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)


enzyme_display_name: laccase

organism: Trametes versicolor

substrate_name: O2; Syringic acid

ki_display: Not reported

kcat_over_km_display: Not reported

mutation_signature: Unknown

source_license: CC BY 4.0; 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.

proof_lines: enzyme conjugated with chitosan of 214 kDa

proof_lines: enzyme conjugated with chitosan of 505 kDa

source_db: brenda

enzyme: laccase

substrate: O2; Syringic acid


organism: Geobacillus thermopakistaniensis

substrate_name: Syringaldazine; O2

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

substrate: Syringaldazine; O2


organism: Klebsiella pneumoniae


organism: Kocuria sp.


organism: Kurthia huakuii

substrate_name: 2,6-Dimethoxyphenol; O2

substrate: 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

kcat_display: Not reported

protein_accession_database: UniProt

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.

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.

mutation_type: substitution

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

substrate: L-homocysteine; L-serine

source: brenda


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

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

source_db: oed

verification_status: unverified

kcat_over_km_origin: calculated

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

data_origin: experimental

enzyme: FAD synthase

substrate: FMN

status: unverified

source: oed


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


verification_status: corrected

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'

status: corrected


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


organism: Methanocaldococcus jannaschii

ki_unit: mM

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

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

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_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

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


organism: Unknown

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


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

km_display: Not reported

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


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

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

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_ec_organism_unique and applied mutation signature L155A with inferred residue-numbering offset -30 to reconstruct the assayed variant sequence.


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

verification_status: verified

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

status: verified


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.

enzyme: tyrosinase


substrate_name: L-Dopa

substrate: L-Dopa


substrate_name: L-alpha-Methyldopa

substrate: L-alpha-Methyldopa


substrate_name: phenol

substrate: phenol


substrate_name: Levisoprenaline

substrate: Levisoprenaline


substrate_name: catechol

substrate: catechol


substrate_name: 3,4-Dihydroxyphenyl propionic acid

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

substrate: tyramine


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


substrate_name: L-alpha-Methyltyrosine

substrate: L-alpha-Methyltyrosine


substrate_name: Dopa methyl ester

substrate: Dopa methyl ester


substrate_name: L-Tyrosine

substrate: L-Tyrosine


substrate_name: 4-Hydroxyphenyl propionic acid

substrate: 4-Hydroxyphenyl propionic acid


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


substrate_name: L-Dopa

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

substrate: L-Dopa


organism: Ralstonia solanacearum

substrate_name: D-Tyrosine

sequence_source: UniProtKB Q8Y2J8 canonical sequence fetched by REST on 2026-05-31; N322S reconstructed after confirming canonical residue 322 is N.

substrate: D-Tyrosine


sequence_source: UniProtKB Q8Y2J8 canonical sequence fetched by REST on 2026-05-31 and matched to NCBI Protein NP_518458.


sequence_source: UniProtKB Q8Y2J8 canonical sequence fetched by REST on 2026-05-31; V153A reconstructed after confirming canonical residue 153 is V.


sequence_source: UniProtKB Q8Y2J8 canonical sequence fetched by REST on 2026-05-31; T359M reconstructed after confirming canonical residue 359 is T.

sequence_variant_note: Mutation signature T359M matches the mutant residue at offset +0 and the reference residue at offset +15; variant reconstruction was not applied because multiple plausible residue-numbering offsets are present.


substrate_name: L-tyrosine

substrate: L-tyrosine


sequence_source: UniProtKB Q8Y2J8 canonical sequence cross-checked against NCBI NP_518458.1/WP_011000301.1.


sequence_source: UniProtKB Q8Y2J8 canonical sequence cross-checked against NCBI NP_518458.1/WP_011000301.1; residue 317 is D and was changed to Y during review.

sequence_variant_note: Mutation signature D317Y matches the mutant residue at offset +0 and the reference residue at offset -6; variant reconstruction was not applied because multiple plausible residue-numbering offsets are present.


sequence_source: UniProtKB Q8Y2J8 canonical sequence cross-checked against NCBI NP_518458.1/WP_011000301.1; residue 183 is T and was changed to I during review.

sequence_variant_note: Mutation signature T183I 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_source: UniProtKB Q8Y2J8 canonical sequence cross-checked against NCBI NP_518458.1/WP_011000301.1; residue 185 is F and was changed to Y during review.

sequence_variant_note: Mutation signature F185Y matches the mutant residue at offset +0 and the reference residue at offset +3; variant reconstruction was not applied because multiple plausible residue-numbering offsets are present.


organism: Saccharomyces cerevisiae

substrate_name: O2; L-Adrenaline

substrate: O2; L-Adrenaline


sequence_source: Reconstructed from the direct UniProtKB Q8Y2J8 / NCBI Protein NP_518458.1 canonical FASTA match after confirming residue 330 is L and applying L330V.


sequence_source: Reconstructed from the direct UniProtKB Q8Y2J8 / NCBI Protein NP_518458.1 canonical FASTA match after confirming residue 119 is Y and applying Y119F.


assay_conditions_summary: pH 7; 298.15 K; buffer phosphate

proof_lines: pH and temperature not specified in the publication


organism: Hahella sp. (strain CCB-MM4)

proof_lines: kcat is 6.12 sec(-1) with L-tyrosine as substrate. kcat is 9.94 sec(-1) with L-dopa as substrate.


substrate_name: L-tyrosine (for latent-HcTyr1)

proof_lines: kcat is 0.71 sec(-1) with L-tyrosine as substrate (for latent-HcTyr1). kcat is 21.8 sec(-1) with L-tyrosine as substrate (for cleaved-HcTyr1). kcat is 1.00 sec(-1) with L-dopa as substrate (for latent-HcTyr1). kcat is 51.3 sec(-1) with L-do

substrate: L-tyrosine (for latent-HcTyr1)


organism: Sarcophaga argyrostoma

substrate_name: L-dopa

substrate: L-dopa


substrate_name: D-Dopa

sequence_source: UniProtKB B2ZB02, linked to PMID 19672047

assay_conditions_summary: pH 7; buffer phosphate

proof_lines: in 50 mM potassium phosphate buffer (pH 7.0), with 30% (v/v) DMSO

substrate: D-Dopa


assay_conditions_summary: pH 7; 301.15 K; buffer Tris-HCl

proof_lines: wild type enzyme, in 50 mM Tris-HCl buffer pH 7.0, 0.01 mM CuSO4, 28°C


organism: Prunus persica

substrate_name: O2; (+)-Catechin

proof_lines: membrane-bound (PPO-B) peach polyphenol oxidase without SDS

proof_lines: membrane-bound (PPO-B) peach polyphenol oxidase with 2 mM SDS

proof_lines: soluble (PPO-S) peach polyphenol oxidase without SDS

substrate: O2; (+)-Catechin


enzyme_display_name: glucuronosyltransferase

substrate_name: Scopoletin; UDPglucuronate

sequence_source: UniProtKB P19224 FASTA checked directly; canonical residue H371 matched and was replaced with A for H371A.

