiacol | condition pH 6.0 | Km 512 uM | kcat 0.32 s^-1 | kcat/Km 0.62 s^-1 mM^-1

normalized_value_unit_evidence: Km: printed Table 1 cell 512 uM; 512 / 1000 = 0.512 mM. kcat: printed Table 1 cell 0.32 s^-1; the printed turnover column is per second, so no unit conversion applies and the value is 0.32 s^(-1). kcat/Km: printed Table 1 cell 0.62 s^-1 mM^-1; the printed efficiency column is already per second per millimolar, so no unit conversion applies and the value is 0.62 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 1, enzyme row E456R: Km = 512 uM; kcat = 0.32 s^-1; kcat/Km = 0.62 s^-1 mM^-1.

row_label: Table 1, enzyme row E456R


sequence_source: UniProtKB:F6DF14 canonical sequence with M460Q reconstructed during exact accession lookup

sequence_variant_note: Mutation signature M460Q matches the mutant residue at offset +0 and the reference residue at offset +1; variant reconstruction was not applied because multiple plausible residue-numbering offsets are present.

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant LacTT M460Q , His-tagged


sequence_source: UniProtKB:F6DF14 canonical sequence with M460H reconstructed during exact accession lookup

sequence_variant_note: Mutation signature M460H matches the mutant residue at offset +0 and the reference residue at offset +1; variant reconstruction was not applied because multiple plausible residue-numbering offsets are present.

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant LacTT M460H, His-tagged


sequence_source: UniProt REST accession F6L7B5, checked 2026-05-30


assay_conditions_summary: pH 5.5; 303.15 K; buffer 100 mM MES

column_label: Table 1 (Specific activities and kinetic constants of ScLac against DMP, ABTS and guaiacol), substrate column head 'DMP'; row head 'Specific activity (U mg^-1)' = 0.540 ± 0.020; row head 'Km (mM)' = 0.2016 ± 0.0348; row head 'Vmax (U)' = 0.0053 ± 0.0007; row head 'kcat (min^-1)' = 57.30 ± 8.10. The table carries no catalytic-efficiency row.

normalized_value_unit_evidence: Km: printed Table 1 cell 0.2016 mM; the printed unit is already mM, so no unit conversion applies; the exported value is 0.2016 mM. kcat: printed Table 1 cell 57.30 min^-1; min^-1 to s^-1 is a division by 60, so 57.30 / 60 = 0.955 s^(-1); the exported value is 0.955 s^(-1). kcat/Km: Table 1 prints no catalytic-efficiency cell, so the ratio is typed as the same-measurement quotient of the two printed cells of this one column, 0.955 / 0.2016 = 4.737103174603174 s^(-1)*mM^(-1); the exported value is 4.737103174603174 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 1, substrate column 'DMP': row 'Km (mM)' = 0.2016 ± 0.0348 mM; row 'kcat (min^-1)' = 57.30 ± 8.10 min^-1. Table 1 prints no catalytic-efficiency cell for this column.

row_label: Table 1, substrate column 'DMP' (2,6-dimethoxyphenol), rows 'Specific activity (U mg^-1)', 'Km (mM)', 'Vmax (U)' and 'kcat (min^-1)'

table_label: Table 1: Specific activities and kinetic constants of ScLac against DMP, ABTS and guaiacol (AMB Express 2017, 7:86)


substrate_name: Guaiacol

substrate: Guaiacol


sequence_variant_note: Resolved canonical sequence from UniProt REST accession A0A0M4FJ81 plus H165A substitution reconstructed against the canonical sequence, checked 2026-05-30 and applied mutation signature H165A to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant H165A


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:A0A0M4FJ81 and applied mutation signature I170A to reconstruct the assayed variant sequence.

column_label: Table 3 (Kinetic parameters of the SgfSL variants), ABTS block, protein row 'I170A'; column 'Km (mM)' = 0.63 ± 0.11; column 'kcat (s-1)' = 0.70 ± 0.20; column 'kcat/Km (mM-1 s-1)' = 1.11; column 'Activity nM O2 min-1 ug-1' = 0.11 ± 0.01.

normalized_value_unit_evidence: kcat: printed Table 3 cell 0.70 under the column head kcat (s-1); the cell is already in s^-1, so no unit conversion applies and the canonical value is 0.7 s^(-1). Km: printed Table 3 cell 0.63 under the column head Km (mM); the cell is already in mM, so no unit conversion applies and the canonical value is 0.63 mM. kcat/Km: printed Table 3 cell 1.11 under the column head kcat/Km (mM-1 s-1); the cell is already in mM^-1 s^-1, so no unit conversion applies and the canonical value is 1.11 s^-1 mM^-1.

raw_value_unit_evidence: Table 3, ABTS block, protein row 'I170A': Km = 0.63 mM; kcat = 0.70 s-1; printed kcat/Km = 1.11 mM-1 s-1.

table_label: Table 3: Kinetic parameters of the SgfSL variants (Int. J. Mol. Sci. 2019, volume 20, article 3184, page 6); printed column heads Substrate | Protein | Km (mM) | kcat (s-1) | kcat/Km (mM-1 s-1) | Activity nM O2 min-1 ug-1


sequence_source: UniProt REST accession A0A0M4FJ81 canonical sequence, checked 2026-05-30


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:A0A0M4FJ81 and applied mutation signature I170F to reconstruct the assayed variant sequence.

column_label: Table 3 (Kinetic parameters of the SgfSL variants), ABTS block, protein row 'I170F'; column 'Km (mM)' = 0.20 ± 0.04; column 'kcat (s-1)' = 0.08 ± 0.01; column 'kcat/Km (mM-1 s-1)' = 0.40; column 'Activity nM O2 min-1 ug-1' = 0.08 ± 0.04.

normalized_value_unit_evidence: kcat: printed Table 3 cell 0.08 under the column head kcat (s-1); the cell is already in s^-1, so no unit conversion applies and the canonical value is 0.08 s^(-1). Km: printed Table 3 cell 0.20 under the column head Km (mM); the cell is already in mM, so no unit conversion applies and the canonical value is 0.2 mM. kcat/Km: printed Table 3 cell 0.40 under the column head kcat/Km (mM-1 s-1); the cell is already in mM^-1 s^-1, so no unit conversion applies and the canonical value is 0.4 s^-1 mM^-1.

raw_value_unit_evidence: Table 3, ABTS block, protein row 'I170F': Km = 0.20 mM; kcat = 0.08 s-1; printed kcat/Km = 0.40 mM-1 s-1.


organism: Toxicodendron vernicifluum

substrate_name: ferrocyanide

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

substrate: ferrocyanide


organism: Trametes cinnabarina

assay_conditions_summary: pH 5

proof_lines: recombinant wild-type enzyme, pH 5.0, temperature not specified in the publication

proof_lines: recombinant wild-type enzyme, pH 3.0, temperature not specified in the publication


sequence_source: UniProtKB Q72HW2 REST FASTA fetched 2026-05-30 as canonical context only; source note says R453T, C-terminal His-tag, but the tag sequence is not specified.

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant R453T, C-terminal His-tag


sequence_source: UniProtKB REST accession Q4H436, checked 2026-05-30


sequence_source: UniProtKB REST accession F6DF14, checked 2026-05-30


organism: Trametes hirsuta

substrate_name: O2; Potassium ferrocyanide

substrate: O2; Potassium ferrocyanide


organism: Trametes sp. C30

sequence_source: UniProtKB:Q6TH77 canonical context only

sequence_variant_note: Canonical sequence is preserved, but mutation context is present; variant sequence was not attached without safely applying the mutation.

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant C-terminal 13 residues deleted, His6-tag

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant C-terminal 13 residues deleted, hyperglycosated, His6-tag


organism: Trametes versicolor

substrate_name: Methyl syringate; O2

sequence_source: UniProtKB:Q12718 direct REST lookup on 2026-05-30 (https://rest.uniprot.org/uniprotkb/Q12718.json)

substrate: Methyl syringate; O2


organism: unknown prokaryotic organism

sequence_source: Canonical parent from GenBank:GQ468313.1 / protein_id ACV83921.1; Lac3T93 row-specific variant sequence unresolved.

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant Lac3T93 (N40S, V55A, F62L, E316V) His-tagged


proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant Lac15D (del323-332)


organism: Pleurotus ostreatus

proof_lines:
pH 7.0, 25°C
pH 6.0, 25°C
25°C, isozyme POXA3a


sequence_variant_note: Resolved canonical sequence from UniProtKB F6DF14 canonical sequence fetched directly; M460L reconstructed after checking residue 460 is methionine. and applied mutation signature M460L to reconstruct the assayed variant sequence.

assay_conditions_summary: pH 6

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant LacTT M460L, His-tagged


sequence_variant_note: Resolved canonical sequence from UniProtKB F6DF14 canonical sequence fetched directly; M460A reconstructed after checking residue 460 is methionine. and applied mutation signature M460A to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant LacTT M460A, His-tagged


sequence_variant_note: Resolved canonical sequence from UniProtKB F6DF14 canonical sequence fetched directly; M460F reconstructed after checking residue 460 is methionine. and applied mutation signature M460F to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant LacTT M460F, His-tagged


organism: Achromobacter xylosoxidans


organism: Acremonium sp.


