substrate_name: Benzamide

column_label: Table 4 (Steady-state kinetic parameters and 15N and 13C KIE for the reaction catalyzed by Pnc1 with alternate substrates), body row 'benzamide': column 'kcat (s-1)' = 0.0088; column 'Km (uM)' = 25; column 'kcat/Km (M-1 s-1)' = 3.5 x 10^2. The number printed after the plus/minus sign beside the kcat and Km cells of this line is the printed uncertainty and is not an exported metric. The 15N KIE and 13C KIE columns of the same line print ND, so the paper reports no isotope effect for this line.

normalized_value_unit_evidence: kcat: printed Table 4 cell 0.0088 in a column headed kcat (s-1); s-1 is already the canonical s^(-1), so no unit conversion applies and the value is 0.0088 s^(-1). Km: printed Table 4 cell 25 in a column headed Km (uM); 1 uM is 0.001 mM, so 25 / 1000 = 0.025 mM. kcat/Km: printed Table 4 cell 3.5 x 10^2 in a column headed kcat/Km (M-1 s-1); 1 M^-1 s^-1 is 0.001 s^-1 mM^-1, so 3.5 x 10^2 / 1000 = 0.35 s^(-1)*mM^(-1). The exported values are kcat 0.0088 s^(-1), Km 0.025 mM and kcat_over_km 0.352 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 4, body row 'benzamide': kcat = 0.0088 s-1; Km = 25 uM; printed kcat/Km = 3.5 x 10^2 in a column headed kcat/Km (M-1 s-1).

row_label: Table 4, row benzamide

substrate: Benzamide


enzyme_display_name: nicotinamidase

organism: Saccharomyces cerevisiae

substrate_name: nicotinamide

source_db: oed

verification_status: unverified

ki_display: Not reported

protein_accession_database: UniProt

kcat_over_km_origin: calculated

source_license: CC BY 4.0

km_unit: mM

sequence_variant_note: Resolved canonical sequence from UniProtKB:P53184 canonical FASTA plus selected mutation_signature D51N and applied mutation signature D51N to reconstruct the assayed variant sequence.

mutation_type: substitution

data_origin: experimental

enzyme: nicotinamidase

substrate: nicotinamide

status: unverified

source: oed


source_db: brenda

kcat_over_km_display: Not reported

sequence_source: UniProtKB:P53184 canonical FASTA plus selected mutation_signature D51N

proof_lines: Km below 0.0018 mM, mutant enzyme D51N, in 50 mM sodium phosphate (pH 7.5), at 25°C

proof_lines: kcat/Km above 35 (1/sec*mM), mutant enzyme D51N, in 50 mM sodium phosphate (pH 7.5), at 25°C

source: brenda


substrate_name: Pyrazinamide

verification_status: corrected

mutation_signature: Unknown

sequence_source: UniProtKB:P53184 reviewed canonical FASTA direct lookup

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

assay_conditions_summary: pH 7.5; 298.15 K; cofactor NADPH

column_label: Table 4 (Steady-state kinetic parameters and 15N and 13C KIE for the reaction catalyzed by Pnc1 with alternate substrates), body row 'pyrazinamide': column 'kcat (s-1)' = 2.56; column 'Km (uM)' = 157; column 'kcat/Km (M-1 s-1)' = 1.6 x 10^4. The number printed after the plus/minus sign beside the kcat and Km cells of this line is the printed uncertainty and is not an exported metric. The 15N KIE and 13C KIE columns of the same line print 1.0231 and 1.0196, which are isotope effects and not kinetic metric fields the export schema carries.

normalized_value_unit_evidence: kcat: printed Table 4 cell 2.56 in a column headed kcat (s-1); s-1 is already the canonical s^(-1), so no unit conversion applies and the value is 2.56 s^(-1). Km: printed Table 4 cell 157 in a column headed Km (uM); 1 uM is 0.001 mM, so 157 / 1000 = 0.157 mM. kcat/Km: printed Table 4 cell 1.6 x 10^4 in a column headed kcat/Km (M-1 s-1); 1 M^-1 s^-1 is 0.001 s^-1 mM^-1, so 1.6 x 10^4 / 1000 = 16.0 s^(-1)*mM^(-1). The exported values are kcat 2.56 s^(-1), Km 0.157 mM and kcat_over_km 16.305732484076433 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 4, body row 'pyrazinamide': kcat = 2.56 s-1; Km = 157 uM; printed kcat/Km = 1.6 x 10^4 in a column headed kcat/Km (M-1 s-1).

row_label: Table 4, row pyrazinamide

table_label: Table 4: Steady-state kinetic parameters and 15N and 13C KIE for the reaction catalyzed by Pnc1 with alternate substrates (Biochemistry 2012, 51(1):243-256). Printed column heads: Substrate; Structure; kcat (s-1); Km (uM); kcat/Km (M-1 s-1); 15N KIE; 13C KIE.

substrate: Pyrazinamide

status: corrected


km_display: Not reported

sequence_variant_note: Resolved canonical sequence from UniProtKB:P53184 canonical FASTA plus selected mutation_signature K122R and applied mutation signature K122R to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme K122R, in 50 mM sodium phosphate (pH 7.5), at 25°C


sequence_variant_note: Resolved canonical sequence from UniProtKB:P53184 canonical FASTA plus selected mutation_signature D8E and applied mutation signature D8E to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme D8E, in 50 mM sodium phosphate (pH 7.5), at 25°C


column_label: Table 4 (Steady-state kinetic parameters and 15N and 13C KIE for the reaction catalyzed by Pnc1 with alternate substrates), body row '5-methylnicotinamide': column 'kcat (s-1)' = 1.75; column 'Km (uM)' = 61; column 'kcat/Km (M-1 s-1)' = 2.9 x 10^4. The number printed after the plus/minus sign beside the kcat and Km cells of this line is the printed uncertainty and is not an exported metric. The 15N KIE and 13C KIE columns of the same line print 1.0151 and 1.0269, which are isotope effects and not kinetic metric fields the export schema carries.

normalized_value_unit_evidence: kcat: printed Table 4 cell 1.75 in a column headed kcat (s-1); s-1 is already the canonical s^(-1), so no unit conversion applies and the value is 1.75 s^(-1). Km: printed Table 4 cell 61 in a column headed Km (uM); 1 uM is 0.001 mM, so 61 / 1000 = 0.061 mM. kcat/Km: printed Table 4 cell 2.9 x 10^4 in a column headed kcat/Km (M-1 s-1); 1 M^-1 s^-1 is 0.001 s^-1 mM^-1, so 2.9 x 10^4 / 1000 = 29.0 s^(-1)*mM^(-1). The exported values are kcat 1.75 s^(-1), Km 0.061 mM and kcat_over_km 28.688524590163937 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 4, body row '5-methylnicotinamide': kcat = 1.75 s-1; Km = 61 uM; printed kcat/Km = 2.9 x 10^4 in a column headed kcat/Km (M-1 s-1).

row_label: Table 4, row 5-methylnicotinamide


sequence_variant_note: Resolved canonical sequence from UniProtKB:P53184 canonical FASTA plus selected mutation_signature H94A and applied mutation signature H94A to reconstruct the assayed variant sequence.

proof_lines: Km below 0.0027 mM, mutant enzyme H94A, in 50 mM sodium phosphate (pH 7.5), at 25°C

proof_lines: kcat/Km above 7.4 (1/sec*mM), mutant enzyme H94A, in 50 mM sodium phosphate (pH 7.5), at 25°C


sequence_variant_note: Resolved canonical sequence from UniProtKB:P53184 canonical FASTA plus selected mutation_signature K122A and applied mutation signature K122A to reconstruct the assayed variant sequence.

proof_lines: Km below 0.0065 mM, mutant enzyme K122A, in 50 mM sodium phosphate (pH 7.5), at 25°C

proof_lines: kcat/Km above 6.7 (1/sec*mM), mutant enzyme K122A, in 50 mM sodium phosphate (pH 7.5), at 25°C


sequence_variant_note: Resolved canonical sequence from UniProtKB:P53184 canonical FASTA plus selected mutation_signature D8A and applied mutation signature D8A to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme D8A, in 50 mM sodium phosphate (pH 7.5), at 25°C


sequence_variant_note: Resolved canonical sequence from UniProtKB:P53184 canonical FASTA plus selected mutation_signature D8N and applied mutation signature D8N to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme D8N, in 50 mM sodium phosphate (pH 7.5), at 25°C


column_label: Table 4 (Steady-state kinetic parameters and 15N and 13C KIE for the reaction catalyzed by Pnc1 with alternate substrates), body row 'nicotinamide': column 'kcat (s-1)' = 0.69; column 'Km (uM)' = 9.6; column 'kcat/Km (M-1 s-1)' = 7.2 x 10^4. The number printed after the plus/minus sign beside the kcat and Km cells of this line is the printed uncertainty and is not an exported metric. The 15N KIE and 13C KIE columns of the same line print 1.0122 and 1.0125, which are isotope effects and not kinetic metric fields the export schema carries.

normalized_value_unit_evidence: kcat: printed Table 4 cell 0.69 in a column headed kcat (s-1); s-1 is already the canonical s^(-1), so no unit conversion applies and the value is 0.69 s^(-1). Km: printed Table 4 cell 9.6 in a column headed Km (uM); 1 uM is 0.001 mM, so 9.6 / 1000 = 0.0096 mM. kcat/Km: printed Table 4 cell 7.2 x 10^4 in a column headed kcat/Km (M-1 s-1); 1 M^-1 s^-1 is 0.001 s^-1 mM^-1, so 7.2 x 10^4 / 1000 = 72.0 s^(-1)*mM^(-1). The exported values are kcat 0.69 s^(-1), Km 0.0096 mM and kcat_over_km 71.875 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 4, body row 'nicotinamide': kcat = 0.69 s-1; Km = 9.6 uM; printed kcat/Km = 7.2 x 10^4 in a column headed kcat/Km (M-1 s-1).

row_label: Table 4, row nicotinamide


sequence_variant_note: Resolved canonical sequence from UniProtKB:P53184 canonical FASTA plus selected mutation_signature D51A and applied mutation signature D51A to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:P53184 and applied mutation signature H53A to reconstruct the assayed variant sequence.

column_label: Table 3 (Steady-state kinetic parameters of Pnc1 mutants), body row 'H53A': column 'kcat (s-1)' = 0.014; column 'Km (uM)' = 6.5; column 'kcat/Km (M-1 s-1)' = 2.2 x 10^3. The number printed after the plus/minus sign beside the kcat and Km cells of this line is the printed uncertainty and is not an exported metric. The 15N KIE and 13C KIE columns of the same line print ND, so the paper reports no isotope effect for this line.

normalized_value_unit_evidence: kcat: printed Table 3 cell 0.014 in a column headed kcat (s-1); s-1 is already the canonical s^(-1), so no unit conversion applies and the value is 0.014 s^(-1). Km: printed Table 3 cell 6.5 in a column headed Km (uM); 1 uM is 0.001 mM, so 6.5 / 1000 = 0.0065 mM. kcat/Km: printed Table 3 cell 2.2 x 10^3 in a column headed kcat/Km (M-1 s-1); 1 M^-1 s^-1 is 0.001 s^-1 mM^-1, so 2.2 x 10^3 / 1000 = 2.2 s^(-1)*mM^(-1). The exported values are kcat 0.014 s^(-1), Km 0.0065 mM and kcat_over_km 2.153846153846154 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 3, body row 'H53A': kcat = 0.014 s-1; Km = 6.5 uM; printed kcat/Km = 2.2 x 10^3 in a column headed kcat/Km (M-1 s-1).

row_label: Table 3, row H53A

table_label: Table 3: Steady-state kinetic parameters of Pnc1 mutants (Biochemistry 2012, 51(1):243-256). Printed column heads: Pnc1 mutant; kcat (s-1); Km (uM); kcat/Km (M-1 s-1); 15N KIE; 13C KIE.


organism: Oceanobacillus iheyensis

sequence_variant_note: Resolved canonical sequence from UniProtKB Q8ESQ6 canonical FASTA direct lookup on 2026-06-16 with Q96A reconstructed after canonical residue-coordinate validation and applied mutation signature Q96A to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from UniProtKB Q8ESQ6 canonical FASTA direct lookup on 2026-06-16 with Q96K reconstructed after canonical residue-coordinate validation and applied mutation signature Q96K to reconstruct the assayed variant sequence.


substrate_name: ethyl nicotinate

sequence_variant_note: Resolved canonical sequence from UniProtKB Q8ESQ6 canonical FASTA direct lookup on 2026-06-16 with C133A reconstructed after canonical residue-coordinate validation and applied mutation signature C133A to reconstruct the assayed variant sequence.

substrate: ethyl nicotinate


sequence_source: UniProtKB Q8ESQ6 canonical FASTA direct lookup on 2026-06-16 with Q96K reconstructed after canonical residue-coordinate validation


substrate_name: methyl nicotinate

sequence_source: UniProtKB Q8ESQ6 canonical FASTA direct lookup on 2026-06-16 with Q96A reconstructed after canonical residue-coordinate validation

substrate: methyl nicotinate


sequence_source: UniProtKB Q8ESQ6 canonical sequence checked on 2026-06-16, but row-specific variant sequence was not accepted.