proof_lines: SABIO-RK modifier species: glucuronosyltransferase(Enzyme) mutant isoenzyme UGT1A6 H371A

enzyme: glucuronosyltransferase

substrate: Scopoletin; UDPglucuronate


sequence_source: UniProtKB O60656 FASTA checked directly; canonical residue H369 matched and was replaced with A for H369A.

proof_lines: SABIO-RK modifier species: glucuronosyltransferase(Enzyme) mutant isoenzyme UGT1A9 H369A


substrate_name: UDPglucuronate; 4-Methylumbelliferone

sequence_source: UniProtKB P22309 FASTA checked directly; canonical residue P229 matched and was replaced with Q for P229Q.

proof_lines: SABIO-RK modifier species: glucuronosyltransferase(Enzyme) mutant UGT1A1*27 (P229Q)

substrate: UDPglucuronate; 4-Methylumbelliferone


substrate_name: 4-Methylumbelliferone; UDPglucuronate

sequence_source: UniProtKB P35503 FASTA checked directly; requested Y486D did not match canonical residue Q486.

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

proof_lines: SABIO-RK modifier species: glucuronosyltransferase(Enzyme) mutant UGT1A3 (Y486D)

substrate: 4-Methylumbelliferone; UDPglucuronate


sequence_source: UniProtKB P22309 FASTA checked directly; canonical residue Y486 matched and was replaced with D for Y486D.

sequence_variant_note: Mutation signature Y486D 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: glucuronosyltransferase(Enzyme) mutant UGT1A1*7 (Y486D)


sequence_source: UniProtKB P19224 FASTA checked directly; requested Y486D did not match canonical residue H486.

proof_lines: SABIO-RK modifier species: glucuronosyltransferase(Enzyme) mutant UGT1A6 (Y486D)


sequence_source: UniProtKB P22309 FASTA checked directly; canonical residue G71 matched and was replaced with R for G71R.

proof_lines: SABIO-RK modifier species: glucuronosyltransferase(Enzyme) mutant UGT1A1*6 (G71R)


sequence_source: UniProtKB REST lookup for P19224; canonical accession sequence.

proof_lines: pH 7.4, 37°C, recombinant UGT1A6, substrate inhibition model


sequence_source: UniProtKB REST lookup for P19224; canonical sequence with E379A applied after residue check.

proof_lines: SABIO-RK modifier species: glucuronosyltransferase(Enzyme) mutant isoenzyme UGT1A6 E379A


substrate_name: UDPglucuronate; Scopoletin

proof_lines:
isozyme UGT1A10, pH 6.4, 37°C
isozyme UGT1A9, pH 7.4, 37°C
UGT1A9/UGT2B7 heterodimer, pH 7.4, 37°C

substrate: UDPglucuronate; Scopoletin


substrate_name: Androstanediol; UDPglucuronate

proof_lines: isozyme UGT2B7 in COS-1 cell microsomes, pH 7.8, 37°C, in presence with CYP2C9

substrate: Androstanediol; UDPglucuronate


sequence_source: UniProt REST Q9HAW8 canonical sequence fetched 2026-05-31; M59I substitution applied after confirming residue 59 is M.

proof_lines: SABIO-RK modifier species: glucuronosyltransferase(Enzyme) mutant UGT1A10 M59I


sequence_source: UniProt REST Q9HAW8 canonical sequence fetched 2026-05-31; T202I substitution applied after confirming residue 202 is T.

proof_lines: SABIO-RK modifier species: glucuronosyltransferase(Enzyme) mutant UGT1A10 T202I


substrate_name: UDPglucuronate; Aldosterone

proof_lines: SABIO-RK modifier species: glucuronosyltransferase(Enzyme) wildtype allelic variant UGT2B7H268

proof_lines: SABIO-RK modifier species: glucuronosyltransferase(Enzyme) wildtype allelic variant UGT2B7Y268

substrate: UDPglucuronate; Aldosterone


substrate_name: UDPglucuronate; Zidovudine

proof_lines: SABIO-RK modifier species: glucuronosyltransferase(Enzyme) wildtype UGT2B7*2 (polymorphism H268Y)

proof_lines: UGT1A9/UGT2B7(mutant H268Y)-CFP heterodimer, pH 7.4, 37°C

proof_lines: pH 7.4, 37°C, recombinant HA-tagged mutant H268Y dimer

substrate: UDPglucuronate; Zidovudine


substrate_name: Entacapone; UDPglucuronate

sequence_source: UniProtKB:P35503 direct REST lookup

proof_lines:
isozyme UGT1A3, pH 7.4, 37°C
pH 7.4, 37°C, isozyme UGT1A3
pH 7.4, 37°C

substrate: Entacapone; UDPglucuronate


substrate_name: 8-Hydroxyquinoline; UDPglucuronate

sequence_source: UniProtKB:P54855 direct REST lookup

proof_lines:
pH 7.4, 37°C, isozyme UGT2B15
37°C, pH 7.4
isoform UGT2B15, at pH 7.4 and 37°C

substrate: 8-Hydroxyquinoline; UDPglucuronate


substrate_name: Testosterone; UDPglucuronate

sequence_source: UniProtKB:O75795 direct REST lookup

proof_lines:
pH 7.4, 37°C, isozyme UGT2B17
37°C, pH 7.5
37°C, pH 7.4

substrate: Testosterone; UDPglucuronate


sequence_source: UniProtKB:P22310 direct REST lookup

proof_lines: imipramine N-glucuronide formation, 1A4 co-expressed with isoform 1A4

proof_lines: imipramine N-glucuronide formation, 1A4 co-expressed with isoform 1A6


sequence_variant_note: Resolved canonical sequence from UniProtKB P16662 direct accession; current canonical residue 268 is Y and UniProt natural variant feature records Y268H for allele UGT2B7*1, matching the source note for wildtype UGT2B7*1/H268H. and applied mutation signature H268H to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: glucuronosyltransferase(Enzyme) wildtype UGT2B7*1 (polymorphism H268H)


substrate_name: UDPglucuronate; Eugenol

sequence_source: UniProtKB Q9HAW8 direct accession sequence; reviewed human UGT1A10 with EC 2.4.1.17.