organism: Agaricus bisporus


organism: Alcaligenes faecalis


organism: Alkalihalobacillus ligniniphilus


organism: Aquifex aeolicus

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant V19A


proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant M449T


proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant 1D11 (K245R-I441L-M449T-R465G-P58S-Y172C-I199T) -F55S


proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant 2B3 (F55S-F17S-V19A-P58S-Y172C-I199T-R465G-K245R-I441L-M449T)


proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant 2G6 (K245R-I441L-M449T-R465G) -I199T-Y172C


proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant 2G6 (K245R-I441L-M449T-R465G) -Y172C


proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant 1D11 (K245R-I441L-M449T-R465G-P58S-Y172C-I199T) -V19A


proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant 2G6 (K245R-I441L-M449T-R465G) -P58S


proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant 2G6 (K245R-I441L-M449T-R465G)


proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant R471G


proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant 1D11 (K245R-I441L-M449T-R465G-P58S-Y172C-I199T)


proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant P58S


proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant I441L


proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant K245R


proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant Y172C


proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant 2G6 (K245R-I441L-M449T-R465G) -I199T


proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant F55S


proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant 2G6 (K245R-I441L-M449T-R465G) -I199T-P58S


proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant 1D11 (K245R-I441L-M449T-R465G-P58S-Y172C-I199T) -F17S


proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant 1B7 (K245R-I441L-M449T)


proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant I199T


organism: Bacillus amyloliquefaciens

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant T317N-L386Y-S427E


proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant T317N-L386Y-A110E


proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant T317N-L386Y


proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant T317N-L386Y-S427E-A110E


organism: Bacillus atrophaeus


organism: Bacillus coagulans

ki_unit: mM


organism: Bacillus pumilus

proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant L2 (L386Q, G417I, +SP)


proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant L9 (L386Q, G417R, V482G)


proof_lines: pH 7.0, 22°C, recombinant enzyme, with 25% v/v DMSO

proof_lines: pH 6.5, 22°C, recombinant enzyme, with 0.8% v/v DMSO


organism: Bacillus sp.


organism: Bacillus subtilis

sequence_source: UniProtKB P07788 canonical CotA sequence used as identity context for the unmutated holoCotA record.

proof_lines:
pH 7, holoCotA
pH 3, holoCotA
pH 7.5, 37°C


organism: Bacillus tequilensis


organism: Carica papaya


organism: Corynebacterium efficiens


organism: Desulfovibrio sp. A2


organism: Enterobacter ludwigii


organism: Citrobacter freundii


organism: Mycothermus thermophilus


organism: Myrioconium sp.


organism: Myrothecium verrucaria


organism: Obba rivulosa

substrate_name: guaiacol; 2,2'-azino-bis(3-ethylbenzathiazoline-6-sulfonate); syringaldazine; 2,6-dimethoxyphenol

proof_lines:
pH 3.0, 25 C, laccase Lac-4.8
pH 3.0, 25 C, laccase Lac-3.5
pH 3.0, 25 C,laccase Lac-4.8


organism: Perenniporia fraxinea


organism: Lysobacter sp. T-15


organism: Lactobacillus plantarum


organism: Lepista nuda


organism: Pseudomonas sp. LBC1

substrate_name: O2; 3,3'-Dimethylbenzidine

substrate: O2; 3,3'-Dimethylbenzidine


organism: Shewanella putrefaciens


organism: Shigella dysenteriae


proof_lines: SABIO-RK modifier species: laccase(Enzyme) mutant E106F, deletion L351-G378


organism: Pleurotus nebrodensis


organism: Sporothrix carnis


organism: Streptomyces anulatus


organism: Streptomyces griseorubens


organism: Thielavia sp.


organism: Thioalkalivibrio sp.


organism: Trametes hirsuta

substrate_name: O2; 2,6-Dimethoxyphenol

proof_lines: pH 4.5, 50°C, enzyme conjugated with 1.1 kDa PEG


organism: Trametes polyzona

proof_lines: pH 5.0, temperature not specified in the publication

proof_lines: pH and temperature not specified in the publication


organism: Trametes sp.

proof_lines: pH 5.0, 30°C, surface display laccase (SDL) biocatalyst (where the enzyme laccase is displayed on the surface of biological cells through synthetic biology)


organism: Trametes versicolor

substrate_name: O2; Syringic acid

proof_lines: enzyme conjugated with chitosan of 214 kDa

proof_lines: enzyme conjugated with chitosan of 505 kDa

substrate: O2; Syringic acid


organism: Geobacillus thermopakistaniensis


organism: Klebsiella pneumoniae


organism: Kocuria sp.


organism: Kurthia huakuii

substrate_name: 2,6-Dimethoxyphenol; O2


enzyme_display_name: cystathionine beta-synthase

organism: Aeropyrum pernix (strain ATCC 700893 / DSM 11879 / JCM 9820 / NBRC 100138 / K1)

substrate_name: O(3)-acetyl-L-serine (in the presence of 1 mM sodium sulfide at pH 6.7 and 60 degrees Celsius)

source_db: uniprot

enzyme: cystathionine beta-synthase

source: uniprot


organism: Caenorhabditis elegans

substrate_name: serine as substrate (at 16 degrees Celsius)


organism: Homo sapiens

substrate_name: L-homocysteine; L-serine

sequence_source: UniProtKB P35520 canonical sequence with P427L substitution reconstructed during review

sequence_variant_note: Mutation signature P427L matches the mutant residue at offset +0 and the reference residue at offset -5; variant reconstruction was not applied because multiple plausible residue-numbering offsets are present.

proof_lines: mutant P427L, 37°C, pH not specified in the publication

substrate: L-homocysteine; L-serine


proof_lines: mutant K523Sfs?18, 37°C, pH not specified in the publication


substrate_name: L-serine; L-homocysteine

sequence_source: UniProtKB P35520 canonical sequence with L540Q substitution reconstructed during review

sequence_variant_note: Mutation signature L540Q matches the mutant residue at offset +0 and the reference residue at offset -1; variant reconstruction was not applied because multiple plausible residue-numbering offsets are present.

proof_lines: mutant L540Q, 37°C, pH not specified in the publication

substrate: L-serine; L-homocysteine


sequence_source: UniProtKB P35520 canonical sequence with V449G substitution reconstructed during review

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

proof_lines: mutant V449G, 37°C, pH not specified in the publication


enzyme_display_name: FAD synthase

organism: Corynebacterium ammoniagenes

substrate_name: FMN

sequence_source: UniProtKB:Q59263 direct FASTA lookup

sequence_variant_note: Resolved canonical sequence from UniProtKB:Q59263 direct FASTA lookup and applied mutation signature Y106W to reconstruct the assayed variant sequence.

assay_conditions_summary: pH 7; 298.15 K; cofactor FMN

enzyme: FAD synthase

substrate: FMN


sequence_variant_note: Resolved canonical sequence from UniProtKB:Q59263 direct FASTA lookup and applied mutation signature F62W to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from UniProtKB:Q59263 direct FASTA lookup and applied mutation signature F128W to reconstruct the assayed variant sequence.


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


organism: Debaryomyces hansenii

sequence_source: UniProtKB:A0A5J6E2Y6 direct FASTA lookup

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

assay_conditions_summary: cofactor FAD

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


substrate_name: ATP; Riboflavin-5-phosphate

sequence_source: UniProtKB:Q8NFF5 direct FASTA lookup

assay_conditions_summary: pH 7.5; 310.15 K; cofactor FAD

proof_lines:
at 37°C, in 50 mM Tris-HCl, pH 7.5
37°C
at 37°C

substrate: ATP; Riboflavin-5-phosphate


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:Q59263 and applied mutation signature D298A to reconstruct the assayed variant sequence.

column_label: Table 2: Steady-state kinetic parameters for the FMNAT activity (FMN to FAD) of the different CaFADS variants; variant row 'D298A'; column 'k cat (min-1)' = 6.1; column 'K m FMN (uM)' = 1.2; column 'K m ATP (uM)' = 20.7; column 'k cat/K m FMN (min-1 uM-1)' = 5.1; column 'k cat/K m ATP (min-1 uM-1)' = 0.30.

raw_value_unit_evidence: Table 2, variant row 'D298A': k cat = 6.1 min-1; K m FMN = 1.2 uM; printed k cat/K m FMN = 5.1 min-1 uM-1. The row's K m ATP cell is 20.7 uM and its k cat/K m ATP cell is 0.30 min-1 uM-1; both describe the co-substrate ATP, not the catalog row's substrate FMN.

row_label: Table 2, variant row 'D298A'


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


sequence_source: UniProt Q59263 canonical sequence with single-substitution reconstruction


sequence_variant_note: Mutation signature F206K matches the mutant residue at offset +0 and the reference residue at offset +7; variant reconstruction was not applied because multiple plausible residue-numbering offsets are present.


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:Q59263 and applied mutation signature E203A to reconstruct the assayed variant sequence.

column_label: Table 2: Steady-state kinetic parameters for the FMNAT activity (FMN to FAD) of the different CaFADS variants; variant row 'E203A'; column 'k cat (min-1)' = 3.2; column 'K m FMN (uM)' = 0.70; column 'K m ATP (uM)' = 10.8; column 'k cat/K m FMN (min-1 uM-1)' = 4.5; column 'k cat/K m ATP (min-1 uM-1)' = 0.30.

raw_value_unit_evidence: Table 2, variant row 'E203A': k cat = 3.2 min-1; K m FMN = 0.70 uM; printed k cat/K m FMN = 4.5 min-1 uM-1. The row's K m ATP cell is 10.8 uM and its k cat/K m ATP cell is 0.30 min-1 uM-1; both describe the co-substrate ATP, not the catalog row's substrate FMN.

row_label: Table 2, variant row 'E203A'


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:Q59263 and applied mutation signature V300K to reconstruct the assayed variant sequence.

column_label: Table 2: Steady-state kinetic parameters for the FMNAT activity (FMN to FAD) of the different CaFADS variants; variant row 'V300K'; column 'k cat (min-1)' = 4.6; column 'K m FMN (uM)' = 8.3; column 'K m ATP (uM)' = 9.5; column 'k cat/K m FMN (min-1 uM-1)' = 0.55; column 'k cat/K m ATP (min-1 uM-1)' = 0.48.

raw_value_unit_evidence: Table 2, variant row 'V300K': k cat = 4.6 min-1; K m FMN = 8.3 uM; printed k cat/K m FMN = 0.55 min-1 uM-1. The row's K m ATP cell is 9.5 uM and its k cat/K m ATP cell is 0.48 min-1 uM-1; both describe the co-substrate ATP, not the catalog row's substrate FMN.