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


sequence_source: UniProtKB Q8ESQ6 canonical FASTA direct lookup on 2026-06-16


substrate_name: 5-methyl nicotinamide

substrate: 5-methyl nicotinamide


sequence_variant_note: Resolved canonical sequence from UniProtKB Q8ESQ6 canonical sequence checked on 2026-06-16, but row-specific variant sequence was not accepted. and applied mutation signature Q96K to reconstruct the assayed variant sequence.


sequence_source: UniProtKB Q8ESQ6 canonical FASTA direct lookup on 2026-06-16 with C133A reconstructed after canonical residue-coordinate validation


organism: uncultured Acidobacteria bacterium

substrate_name: methylnicotinate

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

substrate: methylnicotinate


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


substrate_name: ethylnicotinate

substrate: ethylnicotinate


organism: uncultured Acidobacteriota bacterium


substrate_name: Unknown

kcat_display: Not reported

ki_unit: mM

substrate: Unknown


enzyme_display_name: catechol O-methyltransferase

organism: Homo sapiens

substrate_name: dopamine

enzyme: catechol O-methyltransferase

substrate: dopamine


organism: Mycolicibacterium smegmatis

sequence_source: GenBank AF058285.1 CDS protein_id AAC77368.1 from PMID 9811635; UniProt Q9ZHK8 sequence cross-check


sequence_variant_note: Resolved canonical sequence from NCBI Protein BAL58158.1 canonical sequence plus PMID 28750065 H75E site-directed mutagenesis; reconstructed by replacing residue 75 H with E. and applied mutation signature H75E to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:Q8ESQ6 and applied mutation signature Q96K to reconstruct the assayed variant sequence.

assay_conditions_summary: pH 7.3; 310.15 K; buffer 100 mM phosphate; cofactor NADPH

column_label: Table 1: Kinetic parameters of wild-type OiNIC and its mutants. (PLoS ONE 2013, 8(2): e56727); printed column heads Specific activity (U/mg) | Km (mM) | k cat (s-1) | k cat/Km (mM-1 s-1). Table 1, enzyme block Q96K, substrate row 5-methyl-NAM; column Specific activity (U/mg) = 3.96±0.12; column Km (mM) = 0.62; column k cat (s-1) = 2.16; column k cat/Km (mM-1 s-1) = 3.40. captured primary segment: 5-methyl-NAM | Q96K OiNIC | Km 0.62 mM | kcat 2.16 s^-1 | kcat/Km 3.40 mM^-1 s^-1

normalized_value_unit_evidence: Km: printed Table 1 Km cell 0.62 mM; the printed column is already in millimolar, so no conversion is applied and the value is 0.62 mM. kcat: printed Table 1 k cat cell 2.16 s^-1; the printed turnover column is per second, so no conversion is applied and the value is 2.16 s^(-1). kcat/Km: printed Table 1 k cat/Km cell 3.40 mM^-1 s^-1; the printed efficiency column is already per millimolar per second, so no conversion is applied and the value is 3.4 s^(-1)*mM^(-1). Quotient disclosure for this row: 2.16 / 0.62 = 3.483870967741936 s^-1 mM^-1, against the printed efficiency cell 3.4 s^-1 mM^-1.

raw_value_unit_evidence: Table 1, enzyme block Q96K, substrate row 5-methyl-NAM: Km = 0.62 mM; k cat = 2.16 s^-1; k cat/Km = 3.40 mM^-1 s^-1; Specific activity = 3.96±0.12 U/mg.


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

column_label: Table 1: Kinetic parameters of wild-type OiNIC and its mutants. (PLoS ONE 2013, 8(2): e56727); printed column heads Specific activity (U/mg) | Km (mM) | k cat (s-1) | k cat/Km (mM-1 s-1). Table 1, enzyme block Wild-type, substrate row NAM; column Specific activity (U/mg) = 23.3a/25.0 b±0.5; column Km (mM) = 0.26; column k cat (s-1) = 11.65; column k cat/Km (mM-1 s-1) = 43.48. captured primary segment: NAM | wild-type OiNIC | Km 0.26 mM | kcat 11.65 s^-1 | kcat/Km 43.48 mM^-1 s^-1

normalized_value_unit_evidence: Km: printed Table 1 Km cell 0.26 mM; the printed column is already in millimolar, so no conversion is applied and the value is 0.26 mM. kcat: printed Table 1 k cat cell 11.65 s^-1; the printed turnover column is per second, so no conversion is applied and the value is 11.65 s^(-1). kcat/Km: printed Table 1 k cat/Km cell 43.48 mM^-1 s^-1; the printed efficiency column is already per millimolar per second, so no conversion is applied and the value is 43.48 s^(-1)*mM^(-1). Quotient disclosure for this row: 11.65 / 0.26 = 44.80769230769231 s^-1 mM^-1, against the printed efficiency cell 43.48 s^-1 mM^-1.

raw_value_unit_evidence: Table 1, enzyme block Wild-type, substrate row NAM: Km = 0.26 mM; k cat = 11.65 s^-1; k cat/Km = 43.48 mM^-1 s^-1; Specific activity = 23.3a/25.0 b±0.5 U/mg.


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:Q8ESQ6 and applied mutation signature Q96A to reconstruct the assayed variant sequence.

column_label: Table 1: Kinetic parameters of wild-type OiNIC and its mutants. (PLoS ONE 2013, 8(2): e56727); printed column heads Specific activity (U/mg) | Km (mM) | k cat (s-1) | k cat/Km (mM-1 s-1). Table 1, enzyme block Q96A, substrate row PZA; column Specific activity (U/mg) = 1.80±0.2; column Km (mM) = 1.20; column k cat (s-1) = 1.61; column k cat/Km (mM-1 s-1) = 1.34. captured primary segment: PZA | Q96A OiNIC | Km 1.20 mM | kcat 1.61 s^-1 | kcat/Km 1.34 mM^-1 s^-1

normalized_value_unit_evidence: Km: printed Table 1 Km cell 1.20 mM; the printed column is already in millimolar, so no conversion is applied and the value is 1.2 mM. kcat: printed Table 1 k cat cell 1.61 s^-1; the printed turnover column is per second, so no conversion is applied and the value is 1.61 s^(-1). kcat/Km: printed Table 1 k cat/Km cell 1.34 mM^-1 s^-1; the printed efficiency column is already per millimolar per second, so no conversion is applied and the value is 1.34 s^(-1)*mM^(-1). Quotient disclosure for this row: 1.61 / 1.2 = 1.3416666666666668 s^-1 mM^-1, against the printed efficiency cell 1.34 s^-1 mM^-1.

raw_value_unit_evidence: Table 1, enzyme block Q96A, substrate row PZA: Km = 1.20 mM; k cat = 1.61 s^-1; k cat/Km = 1.34 mM^-1 s^-1; Specific activity = 1.80±0.2 U/mg.


column_label: Table 1: Kinetic parameters of wild-type OiNIC and its mutants. (PLoS ONE 2013, 8(2): e56727); printed column heads Specific activity (U/mg) | Km (mM) | k cat (s-1) | k cat/Km (mM-1 s-1). Table 1, enzyme block Wild-type, substrate row Ethylnicotinate; column Specific activity (U/mg) = 0.13±0.01; column Km (mM) = 1.02; column k cat (s-1) = 0.10; column k cat/Km (mM-1 s-1) = 0.10. captured primary segment: Ethylnicotinate | wild-type OiNIC | Km 1.02 mM | kcat 0.10 s^-1 | kcat/Km 0.10 mM^-1 s^-1

normalized_value_unit_evidence: Km: printed Table 1 Km cell 1.02 mM; the printed column is already in millimolar, so no conversion is applied and the value is 1.02 mM. kcat: printed Table 1 k cat cell 0.10 s^-1; the printed turnover column is per second, so no conversion is applied and the value is 0.1 s^(-1). kcat/Km: printed Table 1 k cat/Km cell 0.10 mM^-1 s^-1; the printed efficiency column is already per millimolar per second, so no conversion is applied and the value is 0.1 s^(-1)*mM^(-1). Quotient disclosure for this row: 0.1 / 1.02 = 0.09803921568627451 s^-1 mM^-1, against the printed efficiency cell 0.1 s^-1 mM^-1.

raw_value_unit_evidence: Table 1, enzyme block Wild-type, substrate row Ethylnicotinate: Km = 1.02 mM; k cat = 0.10 s^-1; k cat/Km = 0.10 mM^-1 s^-1; Specific activity = 0.13±0.01 U/mg.


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:Q8ESQ6 and applied mutation signature E65H to reconstruct the assayed variant sequence.

column_label: Table 1: Kinetic parameters of wild-type OiNIC and its mutants. (PLoS ONE 2013, 8(2): e56727); printed column heads Specific activity (U/mg) | Km (mM) | k cat (s-1) | k cat/Km (mM-1 s-1). Table 1, enzyme block E65H, substrate row PZA; column Specific activity (U/mg) = 0.70±0.03; column Km (mM) = 0.82; column k cat (s-1) = 0.43; column k cat/Km (mM-1 s-1) = 0.52. captured primary segment: PZA | E65H OiNIC | Km 0.82 mM | kcat 0.43 s^-1 | kcat/Km 0.52 mM^-1 s^-1

normalized_value_unit_evidence: Km: printed Table 1 Km cell 0.82 mM; the printed column is already in millimolar, so no conversion is applied and the value is 0.82 mM. kcat: printed Table 1 k cat cell 0.43 s^-1; the printed turnover column is per second, so no conversion is applied and the value is 0.43 s^(-1). kcat/Km: printed Table 1 k cat/Km cell 0.52 mM^-1 s^-1; the printed efficiency column is already per millimolar per second, so no conversion is applied and the value is 0.52 s^(-1)*mM^(-1). Quotient disclosure for this row: 0.43 / 0.82 = 0.524390243902439 s^-1 mM^-1, against the printed efficiency cell 0.52 s^-1 mM^-1.

raw_value_unit_evidence: Table 1, enzyme block E65H, substrate row PZA: Km = 0.82 mM; k cat = 0.43 s^-1; k cat/Km = 0.52 mM^-1 s^-1; Specific activity = 0.70±0.03 U/mg.