proof_lines: isozyme UGT1A10, Michaelis-Menten kinetic, pH 7.4, 37°C

substrate: UDPglucuronate; Eugenol


sequence_source: UniProtKB Q9HAW7 direct accession sequence; reviewed human UGT1A7 with EC 2.4.1.17.

proof_lines: isozyme UGT1A7, Michaelis-Menten kinetic, pH 7.4, 37°C


sequence_source: UniProtKB P22309 direct accession sequence; reviewed human UGT1A1 with EC 2.4.1.17.

proof_lines: recombinant isozyme UGT1A1, pH 7.4, 37°C


substrate_name: UDPglucuronate; Phloretin

sequence_source: UniProtKB Q9HAW9 direct accession sequence; reviewed human UGT1A8 with EC 2.4.1.17.

proof_lines: pH 7.4, temperature not specified in the publication, isozyme UGT1A8

substrate: UDPglucuronate; Phloretin


sequence_source: UniProtKB Q0VAJ1 direct accession sequence; active TrEMBL human UDP-glucuronosyltransferase entry annotated with gene UGT1A4 and EC 2.4.1.17.


sequence_source: UniProtKB Q5DT03 direct accession sequence; active TrEMBL human UDP-glucuronosyltransferase entry annotated with gene UGT1A1 and EC 2.4.1.17.


substrate_name: UDPglucuronate; Clozapine

sequence_source: UniProtKB P22310 direct accession; reviewed human UGT1A4 with EC 2.4.1.17. Direct residue check found canonical L at residue 48 and UniProt records L48V as a natural variant, so the row-specific sequence was reconstructed by substituting V at residue 48.

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: glucuronosyltransferase(Enzyme) mutant isoform UGT1A4 L48V

proof_lines: recombinant mutant isozyme UGT1A4 L48V, pH 8.4, 37°C

substrate: UDPglucuronate; Clozapine


organism: Bellis perennis

sequence_source: UniProtKB Q5NTH0 REST FASTA direct check; accession supplied by OED source snapshot.


sequence_source: UniProtKB Q5NTH0 REST FASTA direct check; accession supplied by SABIO-RK source snapshot.


substrate_name: UDPglucuronate

substrate: UDPglucuronate


organism: Canis familiaris

sequence_source: UniProtKB Q6K1J1 REST FASTA direct check; accession supplied by SABIO-RK source snapshot.


sequence_source: Canonical UniProtKB P19224 checked as context only; not accepted as row sequence because H361A coordinates are unresolved against the current canonical sequence.

proof_lines: SABIO-RK modifier species: glucuronosyltransferase(Enzyme) mutant isoenzyme UGT1A6 H361A


sequence_source: Canonical UniProtKB P19224 checked as context only; not accepted as row sequence because H370Q coordinates are unresolved against the current canonical sequence.

proof_lines: SABIO-RK modifier species: glucuronosyltransferase(Enzyme) mutant isoenzyme UGT1A6 H370Q


sequence_source: Canonical UniProtKB P19224 checked as context only; not accepted as row sequence because H370A coordinates are unresolved against the current canonical sequence.

proof_lines: SABIO-RK modifier species: glucuronosyltransferase(Enzyme) mutant isoenzyme UGT1A6 H370A


proof_lines: SABIO-RK modifier species: glucuronosyltransferase(Enzyme) mutant UGT1A10 (Y486D)


proof_lines: SABIO-RK modifier species: glucuronosyltransferase(Enzyme) mutant UGT1A1*62 (F83L)


substrate_name: Diclofenac; UDPglucuronate

proof_lines: membrane-bound recombinant isoform UGT1A10 from CHO cells, at pH 7.4 and 37°C

proof_lines: membrane-bound recombinant isoform UGT1A10 from HEK-293 cells, at pH 7.4 and 37°C

proof_lines: without phospholipid purified recombinant isoform UGT1A10 from CHO cells, at pH 7.4 and 37°C

substrate: Diclofenac; UDPglucuronate


organism: Macaca mulatta

sequence_source: UniProtKB REST accession lookup for Q95KM4, checked 2026-05-31


organism: Rattus norvegicus

substrate_name: UDPglucuronate; Diclofenac

sequence_source: UniProtKB REST accession lookup for P09875, checked 2026-05-31

substrate: UDPglucuronate; Diclofenac


substrate_name: UDP-alpha-D-glucuronate (when assaying glucuronidation with (1S,2R,4S)-borneol and 4-nitrophenol as substrates)


substrate_name: UDPglucuronate; Bilirubin

sequence_variant_note: Resolved canonical sequence from UniProtKB:P22309 direct sequence; F170del reconstructed from PMID:8226884 and SABIO-RK delta170 note and applied mutation signature Δ170 to reconstruct the assayed variant sequence.

mutation_type: deletion

proof_lines: SABIO-RK modifier species: glucuronosyltransferase(Enzyme) mutant delta170

substrate: UDPglucuronate; Bilirubin


organism: Macaca fascicularis

sequence_source: UniProtKB:Q8WN97 direct accession lookup


sequence_source: UniProtKB:Q9XT55 direct accession lookup


sequence_source: UniProtKB:O97951 direct accession lookup


substrate_name: 17alpha-estradiol/epiestradiol (when assaying glucuronidation at position 3)

proof_lines: Some kinetic parameters were assessed using commercial enzymes, which may represent a mix of both active and inactive protein forms, and therefore modify the kinetic values.