row_label: Table 2, variant row 'V300K'


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:Q59263 and applied mutation signature L304K to reconstruct the assayed variant sequence.

column_label: Table 2: Steady-state kinetic parameters for the FMNAT activity (FMN to FAD) of the different CaFADS variants; variant row 'L304K'; column 'k cat (min-1)' = 5.2; column 'K m FMN (uM)' = 15.2; column 'K m ATP (uM)' = 11.5; column 'k cat/K m FMN (min-1 uM-1)' = 0.34; column 'k cat/K m ATP (min-1 uM-1)' = 0.45.

raw_value_unit_evidence: Table 2, variant row 'L304K': k cat = 5.2 min-1; K m FMN = 15.2 uM; printed k cat/K m FMN = 0.34 min-1 uM-1. The row's K m ATP cell is 11.5 uM and its k cat/K m ATP cell is 0.45 min-1 uM-1; both describe the co-substrate ATP, not the catalog row's substrate FMN.

row_label: Table 2, variant row 'L304K'


sequence_source: UniProtKB Q8NFF5 direct REST/FASTA lookup

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


organism: Methanocaldococcus jannaschii

sequence_source: UniProtKB Q58579 direct REST/FASTA lookup

assay_conditions_summary: pH 7.2; 343.15 K; buffer 35 mM TES (K+; cofactor FAD

proof_lines: apparent value, in 35 mM TES (K+) buffer (pH 7.2) containing 14 mM dithiothreitol, 7 mM MgCl+, at 70°C


organism: Nakaseomyces glabratus

substrate_name: ATP

sequence_source: UniProtKB Q6FNA9 direct REST/FASTA lookup

substrate: ATP


proof_lines: versus ATP, pH and temperature not specified in the publication, wild-type enzyme

proof_lines: versus FMN, pH and temperature not specified in the publication, wild-type enzyme


sequence_source: reconstructed from UniProtKB Q6FNA9 canonical sequence by applying N62S

sequence_variant_note: Mutation signature N62S matches the mutant residue at offset +0 and the reference residue at offset -8; variant reconstruction was not applied because multiple plausible residue-numbering offsets are present.

proof_lines: pH and temperature not specified in the publication, mutant N62S


sequence_source: reconstructed from UniProtKB Q6FNA9 canonical sequence by applying D168A

sequence_variant_note: Mutation signature D168A matches the mutant residue at offset +0 and the reference residue at offset +13; variant reconstruction was not applied because multiple plausible residue-numbering offsets are present.

proof_lines: pH and temperature not specified in the publication, mutant D168A


sequence_source: reconstructed from UniProtKB Q6FNA9 canonical sequence by applying W184A

sequence_variant_note: Mutation signature W184A matches the mutant residue at offset +0 and the reference residue at offset +12; variant reconstruction was not applied because multiple plausible residue-numbering offsets are present.

proof_lines: pH and temperature not specified in the publication, mutant W184A


sequence_source: reconstructed from UniProtKB Q6FNA9 canonical sequence by applying R297A/R300A

proof_lines: pH and temperature not specified in the publication, mutant R297A/R300A


organism: Arabidopsis thaliana

sequence_source: UniProt REST accession Q0WS47 direct check

assay_conditions_summary: pH 8.5; cofactor FAD


sequence_source: UniProt REST accession Q8VZR0 direct check

proof_lines: recombinant enzyme purified under native conditions, at pH 8.5


sequence_source: UniProt REST accession Q9FMW8 direct check


organism: [Candida] glabrata

source_db: skid

source_license: CC BY-NC-ND 4.0

sequence_source: UniProt REST accession Q6FNA9 direct check

source: skid


sequence_source: UniProt REST accession Q59263 direct check


sequence_variant_note: Resolved canonical sequence from UniProt REST accession Q59263 canonical sequence with R66E reconstructed after direct residue check and applied mutation signature R66E to reconstruct the assayed variant sequence.

assay_conditions_summary: pH 7; 298.15 K; cofactor ATP


sequence_variant_note: Resolved canonical sequence from UniProt REST accession Q59263 canonical sequence with R66A reconstructed after direct residue check and applied mutation signature R66A to reconstruct the assayed variant sequence.


sequence_source: UniProt REST accession Q59263 canonical sequence with R66E reconstructed after direct residue check


sequence_source: UniProtKB Q59263 canonical sequence; R66A reconstructed after residue check

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


sequence_source: UniProtKB Q59263 canonical sequence; R66E reconstructed after residue check


sequence_source: UniProtKB reviewed Q59263 canonical sequence


sequence_source: UniProtKB Q59263 canonical sequence; E301A reconstructed after residue check


sequence_source: UniProtKB Q59263 canonical sequence; F206W reconstructed after residue check

sequence_variant_note: Mutation signature F206W matches the mutant residue at offset +0 and the reference residue at offset +7; variant reconstruction was not applied because multiple plausible residue-numbering offsets are present.


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:Q59263 and applied mutation signature E301A to reconstruct the assayed variant sequence.

column_label: Table 2: Steady-state kinetic parameters for the FMNAT activity (FMN to FAD) of the different CaFADS variants; variant row 'E301A'; column 'k cat (min-1)' = 6.9; column 'K m FMN (uM)' = 0.42; column 'K m ATP (uM)' = 19.7; column 'k cat/K m FMN (min-1 uM-1)' = 16.4; column 'k cat/K m ATP (min-1 uM-1)' = 0.35.

raw_value_unit_evidence: Table 2, variant row 'E301A': k cat = 6.9 min-1; K m FMN = 0.42 uM; printed k cat/K m FMN = 16.4 min-1 uM-1. The row's K m ATP cell is 19.7 uM and its k cat/K m ATP cell is 0.35 min-1 uM-1; both describe the co-substrate ATP, not the catalog row's substrate FMN.

row_label: Table 2, variant row 'E301A'


sequence_source: UniProtKB Q59263 canonical sequence; V300A reconstructed after residue check


organism: Unknown


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:Q59263 and applied mutation signature V300A to reconstruct the assayed variant sequence.

assay_conditions_summary: pH 7; 298.15 K; cofactor FAD

column_label: Table 2: Steady-state kinetic parameters for the FMNAT activity (FMN to FAD) of the different CaFADS variants; variant row 'V300A'; column 'k cat (min-1)' = 4.9; column 'K m FMN (uM)' = 1.4; column 'K m ATP (uM)' = 46.2; column 'k cat/K m FMN (min-1 uM-1)' = 3.5; column 'k cat/K m ATP (min-1 uM-1)' = 0.11.

raw_value_unit_evidence: Table 2, variant row 'V300A': k cat = 4.9 min-1; K m FMN = 1.4 uM; printed k cat/K m FMN = 3.5 min-1 uM-1. The row's K m ATP cell is 46.2 uM and its k cat/K m ATP cell is 0.11 min-1 uM-1; both describe the co-substrate ATP, not the catalog row's substrate FMN.

row_label: Table 2, variant row 'V300A'


sequence_variant_note: Resolved canonical sequence from UniProt Q59263 direct REST/FASTA lookup; variant substitution applied only when coordinate validated and applied mutation signature E203A to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:Q59263 and applied mutation signature K202A to reconstruct the assayed variant sequence.

column_label: Table 2: Steady-state kinetic parameters for the FMNAT activity (FMN to FAD) of the different CaFADS variants; variant row 'K202A'; column 'k cat (min-1)' = 2.3; column 'K m FMN (uM)' = 2.9; column 'K m ATP (uM)' = 12.1; column 'k cat/K m FMN (min-1 uM-1)' = 0.80; column 'k cat/K m ATP (min-1 uM-1)' = 0.19.

raw_value_unit_evidence: Table 2, variant row 'K202A': k cat = 2.3 min-1; K m FMN = 2.9 uM; printed k cat/K m FMN = 0.80 min-1 uM-1. The row's K m ATP cell is 12.1 uM and its k cat/K m ATP cell is 0.19 min-1 uM-1; both describe the co-substrate ATP, not the catalog row's substrate FMN.

row_label: Table 2, variant row 'K202A'


sequence_source: UniProt Q59263 direct REST/FASTA lookup retained as canonical context only

sequence_variant_note: Canonical sequence is preserved, but mutation context is present; variant sequence was not attached without safely applying the mutation.


sequence_variant_note: Resolved canonical sequence from UniProt Q59263 direct REST/FASTA lookup; variant substitution applied only when coordinate validated and applied mutation signature E301K to reconstruct the assayed variant sequence.


table_label: Table 2: Steady-state kinetic parameters for the FMNAT activity (FMN to FAD) of the different CaFADS variants


sequence_source: UniProt Q59263 direct REST/FASTA lookup; variant substitution applied only when coordinate validated


sequence_variant_note: Resolved canonical sequence from UniProt Q59263 direct REST/FASTA lookup; variant substitution applied only when coordinate validated and applied mutation signature Y106W to reconstruct the assayed variant sequence.


substrate_name: Mg-ATP (for riboflavin kinase activity)

substrate: Mg-ATP (for riboflavin kinase activity)


organism: Geobacillus thermodenitrificans (strain NG80-2)

substrate_name: Unknown

substrate: Unknown


organism: Methanocaldococcus jannaschii (strain ATCC 43067 / DSM 2661 / JAL-1 / JCM 10045 / NBRC 100440)


assay_conditions_summary: cofactor FMN

proof_lines: kcat is 1.28x10(-1) sec(-1) with FMN as substrate (PubMed:18713732). kcat is 1.28x10(-1) sec(-1) with ATP as substrate (PubMed:18713732).


proof_lines: kcat is 1.08x10(-1) sec(-1) with FMN as substrate (PubMed:18713732). kcat is 1.07x10(-1) sec(-1) with ATP as substrate (PubMed:18713732).