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:Q8ESQ6 and applied mutation signature C133A to reconstruct the assayed variant sequence.

column_label: Table 1: Kinetic parameters of wild-type OiNIC and its mutants. (PLoS ONE 2013, 8(2): e56727); printed column heads Specific activity (U/mg) | Km (mM) | k cat (s-1) | k cat/Km (mM-1 s-1). Table 1, enzyme block C133A, substrate row PZA; column Specific activity (U/mg) = 6.39±0.2; column Km (mM) = 0.36; column k cat (s-1) = 3.31; column k cat/Km (mM-1 s-1) = 9.19. captured primary segment: PZA | C133A OiNIC | Km 0.36 mM | kcat 3.31 s^-1 | kcat/Km 9.19 mM^-1 s^-1

normalized_value_unit_evidence: Km: printed Table 1 Km cell 0.36 mM; the printed column is already in millimolar, so no conversion is applied and the value is 0.36 mM. kcat: printed Table 1 k cat cell 3.31 s^-1; the printed turnover column is per second, so no conversion is applied and the value is 3.31 s^(-1). kcat/Km: printed Table 1 k cat/Km cell 9.19 mM^-1 s^-1; the printed efficiency column is already per millimolar per second, so no conversion is applied and the value is 9.19 s^(-1)*mM^(-1). Quotient disclosure for this row: 3.31 / 0.36 = 9.194444444444445 s^-1 mM^-1, against the printed efficiency cell 9.19 s^-1 mM^-1.

raw_value_unit_evidence: Table 1, enzyme block C133A, substrate row PZA: Km = 0.36 mM; k cat = 3.31 s^-1; k cat/Km = 9.19 mM^-1 s^-1; Specific activity = 6.39±0.2 U/mg.


column_label: Table 1: Kinetic parameters of wild-type OiNIC and its mutants. (PLoS ONE 2013, 8(2): e56727); printed column heads Specific activity (U/mg) | Km (mM) | k cat (s-1) | k cat/Km (mM-1 s-1). Table 1, enzyme block C133A, substrate row Ethylnicotinate; column Specific activity (U/mg) = 0.34±0.02; column Km (mM) = 0.34; column k cat (s-1) = 0.14; column k cat/Km (mM-1 s-1) = 0.41. captured primary segment: Ethylnicotinate | C133A OiNIC | Km 0.34 mM | kcat 0.14 s^-1 | kcat/Km 0.41 mM^-1 s^-1

normalized_value_unit_evidence: Km: printed Table 1 Km cell 0.34 mM; the printed column is already in millimolar, so no conversion is applied and the value is 0.34 mM. kcat: printed Table 1 k cat cell 0.14 s^-1; the printed turnover column is per second, so no conversion is applied and the value is 0.14 s^(-1). kcat/Km: printed Table 1 k cat/Km cell 0.41 mM^-1 s^-1; the printed efficiency column is already per millimolar per second, so no conversion is applied and the value is 0.41 s^(-1)*mM^(-1). Quotient disclosure for this row: 0.14 / 0.34 = 0.411764705882353 s^-1 mM^-1, against the printed efficiency cell 0.41 s^-1 mM^-1.

raw_value_unit_evidence: Table 1, enzyme block C133A, substrate row Ethylnicotinate: Km = 0.34 mM; k cat = 0.14 s^-1; k cat/Km = 0.41 mM^-1 s^-1; Specific activity = 0.34±0.02 U/mg.


column_label: Table 1: Kinetic parameters of wild-type OiNIC and its mutants. (PLoS ONE 2013, 8(2): e56727); printed column heads Specific activity (U/mg) | Km (mM) | k cat (s-1) | k cat/Km (mM-1 s-1). Table 1, enzyme block Q96K, substrate row Methylnicotinate; column Specific activity (U/mg) = 0.31±0.02; column Km (mM) = 0.44; column k cat (s-1) = 0.17; column k cat/Km (mM-1 s-1) = 0.38. captured primary segment: Methylnicotinate | Q96K OiNIC | Km 0.44 mM | kcat 0.17 s^-1 | kcat/Km 0.38 mM^-1 s^-1

normalized_value_unit_evidence: Km: printed Table 1 Km cell 0.44 mM; the printed column is already in millimolar, so no conversion is applied and the value is 0.44 mM. kcat: printed Table 1 k cat cell 0.17 s^-1; the printed turnover column is per second, so no conversion is applied and the value is 0.17 s^(-1). kcat/Km: printed Table 1 k cat/Km cell 0.38 mM^-1 s^-1; the printed efficiency column is already per millimolar per second, so no conversion is applied and the value is 0.38 s^(-1)*mM^(-1). Quotient disclosure for this row: 0.17 / 0.44 = 0.3863636363636364 s^-1 mM^-1, against the printed efficiency cell 0.38 s^-1 mM^-1.

raw_value_unit_evidence: Table 1, enzyme block Q96K, substrate row Methylnicotinate: Km = 0.44 mM; k cat = 0.17 s^-1; k cat/Km = 0.38 mM^-1 s^-1; Specific activity = 0.31±0.02 U/mg.


column_label: Table 1: Kinetic parameters of wild-type OiNIC and its mutants. (PLoS ONE 2013, 8(2): e56727); printed column heads Specific activity (U/mg) | Km (mM) | k cat (s-1) | k cat/Km (mM-1 s-1). Table 1, enzyme block Q96A, substrate row 5-methyl-NAM; column Specific activity (U/mg) = 28.0±0.1; column Km (mM) = 0.55; column k cat (s-1) = 15.90; column k cat/Km (mM-1 s-1) = 29.04. captured primary segment: 5-methyl-NAM | Q96A OiNIC | Km 0.55 mM | kcat 15.90 s^-1 | kcat/Km 29.04 mM^-1 s^-1

normalized_value_unit_evidence: Km: printed Table 1 Km cell 0.55 mM; the printed column is already in millimolar, so no conversion is applied and the value is 0.55 mM. kcat: printed Table 1 k cat cell 15.90 s^-1; the printed turnover column is per second, so no conversion is applied and the value is 15.9 s^(-1). kcat/Km: printed Table 1 k cat/Km cell 29.04 mM^-1 s^-1; the printed efficiency column is already per millimolar per second, so no conversion is applied and the value is 29.04 s^(-1)*mM^(-1). Quotient disclosure for this row: 15.9 / 0.55 = 28.909090909090907 s^-1 mM^-1, against the printed efficiency cell 29.04 s^-1 mM^-1.

raw_value_unit_evidence: Table 1, enzyme block Q96A, substrate row 5-methyl-NAM: Km = 0.55 mM; k cat = 15.90 s^-1; k cat/Km = 29.04 mM^-1 s^-1; Specific activity = 28.0±0.1 U/mg.


verification_status: verified

column_label: Table 1: Kinetic parameters of wild-type OiNIC and its mutants. (PLoS ONE 2013, 8(2): e56727); printed column heads Specific activity (U/mg) | Km (mM) | k cat (s-1) | k cat/Km (mM-1 s-1). Table 1, enzyme block Wild-type, substrate row 5-methyl-NAM; column Specific activity (U/mg) = 9.01±0.1; column Km (mM) = 0.68; column k cat (s-1) = 5.10; column k cat/Km (mM-1 s-1) = 7.50. captured primary segment: 5-methyl-NAM | wild-type OiNIC | Km 0.68 mM | kcat 5.10 s^-1 | kcat/Km 7.50 mM^-1 s^-1

normalized_value_unit_evidence: Km: printed Table 1 Km cell 0.68 mM; the printed column is already in millimolar, so no conversion is applied and the value is 0.68 mM. kcat: printed Table 1 k cat cell 5.10 s^-1; the printed turnover column is per second, so no conversion is applied and the value is 5.1 s^(-1). kcat/Km: printed Table 1 k cat/Km cell 7.50 mM^-1 s^-1; the printed efficiency column is already per millimolar per second, so no conversion is applied and the value is 7.5 s^(-1)*mM^(-1). Quotient disclosure for this row: 5.1 / 0.68 = 7.499999999999999 s^-1 mM^-1, against the printed efficiency cell 7.5 s^-1 mM^-1.

raw_value_unit_evidence: Table 1, enzyme block Wild-type, substrate row 5-methyl-NAM: Km = 0.68 mM; k cat = 5.10 s^-1; k cat/Km = 7.50 mM^-1 s^-1; Specific activity = 9.01±0.1 U/mg.

status: verified


column_label: Table 1: Kinetic parameters of wild-type OiNIC and its mutants. (PLoS ONE 2013, 8(2): e56727); printed column heads Specific activity (U/mg) | Km (mM) | k cat (s-1) | k cat/Km (mM-1 s-1). Table 1, enzyme block C133A, substrate row 5-methyl-NAM; column Specific activity (U/mg) = 11.74±0.2; column Km (mM) = 0.64; column k cat (s-1) = 7.09; column k cat/Km (mM-1 s-1) = 11.07. captured primary segment: 5-methyl-NAM | C133A OiNIC | Km 0.64 mM | kcat 7.09 s^-1 | kcat/Km 11.07 mM^-1 s^-1

normalized_value_unit_evidence: Km: printed Table 1 Km cell 0.64 mM; the printed column is already in millimolar, so no conversion is applied and the value is 0.64 mM. kcat: printed Table 1 k cat cell 7.09 s^-1; the printed turnover column is per second, so no conversion is applied and the value is 7.09 s^(-1). kcat/Km: printed Table 1 k cat/Km cell 11.07 mM^-1 s^-1; the printed efficiency column is already per millimolar per second, so no conversion is applied and the value is 11.07 s^(-1)*mM^(-1). Quotient disclosure for this row: 7.09 / 0.64 = 11.078125 s^-1 mM^-1, against the printed efficiency cell 11.07 s^-1 mM^-1.

raw_value_unit_evidence: Table 1, enzyme block C133A, substrate row 5-methyl-NAM: Km = 0.64 mM; k cat = 7.09 s^-1; k cat/Km = 11.07 mM^-1 s^-1; Specific activity = 11.74±0.2 U/mg.


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

proof_lines: about 3fold higher than with pyrazinamide as substrate

source_databases_merged: brenda skid


sequence_variant_note: Resolved canonical sequence from UniProtKB:P53184 canonical sequence with selected mutation H53A reconstructed by direct residue check and applied mutation signature H53A to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme H53A, in 50 mM sodium phosphate (pH 7.5), at 25°C


sequence_variant_note: Resolved canonical sequence from UniProtKB:P53184 canonical sequence with selected mutation H94A reconstructed by direct residue check and applied mutation signature H94A to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from UniProtKB:P53184 canonical sequence with selected mutation K122A reconstructed by direct residue check and applied mutation signature K122A to reconstruct the assayed variant sequence.


organism: Rattus norvegicus

substrate_name: S-adenosyl-L-methionine

source_db: uniprot

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

substrate: S-adenosyl-L-methionine

source: uniprot


enzyme_display_name: anhydro-N-acetylmuramic acid kinase

organism: Escherichia coli (strain K12)

substrate_name: anhMurNAc

assay_conditions_summary: cofactor ATP

enzyme: anhydro-N-acetylmuramic acid kinase

substrate: anhMurNAc


enzyme_display_name: glycoprotein-mannosyl O(6)-kinase

substrate_name: ATP

enzyme: glycoprotein-mannosyl O(6)-kinase

substrate: ATP


enzyme_display_name: tRNA uracil 4-sulfurtransferase

enzyme: tRNA uracil 4-sulfurtransferase


enzyme_display_name: keratan sulfotransferase

enzyme: keratan sulfotransferase


enzyme_display_name: (3S)-malyl-CoA thioesterase

organism: Cereibacter sphaeroides (strain ATCC 17023 / DSM 158 / JCM 6121 / CCUG 31486 / LMG 2827 / NBRC 12203 / NCIMB 8253 / ATH 2.4.1.)

substrate_name: (3S)-malyl-CoA (in the presence of Mg(2+) or Mn(2+))

assay_conditions_summary: cofactor CoA

enzyme: (3S)-malyl-CoA thioesterase


organism: Haloarcula marismortui (strain ATCC 43049 / DSM 3752 / JCM 8966 / VKM B-1809)

substrate_name: (S)-malyl-CoA (cleavage reaction)

substrate: (S)-malyl-CoA (cleavage reaction)


enzyme_display_name: hydroxyacylglutathione hydrolase

organism: Arabidopsis thaliana

substrate_name: S-D-lactoylglutathion

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

enzyme: hydroxyacylglutathione hydrolase

substrate: S-D-lactoylglutathion


substrate_name: S-D-lactoylglutathione

substrate: S-D-lactoylglutathione


enzyme_display_name: glutamyl aminopeptidase

organism: Bitis rhinoceros

substrate_name: Glu-AMC (in presence of 1.2 mM Ca(2+))

enzyme: glutamyl aminopeptidase

substrate: Glu-AMC (in presence of 1.2 mM Ca(2+))


substrate_name: nicotinamide


organism: Saccharomyces cerevisiae (strain ATCC 204508 / S288c)


sequence_variant_note: Resolved canonical sequence from Direct UniProt REST FASTA lookup for P53184; C167A reconstructed from residue 167 of the canonical sequence. and applied mutation signature C167A to reconstruct the assayed variant sequence.

proof_lines: kcat below 0.0005 s-1, mutant enzyme C167A, in 50 mM sodium phosphate (pH 7.5), at 25°C


organism: Thermus thermophilus

proof_lines: pH and temperature not specified in the publication


substrate_name: Pyrazinamide


sequence_variant_note: Resolved canonical sequence from UniProt P21964-2 soluble isoform FASTA; mutation positions match selected COMT mutation signature directly and applied mutation signature Y68A to reconstruct the assayed variant sequence.