substrate_name: 17beta-estradiol/estradiol (when assaying glucuronidation at position 17)


substrate_name: 17alpha-estradiol/epiestradiol (when assaying glucuronidation at position 17)


substrate_name: calcidiol (when assaying glucuronidation at position 25)


substrate_name: estrone (when assaying glucuronidation at position 3)


substrate_name: 17beta-estradiol/estradiol (when assaying glucuronidation at position 3)


substrate: 17alpha-estradiol/epiestradiol (when assaying glucuronidation at position 3)


substrate_name: (E)-ferulate (when assaying glucuronidation at the phenolic group)


substrate_name: androsterone

substrate: androsterone


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

substrate: 17beta-estradiol/estradiol (when assaying glucuronidation at position 17)


substrate: 17alpha-estradiol/epiestradiol (when assaying glucuronidation at position 17)


substrate_name: UDPglucuronate; Denopamine

substrate: UDPglucuronate; Denopamine


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

substrate: 17beta-estradiol/estradiol (when assaying glucuronidation at position 3)


substrate: estrone (when assaying glucuronidation at position 3)


substrate_name: bilirubin

substrate: bilirubin


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

proof_lines: isoform UGT2B9, at pH 7.4, temperature not specified in the publication


organism: Oryctolagus cuniculus

proof_lines: liver microsomes, substrate denopamine, pH 7.5, 37°C

proof_lines: 1.5 mM morphine added, ethanol-induced enzyme


proof_lines: 17beta-hydroxysteroid UDP-glucuronosyltransferase activity, Wistar strain


substrate_name: lamotrigine

substrate: lamotrigine


substrate_name: propofol

substrate: propofol


substrate_name: Mycophenolic acid

substrate: Mycophenolic acid


substrate_name: sulfinpyrazone

substrate: sulfinpyrazone


substrate_name: retigabine

substrate: retigabine


enzyme_display_name: beta-glucosidase

substrate_name: daidzin

sequence_source: UniProtKB Q9H227 canonical sequence, direct REST/FASTA check 2026-05-31.

enzyme: beta-glucosidase

substrate: daidzin


substrate_name: p-Nitrophenyl-beta-D-glucoside

substrate: p-Nitrophenyl-beta-D-glucoside


substrate_name: p-Nitrophenyl-beta-D-galactoside

substrate: p-Nitrophenyl-beta-D-galactoside


substrate_name: Naringenin 7-O-beta-D-glucoside

substrate: Naringenin 7-O-beta-D-glucoside


substrate_name: Eriodictyol 7-O-beta-D-glucoside

substrate: Eriodictyol 7-O-beta-D-glucoside


substrate_name: Genistein 7-O-beta-D-glucoside

substrate: Genistein 7-O-beta-D-glucoside


sequence_source: UniProtKB Q9H227 canonical sequence, direct REST/FASTA check 2026-05-31; Y308F reconstructed by 1-based coordinate substitution after confirming residue 308 is Y.


sequence_source: UniProtKB REST Q9H227 canonical sequence; local residue check confirmed V168; row-specific V168Y sequence reconstructed by substituting Y at position 168.


substrate_name: p-Nitrophenyl-beta-D-fucopyranoside

substrate: p-Nitrophenyl-beta-D-fucopyranoside


sequence_source: UniProtKB REST Q9H227 canonical sequence; local residue check confirmed Y308; row-specific Y308F sequence reconstructed by substituting F at position 308.


substrate_name: Quercetin 7-O-beta-D-glucoside

sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:Q9H227 and applied mutation signature V168Y to reconstruct the assayed variant sequence.

assay_conditions_summary: pH 6.5; 310.15 K; buffer the substrate at 10-500 uM in 50 mM sodium phosphate

column_label: Table 3 prints one Substrate column and then one column block per enzyme, each block headed K m (uM), k cat (s-1 ) and k cat /K m (mM-1 s-1 ). The cited cell set is the Val168 to Tyr block (V168Y), substrate row Quercetin-7-Glc, where the printed cells are K m = 158, k cat = 0.12 and k cat /K m = 0.75. The printed K m number of this flavonoid glucoside row is micromolar on the paper's own arithmetic, because 158 uM is 0.158 mM and the quotient of the two printed component cells is then 0.76 mM^-1 s^-1, alongside the printed efficiency cell 0.75 mM^-1 s^-1.

normalized_value_unit_evidence: Km: printed Table 3 cell 158 under the head K m (uM) in the Val168 to Tyr block (V168Y), and 158 / 1000 = 0.158 mM. kcat: printed Table 3 cell 0.12 under the head k cat (s-1 ), already per second, so no unit conversion applies and the paper value is 0.12 s^(-1). kcat/Km: printed Table 3 cell 0.75 under the head k cat /K m (mM-1 s-1 ), and 1 mM^-1 s^-1 is 1 s^-1 mM^-1, so no unit conversion applies and the paper value is 0.75 s^(-1)*mM^(-1). Quotient disclosure for this row: 0.12 / 0.158 = 0.7594936708860759 s^-1 mM^-1, alongside the printed efficiency cell 0.75 s^-1 mM^-1.

raw_value_unit_evidence: Table 3, Val168 to Tyr block (V168Y), substrate row Quercetin-7-Glc: K m = 158 uM, k cat = 0.12 s^-1, printed k cat /K m = 0.75 mM^-1 s^-1.

table_label: Table 3: Kinetic parameters of wild-type and mutant hCBGs on pNP-glycosides and flavonoid glucosides (Biochem. J. 2003, 373:41-48)

substrate: Quercetin 7-O-beta-D-glucoside


substrate_name: Luteolin 7-O-beta-D-glucoside

substrate: Luteolin 7-O-beta-D-glucoside


column_label: Table 3 prints one Substrate column and then one column block per enzyme, each block headed K m (uM), k cat (s-1 ) and k cat /K m (mM-1 s-1 ). The cited cell set is the Val168 to Tyr block (V168Y), substrate row Naringenin-7-Glc, where the printed cells are K m = 1240, k cat = 0.33 and k cat /K m = 0.26. The printed K m number of this flavonoid glucoside row is micromolar on the paper's own arithmetic, because 1240 uM is 1.24 mM and the quotient of the two printed component cells is then 0.27 mM^-1 s^-1, alongside the printed efficiency cell 0.26 mM^-1 s^-1.