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

assay_conditions_summary: pH 7; cofactor FMN

proof_lines: kcat is 0.393 sec(-1) with FMN as substrate (at 30 degrees Celsius and at pH 7.0) (PubMed:36273586). kcat is 0.287 sec(-1) with ATP as substrate (at 30 degrees Celsius and at pH 7.0) (PubMed:36273586).


organism: Candida glabrata (strain ATCC 2001 / BCRC 20586 / JCM 3761 / NBRC 0622 / NRRL Y-65 / CBS 138)

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

assay_conditions_summary: pH 7.5; cofactor FMN

proof_lines: kcat is 0.087 sec(-1) (at 25 degrees Celsius and at pH 7.5) (PubMed:19375431). kcat is 0.089 sec(-1) (at 25 degrees Celsius and at pH 7.5) (PubMed:23663086).


organism: Chlamydomonas reinhardtii

assay_conditions_summary: pH 7; cofactor FAD

proof_lines: kcat is 0.103 sec(-1) with FMN as substrate (at 30 degrees Celsius and at pH 7.0) (PubMed:36273586). kcat is 0.395 sec(-1) with FAD as substrate (at 30 degrees Celsius and at pH 7.0) (PubMed:36273586).

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


substrate_name: FMN using enzyme purified from E.coli

assay_conditions_summary: pH 7.5; cofactor FAD

proof_lines: kcat is 0.69 sec(-1) for FAD synthase activity using enzyme purified from E.coli (at 37 degrees Celsius and at pH 7.5) (PubMed:21951714). kcat is 0.69 sec(-1) for FAD synthase activity using enzyme purified from E.coli (at 37 degrees Celsiu

proof_lines: kcat is 0.042 sec(-1) for FAD synthase activity using C-terminal region (329-587) purified from E.coli (at 37 degrees Celsius and at pH 7.5) (PubMed:23443125). kcat is 0.0037 sec(-1) for the reverse pyrophosphorolytic reaction using C-termi

substrate: FMN using enzyme purified from E.coli


organism: Bos taurus

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


assay_conditions_summary: pH 8; 310.15 K; cofactor FAD

proof_lines: FADS trimer, 10 mM MgCl2 in 50 mM Tris–HCl (pH 8.0), at 37°C

proof_lines: FADS monomer, 10 mM MgCl2 in 50 mM Tris–HCl (pH 8.0), at 37°C


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

assay_conditions_summary: pH 7; 310.15 K; cofactor FMN

proof_lines: mutant enzyme E268A, at 37°C in 20 mM PIPES pH 7.0, 10 mM MgCl2


proof_lines: mutant enzyme T208A, at 37°C in 20 mM PIPES pH 7.0, 10 mM MgCl2


proof_lines: mutant enzyme N210D, at 37°C in 20 mM PIPES pH 7.0, 10 mM MgCl2


proof_lines: mutant enzyme T208D, at 37°C in 20 mM PIPES pH 7.0, 10 mM MgCl2


proof_lines: mutant enzyme E268D, at 37°C in 20 mM PIPES pH 7.0, 10 mM MgCl2


proof_lines: wild type enzyme, at 37°C in 20 mM PIPES pH 7.0, 10 mM MgCl2


proof_lines: mutant enzyme N210A, at 37°C in 20 mM PIPES pH 7.0, 10 mM MgCl2


proof_lines: with 24 mM Na2S2O4, in 50 mM potassium phosphate (pH 7.5), at 37°C

proof_lines: without Na2S2O4, in 50 mM potassium phosphate (pH 7.5), at 37°C


organism: Rattus norvegicus

substrate_name: Riboflavin-5-phosphate; ATP

assay_conditions_summary: pH 7.1; 310.15 K; cofactor FAD

proof_lines:
pH 7.1, 37°C
pH 7.1, 37°C, direct assay
pH 8.0, 37°C

substrate: Riboflavin-5-phosphate; ATP


organism: Streptococcus pneumoniae


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:Q59263 and applied mutation signature E301K to reconstruct the assayed variant sequence.

column_label: Table 2: Steady-state kinetic parameters for the FMNAT activity (FMN to FAD) of the different CaFADS variants; variant row 'E301K'; column 'k cat (min-1)' = 6.3; column 'K m FMN (uM)' = 0.85; column 'K m ATP (uM)' = 15.3; column 'k cat/K m FMN (min-1 uM-1)' = 7.4; column 'k cat/K m ATP (min-1 uM-1)' = 0.41.

raw_value_unit_evidence: Table 2, variant row 'E301K': k cat = 6.3 min-1; K m FMN = 0.85 uM; printed k cat/K m FMN = 7.4 min-1 uM-1. The row's K m ATP cell is 15.3 uM and its k cat/K m ATP cell is 0.41 min-1 uM-1; both describe the co-substrate ATP, not the catalog row's substrate FMN.

row_label: Table 2, variant row 'E301K'


sequence_variant_note: Mutation signature F206A matches the mutant residue at offset +0 and the reference residue at offset +7; variant reconstruction was not applied because multiple plausible residue-numbering offsets are present.


sequence_variant_note: Mutation signature F206K matches the mutant residue at offset +0 and the reference residue at offset +7; variant reconstruction was not applied because multiple plausible residue-numbering offsets are present.


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:Q59263 and applied mutation signature D298E to reconstruct the assayed variant sequence.

column_label: Table 2: Steady-state kinetic parameters for the FMNAT activity (FMN to FAD) of the different CaFADS variants; variant row 'D298E'; column 'k cat (min-1)' = 3.3; column 'K m FMN (uM)' = 0.95; column 'K m ATP (uM)' = 10.4; column 'k cat/K m FMN (min-1 uM-1)' = 3.47; column 'k cat/K m ATP (min-1 uM-1)' = 0.31.

raw_value_unit_evidence: Table 2, variant row 'D298E': k cat = 3.3 min-1; K m FMN = 0.95 uM; printed k cat/K m FMN = 3.47 min-1 uM-1. The row's K m ATP cell is 10.4 uM and its k cat/K m ATP cell is 0.31 min-1 uM-1; both describe the co-substrate ATP, not the catalog row's substrate FMN.

row_label: Table 2, variant row 'D298E'


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:Q59263 and applied mutation signature D298A to reconstruct the assayed variant sequence.


organism: Listeria monocytogenes serovar 1/2a (strain ATCC BAA-679 / EGD-e)

substrate_name: riboflavin

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

proof_lines: kcat is 0.40 sec(-1) with riboflavin as substrate. kcat is 4.39 sec(-1) with FMN as substrate.

substrate: riboflavin


enzyme_display_name: FAD synthase / FMN adenylyltransferase

sequence_variant_note: Resolved canonical sequence from UniProtKB Q6FNA9 canonical sequence with row-specific single-substitution reconstruction and applied mutation signature R297A to reconstruct the assayed variant sequence.

proof_lines: pH and temperature not specified in the publication, mutant R297A

enzyme: FAD synthase / FMN adenylyltransferase


sequence_variant_note: Resolved canonical sequence from UniProtKB Q6FNA9 canonical sequence with row-specific single-substitution reconstruction and applied mutation signature D181A to reconstruct the assayed variant sequence.

proof_lines: pH and temperature not specified in the publication, mutant D181A


sequence_variant_note: Resolved canonical sequence from UniProtKB Q6FNA9 canonical sequence with row-specific single-substitution reconstruction and applied mutation signature N62A to reconstruct the assayed variant sequence.

proof_lines: pH and temperature not specified in the publication, mutant N62A


sequence_variant_note: Resolved canonical sequence from UniProtKB Q6FNA9 canonical sequence with row-specific single-substitution reconstruction and applied mutation signature R300A to reconstruct the assayed variant sequence.

proof_lines: pH and temperature not specified in the publication, mutant R300A


enzyme_display_name: thermolysin

enzyme: thermolysin


organism: Bacillus sp. (in: firmicutes)

substrate_name: N-benzyloxycarbonyl-Gly-L-Leu amide

sequence_variant_note: Resolved canonical sequence from unresolved_mature_coordinate_conflict and applied mutation signature L155S/I156V with inferred residue-numbering offset -30 to reconstruct the assayed variant sequence.

proof_lines: recombinant mutant L155S/I156V, pH 7.0, 37°C

substrate: N-benzyloxycarbonyl-Gly-L-Leu amide


sequence_variant_note: Mutation signature L155A/I156V matches the mutant residue at offset +1 and the reference residue at offset -30; variant reconstruction was not applied because multiple plausible residue-numbering offsets are present.

proof_lines: recombinant mutant L155A/I156V, pH 7.0, 37°C


organism: Bacillus thermoproteolyticus

sequence_variant_note: Resolved sequence already matches the mutant residue(s) after applying an inferred residue-numbering offset -13, so it is treated as a source-provided variant sequence.


sequence_variant_note: Mutation signature L155S could not be applied safely because the resolved sequence does not match the expected reference residue(s).


sequence_variant_note: Mutation signature L155F could not be applied safely because the resolved sequence does not match the expected reference residue(s).


substrate_name: N-carbobenzoxy-L-Asp-L-Phe methyl ester

sequence_variant_note: Resolved canonical sequence from uniprot_p00800_mature_chain_coordinate_reconstruction and applied mutation signature Q225K with inferred residue-numbering offset -4 to reconstruct the assayed variant sequence.

substrate: N-carbobenzoxy-L-Asp-L-Phe methyl ester


sequence_variant_note: Mutation signature Q128K could not be applied safely because the resolved sequence does not match the expected reference residue(s).


sequence_variant_note: Mutation signature Q128K/Q225R could not be applied safely because the resolved sequence does not match the expected reference residue(s).

proof_lines: pH 7.5, 25°C, recombinant mutants Q128K and Q225R


sequence_variant_note: Mutation signature Q128E/Q225E could not be applied safely because the resolved sequence does not match the expected reference residue(s).

proof_lines: pH 7.5, 25°C, recombinant mutants Q128E and Q225E


sequence_variant_note: Mutation signature Q225A matches the mutant residue at offset +0 and the reference residue at offset -4; variant reconstruction was not applied because multiple plausible residue-numbering offsets are present.


sequence_variant_note: Mutation signature G8C/N60C/S65P/L144S could not be applied safely because the resolved sequence does not match the expected reference residue(s).