proof_lines:
mutant enzyme Y68A, at pH 7.5 and 10°C
mutant enzyme Y68A, at pH 7.4 and 37°C
mutant enzyme Y68A, at pH 7.5 and 20°C


sequence_source: UniProt P21964 canonical FASTA direct accession lookup

proof_lines:
wild type enzyme, at pH 7.5 and 5°C
wild type enzyme, at pH 7.5 and 29°C
wild type enzyme, at pH 7.5 and 33°C


sequence_variant_note: Resolved canonical sequence from UniProt P21964-2 soluble isoform FASTA; mutation positions match selected COMT mutation signature directly and applied mutation signature W143F to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from UniProt P21964-2 soluble isoform FASTA; mutation positions match selected COMT mutation signature directly and applied mutation signature W143F/Y68A to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme W143F/Y68A, at pH 7.4 and 37°C


proof_lines: enzyme from liver microsomal fraction, at pH 7.4 and 37°C


substrate_name: epinephrine

sequence_source: UniProt P22734 canonical FASTA direct accession lookup

proof_lines: enzyme from pooled liver S9, at pH 7.4 and 37°C

substrate: epinephrine


organism: Schizosaccharomyces pombe

substrate_name: esculetin

sequence_source: UniProt O42898 canonical FASTA direct accession lookup

proof_lines: pH 7.5, temperature not specified in the publication

substrate: esculetin


organism: Sus scrofa

substrate_name: (-)-norepinephrine

sequence_source: UniProt Q99028 canonical FASTA direct accession lookup

substrate: (-)-norepinephrine


organism: Aspergillus oryzae


organism: Lactobacillus delbrueckii subsp. lactis


organism: Mus musculus

substrate_name: alpha-L-Glu-beta-naphthylamide

sequence_source: UniProt P16406 canonical FASTA direct accession lookup

substrate: alpha-L-Glu-beta-naphthylamide


sequence_variant_note: Resolved canonical sequence from UniProt P16406 canonical FASTA; mutation positions match selected mutation signature directly and applied mutation signature H450F to reconstruct the assayed variant sequence.

proof_lines: pH 7.4, 37°C, mutant enzyme H450F enzyme, in presence of 4 mM Ca2+

proof_lines: pH 7.4, 37°C, mutant enzyme H450F, in presence of 0.025 mM Ca2+

proof_lines: pH 7.4, 37°C, mutant enzyme H450F, 0.025 mM Ca2+


substrate_name: angiotensin II

assay_conditions_summary: pH 7.4; 310.15 K; buffer 50 mM Tris/HCl

proof_lines: pH 7.4, 37°C, mutant enzyme wild-type enzyme N353Q

proof_lines: pH 7.4, 37°C, mutant enzyme wild-type enzyme N353A

substrate: angiotensin II


substrate_name: Unknown

sequence_variant_note: Resolved canonical sequence from UniProt P16406 canonical FASTA; mutation positions match selected mutation signature directly and applied mutation signature N363A to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from UniProt P16406 canonical FASTA; mutation positions match selected mutation signature directly and applied mutation signature N363Q to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from UniProt P16406 canonical FASTA; mutation positions match selected mutation signature directly and applied mutation signature E215D to reconstruct the assayed variant sequence.


enzyme_display_name: (R)-2-methylmalate dehydratase

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

enzyme: (R)-2-methylmalate dehydratase


organism: Candida tropicalis

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

proof_lines:
cosubstrate esculetin
cosubstrate S-adenosyl-L-methionine


organism: Clarias gariepinus

substrate_name: dopamine


organism: Cricetinae

substrate_name: quercetin

substrate: quercetin


organism: Homo sapiens

assay_conditions_summary: pH 7.8; 310.15 K; buffer phosphate

proof_lines:
S-COMT, 0.1 mM S-adenosyl-L-methionine
pH 7.8
S-COMT, 0.25 mM S-adenosyl-L-methionine


proof_lines:
membrane-bound enzyme form
soluble enzyme form


organism: Neotoma albigula

substrate_name: catechol

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

substrate: catechol


organism: Neotoma stephensi


organism: Oryctolagus cuniculus

proof_lines:
soluble enzyme form
membrane-bound enzyme form


organism: Rattus norvegicus

substrate_name: adrenaline

proof_lines:
brain MB-COMT, native
brain MB-COMT, solubilized
S-COMT, recombinant fusion protein

substrate: adrenaline


organism: Solanum lycopersicum


organism: Streptomyces griseus

substrate_name: S-adenosyl-L-methionine


substrate_name: epigallocatechin gallate

substrate: epigallocatechin gallate


sequence_variant_note: Resolved canonical sequence from UniProt REST P16406 (2026-06-16) and applied mutation signature E215Q to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from UniProt REST P16406 (2026-06-16) and applied mutation signature E215A to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from UniProt REST P16406 (2026-06-16) and applied mutation signature T348Y to reconstruct the assayed variant sequence.

assay_conditions_summary: pH 7.4; buffer 50 mM Tris-HCl

proof_lines: recombinant mutant enzyme T348Y, in 50 mM Tris-HCl buffer, pH 7.4, in the absence of Ca2+

proof_lines: recombinant mutant enzyme T348Y, in 50 mM Tris-HCl buffer, pH 7.4, in the presence of 4 mM Ca2+


sequence_variant_note: Resolved canonical sequence from UniProt REST P16406 (2026-06-16) and applied mutation signature T348S to reconstruct the assayed variant sequence.

proof_lines: recombinant mutant enzyme T348S, in 50 mM Tris-HCl buffer, pH 7.4, in the absence of Ca2+

proof_lines: recombinant mutant enzyme T348S, in 50 mM Tris-HCl buffer, pH 7.4, in the presence of 4 mM Ca2+


sequence_variant_note: Resolved canonical sequence from UniProt REST P16406 (2026-06-16) and applied mutation signature T348D to reconstruct the assayed variant sequence.

proof_lines: recombinant mutant enzyme T348D, in 50 mM Tris-HCl buffer, pH 7.4, in the presence of 4 mM Ca2+

proof_lines: recombinant mutant enzyme T348D, in 50 mM Tris-HCl buffer, pH 7.4, in the absence of Ca2+


substrate_name: alpha-L-glutamyl-beta-naphthylamide

sequence_variant_note: Resolved canonical sequence from UniProt REST P16406 (2026-06-16) and applied mutation signature R878K to reconstruct the assayed variant sequence.

proof_lines: pH 7.4, temperature not specified in the publication, recombinant His-tagged mutant R878K, with 4 mM Ca2+

substrate: alpha-L-glutamyl-beta-naphthylamide


sequence_variant_note: Resolved canonical sequence from UniProt REST P16406 (2026-06-16) and applied mutation signature R878A to reconstruct the assayed variant sequence.

proof_lines: pH 7.4, temperature not specified in the publication, recombinant His-tagged mutant R878A, with 4 mM Ca2+


organism: Plasmodium falciparum

proof_lines:
in the absence of cobalt
in the presence of 1 mM cobalt


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

proof_lines: pH and temperature not specified in the publication

substrate: Unknown


organism: Acidilobus saccharovorans

substrate: Pyrazinamide


substrate: nicotinamide


organism: Oceanobacillus iheyensis

substrate_name: methyl nicotinate

sequence_source: UniProt REST Q8ESQ6 (2026-06-16)

column_label: Table 1: Kinetic parameters of wild-type OiNIC and its mutants. (PLoS ONE 2013, 8(2): e56727); printed column heads Specific activity (U/mg) | Km (mM) | k cat (s-1) | k cat/Km (mM-1 s-1). Table 1, enzyme block Wild-type, substrate row Methylnicotinate; column Specific activity (U/mg) = 0.45±0.02; column Km (mM) = 1.03; column k cat (s-1) = 0.32; column k cat/Km (mM-1 s-1) = 0.31. captured primary segment: Methylnicotinate | wild-type OiNIC | Km 1.03 mM | kcat 0.32 s^-1 | kcat/Km 0.31 mM^-1 s^-1

normalized_value_unit_evidence: Km: printed Table 1 Km cell 1.03 mM; the printed column is already in millimolar, so no conversion is applied and the value is 1.03 mM. kcat: printed Table 1 k cat cell 0.32 s^-1; the printed turnover column is per second, so no conversion is applied and the value is 0.32 s^(-1). kcat/Km: printed Table 1 k cat/Km cell 0.31 mM^-1 s^-1; the printed efficiency column is already per millimolar per second, so no conversion is applied and the value is 0.31 s^(-1)*mM^(-1). Quotient disclosure for this row: 0.32 / 1.03 = 0.3106796116504854 s^-1 mM^-1, against the printed efficiency cell 0.31 s^-1 mM^-1.

raw_value_unit_evidence: Table 1, enzyme block Wild-type, substrate row Methylnicotinate: Km = 1.03 mM; k cat = 0.32 s^-1; k cat/Km = 0.31 mM^-1 s^-1; Specific activity = 0.45±0.02 U/mg.

substrate: methyl nicotinate


sequence_source: UniProt REST Q8ESQ6 (2026-06-16); variant sequence unresolved

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

proof_lines: recombinant mutant F68W/C133A, pH 7.3, 37°C


substrate_name: ethyl nicotinate

sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:Q8ESQ6 and applied mutation signature Q96A to reconstruct the assayed variant sequence.

column_label: Table 1: Kinetic parameters of wild-type OiNIC and its mutants. (PLoS ONE 2013, 8(2): e56727); printed column heads Specific activity (U/mg) | Km (mM) | k cat (s-1) | k cat/Km (mM-1 s-1). Table 1, enzyme block Q96A, substrate row Ethylnicotinate; column Specific activity (U/mg) = 0.22±0.02; column Km (mM) = 1.43; column k cat (s-1) = 0.19; column k cat/Km (mM-1 s-1) = 0.13. captured primary segment: Ethylnicotinate | Q96A OiNIC | Km 1.43 mM | kcat 0.19 s^-1 | kcat/Km 0.13 mM^-1 s^-1

normalized_value_unit_evidence: Km: printed Table 1 Km cell 1.43 mM; the printed column is already in millimolar, so no conversion is applied and the value is 1.43 mM. kcat: printed Table 1 k cat cell 0.19 s^-1; the printed turnover column is per second, so no conversion is applied and the value is 0.19 s^(-1). kcat/Km: printed Table 1 k cat/Km cell 0.13 mM^-1 s^-1; the printed efficiency column is already per millimolar per second, so no conversion is applied and the value is 0.13 s^(-1)*mM^(-1). Quotient disclosure for this row: 0.19 / 1.43 = 0.13286713286713286 s^-1 mM^-1, against the printed efficiency cell 0.13 s^-1 mM^-1.

raw_value_unit_evidence: Table 1, enzyme block Q96A, substrate row Ethylnicotinate: Km = 1.43 mM; k cat = 0.19 s^-1; k cat/Km = 0.13 mM^-1 s^-1; Specific activity = 0.22±0.02 U/mg.

substrate: ethyl nicotinate


substrate_name: nicotinamide

sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:Q8ESQ6 and applied mutation signature E65H to reconstruct the assayed variant sequence.

column_label: Table 1: Kinetic parameters of wild-type OiNIC and its mutants. (PLoS ONE 2013, 8(2): e56727); printed column heads Specific activity (U/mg) | Km (mM) | k cat (s-1) | k cat/Km (mM-1 s-1). Table 1, enzyme block E65H, substrate row NAM; column Specific activity (U/mg) = 13.50±0.1; column Km (mM) = 0.13; column k cat (s-1) = 5.63; column k cat/Km (mM-1 s-1) = 42.65. captured primary segment: NAM | E65H OiNIC | Km 0.13 mM | kcat 5.63 s^-1 | kcat/Km 42.65 mM^-1 s^-1

normalized_value_unit_evidence: Km: printed Table 1 Km cell 0.13 mM; the printed column is already in millimolar, so no conversion is applied and the value is 0.13 mM. kcat: printed Table 1 k cat cell 5.63 s^-1; the printed turnover column is per second, so no conversion is applied and the value is 5.63 s^(-1). kcat/Km: printed Table 1 k cat/Km cell 42.65 mM^-1 s^-1; the printed efficiency column is already per millimolar per second, so no conversion is applied and the value is 42.65 s^(-1)*mM^(-1). Quotient disclosure for this row: 5.63 / 0.13 = 43.30769230769231 s^-1 mM^-1, against the printed efficiency cell 42.65 s^-1 mM^-1.

raw_value_unit_evidence: Table 1, enzyme block E65H, substrate row NAM: Km = 0.13 mM; k cat = 5.63 s^-1; k cat/Km = 42.65 mM^-1 s^-1; Specific activity = 13.50±0.1 U/mg.


enzyme_display_name: 1-deoxy-D-xylulose-5-phosphate synthase

organism: Escherichia coli

substrate_name: D-glyceraldehyde 3-phosphate

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

proof_lines: apparent value, mutant enzyme H299A, at pH 8.0 and 25-28°C

enzyme: 1-deoxy-D-xylulose-5-phosphate synthase

substrate: D-glyceraldehyde 3-phosphate


proof_lines: apparent value, mutant enzyme H49N, at pH 8.0 and 25-28°C


proof_lines: apparent value, wild type enzyme, at pH 8.0 and 25-28°C


proof_lines: apparent value, mutant enzyme H49A, at pH 8.0 and 25-28°C


proof_lines: apparent value, mutant enzyme H299N, at pH 8.0 and 25-28°C


organism: Mycobacterium tuberculosis

substrate_name: D-erythrose 4-phosphate

substrate: D-erythrose 4-phosphate


substrate_name: pyruvate

substrate: pyruvate


organism: Plasmodium vivax


organism: Populus tremula x Populus alba


organism: Populus trichocarpa


organism: Streptomyces sp.