normalized_value_unit_evidence: Km: printed Table 3 cell 1240 under the head K m (uM) in the Val168 to Tyr block (V168Y), and 1240 / 1000 = 1.24 mM. kcat: printed Table 3 cell 0.33 under the head k cat (s-1 ), already per second, so no unit conversion applies and the paper value is 0.33 s^(-1). kcat/Km: printed Table 3 cell 0.26 under the head k cat /K m (mM-1 s-1 ), and 1 mM^-1 s^-1 is 1 s^-1 mM^-1, so no unit conversion applies and the paper value is 0.26 s^(-1)*mM^(-1). Quotient disclosure for this row: 0.33 / 1.24 = 0.26612903225806456 s^-1 mM^-1, alongside the printed efficiency cell 0.26 s^-1 mM^-1.

raw_value_unit_evidence: Table 3, Val168 to Tyr block (V168Y), substrate row Naringenin-7-Glc: K m = 1240 uM, k cat = 0.33 s^-1, printed k cat /K m = 0.26 mM^-1 s^-1.


organism: Bacteroides ovatus

assay_conditions_summary: pH 6; 310.15 K; buffer and pH optimum

column_label: Extended Data Table ED1; row Bo GH3B / Glc-β-PNP; columns k cat (s -1 ), K m (mM), and same-row kcat/Km quotient; raw kcat 1.74 ± 0.21; raw Km 0.155 ± 0.039.

row_label: Bo GH3B / Glc-β-PNP

table_label: Extended Data Table ED1: Summary of kinetic analyses of XyGUL glycoside hydrolases


organism: Bifidobacterium breve


organism: Caldicellulosiruptor saccharolyticus


organism: Halothermothrix orenii

substrate_name: cellobiose

sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:B8CYA8 and applied mutation signature F417Y to reconstruct the assayed variant sequence.

assay_conditions_summary: pH 7; 343.15 K; buffer the coupled glucose oxidase/peroxidase method in 20 mM HEPES

column_label: Table 2 (Kinetic parameters for beta-glucosidase and beta-galactosidase activities). Column 1 is the enzyme form; columns 2-4 are the D-cellobiose block with heads 'k_cat,app (s^-1)', 'K_m (mM)', 'k_cat,app/K_m (mM^-1 s^-1)'; columns 5-7 are the D-lactose block with heads 'k_cat,app (s^-1)', 'K_m (mM)', 'k_cat,app/K_m (s^-1 mM^-1)'; column 8 is the selectivity ratio. Body row 'F417Y', D-cellobiose block: k_cat,app = 38.7 ± 0.6; K_m = 3.5 ± 0.2; k_cat,app/K_m = 11.1.

normalized_value_unit_evidence: k_cat: printed Table 2 cell 38.7 ± 0.6 in column 'k_cat,app (s^-1)' of the D-cellobiose block; the value is already s^-1, so no conversion is applied and the paper value is 38.7 s^(-1). K_m: printed Table 2 cell 3.5 ± 0.2 in column 'K_m (mM)' of the D-cellobiose block; already mM, no conversion, paper value 3.5 mM. k_cat/K_m: printed Table 2 cell 11.1 in column 'k_cat,app/K_m (mM^-1 s^-1)'; mM^-1 s^-1 and s^-1 mM^-1 are the same unit, already the export unit, so no conversion and the paper value is 11.1 s^(-1)*mM^(-1). Closure check: 38.7 / 3.5 = 11.0571 s^-1 mM^-1, which rounds to the printed efficiency cell 11.1 at the precision the table prints, so the printed ratio and the quotient of the printed components agree.

raw_value_unit_evidence: Table 2: Kinetic parameters for β-glucosidase and β-galactosidase activities (Applied microbiology and biotechnology 2016, 100(8):3533-3543), body row 'F417Y', D-cellobiose block: column 'k_cat,app (s^-1)' prints 38.7 ± 0.6; column 'K_m (mM)' prints 3.5 ± 0.2; column 'k_cat,app/K_m (mM^-1 s^-1)' prints 11.1.

row_label: Table 2, body row 'F417Y' (variant F417Y), D-cellobiose block

substrate: cellobiose


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:B8CYA8 and applied mutation signature Y296F to reconstruct the assayed variant sequence.

column_label: Table 2 (Kinetic parameters for beta-glucosidase and beta-galactosidase activities). Column 1 is the enzyme form; columns 2-4 are the D-cellobiose block with heads 'k_cat,app (s^-1)', 'K_m (mM)', 'k_cat,app/K_m (mM^-1 s^-1)'; columns 5-7 are the D-lactose block with heads 'k_cat,app (s^-1)', 'K_m (mM)', 'k_cat,app/K_m (s^-1 mM^-1)'; column 8 is the selectivity ratio. Body row 'Y296F', D-cellobiose block: k_cat,app = 8.5 ± 0.3; K_m = 20.2 ± 1.6; k_cat,app/K_m = 0.42.

normalized_value_unit_evidence: k_cat: printed Table 2 cell 8.5 ± 0.3 in column 'k_cat,app (s^-1)' of the D-cellobiose block; the value is already s^-1, so no conversion is applied and the paper value is 8.5 s^(-1). K_m: printed Table 2 cell 20.2 ± 1.6 in column 'K_m (mM)' of the D-cellobiose block; already mM, no conversion, paper value 20.2 mM. k_cat/K_m: printed Table 2 cell 0.42 in column 'k_cat,app/K_m (mM^-1 s^-1)'; mM^-1 s^-1 and s^-1 mM^-1 are the same unit, already the export unit, so no conversion and the paper value is 0.42 s^(-1)*mM^(-1). Closure check: 8.5 / 20.2 = 0.420792 s^-1 mM^-1, which rounds to the printed efficiency cell 0.42 at the precision the table prints, so the printed ratio and the quotient of the printed components agree.

raw_value_unit_evidence: Table 2: Kinetic parameters for β-glucosidase and β-galactosidase activities (Applied microbiology and biotechnology 2016, 100(8):3533-3543), body row 'Y296F', D-cellobiose block: column 'k_cat,app (s^-1)' prints 8.5 ± 0.3; column 'K_m (mM)' prints 20.2 ± 1.6; column 'k_cat,app/K_m (mM^-1 s^-1)' prints 0.42.