proof_lines: pH 7.0, 25°C, recombinant mutant G8C/N60C/S65P/L144S


sequence_variant_note: Mutation signature G8C/N60C/S65P could not be applied safely because the resolved sequence does not match the expected reference residue(s).

assay_conditions_summary: pH 7.5; 298.15 K; buffer 40 mM HEPES-NaOH

proof_lines: pH 7.0, 25°C, recombinant mutant G8C/N60C/S65P

proof_lines: mutant enzyme G8C/N60C/S65P, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C

proof_lines: mutant enzyme G8C/N60C/S65P, in the presence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C


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

proof_lines: pH 7.0, 37°C, Co2+-bound enzyme, in presence of 1 M NaSCN


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature L155A/I156N with inferred residue-numbering offset -30 to reconstruct the assayed variant sequence.

proof_lines: recombinant mutant L155A/I156N, pH 7.0, 37°C


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:Q59223 and applied mutation signature L155A with inferred residue-numbering offset +230 to reconstruct the assayed variant sequence.

assay_conditions_summary: pH 7; 310.15 K; buffer containing 5 mM CaCl2

column_label: Table 1 (Specific activity and kinetic parameters of each mutant TLN), enzyme-form row 'L155A'; column 'Km (mM)' = 20; column 'kcat (min^-1)' = 4,800; column 'kcat/Km (min^-1 mM^-1)' = 240; column 'Half-life at 80 C (min)' = 25.0.

normalized_value_unit_evidence: Km: printed Table 1 cell 20 mM; already in mM, so no unit conversion applies; the exported value is 20.0 mM. kcat: printed Table 1 cell 4,800 min^-1; 4800 / 60 = 80.0 s^-1; the exported value is 80.0 s^(-1). kcat/Km: printed Table 1 cell 240 min^-1 mM^-1; 240 / 60 = 4.0 s^-1 mM^-1; the exported value is 4.0 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 1, row 'L155A': Km = 20 mM; kcat = 4,800 min^-1; printed kcat/Km = 240 min^-1 mM^-1; half-life at 80 C = 25.0 min.

row_label: Table 1, row 'L155A'

table_label: Table 1: Specific activity and kinetic parameters of each mutant TLN (Appl Biochem Biotechnol 2012, 166(3):735-743)


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature L155S/I156N with inferred residue-numbering offset -30 to reconstruct the assayed variant sequence.

proof_lines: recombinant mutant L155S/I156N, pH 7.0, 37°C


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


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature L155S with inferred residue-numbering offset -30 to reconstruct the assayed variant sequence.


substrate_name: H2O; Z-Phe-Leu-Ala-OH

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

sequence_source: multiple

proof_lines: pH 8.0, 70°C, recombinant enzyme, autoactivation, without radiofrequency

proof_lines: pH 8.0, 70°C, recombinant enzyme, autoactivation, with radiofrequency

source_databases_merged: sabio_rk brenda skid

substrate: H2O; Z-Phe-Leu-Ala-OH


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature L155A/G8C/N60C/S65P with inferred residue-numbering offset +232 to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme L155A/G8C/N60C/S65P, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C

proof_lines: mutant enzyme L155A/G8C/N60C/S65P, in the presence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C

proof_lines: mutant enzyme L155A/G8C/N60C/S65P, in the absence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature L155A with inferred residue-numbering offset +232 to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme L155A, in the absence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C

proof_lines: mutant enzyme L155A, in the presence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature S53D/G8C/N60C/S65P with inferred residue-numbering offset +232 to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme S53D/G8C/N60C/S65P, in the absence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C

proof_lines: mutant enzyme S53D/G8C/N60C/S65P, in the presence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature L144S with inferred residue-numbering offset +232 to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme L144S, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C

proof_lines: mutant enzyme L144S, in the presence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature L144S/D150E with inferred residue-numbering offset +232 to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme L144S/D150E, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C

proof_lines: mutant enzyme L144S/D150E, in the presence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C

proof_lines: mutant enzyme L144S/D150E, in the absence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature Q225R with inferred residue-numbering offset -4 to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature Q225V with inferred residue-numbering offset -4 to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature Q225E with inferred residue-numbering offset -4 to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved sequence already matches the mutant residue(s) after applying an inferred residue-numbering offset -4, so it is treated as a source-provided variant sequence.


sequence_variant_note: Mutation signature Q128E could not be applied safely because the resolved sequence does not match the expected reference residue(s).


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature Q225D with inferred residue-numbering offset -4 to reconstruct the assayed variant sequence.


sequence_variant_note: Mutation signature L144S/D150E/S53D could not be applied safely because the resolved sequence does not match the expected reference residue(s).

assay_conditions_summary: pH 7.5; 298.15 K; buffer HEPES/CaCl2

proof_lines: mutant enzyme L144S/D150E/S53D, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C

proof_lines: mutant enzyme L144S/D150E/S53D, in the absence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C

proof_lines: mutant enzyme L144S/D150E/S53D, in the presence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C


sequence_variant_note: Mutation signature G117E could not be applied safely because the resolved sequence does not match the expected reference residue(s).


sequence_variant_note: Mutation signature G117K could not be applied safely because the resolved sequence does not match the expected reference residue(s).


sequence_variant_note: Mutation signature G117R could not be applied safely because the resolved sequence does not match the expected reference residue(s).


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

proof_lines: purified recombinant mutant N116D, pH 7.5, 25°C


sequence_variant_note: Mutation signature N116A could not be applied safely because the resolved sequence does not match the expected reference residue(s).

proof_lines: purified recombinant mutant N116A, pH 7.5, 25°C


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature S53D/L155A/G8C/N60C/S65P with inferred residue-numbering offset +232 to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme S53D/L155A/G8C/N60C/S65P, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C

proof_lines: mutant enzyme S53D/L155A/G8C/N60C/S65P, in the presence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C

proof_lines: mutant enzyme S53D/L155A/G8C/N60C/S65P, in the absence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature D150E/I168A with inferred residue-numbering offset +232 to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme D150E/I168A, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature S53D/L155A with inferred residue-numbering offset +232 to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme S53D/L155A, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C

proof_lines: mutant enzyme S53D/L155A, in the absence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C

proof_lines: mutant enzyme S53D/L155A, in the presence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C


substrate_name: N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide

sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature S53D with inferred residue-numbering offset +232 to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme S53D, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C

proof_lines: mutant enzyme S53D, in the presence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C

proof_lines: mutant enzyme S53D, in the absence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C

substrate: N-[3-(2-furyl)acryloyl]-Gly-L-Leu amide


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature I168A with inferred residue-numbering offset +232 to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme I168A, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C

proof_lines: mutant enzyme I168A, in the presence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C


substrate_name: N-carbobenzoxy-L-Asp-L-Phe-methyl ester

sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature D150E with inferred residue-numbering offset +232 to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme D150E, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C

proof_lines: mutant enzyme D150E, in the presence of 4 M NaCl, in 40 mM HEPES-NaOH buffer at pH 7.5 containing 10 mM CaCl2, at 25°C

substrate: N-carbobenzoxy-L-Asp-L-Phe-methyl ester


substrate_name: leucine enkephalin

sequence_variant_note: Resolved sequence already matches the mutant residue(s) after applying an inferred residue-numbering offset -10, so it is treated as a source-provided variant sequence.

substrate: leucine enkephalin


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

proof_lines:
pH 6.8, 37°C, wild-type
pH 7.4, 37°C, wild-type


sequence_variant_note: Mutation signature R203M could not be applied safely because the resolved sequence does not match the expected reference residue(s).


sequence_variant_note: Mutation signature N116T could not be applied safely because the resolved sequence does not match the expected reference residue(s).

proof_lines: purified recombinant mutant N116T, pH 7.5, 25°C


assay_conditions_summary: pH 7; 298.15 K; buffer 40 mM HEPES-NaOH

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

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

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


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

proof_lines: purified recombinant mutant N116Q, pH 7.5, 25°C


sequence_variant_note: Mutation signature D226A could not be applied safely because the resolved sequence does not match the expected reference residue(s).


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature Y157F with inferred residue-numbering offset +12 to reconstruct the assayed variant sequence.


sequence_variant_note: Mutation signature N227H could not be applied safely because the resolved sequence does not match the expected reference residue(s).


sequence_variant_note: Mutation signature D150W could not be applied safely because the resolved sequence does not match the expected reference residue(s).


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


sequence_variant_note: Mutation signature F114A could not be applied safely because the resolved sequence does not match the expected reference residue(s).


substrate_name: L-phenylalanine methyl ester

substrate: L-phenylalanine methyl ester


substrate_name: N-carbobenzoxy-L-aspartic acid

substrate: N-carbobenzoxy-L-aspartic acid


sequence_variant_note: Mutation signature D150E could not be applied safely because the resolved sequence does not match the expected reference residue(s).