organism: Zea mays

proof_lines: kcat is 2.1 sec(-1) for DIBOA-glucoside.


enzyme_display_name: geraniol 8-hydroxylase

organism: Catharanthus roseus

substrate_name: geraniol

proof_lines: kcat is 3.15 min(-1) for naringenin. kcat is 7.86 min(-1) for geraniol.

enzyme: geraniol 8-hydroxylase

substrate: geraniol


enzyme_display_name: 4-aminobenzoate N-oxygenase

organism: Streptomyces thioluteus

substrate_name: p-aminobenzoate

proof_lines: kcat is 6.21 min(-1) with p-aminobenzoate as substrate.

enzyme: 4-aminobenzoate N-oxygenase

substrate: p-aminobenzoate


enzyme_display_name: L-methionine (S)-S-oxide reductase

organism: Escherichia coli (strain K12)

substrate_name: L-methionine (S)-S-oxide

proof_lines: kcat is 3.2 min(-1) for the reduction of Met-S-SO using benzyl viologen as an artificial electron donor.

enzyme: L-methionine (S)-S-oxide reductase

substrate: L-methionine (S)-S-oxide


enzyme_display_name: mitomycin 6-O-methyltransferase

organism: Streptomyces lavendulae

proof_lines: kcat is 0.110 min(-1) for methyltransferase activity with S-adenosyl-L-methionine as substrate. kcat is 0.103 min(-1) for methyltransferase activity with 6-demethylmitomycin A as substrate.

enzyme: mitomycin 6-O-methyltransferase

substrate: S-adenosyl-L-methionine


organism: Homo sapiens

substrate_name: 2-hydroxy-17beta-estradiol (at 37 degrees Celsius)

proof_lines: kcats are 6.8 min(-1), 3.4 min(-1), 3.3 min(-1) and 6.7 min(-1) with 2-hydroxy-17beta-estradiol, 4-hydroxy-17beta-estradiol, 2-hydroxyestrone and 4-hydroxyestrone as substrates, respectively.


organism: Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)

proof_lines: kcat is 0.080 min(-1) with 5OMeBA as substrate. kcat is 0.22 min(-1) with PCA as substrate. kcat is 0.43 min(-1) with CA as substrate.


organism: Salvia splendens

substrate_name: malonyl-CoA (with native enzyme)

proof_lines: kcat is 0.55 sec(-1) (with native enzyme). kcat is 0.76 sec(-1) (with recombinant enzyme).

substrate: malonyl-CoA (with native enzyme)


proof_lines: kcat is 0.4 min(-1) with trehalose-2-sulfate as substrate. kcat is 0.19 min(-1) with palmitoyl-CoA as substrate.


substrate_name: benzyloxycarbonyl-Gln-Gly (for protein-glutamine deamidase activity)

proof_lines: kcat is 11.2 min(-1) with benzyloxycarbonyl-Gln-Gly substrate for protein-glutamine deamidase activity.


organism: Lachnoclostridium phytofermentans (strain ATCC 700394 / DSM 18823 / ISDg)

substrate_name: L-rhamnose

proof_lines: kcat is 16 sec(-1) with L-rhamnose. kcat is 2.5 sec(-1) with beta-D-glucose 1-phosphate.

substrate: L-rhamnose


enzyme_display_name: sym-norspermidine synthase

organism: Pyrococcus furiosus (strain ATCC 43587 / DSM 3638 / JCM 8422 / Vc1)

substrate_name: agmatine

proof_lines: kcat is 2.09 sec(-1) for aminopropyl transferase activity with cadaverine as amine acceptor. kcat is 0.18 sec(-1) for aminopropyl transferase activity with putrescine as amine acceptor. kcat is 0.1 sec(-1) for aminopropyl transferase activi

enzyme: sym-norspermidine synthase

substrate: agmatine


enzyme_display_name: fumigaclavine B O-acetyltransferase

organism: Aspergillus fumigatus

substrate_name: Fumigaclavine B

sequence_source: UniProtKB:Q4WZ64 direct lookup 2026-06-16

enzyme: fumigaclavine B O-acetyltransferase

substrate: Fumigaclavine B


substrate_name: Acetyl-CoA

assay_conditions_summary: pH 7.5; 310.15 K; cofactor CoA

substrate: Acetyl-CoA


enzyme_display_name: protein-serine/threonine phosphatase

organism: Escherichia coli

substrate_name: 4-Nitrophenyl phosphate; H2O

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

sequence_source: UniProtKB:P76395 direct lookup 2026-06-16

enzyme: protein-serine/threonine phosphatase

substrate: 4-Nitrophenyl phosphate; H2O


substrate_name: O-Phospho-tau-protein; H2O

sequence_source: UniProtKB:P53041 direct lookup 2026-06-16

substrate: O-Phospho-tau-protein; H2O


substrate_name: Casein kinase I epsilon phosphorylated; H2O


substrate: Casein kinase I epsilon phosphorylated; H2O


substrate_name: 4-Nitrophenyl phosphate

sequence_source: UniProtKB:O95476 direct lookup 2026-06-16

substrate: 4-Nitrophenyl phosphate


substrate_name: 4-nitrophenyl phosphate

sequence_source: UniProtKB:P62136 direct lookup 2026-06-16

substrate: 4-nitrophenyl phosphate


substrate_name: S6K1p; H2O

substrate: S6K1p; H2O


sequence_source: UniProtKB:O60346 direct lookup 2026-06-16


sequence_source: UniProtKB:Q6ZVD8 direct lookup 2026-06-16


substrate_name: RRApTVA

substrate: RRApTVA


substrate_name: SQLHIpTPGTAY

sequence_source: UniProtKB:Q96HS1 direct REST lookup 2026-06-16

proof_lines: at pH 7.5 and, temperature not specified in the publication

substrate: SQLHIpTPGTAY


proof_lines: recombinant mutant S39E, pH and temperature not specified in the publication


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


proof_lines: recombinant mutant S19A, pH and temperature not specified in the publication


proof_lines: recombinant mutant S19E, pH and temperature not specified in the publication


organism: Listeria monocytogenes

substrate_name: [Myelin basic protein]-serine/threonine phosphate; H2O

sequence_source: UniProtKB:Q8Y678 direct REST lookup 2026-06-16


organism: Mycoplasmopsis synoviae

sequence_source: UniProtKB:Q4A6S8 direct REST lookup 2026-06-16

proof_lines: pH 9.0, 30°C, recombinant wild-type enzyme


substrate_name: RRA(pT)VA

substrate: RRA(pT)VA


organism: Opisthokonta

substrate_name: cdc2k-P; H2O

substrate: cdc2k-P; H2O


organism: Rattus norvegicus

substrate_name: H2O; 4-Nitrophenyl phosphate

sequence_source: UniProtKB:P53042 direct REST lookup 2026-06-16

substrate: H2O; 4-Nitrophenyl phosphate


sequence_source: UniProtKB:P53042 direct REST lookup 2026-06-16; selected from source accession bundle after P10636 checked as tau substrate


organism: Saccharomyces cerevisiae

substrate_name: IERPKpSerFSTEK; H2O

sequence_source: UniProtKB:P38797 direct REST lookup 2026-06-16 plus selected SABIO-RK Nd38 N-terminal deletion note

sequence_variant_note: Mutation context was captured, but the signature could not be translated into an exact residue substitution safely.

mutation_type: truncation

proof_lines: SABIO-RK modifier species: phosphoprotein phosphatase(Enzyme) wildtype Nd38 (N-terminal truncated 1-38)

substrate: IERPKpSerFSTEK; H2O


sequence_variant_note: Resolved canonical sequence from UniProtKB:P38797 direct REST lookup 2026-06-16 plus selected SABIO-RK D109A Nd38 note and applied mutation signature D109A with inferred residue-numbering offset -14 to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: phosphoprotein phosphatase(Enzyme) mutant D109A Nd38 (N-terminal truncated 1-38)


organism: Streptococcus pyogenes

sequence_source: UniProtKB:Q99YM9 direct REST lookup 2026-06-16


organism: Sulfolobus acidocaldarius

substrate_name: ETTYERW(pT)TITQRER

substrate: ETTYERW(pT)TITQRER


substrate_name: 4-nitrophenyl phosphate; RRA(pT)VA

sequence_source: UniProtKB Q4JAD1 REST FASTA checked 2026-06-16

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

substrate: 4-nitrophenyl phosphate; RRA(pT)VA


organism: Thermococcus kodakarensis

sequence_variant_note: Resolved canonical sequence from UniProtKB Q8X270 REST FASTA checked 2026-06-16; reconstructed E132A and applied mutation signature E132A to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from UniProtKB Q8X270 REST FASTA checked 2026-06-16; reconstructed E132Q and applied mutation signature E132Q to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from UniProtKB Q8X270 REST FASTA checked 2026-06-16; reconstructed R124A and applied mutation signature R124A to reconstruct the assayed variant sequence.


sequence_source: UniProtKB Q8X270 REST FASTA checked 2026-06-16; reconstructed E132Q

proof_lines:
pH 5.0, 60°C, mutant E132Q
pH 5.0, 20°C, mutant E132Q


sequence_variant_note: Resolved canonical sequence from UniProtKB Q8X270 REST FASTA checked 2026-06-16; reconstructed D63N and applied mutation signature D63N to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from UniProtKB Q8X270 REST FASTA checked 2026-06-16; reconstructed R124E and applied mutation signature R124E to reconstruct the assayed variant sequence.

proof_lines:
pH 5.0, 60°C, mutant R124E
pH 5.0, 20°C, mutant R124E


sequence_variant_note: Resolved canonical sequence from UniProtKB Q8X270 REST FASTA checked 2026-06-16; reconstructed G95A and applied mutation signature G95A to reconstruct the assayed variant sequence.

proof_lines:
pH 5.0, 60°C, mutant G95A
pH 5.0, 20°C, mutant G95A


sequence_source: UniProtKB Q8X270 REST FASTA checked 2026-06-16; reconstructed R124E


sequence_variant_note: Resolved canonical sequence from UniProtKB Q8X270 REST FASTA checked 2026-06-16; reconstructed E132L and applied mutation signature E132L to reconstruct the assayed variant sequence.


sequence_source: UniProtKB Q8X270 REST FASTA checked 2026-06-16; reconstructed E132A

proof_lines:
pH 5.0, 60°C, mutant E132A
pH 5.0, 20°C, mutant E132A


sequence_variant_note: Resolved canonical sequence from UniProtKB Q8X270 REST FASTA checked 2026-06-16; reconstructed Q136A and applied mutation signature Q136A to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from UniProtKB Q8X270 REST FASTA checked 2026-06-16; reconstructed D63N/E132A and applied mutation signature D63N/E132A to reconstruct the assayed variant sequence.

proof_lines:
pH 5.0, 20°C, mutant D63N/E132A
pH 5.0, 60°C, mutant D63N/E132A


sequence_source: UniProtKB Q8X270 REST FASTA checked 2026-06-16; reconstructed G95A


sequence_source: UniProtKB Q8X270 REST FASTA checked 2026-06-16

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


proof_lines:
pH 5.0, 60°C, wild-type enzyme
pH 5.0, 20°C, wild-type enzyme


substrate_name: p-nitrophenyl phosphate

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

substrate: p-nitrophenyl phosphate


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

substrate_name: MBP; EF-Tu

substrate: MBP; EF-Tu


organism: Mus musculus

substrate_name: a peptide

substrate: a peptide


organism: Oryctolagus cuniculus

substrate_name: phosphorylase

substrate: phosphorylase


substrate_name: MAPT/TAU (at pH 7.4 and 30 degrees Celsius)


organism: Saccharomyces cerevisiae (strain ATCC 204508 / S288c)

substrate_name: p-nitrophenylphosphate

substrate: p-nitrophenylphosphate


substrate_name: phosphorylated CIT1

substrate: phosphorylated CIT1


organism: Trypanosoma brucei brucei (strain 927/4 GUTat10.1)

substrate_name: p-nitrophenylphosphate (at 30 degrees Celsius and at pH 7.4)


enzyme_display_name: cysteate synthase

organism: Capnocytophaga ochracea (strain ATCC 27872 / DSM 7271 / CCUG 9716 / JCM 12966 / NCTC 12371 / SS31 / VPI 2845)

substrate_name: O-phospho-L-serine

sequence_source: source_record; accession cache; direct UniProt REST check 2026-06-16

proof_lines: kcat is 7.4 sec(-1) with O-phospho-L-serine as substrate. kcat is 11.9 sec(-1) with sulfite as substrate.