row_label: Table 2, body row 'Y296F' (variant Y296F), D-cellobiose block


substrate_name: lactose

sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:B8CYA8 and applied mutation signature N222F to reconstruct the assayed variant sequence.

column_label: Table 2 (Kinetic parameters for beta-glucosidase and beta-galactosidase activities). Column 1 is the enzyme form; columns 2-4 are the D-cellobiose block with heads 'k_cat,app (s^-1)', 'K_m (mM)', 'k_cat,app/K_m (mM^-1 s^-1)'; columns 5-7 are the D-lactose block with heads 'k_cat,app (s^-1)', 'K_m (mM)', 'k_cat,app/K_m (s^-1 mM^-1)'; column 8 is the selectivity ratio. Body row 'N222F', D-lactose block: k_cat,app = 1.47 ± 0.05; K_m = 41.3 ± 3.1; k_cat,app/K_m = 0.036.

normalized_value_unit_evidence: k_cat: printed Table 2 cell 1.47 ± 0.05 in column 'k_cat,app (s^-1)' of the D-lactose block; the value is already s^-1, so no conversion is applied and the paper value is 1.47 s^(-1). K_m: printed Table 2 cell 41.3 ± 3.1 in column 'K_m (mM)' of the D-lactose block; already mM, no conversion, paper value 41.3 mM. k_cat/K_m: printed Table 2 cell 0.036 in column 'k_cat,app/K_m (s^-1 mM^-1)'; mM^-1 s^-1 and s^-1 mM^-1 are the same unit, already the export unit, so no conversion and the paper value is 0.036 s^(-1)*mM^(-1). Closure check: 1.47 / 41.3 = 0.0355932 s^-1 mM^-1, which rounds to the printed efficiency cell 0.036 at the precision the table prints, so the printed ratio and the quotient of the printed components agree.

raw_value_unit_evidence: Table 2: Kinetic parameters for β-glucosidase and β-galactosidase activities (Applied microbiology and biotechnology 2016, 100(8):3533-3543), body row 'N222F', D-lactose block: column 'k_cat,app (s^-1)' prints 1.47 ± 0.05; column 'K_m (mM)' prints 41.3 ± 3.1; column 'k_cat,app/K_m (s^-1 mM^-1)' prints 0.036.

row_label: Table 2, body row 'N222F' (variant N222F), D-lactose block

substrate: lactose


organism: Homo sapiens

sequence_source: UniProtKB reviewed accession Q9H227 canonical sequence; worker confirmed canonical residue Y at position 308 and reconstructed Y308F on 2026-05-31.


sequence_source: UniProtKB reviewed accession Q9H227 canonical sequence; worker confirmed canonical residue V at position 168 and reconstructed V168Y on 2026-05-31.


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:Q9H227 and applied mutation signature Y308F to reconstruct the assayed variant sequence.

column_label: Table 3 prints one Substrate column and then one column block per enzyme, each block headed K m (uM), k cat (s-1 ) and k cat /K m (mM-1 s-1 ). The cited cell set is the Tyr308 to Phe block (Y308F), substrate row pNPFuc, where the printed cells are K m = 0.18, k cat = 1.18 and k cat /K m = 6.55. The printed K m number of this pNP-glycoside row is millimolar on the paper's own arithmetic, because reading 0.18 in mM puts the quotient of the two printed component cells at 6.56 mM^-1 s^-1, alongside the printed efficiency cell 6.55 mM^-1 s^-1, while reading it in uM would put that quotient at 6555556 mM^-1 s^-1.

normalized_value_unit_evidence: Km: printed Table 3 cell 0.18 under the head K m (uM) in the Tyr308 to Phe block (Y308F), read in mM as the paper's own printed efficiency cell for this row requires, so no unit conversion applies and the paper value is 0.18 mM. kcat: printed Table 3 cell 1.18 under the head k cat (s-1 ), already per second, so no unit conversion applies and the paper value is 1.18 s^(-1). kcat/Km: printed Table 3 cell 6.55 under the head k cat /K m (mM-1 s-1 ), and 1 mM^-1 s^-1 is 1 s^-1 mM^-1, so no unit conversion applies and the paper value is 6.55 s^(-1)*mM^(-1). Quotient disclosure for this row: 1.18 / 0.18 = 6.555555555555555 s^-1 mM^-1, alongside the printed efficiency cell 6.55 s^-1 mM^-1.

raw_value_unit_evidence: Table 3, Tyr308 to Phe block (Y308F), substrate row pNPFuc: K m = 0.18 mM, k cat = 1.18 s^-1, printed k cat /K m = 6.55 mM^-1 s^-1.


column_label: Table 3 prints one Substrate column and then one column block per enzyme, each block headed K m (uM), k cat (s-1 ) and k cat /K m (mM-1 s-1 ). The cited cell set is the Val168 to Tyr block (V168Y), substrate row Daidzein-7-Glc, where the printed cells are K m = 470, k cat = 0.69 and k cat /K m = 1.46. The printed K m number of this flavonoid glucoside row is micromolar on the paper's own arithmetic, because 470 uM is 0.47 mM and the quotient of the two printed component cells is then 1.47 mM^-1 s^-1, alongside the printed efficiency cell 1.46 mM^-1 s^-1.

normalized_value_unit_evidence: Km: printed Table 3 cell 470 under the head K m (uM) in the Val168 to Tyr block (V168Y), and 470 / 1000 = 0.47 mM. kcat: printed Table 3 cell 0.69 under the head k cat (s-1 ), already per second, so no unit conversion applies and the paper value is 0.69 s^(-1). kcat/Km: printed Table 3 cell 1.46 under the head k cat /K m (mM-1 s-1 ), and 1 mM^-1 s^-1 is 1 s^-1 mM^-1, so no unit conversion applies and the paper value is 1.46 s^(-1)*mM^(-1). Quotient disclosure for this row: 0.69 / 0.47 = 1.4680851063829787 s^-1 mM^-1, alongside the printed efficiency cell 1.46 s^-1 mM^-1.