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_unique and applied mutation signature I168A with inferred residue-numbering offset -15 to reconstruct the assayed variant sequence.


substrate_name: L-Phe methyl ester

substrate: L-Phe methyl ester


substrate_name: Benzyloxycarbonyl-Phe-Ala-Ala

substrate: Benzyloxycarbonyl-Phe-Ala-Ala


substrate_name: Benzyloxycarbonyl-Gly-Pro-Leu-Gly

assay_conditions_summary: pH 7; 313.15 K; buffer pH 7

column_label: Results text, page 378, left column, the sentence '2-Gly-Pro-Leu-Gly were 12.8 mM and 11.2, respectively': Km = 12.8 mM and kcat = 11.2 sec^-1 for Z-Gly-Pro-Leu-Gly; the sentence fixes the order as 'the K , and kcat (sec-l)'; the article prints no kcat/Km value for this peptide.

normalized_value_unit_evidence: Km: stated value 12.8 mM; already in mM, no unit conversion applies; the exported value is 12.8 mM. kcat: stated value 11.2 sec^-1; 1 sec^-1 is 1 s^-1, no unit conversion applies; the exported value is 11.2 s^(-1). kcat/Km: not printed; the quotient of the two stated values is 11.2 / 12.8 = 0.8749999999999999 s^-1 mM^-1, which is the exported value 0.8749999999999999 s^(-1)*mM^(-1).

raw_value_unit_evidence: Results text, page 378, sentence '2-Gly-Pro-Leu-Gly were 12.8 mM and 11.2, respectively': Km = 12.8 mM; kcat = 11.2 sec^-1; the article prints no kcat/Km value for this peptide.

row_label: Results sentence on page 378 giving the Km and kcat of Z-Gly-Pro-Leu-Gly

table_label: Results text, page 378 (Eur. J. Biochem. 15 (1970) 374-380): kinetic parameters of Z-Gly-Pro-Leu-Gly and Z-Gly-Pro-Leu-Gly-Pro stated in prose

substrate: Benzyloxycarbonyl-Gly-Pro-Leu-Gly


substrate_name: Benzyloxycarbonyl-Ala-Gly-Gly-Leu-Ala

column_label: Table 1: Kinetics of Z-Gly-Leu-(Gly)n-B and Z-A-(Gly)n-Leu-Ala hydrolysis (Eur. J. Biochem. 15 (1970) 374-380, page 376), group V, peptide row Z-Ala-Gly-Gly-Leu-Ala (text-layer line 'v        Z---Ala--Gly--Gly---Leu--Ala                                        7.5                     50.0                    6.6'); column Km (mM) = 7.5; column kcat (sec^-1) = 50.0; column kcat/Km (sec^-1 x mM^-1) = 6.6.

normalized_value_unit_evidence: Km: printed Table 1 cell 7.5 mM; already in mM, no unit conversion applies; the exported value is 7.5 mM. kcat: printed Table 1 cell 50.0 sec^-1; 1 sec^-1 is 1 s^-1, no unit conversion applies; the exported value is 50.0 s^(-1). kcat/Km: printed Table 1 cell 6.6 sec^-1 x mM^-1; 1 sec^-1 x mM^-1 is 1 s^-1 mM^-1, no unit conversion applies, so the printed cell is 6.6 s^-1 mM^-1; the exported value is 6.666666666666667 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 1, group V row Z-Ala-Gly-Gly-Leu-Ala (text-layer line 163, 'v        Z---Ala--Gly--Gly---Leu--Ala                                        7.5                     50.0                    6.6'): Km = 7.5 mM; kcat = 50.0 sec^-1; printed kcat/Km = 6.6 sec^-1 x mM^-1.

row_label: Table 1, group V row Z---Ala--Gly--Gly---Leu--Ala

substrate: Benzyloxycarbonyl-Ala-Gly-Gly-Leu-Ala


substrate_name: Benzyloxycarbonyl-Ala-Gly-Leu-Ala

substrate: Benzyloxycarbonyl-Ala-Gly-Leu-Ala


substrate_name: Benzyloxycarbonyl-Ala-Leu-Ala

substrate: Benzyloxycarbonyl-Ala-Leu-Ala


substrate_name: Benzyloxycarbonyl-D-Ala-Gly-Leu-Ala

column_label: Table 1: Kinetics of Z-Gly-Leu-(Gly)n-B and Z-A-(Gly)n-Leu-Ala hydrolysis (Eur. J. Biochem. 15 (1970) 374-380, page 376), group IV, peptide row Z-D-Ala-Gly-Leu-Ala (text-layer line 'Z--D-Ala-Gly---Leu--Ala                                      16.0                    625.0                  39.0'); column Km (mM) = 16.0; column kcat (sec^-1) = 625.0; column kcat/Km (sec^-1 x mM^-1) = 39.0.

normalized_value_unit_evidence: Km: printed Table 1 cell 16.0 mM; already in mM, no unit conversion applies; the exported value is 16.0 mM. kcat: printed Table 1 cell 625.0 sec^-1; 1 sec^-1 is 1 s^-1, no unit conversion applies; the exported value is 625.0 s^(-1). kcat/Km: printed Table 1 cell 39.0 sec^-1 x mM^-1; 1 sec^-1 x mM^-1 is 1 s^-1 mM^-1, no unit conversion applies, so the printed cell is 39.0 s^-1 mM^-1; the exported value is 39.0625 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 1, group IV row Z-D-Ala-Gly-Leu-Ala (text-layer line 160, 'Z--D-Ala-Gly---Leu--Ala                                      16.0                    625.0                  39.0'): Km = 16.0 mM; kcat = 625.0 sec^-1; printed kcat/Km = 39.0 sec^-1 x mM^-1.

row_label: Table 1, group IV row Z--D-Ala-Gly---Leu--Ala

substrate: Benzyloxycarbonyl-D-Ala-Gly-Leu-Ala


substrate_name: Benzyloxycarbonyl-Gly-Gly-Leu-Ala

substrate: Benzyloxycarbonyl-Gly-Gly-Leu-Ala


substrate_name: GFS

assay_conditions_summary: pH 8; 310.15 K; buffer and 5 M glycerol contexts are separate measurements

proof_lines:
pH 8.0, 37°C, 5 M glycerol
pH 8.0, 37°C

substrate: GFS


substrate_name: Benzyloxycarbonyl-Gly-Leu-Ala

column_label: Table 1: Kinetics of Z-Gly-Leu-(Gly)n-B and Z-A-(Gly)n-Leu-Ala hydrolysis (Eur. J. Biochem. 15 (1970) 374-380, page 376), group I, peptide row Z-Gly-Leu-Ala (text-layer line 'Z---Gly---Lcu---Ala                                   10.6                    780.6                  78.1'); column Km (mM) = 10.6; column kcat (sec^-1) = 780.6; column kcat/Km (sec^-1 x mM^-1) = 78.1.

normalized_value_unit_evidence: Km: printed Table 1 cell 10.6 mM; already in mM, no unit conversion applies; the exported value is 10.6 mM. kcat: printed Table 1 cell 780.6 sec^-1; 1 sec^-1 is 1 s^-1, no unit conversion applies; the exported value is 780.6 s^(-1). kcat/Km: printed Table 1 cell 78.1 sec^-1 x mM^-1; 1 sec^-1 x mM^-1 is 1 s^-1 mM^-1, no unit conversion applies, so the printed cell is 78.1 s^-1 mM^-1; the exported value is 73.64150943396227 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 1, group I row Z-Gly-Leu-Ala (text-layer line 146, 'Z---Gly---Lcu---Ala                                   10.6                    780.6                  78.1'): Km = 10.6 mM; kcat = 780.6 sec^-1; printed kcat/Km = 78.1 sec^-1 x mM^-1.

row_label: Table 1, group I row Z---Gly---Lcu---Ala

substrate: Benzyloxycarbonyl-Gly-Leu-Ala


substrate_name: Benzyloxycarbonyl-Gly-Leu-D-Ala

substrate: Benzyloxycarbonyl-Gly-Leu-D-Ala


substrate_name: Benzyloxycarbonyl-Gly-Leu-Gly

substrate: Benzyloxycarbonyl-Gly-Leu-Gly


substrate_name: Benzyloxycarbonyl-Gly-Leu-Gly-Ala

substrate: Benzyloxycarbonyl-Gly-Leu-Gly-Ala


substrate_name: Benzyloxycarbonyl-Gly-Leu-Gly-Gly

substrate: Benzyloxycarbonyl-Gly-Leu-Gly-Gly


substrate_name: Benzyloxycarbonyl-Gly-Leu-Leu

column_label: Table 1: Kinetics of Z-Gly-Leu-(Gly)n-B and Z-A-(Gly)n-Leu-Ala hydrolysis (Eur. J. Biochem. 15 (1970) 374-380, page 376), group I, peptide row Z-Gly-Leu-Leu (text-layer line '2---Gly---Leu---Leu                                    2.6                    374.0                 143.9'); column Km (mM) = 2.6; column kcat (sec^-1) = 374.0; column kcat/Km (sec^-1 x mM^-1) = 143.9.

normalized_value_unit_evidence: Km: printed Table 1 cell 2.6 mM; already in mM, no unit conversion applies; the exported value is 2.6 mM. kcat: printed Table 1 cell 374.0 sec^-1; 1 sec^-1 is 1 s^-1, no unit conversion applies; the exported value is 374.0 s^(-1). kcat/Km: printed Table 1 cell 143.9 sec^-1 x mM^-1; 1 sec^-1 x mM^-1 is 1 s^-1 mM^-1, no unit conversion applies, so the printed cell is 143.9 s^-1 mM^-1; the exported value is 143.84615384615384 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 1, group I row Z-Gly-Leu-Leu (text-layer line 148, '2---Gly---Leu---Leu                                    2.6                    374.0                 143.9'): Km = 2.6 mM; kcat = 374.0 sec^-1; printed kcat/Km = 143.9 sec^-1 x mM^-1.

row_label: Table 1, group I row 2---Gly---Leu---Leu

substrate: Benzyloxycarbonyl-Gly-Leu-Leu


substrate_name: Benzyloxycarbonyl-Gly-Leu-Phe

assay_conditions_summary: pH 7; 313.15 K; buffer of pH 7

column_label: Table 1 (PDF page 3 of 7, journal page 376), peptide row `Z---Gly---Leu---Phe`: Km (mM) = 2.4; kcat (sec^-1) = 120.0; kcat/Km (sec^-1 x mM^-1) = 50.0.

normalized_value_unit_evidence: kcat: printed cell 120.0 sec^-1 is already in the export unit -> 120.0 s^(-1); the exported value is 120.0 s^(-1). Km: printed cell 2.4 mM is already in the export unit -> 2.4 mM; the exported value is 2.4 mM. kcat/Km: the paper PRINTS the kcat/Km cell 50.0 sec^-1 x mM^-1 for this peptide row, equal to 50.0 s^(-1)*mM^(-1) in the export unit; the exported value is 50.0 s^(-1)*mM^(-1).

row_label: Table 1, decoded plain-numeral transcription, peptide row `Z---Gly---Leu---Phe`

table_label: Table 1 (PDF page 3 of 7, journal page 376) 'Kinetics of Z-Gly-Leu-(Gly)n-B and Z-A-(Gly)n-Leu-Ala hydrolysis'

substrate: Benzyloxycarbonyl-Gly-Leu-Phe


organism: Geobacillus stearothermophilus

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


sequence_variant_note: Mutation signature N116A could not be applied safely because the resolved sequence does not match the expected reference residue(s).