enzyme: cysteate synthase

substrate: O-phospho-L-serine


enzyme_display_name: thermospermine synthase

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

substrate_name: norspermidine (at pH 9 and 37 degrees Celsius)

assay_conditions_summary: pH 9

proof_lines: kcat is 0.73 sec(-1) for aminopropyl transferase activity with S-adenosylmethioninamine as substrate (at pH 9 and 37 degrees Celsius). kcat is 0.65 sec(-1) for aminopropyl transferase activity with spermidine as substrate (at pH 9 and 37 de

enzyme: thermospermine synthase


organism: Arabidopsis thaliana

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

proof_lines: Experiments have been done in the presence of 100 uM arachidonic acid (AA).

substrate: pNPP (at pH 7.5 and 30 degrees Celsius)


organism: Aspergillus oryzae (strain ATCC 42149 / RIB 40)

substrate_name: p-nitrophenylphosphate (at 30 degrees Celsius and at pH 7)

assay_conditions_summary: pH 7

proof_lines: kcat is 5.4 sec(-1) with p-nitrophenylphosphate as substrate (at 30 degrees Celsius and at pH 7).


organism: Escherichia coli (strain K12)

substrate_name: pNPP

proof_lines: kcat is 0.093 sec(-1) with pNPP as substrate.

substrate: pNPP


organism: Homo sapiens

proof_lines: kcat is 1.52 sec(-1) for the dephosphorylation of pyridoxal 5'-phosphate (at pH 7.4 and 37 degrees Celsius). kcat is 1.25 sec(-1) for the dephosphorylation of pyridoxine 5'-phosphate (at pH 7.4 and 37 degrees Celsius). kcat is 0.45 sec(-1)


substrate_name: para-nitrophenylphosphate (at pH 5.5)

proof_lines: kcat is 0.330 sec(-1) for para-nitrophenylphosphate.

substrate: para-nitrophenylphosphate (at pH 5.5)


substrate_name: BCKDHA

proof_lines: kcat is 17.2 min(-1) for BCKDHA. kcat is 43.2 sec(-1) for p-nitrophenylphosphate.

substrate: BCKDHA


substrate_name: pyridoxal 5'-phosphate

proof_lines: kcat varies between 1.5 and 2.0 sec(-1) for the dephosphorylation of pyridoxal 5'-phosphate (at pH 7.4 and 22 degrees Celsius).

substrate: pyridoxal 5'-phosphate


organism: Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)

substrate_name: P-tyr

proof_lines: kcat is 60.0 sec(-1) with P-Tyr as substrate. kcat is 18.32 sec(-1) with P-Ser as substrate. kcat is 12.5 sec(-1) with P-Thr as substrate. kcat is 159.57 sec(-1) with P-Tyr insulin as substrate. kcat is 158.31 sec(-1) with P-Tyr EGFR as sub

substrate: P-tyr


proof_lines: kcat is 14.2 min(-1) for pNPP hydrolysis.


enzyme_display_name: polynucleotide adenylyltransferase

organism: Bos taurus

substrate_name: ATP

sequence_variant_note: Resolved canonical sequence from UniProtKB P25500 direct REST/FASTA lookup on 2026-06-16 and applied mutation signature R225Y to reconstruct the assayed variant sequence.

assay_conditions_summary: pH 8.3; 310.15 K; cofactor ATP

enzyme: polynucleotide adenylyltransferase

substrate: ATP


sequence_variant_note: Resolved canonical sequence from UniProtKB P25500 direct REST/FASTA lookup on 2026-06-16 and applied mutation signature L213K to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: polynucleotide adenylyltransferase(Enzyme) mutant bPAP-513 L213K


sequence_variant_note: Resolved canonical sequence from UniProtKB P25500 direct REST/FASTA lookup on 2026-06-16 and applied mutation signature W230A to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from UniProtKB P25500 direct REST/FASTA lookup on 2026-06-16 and applied mutation signature H212A to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from UniProtKB P25500 direct REST/FASTA lookup on 2026-06-16 and applied mutation signature N216A to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from UniProtKB P25500 direct REST/FASTA lookup on 2026-06-16 and applied mutation signature D218A to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: polynucleotide adenylyltransferase(Enzyme) mutant bPAP-513 D218A


sequence_variant_note: Resolved canonical sequence from UniProtKB P25500 direct REST/FASTA lookup on 2026-06-16 and applied mutation signature R172G to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: polynucleotide adenylyltransferase(Enzyme) mutant bPAP-513 R172G


sequence_variant_note: Resolved canonical sequence from UniProtKB P25500 direct REST/FASTA lookup on 2026-06-16 and applied mutation signature E209A to reconstruct the assayed variant sequence.


sequence_source: UniProtKB P25500 direct REST/FASTA lookup on 2026-06-16

proof_lines: SABIO-RK modifier species: polynucleotide adenylyltransferase(Enzyme) mutant bPAP-513 N216A


sequence_variant_note: Resolved canonical sequence from UniProtKB P25500 direct REST/FASTA lookup on 2026-06-16 and applied mutation signature R104A to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from UniProtKB P25500 direct REST/FASTA lookup on 2026-06-16 and applied mutation signature Y103G to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from UniProtKB P25500 direct REST/FASTA lookup on 2026-06-16 and applied mutation signature F220S to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: polynucleotide adenylyltransferase(Enzyme) mutant bPAP-513 F220S


sequence_variant_note: Resolved canonical sequence from UniProtKB P25500 direct REST/FASTA lookup on 2026-06-16 and applied mutation signature L105A to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from UniProtKB P25500 direct REST/FASTA lookup on 2026-06-16 and applied mutation signature I217T to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from UniProtKB P25500 direct REST/FASTA lookup on 2026-06-16 and applied mutation signature F220R to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: polynucleotide adenylyltransferase(Enzyme) mutant bPAP-513 F220R


organism: Aspergillus fumigatus (strain ATCC MYA-4609 / CBS 101355 / FGSC A1100 / Af293)

substrate_name: fumigaclavine B

substrate: fumigaclavine B


substrate_name: pNPP (at pH 7.8 and 25 degrees Celsius)

substrate: pNPP (at pH 7.8 and 25 degrees Celsius)


substrate_name: p-nitrophenol phosphate (at pH 6.0)

assay_conditions_summary: pH 6

substrate: p-nitrophenol phosphate (at pH 6.0)


substrate_name: CSNK1E (at 30 degrees Celsius)

substrate: CSNK1E (at 30 degrees Celsius)


substrate_name: p-nitrophenylphosphate


substrate_name: p-nitrophenyl phosphate

substrate: p-nitrophenyl phosphate


sequence_variant_note: Resolved canonical sequence from UniProt REST/FASTA direct accession check; variant sequence reconstructed from canonical accession sequence and applied mutation signature D63A to reconstruct the assayed variant sequence.

proof_lines:
pH 5.0, 60°C, mutant D63A
pH 5.0, 20°C, mutant D63A


sequence_variant_note: Resolved canonical sequence from UniProt REST/FASTA direct accession check; variant sequence reconstructed from canonical accession sequence and applied mutation signature Q136A to reconstruct the assayed variant sequence.

proof_lines:
pH 5.0, 60°C, mutant Q136A
pH 5.0, 20°C, mutant Q136A


sequence_variant_note: Resolved canonical sequence from UniProt REST/FASTA direct accession check; variant sequence reconstructed from canonical accession sequence and applied mutation signature E132L to reconstruct the assayed variant sequence.

proof_lines:
pH 5.0, 60°C, mutant E132L
pH 5.0, 20°C, mutant E132L


sequence_variant_note: Resolved canonical sequence from UniProt REST/FASTA direct accession check; variant sequence reconstructed from canonical accession sequence and applied mutation signature D63N/E132A to reconstruct the assayed variant sequence.


sequence_source: UniProt REST/FASTA direct accession check; variant sequence reconstructed from canonical accession sequence


sequence_variant_note: Resolved canonical sequence from UniProt REST/FASTA direct accession check; variant sequence reconstructed from canonical accession sequence and applied mutation signature D63N to reconstruct the assayed variant sequence.

proof_lines:
pH 5.0, 60°C, mutant D63N
pH 5.0, 20°C, mutant D63N


sequence_variant_note: Resolved canonical sequence from UniProt REST/FASTA direct accession check; variant sequence reconstructed from canonical accession sequence and applied mutation signature R124A to reconstruct the assayed variant sequence.

proof_lines:
pH 5.0, 60°C, mutant R124A
pH 5.0, 20°C, mutant R124A


source_db: skid

source_license: CC BY-NC-ND 4.0

sequence_source: UniProt REST/FASTA direct accession check

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

proof_lines:
Raw SKiD Env: Temp=60°C, pH=5
Raw SKiD Env: Temp=20°C, pH=5

source: skid


enzyme_display_name: nicotinamidase

organism: Oceanobacillus iheyensis

substrate_name: Pyrazinamide

sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:Q8ESQ6 and applied mutation signature E65H to reconstruct the assayed variant sequence.

row_label: Table 1, enzyme block E65H, substrate row PZA

table_label: Table 1: Kinetic parameters of wild-type OiNIC and its mutants. (PLoS ONE 2013, 8(2): e56727); printed column heads Specific activity (U/mg) | Km (mM) | k cat (s-1) | k cat/Km (mM-1 s-1)

enzyme: nicotinamidase

substrate: Pyrazinamide


sequence_variant_note: Resolved canonical sequence from UniProtKB:P25500 displayed sequence; mutation L213K reconstructed from batch signature and applied mutation signature L213K to reconstruct the assayed variant sequence.


substrate_name: ATP; n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n RNA (poly(A))

sequence_variant_note: Resolved canonical sequence from UniProtKB:P25500 residues 1-513; selected source note names bPAP-513; mutation L229R reconstructed from batch signature and applied mutation signature L229R to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: polynucleotide adenylyltransferase(Enzyme) mutant bPAP-513 L229R


substrate_name: n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n RNA (poly(A)); ATP

sequence_variant_note: Resolved canonical sequence from UniProtKB:P25500 residues 1-513; selected source note names bPAP-513; mutation K228A reconstructed from batch signature and applied mutation signature K228A to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: polynucleotide adenylyltransferase(Enzyme) mutant bPAP-513 K228A

proof_lines: SABIO-RK modifier species: polynucleotide adenylyltransferase(Enzyme) mutant K228A


sequence_variant_note: Resolved canonical sequence from UniProtKB:P25500 displayed sequence; mutation K191A reconstructed from batch signature and applied mutation signature K191A to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from UniProtKB:P25500 residues 1-513; selected source note names bPAP-513; mutation W230A reconstructed from batch signature and applied mutation signature W230A to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: polynucleotide adenylyltransferase(Enzyme) mutant bPAP-513 W230A

substrate: ATP; n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n RNA (poly(A))


sequence_variant_note: Resolved canonical sequence from UniProtKB:P25500 residues 1-513; selected source note names bPAP-513; mutation T177M reconstructed from batch signature and applied mutation signature T177M to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: polynucleotide adenylyltransferase(Enzyme) mutant bPAP-513 T177M

substrate: n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n RNA (poly(A)); ATP


sequence_variant_note: Resolved canonical sequence from UniProtKB:P25500 displayed sequence; mutation S102A reconstructed from batch signature and applied mutation signature S102A to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from UniProtKB:P25500 displayed sequence; mutation L193M reconstructed from batch signature and applied mutation signature L193M to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from UniProtKB:P25500 residues 1-513; selected source note names bPAP-513; mutation L105A reconstructed from batch signature and applied mutation signature L105A to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: polynucleotide adenylyltransferase(Enzyme) mutant bPAP-513 L105A


sequence_variant_note: Resolved canonical sequence from UniProtKB:P25500 residues 1-513; selected source note names bPAP-513; mutation D208A reconstructed from batch signature and applied mutation signature D208A to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: polynucleotide adenylyltransferase(Enzyme) mutant bPAP-513 D208A


sequence_variant_note: Resolved canonical sequence from UniProtKB:P25500 residues 1-513; selected source note names bPAP-513; mutation Y103G reconstructed from batch signature and applied mutation signature Y103G to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: polynucleotide adenylyltransferase(Enzyme) mutant bPAP-513 Y103G


sequence_variant_note: Resolved canonical sequence from UniProtKB:P25500 residues 1-513; selected source note names bPAP-513; mutation S102A reconstructed from batch signature and applied mutation signature S102A to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: polynucleotide adenylyltransferase(Enzyme) mutant bPAP-513 S102A


sequence_variant_note: Resolved canonical sequence from UniProtKB:P25500 residues 1-513; selected source note names bPAP-513; mutation Y237A reconstructed from batch signature and applied mutation signature Y237A to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: polynucleotide adenylyltransferase(Enzyme) mutant bPAP-513 Y237A

proof_lines: SABIO-RK modifier species: polynucleotide adenylyltransferase(Enzyme) mutant Y237A


sequence_variant_note: Resolved canonical sequence from UniProtKB:P25500 residues 1-513; selected source note names bPAP-513; mutation R225Y reconstructed from batch signature and applied mutation signature R225Y to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: polynucleotide adenylyltransferase(Enzyme) mutant bPAP-513 R225Y


sequence_variant_note: Resolved canonical sequence from UniProtKB:P25500 residues 1-513; selected source note names bPAP-513; mutation I217T reconstructed from batch signature and applied mutation signature I217T to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: polynucleotide adenylyltransferase(Enzyme) mutant bPAP-513 I217T


sequence_variant_note: Resolved canonical sequence from UniProtKB:P25500 displayed sequence; mutation G203H reconstructed from batch signature and applied mutation signature G203H to reconstruct the assayed 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: Sequence is treated as wild-type because the source explicitly marked the record as wild-type.