raw_value_unit_evidence: Table 3, Val168 to Tyr block (V168Y), substrate row Daidzein-7-Glc: K m = 470 uM, k cat = 0.69 s^-1, printed k cat /K m = 1.46 mM^-1 s^-1.


substrate_name: p-Nitrophenyl-beta-L-arabinopyranoside

column_label: Table 3 prints one Substrate column and then one column block per enzyme, each block headed K m (uM), k cat (s-1 ) and k cat /K m (mM-1 s-1 ). The cited cell set is the Val168 to Tyr block (V168Y), substrate row pNPAra, where the printed cells are K m = 0.18, k cat = 0.52 and k cat /K m = 2.96. The printed K m number of this pNP-glycoside row is millimolar on the paper's own arithmetic, because reading 0.18 in mM puts the quotient of the two printed component cells at 2.89 mM^-1 s^-1, alongside the printed efficiency cell 2.96 mM^-1 s^-1, while reading it in uM would put that quotient at 2888889 mM^-1 s^-1.

normalized_value_unit_evidence: Km: printed Table 3 cell 0.18 under the head K m (uM) in the Val168 to Tyr block (V168Y), read in mM as the paper's own printed efficiency cell for this row requires, so no unit conversion applies and the paper value is 0.18 mM. kcat: printed Table 3 cell 0.52 under the head k cat (s-1 ), already per second, so no unit conversion applies and the paper value is 0.52 s^(-1). kcat/Km: printed Table 3 cell 2.96 under the head k cat /K m (mM-1 s-1 ), and 1 mM^-1 s^-1 is 1 s^-1 mM^-1, so no unit conversion applies and the paper value is 2.96 s^(-1)*mM^(-1). Quotient disclosure for this row: 0.52 / 0.18 = 2.8888888888888893 s^-1 mM^-1, alongside the printed efficiency cell 2.96 s^-1 mM^-1.

raw_value_unit_evidence: Table 3, Val168 to Tyr block (V168Y), substrate row pNPAra: K m = 0.18 mM, k cat = 0.52 s^-1, printed k cat /K m = 2.96 mM^-1 s^-1.

substrate: p-Nitrophenyl-beta-L-arabinopyranoside


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:B8CYA8 and applied mutation signature F417S to reconstruct the assayed variant sequence.

column_label: Table 2 (Kinetic parameters for beta-glucosidase and beta-galactosidase activities). Column 1 is the enzyme form; columns 2-4 are the D-cellobiose block with heads 'k_cat,app (s^-1)', 'K_m (mM)', 'k_cat,app/K_m (mM^-1 s^-1)'; columns 5-7 are the D-lactose block with heads 'k_cat,app (s^-1)', 'K_m (mM)', 'k_cat,app/K_m (s^-1 mM^-1)'; column 8 is the selectivity ratio. Body row 'F417S', D-lactose block: k_cat,app = 7.8 ± 0.4; K_m = 23.2 ± 2.7; k_cat,app/K_m = 0.34.

normalized_value_unit_evidence: k_cat: printed Table 2 cell 7.8 ± 0.4 in column 'k_cat,app (s^-1)' of the D-lactose block; the value is already s^-1, so no conversion is applied and the paper value is 7.8 s^(-1). K_m: printed Table 2 cell 23.2 ± 2.7 in column 'K_m (mM)' of the D-lactose block; already mM, no conversion, paper value 23.2 mM. k_cat/K_m: printed Table 2 cell 0.34 in column 'k_cat,app/K_m (s^-1 mM^-1)'; mM^-1 s^-1 and s^-1 mM^-1 are the same unit, already the export unit, so no conversion and the paper value is 0.34 s^(-1)*mM^(-1). Closure check: 7.8 / 23.2 = 0.336207 s^-1 mM^-1, which rounds to the printed efficiency cell 0.34 at the precision the table prints, so the printed ratio and the quotient of the printed components agree.

raw_value_unit_evidence: Table 2: Kinetic parameters for β-glucosidase and β-galactosidase activities (Applied microbiology and biotechnology 2016, 100(8):3533-3543), body row 'F417S', D-lactose block: column 'k_cat,app (s^-1)' prints 7.8 ± 0.4; column 'K_m (mM)' prints 23.2 ± 2.7; column 'k_cat,app/K_m (s^-1 mM^-1)' prints 0.34.

row_label: Table 2, body row 'F417S' (variant F417S), D-lactose block


sequence_source: UniProtKB Q9H227 live REST direct lookup; reviewed Swiss-Prot GBA3_HUMAN canonical sequence.

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


substrate_name: p-Nitrophenyl-beta-D-xylopyranoside

substrate: p-Nitrophenyl-beta-D-xylopyranoside


sequence_source: UniProt Q9H227 canonical sequence reconstructed with Q307N after validating residue Q307

sequence_variant_note: Mutation signature Q307N matches the mutant residue at offset +0 and the reference residue at offset +10; variant reconstruction was not applied because multiple plausible residue-numbering offsets are present.


sequence_source: UniProt Q9H227 canonical sequence reconstructed with W345M after validating residue W345


sequence_source: UniProt Q9H227 canonical sequence reconstructed with W345A after validating residue W345


sequence_source: UniProt Q9H227 canonical sequence reconstructed with M172L after validating residue M172

sequence_variant_note: Mutation signature M172L matches the mutant residue at offset +0 and the reference residue at offset +6; variant reconstruction was not applied because multiple plausible residue-numbering offsets are present.


sequence_variant_note: Resolved canonical sequence from reconstructed_from_uniprot_Q9H227_canonical_sequence and applied mutation signature W345A to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from reconstructed_from_uniprot_Q9H227_canonical_sequence and applied mutation signature M172L to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from reconstructed_from_uniprot_Q9H227_canonical_sequence and applied mutation signature Q307N to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from reconstructed_from_uniprot_Q9H227_canonical_sequence and applied mutation signature W345M to reconstruct the assayed variant sequence.


sequence_source: UniProtKB Q9H227 canonical FASTA checked directly; reconstructed Q307N from canonical Q307


sequence_source: UniProtKB Q9H227 canonical FASTA checked directly; reconstructed W345M from canonical W345