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


substrate_name: Z-Phe-Leu-Ala-OH

substrate: Z-Phe-Leu-Ala-OH


substrate_name: Benzyloxycarbonyl-Phe-Gly-Ala

substrate: Benzyloxycarbonyl-Phe-Gly-Ala


substrate_name: Benzyloxycarbonyl-Phe-Gly-Leu-Ala

substrate: Benzyloxycarbonyl-Phe-Gly-Leu-Ala


substrate_name: Benzyloxycarbonyl-Phe-Leu-Ala

substrate: Benzyloxycarbonyl-Phe-Leu-Ala


substrate_name: Benzyloxycarbonyl-Phe-Phe-Ala

column_label: Table 2 (PDF page 4 of 7, journal page 377), peptide row `Z---Phe---Phe--Ala`: Km (mM) = 0.45; kcat (sec^-1) = 162.0; kcat/Km (sec^-1 x mM^-1) = 360.0.

normalized_value_unit_evidence: kcat: printed cell 162.0 sec^-1 is already in the export unit -> 162.0 s^(-1); the exported value is 162.0 s^(-1). Km: printed cell 0.45 mM is already in the export unit -> 0.45 mM; the exported value is 0.45 mM. kcat/Km: the paper PRINTS the kcat/Km cell 360.0 sec^-1 x mM^-1 for this peptide row, equal to 360.0 s^(-1)*mM^(-1) in the export unit; the exported value is 360.0 s^(-1)*mM^(-1).

row_label: Table 2, decoded plain-numeral transcription, peptide row `Z---Phe---Phe--Ala`

table_label: Table 2 (PDF page 4 of 7, journal page 377) 'Kinetics of Z-Gly-X-NH2, Z-Tyr-X-NH2 and Z-Phe-X-Ala'

substrate: Benzyloxycarbonyl-Phe-Phe-Ala


substrate_name: Benzyloxycarbonyl-Phe-Tyr-Ala

substrate: Benzyloxycarbonyl-Phe-Tyr-Ala


substrate_name: Phe-Gly-Leu-Ala

substrate: Phe-Gly-Leu-Ala


substrate_name: Z-Gly-Pro-Leu-Gly-Pro

column_label: Results text, page 378, left column, the sentence 'those of Z-Gly-Pro-Leu-Gly-Pro were 13.3 mM and 41.6, respectively': Km = 13.3 mM and kcat = 41.6 sec^-1 for Z-Gly-Pro-Leu-Gly-Pro; the sentence fixes the order as 'the K , and kcat (sec-l)'; the article prints no kcat/Km value for this peptide.

normalized_value_unit_evidence: Km: stated value 13.3 mM; already in mM, no unit conversion applies; the exported value is 13.3 mM. kcat: stated value 41.6 sec^-1; 1 sec^-1 is 1 s^-1, no unit conversion applies; the exported value is 41.6 s^(-1). kcat/Km: not printed; the quotient of the two stated values is 41.6 / 13.3 = 3.1278195488721803 s^-1 mM^-1, which is the exported value 3.1278195488721803 s^(-1)*mM^(-1).

raw_value_unit_evidence: Results text, page 378, sentence 'those of Z-Gly-Pro-Leu-Gly-Pro were 13.3 mM and 41.6, respectively': Km = 13.3 mM; kcat = 41.6 sec^-1; the article prints no kcat/Km value for this peptide.

row_label: Results sentence on page 378 giving the Km and kcat of Z-Gly-Pro-Leu-Gly-Pro

substrate: Z-Gly-Pro-Leu-Gly-Pro


substrate_name: leucine enkephalin

sequence_variant_note: Resolved canonical sequence from candidate_reconstruction_from_uniprot_P00800_mature_chain_and_pmid_9827548_mutation and applied mutation signature Y157F with inferred residue-numbering offset +232 to reconstruct the assayed variant sequence.

substrate: leucine enkephalin


sequence_variant_note: Resolved canonical sequence from candidate_reconstruction_from_uniprot_P00800_mature_chain_and_pmid_9827548_mutation and applied mutation signature D226A with inferred residue-numbering offset +232 to reconstruct the assayed variant sequence.


enzyme_display_name: tyrosinase

organism: Agaricus bisporus

sequence_source: UniProtKB O42713 FASTA direct lookup, accessed 2026-05-31

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

column_label: Table 4 (Kinetic constants for the monophenolase activity of mushroom tyrosinase) prints one substrate column, Monophenol, followed by four numeric columns whose heads are Vmax^D (uM min^-1), Kcat^M (s^-1), Km^M (mM) and kcat^M/Km^M (s^-1 mM^-1) in that printed order. The cited cell set is Table 4, monophenol row 4-Ethoxyphenol: Kcat^M (s^-1) = 132.0; Km^M (mM) = 0.17; kcat^M/Km^M (s^-1 mM^-1) = 776. The printed uncertainties of that row are Vmax 30.1 +/- 2.2, kcat 132.0 +/- 4.4, Km 0.17 +/- 0.005 and kcat/Km 776 +/- 48.

normalized_value_unit_evidence: kcat: printed Table 4 cell 132.0 s-1; the turnover column of this paper is already per second, so no unit conversion applies and the paper value is 132.0 s^(-1). Km: printed Table 4 cell 0.17 under the millimolar column head, so no unit conversion applies and the paper value is 0.17 mM. kcat/Km: printed Table 4 cell 776 under the s^-1 mM^-1 column head, so no unit conversion applies and the paper value is 776.0 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 4, monophenol row 4-Ethoxyphenol: kcat = 132.0 s-1; Km = 0.17 mM; printed kcat/Km = 776 s-1 mM-1.

row_label: Table 4, monophenol row 4-Ethoxyphenol

table_label: Table 4: Kinetic constants for the monophenolase activity of mushroom tyrosinase (Eur J Biochem 2000, 267(5):1270-1279)

enzyme: tyrosinase


substrate_name: L-Dopa

column_label: Table 2 (Kinetic constants for the diphenolase activity of mushroom tyrosinase) prints one substrate column, o-Diphenol, followed by four numeric columns whose heads are Vmax^D (uM min^-1), kcat^D (s^-1), Km^D (mM) and kcat^D/Km^D (s^-1 mM^-1) in that printed order. The cited cell set is Table 2, o-diphenol row L-Dopa: kcat^D (s^-1) = 107.4; Km^D (mM) = 0.8; kcat^D/Km^D (s^-1 mM^-1) = 134. The printed uncertainties of that row are Vmax 24.5 +/- 1.0, kcat 107.4 +/- 3.1, Km 0.8 +/- 0.03 and kcat/Km 134 +/- 9.

normalized_value_unit_evidence: kcat: printed Table 2 cell 107.4 s-1; the turnover column of this paper is already per second, so no unit conversion applies and the paper value is 107.4 s^(-1). Km: printed Table 2 cell 0.8 under the millimolar column head, so no unit conversion applies and the paper value is 0.8 mM. kcat/Km: printed Table 2 cell 134 under the s^-1 mM^-1 column head, so no unit conversion applies and the paper value is 134.0 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 2, o-diphenol row L-Dopa: kcat = 107.4 s-1; Km = 0.8 mM; printed kcat/Km = 134 s-1 mM-1.

row_label: Table 2, o-diphenol row L-Dopa

table_label: Table 2: Kinetic constants for the diphenolase activity of mushroom tyrosinase (Eur J Biochem 2000, 267(5):1270-1279)

substrate: L-Dopa


substrate_name: L-alpha-Methyldopa

column_label: Table 2 (Kinetic constants for the diphenolase activity of mushroom tyrosinase) prints one substrate column, o-Diphenol, followed by four numeric columns whose heads are Vmax^D (uM min^-1), kcat^D (s^-1), Km^D (mM) and kcat^D/Km^D (s^-1 mM^-1) in that printed order. The cited cell set is Table 2, o-diphenol row L-alpha-Methyldopa: kcat^D (s^-1) = 44.3; Km^D (mM) = 6.8; kcat^D/Km^D (s^-1 mM^-1) = 6.5. The printed uncertainties of that row are Vmax 10.1 +/- 0.5, kcat 44.3 +/- 1.8, Km 6.8 +/- 0.2 and kcat/Km 6.5 +/- 0.5.

normalized_value_unit_evidence: kcat: printed Table 2 cell 44.3 s-1; the turnover column of this paper is already per second, so no unit conversion applies and the paper value is 44.3 s^(-1). Km: printed Table 2 cell 6.8 under the millimolar column head, so no unit conversion applies and the paper value is 6.8 mM. kcat/Km: printed Table 2 cell 6.5 under the s^-1 mM^-1 column head, so no unit conversion applies and the paper value is 6.5 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 2, o-diphenol row L-alpha-Methyldopa: kcat = 44.3 s-1; Km = 6.8 mM; printed kcat/Km = 6.5 s-1 mM-1.

substrate: L-alpha-Methyldopa


substrate_name: phenol

column_label: Table 4 (Kinetic constants for the monophenolase activity of mushroom tyrosinase) prints one substrate column, Monophenol, followed by four numeric columns whose heads are Vmax^D (uM min^-1), Kcat^M (s^-1), Km^M (mM) and kcat^M/Km^M (s^-1 mM^-1) in that printed order. The cited cell set is Table 4, monophenol row Phenol: Kcat^M (s^-1) = 12.7; Km^M (mM) = 0.70; kcat^M/Km^M (s^-1 mM^-1) = 18. The printed uncertainties of that row are Vmax 2.9 +/- 0.1, kcat 12.7 +/- 0.6, Km 0.70 +/- 0.04 and kcat/Km 18 +/- 2.