substrate_name: ATP; UTP

substrate: ATP; UTP


organism: Semliki forest virus

substrate_name: nsP2 RNA triphosphatase activity (at pH 8.0); nsP2 NTPase activity (at pH 7.5)

assay_conditions_summary: pH 8


organism: Venezuelan equine encephalitis virus (strain 3880)

substrate_name: a 12 residues substrate for nsp2 protease activity


organism: Venezuelan equine encephalitis virus (strain Trinidad donkey)

substrate: a 12 residues substrate for nsp2 protease activity


enzyme_display_name: phosphite dehydrogenase

organism: Pseudomonas stutzeri

substrate_name: NAD+; H2O; Phosphonate

sequence_variant_note: Resolved canonical sequence from UniProt O69054 FASTA plus direct residue-coordinate check and applied mutation signature K76A to reconstruct the assayed variant sequence.

assay_conditions_summary: cofactor NAD+

proof_lines: SABIO-RK modifier species: phosphonate dehydrogenase(Enzyme) mutant Lys76Ala (N-terminal His6 tag)

enzyme: phosphite dehydrogenase

substrate: NAD+; H2O; Phosphonate


substrate_name: Phosphonate; H2O; NAD+

sequence_variant_note: Resolved canonical sequence from UniProt O69054 FASTA plus direct residue-coordinate check and applied mutation signature K76C to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: phosphonate dehydrogenase(Enzyme) mutant Lys76Cys (N-terminal His6 tag)

substrate: Phosphonate; H2O; NAD+


sequence_variant_note: Resolved canonical sequence from UniProt O69054 FASTA plus direct residue-coordinate check and applied mutation signature R237K to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: phosphonate dehydrogenase(Enzyme) mutant Arg237Lys (N-terminal His6 tag)


substrate_name: H2O; NAD+; Phosphonate

sequence_variant_note: Resolved canonical sequence from UniProt O69054 FASTA plus direct residue-coordinate check and applied mutation signature K76M to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: phosphonate dehydrogenase(Enzyme) mutant Lys76Met (N-terminal His6 tag)

substrate: H2O; NAD+; Phosphonate


substrate_name: phosphonate

sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:O69054 and applied mutation signature S295A to reconstruct the assayed variant sequence.

smiles: O=[PH](O)O

assay_conditions_summary: pH 7.3; 298.15 K; cofactor NADH

column_label: Table 2 (Steady State Parameters of 17X-PTDH Mutants), enzyme-form row 'S295A'; column 'kcat (s-1)' = 2.53; column 'Km,PT (mM)' = 0.103; column 'Km,NAD (mM)' = 0.250; column 'Kia,NAD (mM)' = nd; column 'kcat/Km,PT (M-1 s-1)' = 2.46 x 10^4; column 'Dkcat' = nd; column 'D(kcat/Km,PT)' = nd.

normalized_value_unit_evidence: kcat: printed Table 2 cell 2.53 s^-1; already in s^-1, no conversion; the paper value is 2.53 s^(-1). Km: printed Table 2 K_m,PT cell 0.103 mM; already in mM, no conversion; the paper value is 0.103 mM. kcat/Km: printed Table 2 K_cat/K_m,PT cell 2.46 x 10^4 M^-1 s^-1; 1 M^-1 s^-1 is 0.001 s^-1 mM^-1, so 24600 / 1000 = 24.6 s^-1 mM^-1.

raw_value_unit_evidence: Table 2, enzyme-form row 'S295A': k_cat = 2.53 s^-1 (column 'kcat (s-1)'); K_m,PT = 0.103 mM (column 'Km,PT (mM)'); printed k_cat/K_m,PT = 2.46 x 10^4 M^-1 s^-1 (column 'kcat/Km,PT (M-1 s-1)').

row_label: Table 2, enzyme-form row 'S295A'

table_label: Table 2: Steady State Parameters of 17X-PTDH Mutants (Biochemistry 2012, 51(21):4254-4262)

substrate: phosphonate


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:O69054 and applied mutation signature R301K to reconstruct the assayed variant sequence.

column_label: Table 2 (Steady State Parameters of 17X-PTDH Mutants), enzyme-form row 'R301K'; column 'kcat (s-1)' = 4.50; column 'Km,PT (mM)' = 1.11; column 'Km,NAD (mM)' = 0.074; column 'Kia,NAD (mM)' = 0.97; column 'kcat/Km,PT (M-1 s-1)' = 4050; column 'Dkcat' = 1.90; column 'D(kcat/Km,PT)' = 2.19.

normalized_value_unit_evidence: kcat: printed Table 2 cell 4.50 s^-1; already in s^-1, no conversion; the paper value is 4.5 s^(-1). Km: printed Table 2 K_m,PT cell 1.11 mM; already in mM, no conversion; the paper value is 1.11 mM. kcat/Km: printed Table 2 K_cat/K_m,PT cell 4050 M^-1 s^-1; 1 M^-1 s^-1 is 0.001 s^-1 mM^-1, so 4050 / 1000 = 4.05 s^-1 mM^-1.

raw_value_unit_evidence: Table 2, enzyme-form row 'R301K': k_cat = 4.50 s^-1 (column 'kcat (s-1)'); K_m,PT = 1.11 mM (column 'Km,PT (mM)'); printed k_cat/K_m,PT = 4050 M^-1 s^-1 (column 'kcat/Km,PT (M-1 s-1)').

row_label: Table 2, enzyme-form row 'R301K'


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:O69054 and applied mutation signature Y139F to reconstruct the assayed variant sequence.

column_label: Table 2 (Steady State Parameters of 17X-PTDH Mutants), enzyme-form row 'Y139F'; column 'kcat (s-1)' = 3.68; column 'Km,PT (mM)' = 0.039; column 'Km,NAD (mM)' = 0.036; column 'Kia,NAD (mM)' = nd; column 'kcat/Km,PT (M-1 s-1)' = 9.44 x 10^4; column 'Dkcat' = nd; column 'D(kcat/Km,PT)' = nd.

normalized_value_unit_evidence: kcat: printed Table 2 cell 3.68 s^-1; already in s^-1, no conversion; the paper value is 3.68 s^(-1). Km: printed Table 2 K_m,PT cell 0.039 mM; already in mM, no conversion; the paper value is 0.039 mM. kcat/Km: printed Table 2 K_cat/K_m,PT cell 9.44 x 10^4 M^-1 s^-1; 1 M^-1 s^-1 is 0.001 s^-1 mM^-1, so 94400 / 1000 = 94.4 s^-1 mM^-1.

raw_value_unit_evidence: Table 2, enzyme-form row 'Y139F': k_cat = 3.68 s^-1 (column 'kcat (s-1)'); K_m,PT = 0.039 mM (column 'Km,PT (mM)'); printed k_cat/K_m,PT = 9.44 x 10^4 M^-1 s^-1 (column 'kcat/Km,PT (M-1 s-1)').

row_label: Table 2, enzyme-form row 'Y139F'


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:O69054 and applied mutation signature W134A to reconstruct the assayed variant sequence.

column_label: Table 2 (Steady State Parameters of 17X-PTDH Mutants), enzyme-form row 'W134A'; column 'kcat (s-1)' = 1.73; column 'Km,PT (mM)' = 1.91; column 'Km,NAD (mM)' = 0.187; column 'Kia,NAD (mM)' = nd; column 'kcat/Km,PT (M-1 s-1)' = 901; column 'Dkcat' = 2.16; column 'D(kcat/Km,PT)' = 2.09.

normalized_value_unit_evidence: kcat: printed Table 2 cell 1.73 s^-1; already in s^-1, no conversion; the paper value is 1.73 s^(-1). Km: printed Table 2 K_m,PT cell 1.91 mM; already in mM, no conversion; the paper value is 1.91 mM. kcat/Km: printed Table 2 K_cat/K_m,PT cell 901 M^-1 s^-1; 1 M^-1 s^-1 is 0.001 s^-1 mM^-1, so 901 / 1000 = 0.901 s^-1 mM^-1.

raw_value_unit_evidence: Table 2, enzyme-form row 'W134A': k_cat = 1.73 s^-1 (column 'kcat (s-1)'); K_m,PT = 1.91 mM (column 'Km,PT (mM)'); printed k_cat/K_m,PT = 901 M^-1 s^-1 (column 'kcat/Km,PT (M-1 s-1)').

row_label: Table 2, enzyme-form row 'W134A'


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:O69054 and applied mutation signature W134F to reconstruct the assayed variant sequence.

column_label: Table 2 (Steady State Parameters of 17X-PTDH Mutants), enzyme-form row 'W134F'; column 'kcat (s-1)' = 2.96; column 'Km,PT (mM)' = 0.238; column 'Km,NAD (mM)' = 0.102; column 'Kia,NAD (mM)' = nd; column 'kcat/Km,PT (M-1 s-1)' = 1.24 x 10^4; column 'Dkcat' = 2.22; column 'D(kcat/Km,PT)' = 2.68.

normalized_value_unit_evidence: kcat: printed Table 2 cell 2.96 s^-1; already in s^-1, no conversion; the paper value is 2.96 s^(-1). Km: printed Table 2 K_m,PT cell 0.238 mM; already in mM, no conversion; the paper value is 0.238 mM. kcat/Km: printed Table 2 K_cat/K_m,PT cell 1.24 x 10^4 M^-1 s^-1; 1 M^-1 s^-1 is 0.001 s^-1 mM^-1, so 12400 / 1000 = 12.4 s^-1 mM^-1.

raw_value_unit_evidence: Table 2, enzyme-form row 'W134F': k_cat = 2.96 s^-1 (column 'kcat (s-1)'); K_m,PT = 0.238 mM (column 'Km,PT (mM)'); printed k_cat/K_m,PT = 1.24 x 10^4 M^-1 s^-1 (column 'kcat/Km,PT (M-1 s-1)').

row_label: Table 2, enzyme-form row 'W134F'


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:O69054 and applied mutation signature R301A to reconstruct the assayed variant sequence.

column_label: Table 2 (Steady State Parameters of 17X-PTDH Mutants), enzyme-form row 'R301A'; column 'kcat (s-1)' = 0.041; column 'Km,PT (mM)' = 19.4; column 'Km,NAD (mM)' = 0.216; column 'Kia,NAD (mM)' = 0.48; column 'kcat/Km,PT (M-1 s-1)' = 2.11; column 'Dkcat' = 2.7; column 'D(kcat/Km,PT)' = 2.1.

normalized_value_unit_evidence: kcat: printed Table 2 cell 0.041 s^-1; already in s^-1, no conversion; the paper value is 0.041 s^(-1). Km: printed Table 2 K_m,PT cell 19.4 mM; already in mM, no conversion; the paper value is 19.4 mM. kcat/Km: printed Table 2 K_cat/K_m,PT cell 2.11 M^-1 s^-1; 1 M^-1 s^-1 is 0.001 s^-1 mM^-1, so 2.11 / 1000 = 0.00211 s^-1 mM^-1.

raw_value_unit_evidence: Table 2, enzyme-form row 'R301A': k_cat = 0.041 s^-1 (column 'kcat (s-1)'); K_m,PT = 19.4 mM (column 'Km,PT (mM)'); printed k_cat/K_m,PT = 2.11 M^-1 s^-1 (column 'kcat/Km,PT (M-1 s-1)').