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


sequence_source: UniProtKB Q9H227 canonical FASTA checked directly; reconstructed W345A from canonical W345


organism: Arabidopsis thaliana

assay_conditions_summary: pH 4.5; 310.15 K; buffer 50 mM sodium citrate

column_label: Table label: Table 2 Kinetic parameters of recombinant TGG1, TGG4 and TGG5. Row: TGG1 | pNPG. Columns read: Km (mM) cell 34, Kcat (s^-1) cell 1.2, Kcat/Km (s^-1 mM^-1) cell 0.035. metric: kcat; raw value: 1.2; raw unit: s^-1; normalized value: 1.2 s^(-1); metric: Km; raw value: 34; raw unit: mM; normalized value: 34.0 mM; metric: kcat/Km; raw unit: s^-1 mM^-1; normalized value: 0.03529411764705882 s^-1 mM^-1; assay condition: Recombinant Arabidopsis thaliana myrosinase expressed in Pichia pastoris with a 6xHis-tag; O-b-glucosidase activity assay (Methods 5.6): myrosinase was dissolved in 50 mM sodium citrate buffer, pH 4.5, and p-Nitrophenyl b-D-glucopyranoside (PNPG) was dissolved in 50 mM sodium citrate buffer, pH 4.5; the PNPG solution was incubated in an Eppendorf Thermomixer (37 °C, 350 rpm) for 5 min prior to the experiment; the reaction was terminated with 1 M sodium carbonate and the product read at 405 nm. Export row conditions: pH 4.5; 310.15 K.

normalized_value_unit_evidence: Table 2 row TGG1 | pNPG: kcat 1.2 s^-1 = 1.2 s^(-1) (no conversion needed); Km 34 mM = 34.0 mM (already in mM, no conversion needed); kcat/Km 0.03529411764705882 s^-1 mM^-1, the quotient of this row's own printed kcat and Km cells (1.2 / 34). In the PDF text extraction the micro sign of µmol renders as the letter l (`lmol`) and superscript minus-one exponents are flattened to a control character (U+0002) followed by 1, so the header cell `Kcat (s1)` is the paper's Kcat (s^-1) and the header cell `Kcat/Km (s1 mM1)` is the paper's Kcat/Km (s^-1 mM^-1). The printed Kcat/Km cell 0.035 is already in s^-1 mM^-1, the export's ratio unit.

raw_value_unit_evidence: Table 2 Kinetic parameters of recombinant TGG1, TGG4 and TGG5. | metric header row of Table 2 (as extracted): Enzyme Substrate Km (mM) Vmax (lmol min1 g1) Kcat (s1) Kcat/Km (s1 mM1) | row: TGG1 pNPG 34 ± 0 1.2 ± 0 1.2 ± 0 0.035 | cells read: Km = 34 mM; Vmax = 1.2 ± 0 lmol min1 mg1; Kcat = 1.2 s^-1; Kcat/Km = 0.035 s^-1 mM^-1. Table 2 footnote: The data represent the means and standard deviations of three (sinigrin) or two experiments (pNPG) with values calculated from Hanes plots. Sinigrin concentrations varied from 0.03 to 7.2 mM while p-nitrophenyl b-D-glucopyranoside (pNPG) concentrations varied from 1.5 to 99 mM depending on enzyme.

row_label: TGG1 | pNPG


column_label: Table label: Table 2 Kinetic parameters of recombinant TGG1, TGG4 and TGG5. Row: TGG4 | pNPG. Columns read: Km (mM) cell 28, Kcat (s^-1) cell 7.3, Kcat/Km (s^-1 mM^-1) cell 0.26. metric: kcat; raw value: 7.3; raw unit: s^-1; normalized value: 7.3 s^(-1); metric: Km; raw value: 28; raw unit: mM; normalized value: 28.0 mM; metric: kcat/Km; raw unit: s^-1 mM^-1; normalized value: 0.26071428571428573 s^-1 mM^-1; assay condition: Recombinant Arabidopsis thaliana myrosinase expressed in Pichia pastoris with a 6xHis-tag; O-b-glucosidase activity assay (Methods 5.6): myrosinase was dissolved in 50 mM sodium citrate buffer, pH 4.5, and p-Nitrophenyl b-D-glucopyranoside (PNPG) was dissolved in 50 mM sodium citrate buffer, pH 4.5; the PNPG solution was incubated in an Eppendorf Thermomixer (37 °C, 350 rpm) for 5 min prior to the experiment; the reaction was terminated with 1 M sodium carbonate and the product read at 405 nm. Export row conditions: pH 4.5; 310.15 K.

normalized_value_unit_evidence: Table 2 row TGG4 | pNPG: kcat 7.3 s^-1 = 7.3 s^(-1) (no conversion needed); Km 28 mM = 28.0 mM (already in mM, no conversion needed); kcat/Km 0.26071428571428573 s^-1 mM^-1, the quotient of this row's own printed kcat and Km cells (7.3 / 28). In the PDF text extraction the micro sign of µmol renders as the letter l (`lmol`) and superscript minus-one exponents are flattened to a control character (U+0002) followed by 1, so the header cell `Kcat (s1)` is the paper's Kcat (s^-1) and the header cell `Kcat/Km (s1 mM1)` is the paper's Kcat/Km (s^-1 mM^-1). The printed Kcat/Km cell 0.26 is already in s^-1 mM^-1, the export's ratio unit.

raw_value_unit_evidence: Table 2 Kinetic parameters of recombinant TGG1, TGG4 and TGG5. | metric header row of Table 2 (as extracted): Enzyme Substrate Km (mM) Vmax (lmol min1 g1) Kcat (s1) Kcat/Km (s1 mM1) | row: TGG4 pNPG 28 ± 0 7.6 ± 0 7.3 ± 0 0.26 | cells read: Km = 28 mM; Vmax = 7.6 ± 0 lmol min1 mg1; Kcat = 7.3 s^-1; Kcat/Km = 0.26 s^-1 mM^-1. Table 2 footnote: The data represent the means and standard deviations of three (sinigrin) or two experiments (pNPG) with values calculated from Hanes plots. Sinigrin concentrations varied from 0.03 to 7.2 mM while p-nitrophenyl b-D-glucopyranoside (pNPG) concentrations varied from 1.5 to 99 mM depending on enzyme.

row_label: TGG4 | pNPG