normalized_value_unit_evidence: kcat: printed Table 4 cell 12.7 s-1; the turnover column of this paper is already per second, so no unit conversion applies and the paper value is 12.7 s^(-1). Km: printed Table 4 cell 0.70 under the millimolar column head, so no unit conversion applies and the paper value is 0.7 mM. kcat/Km: printed Table 4 cell 18 under the s^-1 mM^-1 column head, so no unit conversion applies and the paper value is 18.0 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 4, monophenol row Phenol: kcat = 12.7 s-1; Km = 0.70 mM; printed kcat/Km = 18 s-1 mM-1.

row_label: Table 4, monophenol row Phenol

substrate: phenol


substrate_name: Levisoprenaline

column_label: Table 2 (Kinetic constants for the diphenolase activity of mushroom tyrosinase) prints one substrate column, o-Diphenol, followed by four numeric columns whose heads are Vmax^D (uM min^-1), kcat^D (s^-1), Km^D (mM) and kcat^D/Km^D (s^-1 mM^-1) in that printed order. The cited cell set is Table 2, o-diphenol row L-Isoproterenol: kcat^D (s^-1) = 29.4; Km^D (mM) = 7.1; kcat^D/Km^D (s^-1 mM^-1) = 4. The printed uncertainties of that row are Vmax 6.7 +/- 0.3, kcat 29.4 +/- 1.3, Km 7.1 +/- 0.3 and kcat/Km 4 +/- 0.3.

normalized_value_unit_evidence: kcat: printed Table 2 cell 29.4 s-1; the turnover column of this paper is already per second, so no unit conversion applies and the paper value is 29.4 s^(-1). Km: printed Table 2 cell 7.1 under the millimolar column head, so no unit conversion applies and the paper value is 7.1 mM. kcat/Km: printed Table 2 cell 4 under the s^-1 mM^-1 column head, so no unit conversion applies and the paper value is 4.0 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 2, o-diphenol row L-Isoproterenol: kcat = 29.4 s-1; Km = 7.1 mM; printed kcat/Km = 4 s-1 mM-1.

row_label: Table 2, o-diphenol row L-Isoproterenol

substrate: Levisoprenaline


substrate_name: catechol

assay_conditions_summary: pH 6.8; 298.15 K; cofactor NADH

column_label: Table 2 (Kinetic constants for the diphenolase activity of mushroom tyrosinase) prints one substrate column, o-Diphenol, followed by four numeric columns whose heads are Vmax^D (uM min^-1), kcat^D (s^-1), Km^D (mM) and kcat^D/Km^D (s^-1 mM^-1) in that printed order. The cited cell set is Table 2, o-diphenol row Catechol: kcat^D (s^-1) = 877.6; Km^D (mM) = 0.3; kcat^D/Km^D (s^-1 mM^-1) = 2925. The printed uncertainties of that row are Vmax 200.1 +/- 4.6, kcat 877.6 +/- 20.2, Km 0.3 +/- 0.02 and kcat/Km 2925 +/- 301.

normalized_value_unit_evidence: kcat: printed Table 2 cell 877.6 s-1; the turnover column of this paper is already per second, so no unit conversion applies and the paper value is 877.6 s^(-1). Km: printed Table 2 cell 0.3 under the millimolar column head, so no unit conversion applies and the paper value is 0.3 mM. kcat/Km: printed Table 2 cell 2925 under the s^-1 mM^-1 column head, so no unit conversion applies and the paper value is 2925.0 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 2, o-diphenol row Catechol: kcat = 877.6 s-1; Km = 0.3 mM; printed kcat/Km = 2925 s-1 mM-1.

row_label: Table 2, o-diphenol row Catechol

substrate: catechol


substrate_name: 3,4-Dihydroxyphenyl propionic acid

column_label: Table 2 (Kinetic constants for the diphenolase activity of mushroom tyrosinase) prints one substrate column, o-Diphenol, followed by four numeric columns whose heads are Vmax^D (uM min^-1), kcat^D (s^-1), Km^D (mM) and kcat^D/Km^D (s^-1 mM^-1) in that printed order. The cited cell set is Table 2, o-diphenol row DHPPA: kcat^D (s^-1) = 553.5; Km^D (mM) = 1.89; kcat^D/Km^D (s^-1 mM^-1) = 294. The printed uncertainties of that row are Vmax 126.2 +/- 6.1, kcat 553.5 +/- 25.6, Km 1.89 +/- 0.1 and kcat/Km 294 +/- 29.

normalized_value_unit_evidence: kcat: printed Table 2 cell 553.5 s-1; the turnover column of this paper is already per second, so no unit conversion applies and the paper value is 553.5 s^(-1). Km: printed Table 2 cell 1.89 under the millimolar column head, so no unit conversion applies and the paper value is 1.89 mM. kcat/Km: printed Table 2 cell 294 under the s^-1 mM^-1 column head, so no unit conversion applies and the paper value is 294.0 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 2, o-diphenol row DHPPA: kcat = 553.5 s-1; Km = 1.89 mM; printed kcat/Km = 294 s-1 mM-1.

row_label: Table 2, o-diphenol row DHPPA

substrate: 3,4-Dihydroxyphenyl propionic acid


organism: Priestia megaterium

sequence_source: UniProtKB B2ZB02 canonical FASTA fetched by REST on 2026-05-31.


sequence_source: UniProtKB B2ZB02 canonical sequence fetched by REST on 2026-05-31; V218F reconstructed after confirming residue 218 is V.


sequence_source: UniProtKB B2ZB02 canonical sequence fetched by REST on 2026-05-31; V218G reconstructed after confirming residue 218 is V.


substrate_name: L-tyrosine

substrate: L-tyrosine


substrate_name: L-Adrenaline

sequence_source: UniProtKB direct accession lookup: P02638, reviewed S100B_BOVIN

substrate: L-Adrenaline


sequence_source: UniProtKB direct accession lookup: Q8MIU0, reviewed TYRO_BOVIN


substrate_name: D-Dopa

substrate: D-Dopa


substrate_name: L-Noradrenaline

substrate: L-Noradrenaline


organism: Concholepas concholepas


substrate_name: O2; L-Noradrenaline

substrate: O2; L-Noradrenaline


organism: Homo sapiens

substrate_name: L-Tyrosine; O2

sequence_source: UniProtKB REST lookup https://rest.uniprot.org/uniprotkb/P14679.json on 2026-05-31

substrate: L-Tyrosine; O2


organism: Pholiota nameko

substrate_name: O2; 4-Cresol

sequence_source: UniProtKB REST lookup https://rest.uniprot.org/uniprotkb/A7BHQ9.json on 2026-05-31

substrate: O2; 4-Cresol


substrate_name: tyramine

sequence_source: UniProtKB reviewed canonical sequence O42713 (PPO2_AGABI), direct REST lookup on 2026-05-31

column_label: Table 4 (Kinetic constants for the monophenolase activity of mushroom tyrosinase) prints one substrate column, Monophenol, followed by four numeric columns whose heads are Vmax^D (uM min^-1), Kcat^M (s^-1), Km^M (mM) and kcat^M/Km^M (s^-1 mM^-1) in that printed order. The cited cell set is Table 4, monophenol row Tyramine: Kcat^M (s^-1) = 25.9; Km^M (mM) = 0.51; kcat^M/Km^M (s^-1 mM^-1) = 51. The printed uncertainties of that row are Vmax 5.9 +/- 0.3, kcat 25.9 +/- 1.1, Km 0.51 +/- 0.02 and kcat/Km 51 +/- 4.

normalized_value_unit_evidence: kcat: printed Table 4 cell 25.9 s-1; the turnover column of this paper is already per second, so no unit conversion applies and the paper value is 25.9 s^(-1). Km: printed Table 4 cell 0.51 under the millimolar column head, so no unit conversion applies and the paper value is 0.51 mM. kcat/Km: printed Table 4 cell 51 under the s^-1 mM^-1 column head, so no unit conversion applies and the paper value is 51.0 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 4, monophenol row Tyramine: kcat = 25.9 s-1; Km = 0.51 mM; printed kcat/Km = 51 s-1 mM-1.

row_label: Table 4, monophenol row Tyramine

substrate: tyramine


column_label: Table 2 (Kinetic constants for the diphenolase activity of mushroom tyrosinase) prints one substrate column, o-Diphenol, followed by four numeric columns whose heads are Vmax^D (uM min^-1), kcat^D (s^-1), Km^D (mM) and kcat^D/Km^D (s^-1 mM^-1) in that printed order. The cited cell set is Table 2, o-diphenol row TBC: kcat^D (s^-1) = 641.8; Km^D (mM) = 2.8; kcat^D/Km^D (s^-1 mM^-1) = 229. The printed uncertainties of that row are Vmax 146.35 +/- 6.7, kcat 641.8 +/- 29.5, Km 2.8 +/- 0.3 and kcat/Km 229 +/- 36.

normalized_value_unit_evidence: kcat: printed Table 2 cell 641.8 s-1; the turnover column of this paper is already per second, so no unit conversion applies and the paper value is 641.8 s^(-1). Km: printed Table 2 cell 2.8 under the millimolar column head, so no unit conversion applies and the paper value is 2.8 mM. kcat/Km: printed Table 2 cell 229 under the s^-1 mM^-1 column head, so no unit conversion applies and the paper value is 229.0 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 2, o-diphenol row TBC: kcat = 641.8 s-1; Km = 2.8 mM; printed kcat/Km = 229 s-1 mM-1.

row_label: Table 2, o-diphenol row TBC


substrate_name: Dopamine quinone

substrate: Dopamine quinone


substrate_name: quercetin

substrate: quercetin


substrate_name: L-Tyrosine; O2; L-Dopa

proof_lines: Sigma mushroom tyrosinase (SMT, crude powder, lot 105k7026)

proof_lines: Worthington mushroom tyrosinase (WMT, crude powder, lot 33H6588Q)

substrate: L-Tyrosine; O2; L-Dopa


substrate_name: O2; L-Dopa

sequence_source: UniProtKB reviewed canonical sequence Q00024 (PPO1_AGABI), direct REST lookup on 2026-05-31

substrate: O2; L-Dopa