row_label: Table 2, enzyme-form row 'R301A'


sequence_variant_note: Resolved canonical sequence from UniProt O69054 FASTA plus direct residue-coordinate check and applied mutation signature E175A to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from UniProt O69054 FASTA plus direct residue-coordinate check and applied mutation signature M53N to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from UniProt O69054 FASTA plus direct residue-coordinate check and applied mutation signature M53A to reconstruct the assayed variant sequence.


organism: Ralstonia sp.

substrate_name: NAD+

sequence_source: UniProt G4XDR8 FASTA

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

assay_conditions_summary: pH 7.2; 303.15 K; cofactor NAD+

proof_lines:
pH 7.2, 30°C
pH 7.2, 40°C
Raw SKiD Env: Temp=30°C, pH=7.2

substrate: NAD+


organism: Ralstonia sp. 4506

proof_lines: recombinant wild-type enzyme, pH 7.0, 30°C


sequence_variant_note: Resolved canonical sequence from UniProt G4XDR8 FASTA plus direct residue-coordinate check and applied mutation signature Y139F to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from UniProt REST exact accession lookup; residue-coordinate reconstruction for substitution rows and applied mutation signature Y139W to reconstruct the assayed variant sequence.

assay_conditions_summary: pH 7; 303.15 K; cofactor NAD+


sequence_variant_note: Resolved canonical sequence from UniProt REST exact accession lookup; residue-coordinate reconstruction for substitution rows and applied mutation signature Y139R to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from UniProt REST exact accession lookup; residue-coordinate reconstruction for substitution rows and applied mutation signature Y139L to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from UniProt REST exact accession lookup; residue-coordinate reconstruction for substitution rows and applied mutation signature Y139M to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from UniProt REST exact accession lookup; residue-coordinate reconstruction for substitution rows and applied mutation signature Y139Q to reconstruct the assayed variant sequence.


organism: Stutzerimonas stutzeri

substrate_name: phosphite

sequence_source: UniProt REST exact accession lookup; residue-coordinate reconstruction for substitution rows

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

substrate: phosphite


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

assay_conditions_summary: pH 7.3; 298.15 K; cofactor NAD+

proof_lines:
Opt14 variant
mutant 17X-PTDH, pH 7.3, 25°C
Opt12 and Opt13 variants


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:O69054 and applied mutation signature R301K to reconstruct the assayed variant sequence.


proof_lines:
wild-type, pH 7.3, 25°C
wild type enzyme, at pH 7.3 and 25°C


sequence_variant_note: Resolved canonical sequence from UniProt REST exact accession lookup; residue-coordinate reconstruction for substitution rows and applied mutation signature S295A to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from UniProt REST exact accession lookup; residue-coordinate reconstruction for substitution rows and applied mutation signature M53A to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from UniProt REST exact accession lookup; residue-coordinate reconstruction for substitution rows and applied mutation signature E175A to reconstruct the assayed variant sequence.


substrate_name: H2O; Phosphonate; NAD+

sequence_variant_note: Resolved canonical sequence from UniProtKB O69054 canonical sequence reconstructed with selected-batch mutation_signature E175A and applied mutation signature E175A to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: phosphonate dehydrogenase(Enzyme) mutant E175A

substrate: H2O; Phosphonate; NAD+


sequence_variant_note: Resolved canonical sequence from UniProtKB O69054 canonical sequence reconstructed with selected-batch mutation_signature E175A/A176R and applied mutation signature E175A/A176R to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: phosphonate dehydrogenase(Enzyme) mutant E175A, A176R


sequence_source: UniProtKB O69054 canonical sequence from direct REST/FASTA lookup on 2026-06-16


substrate_name: NADP+

sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:O69054 and applied mutation signature E175A/A176R to reconstruct the assayed variant sequence.

assay_conditions_summary: pH 7.25; 298.15 K; cofactor NADP+

column_label: Table 1: Kinetic Parameters for Recombinant WT Phosphite Dehydrogenase and Mutants Using NADP+ and NAD+ as Substrates (Biochemistry 2003, 42, 11604-11614); enzyme row 'E175A, A176R'; column 'KM (µM, NADP+)' = 3.5; column 'kcat (min-1)' (the NADP+ block) = 114.0; column 'kcat/KM,NADP (µM-1 min-1)' = 32.6. The KM (µM, Pt-H) cell of the same block is the phosphite Michaelis constant, and the remaining four cells of the printed row belong to the NAD+ block.

raw_value_unit_evidence: Table 1, enzyme row 'E175A, A176R', NADP+ block: KM = 3.5 uM; kcat = 114.0 min^-1; kcat/KM = 32.6 uM^-1 min^-1.

row_label: Table 1, enzyme row 'E175A, A176R', the three NADP+ cells (KM for NADP+, kcat, kcat/KM)

substrate: NADP+


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

proof_lines:
Raw SKiD Env: Temp=25°C, pH=7.3
Raw SKiD Env: Temp=-----, pH=-----


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:O69054 and applied mutation signature E175A to reconstruct the assayed variant sequence.

column_label: Table 1: Kinetic Parameters for Recombinant WT Phosphite Dehydrogenase and Mutants Using NADP+ and NAD+ as Substrates (Biochemistry 2003, 42, 11604-11614); enzyme row 'E175A'; column 'KM (µM, NADP+)' = 144; column 'kcat (min-1)' (the NADP+ block) = 130.8; column 'kcat/KM,NADP (µM-1 min-1)' = 0.91. The KM (µM, Pt-H) cell of the same block is the phosphite Michaelis constant, and the remaining four cells of the printed row belong to the NAD+ block.

raw_value_unit_evidence: Table 1, enzyme row 'E175A', NADP+ block: KM = 144 uM; kcat = 130.8 min^-1; kcat/KM = 0.91 uM^-1 min^-1.

row_label: Table 1, enzyme row 'E175A', the three NADP+ cells (KM for NADP+, kcat, kcat/KM)


substrate_name: Phosphonate; NAD+; H2O

sequence_variant_note: Resolved canonical sequence from UniProtKB O69054 canonical sequence reconstructed with selected-batch mutation_signature A176R and applied mutation signature A176R to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: phosphonate dehydrogenase(Enzyme) mutant A176R

substrate: Phosphonate; NAD+; H2O


column_label: Table 1: Kinetic Parameters for Recombinant WT Phosphite Dehydrogenase and Mutants Using NADP+ and NAD+ as Substrates (Biochemistry 2003, 42, 11604-11614); enzyme row 'WT'; column 'KM (µM, NADP+)' = 2510; column 'kcat (min-1)' (the NADP+ block) = 84.6; column 'kcat/KM,NADP (µM-1 min-1)' = 0.0337. The KM (µM, Pt-H) cell of the same block is the phosphite Michaelis constant, and the remaining four cells of the printed row belong to the NAD+ block.

raw_value_unit_evidence: Table 1, enzyme row 'WT', NADP+ block: KM = 2510 uM; kcat = 84.6 min^-1; kcat/KM = 0.0337 uM^-1 min^-1.

row_label: Table 1, enzyme row 'WT', the three NADP+ cells (KM for NADP+, kcat, kcat/KM)


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:O69054 and applied mutation signature A176R to reconstruct the assayed variant sequence.

assay_conditions_summary: pH 7.25; 298.15 K; cofactor NAD+

column_label: Table 1: Kinetic Parameters for Recombinant WT Phosphite Dehydrogenase and Mutants Using NADP+ and NAD+ as Substrates (Biochemistry 2003, 42, 11604-11614); enzyme row 'A176R'; column 'KM (µM, NAD+)' = 60; column 'kcat (min-1)' (the NAD+ block) = 256.8; column 'kcat/KM,NAD (µM-1 min-1)' = 4.3. The KM (µM, Pt-H) cell of the same block is the phosphite Michaelis constant, and the remaining four cells of the printed row belong to the NADP+ block.

raw_value_unit_evidence: Table 1, enzyme row 'A176R', NAD+ block: KM = 60 uM; kcat = 256.8 min^-1; kcat/KM = 4.3 uM^-1 min^-1.

row_label: Table 1, enzyme row 'A176R', the three NAD+ cells (KM for NAD+, kcat, kcat/KM)


column_label: Table 1: Kinetic Parameters for Recombinant WT Phosphite Dehydrogenase and Mutants Using NADP+ and NAD+ as Substrates (Biochemistry 2003, 42, 11604-11614); enzyme row 'E175A'; column 'KM (µM, NAD+)' = 16; column 'kcat (min-1)' (the NAD+ block) = 210.0; column 'kcat/KM,NAD (µM-1 min-1)' = 13.1. The KM (µM, Pt-H) cell of the same block is the phosphite Michaelis constant, and the remaining four cells of the printed row belong to the NADP+ block.

raw_value_unit_evidence: Table 1, enzyme row 'E175A', NAD+ block: KM = 16 uM; kcat = 210.0 min^-1; kcat/KM = 13.1 uM^-1 min^-1.

row_label: Table 1, enzyme row 'E175A', the three NAD+ cells (KM for NAD+, kcat, kcat/KM)


column_label: Table 1: Kinetic Parameters for Recombinant WT Phosphite Dehydrogenase and Mutants Using NADP+ and NAD+ as Substrates (Biochemistry 2003, 42, 11604-11614); enzyme row 'A176R'; column 'KM (µM, NADP+)' = 77; column 'kcat (min-1)' (the NADP+ block) = 130.8; column 'kcat/KM,NADP (µM-1 min-1)' = 1.7. The KM (µM, Pt-H) cell of the same block is the phosphite Michaelis constant, and the remaining four cells of the printed row belong to the NAD+ block.

raw_value_unit_evidence: Table 1, enzyme row 'A176R', NADP+ block: KM = 77 uM; kcat = 130.8 min^-1; kcat/KM = 1.7 uM^-1 min^-1.

row_label: Table 1, enzyme row 'A176R', the three NADP+ cells (KM for NADP+, kcat, kcat/KM)


column_label: Table 1: Kinetic Parameters for Recombinant WT Phosphite Dehydrogenase and Mutants Using NADP+ and NAD+ as Substrates (Biochemistry 2003, 42, 11604-11614); enzyme row 'E175A, A176R'; column 'KM (µM, NAD+)' = 20; column 'kcat (min-1)' (the NAD+ block) = 236.4; column 'kcat/KM,NAD (µM-1 min-1)' = 11.8. The KM (µM, Pt-H) cell of the same block is the phosphite Michaelis constant, and the remaining four cells of the printed row belong to the NADP+ block.

raw_value_unit_evidence: Table 1, enzyme row 'E175A, A176R', NAD+ block: KM = 20 uM; kcat = 236.4 min^-1; kcat/KM = 11.8 uM^-1 min^-1.

row_label: Table 1, enzyme row 'E175A, A176R', the three NAD+ cells (KM for NAD+, kcat, kcat/KM)


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

proof_lines: SABIO-RK modifier species: phosphonate dehydrogenase(Enzyme) mutant Glu266Gln (N-terminal His6 tag)


proof_lines: SABIO-RK modifier species: phosphonate dehydrogenase(Enzyme) mutant Lys76Arg (N-terminal His6 tag)


enzyme_display_name: thymidine phosphorylase

organism: Escherichia coli

sequence_source: UniProtKB P07650 / TYPH_ECOLI, Escherichia coli K12 thymidine phosphorylase, EC 2.4.2.4; direct UniProt REST accession check on 2026-06-16

assay_conditions_summary: pH 6.5; 298.15 K; buffer 50 mM potassium phosphate

proof_lines: in 50 mM potassium phosphate buffer (pH 6.5), at 25°C

proof_lines: in 50 mM potassium phosphate buffer, pH 6.5, 25°C

enzyme: thymidine phosphorylase


substrate_name: thymidine

substrate: thymidine


substrate_name: Unknown

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

proof_lines: pH not specified in the publication, 30°C

substrate: Unknown


substrate_name: thymine

substrate: thymine


sequence_source: UniProtKB P19971 / TYPH_HUMAN, Homo sapiens thymidine phosphorylase, EC 2.4.2.4; direct UniProt REST accession check on 2026-06-16

proof_lines: ph 5.5, 37°C, recombinant wild-type enzyme


substrate_name: Phosphate; Uridine

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

assay_conditions_summary: pH 7.5; 310.15 K; buffer 0.1 M potassium phosphate

proof_lines: in 0.1 M potassium phosphate buffer (pH 7.4), at 25°C

source_databases_merged: sabio_rk brenda skid

substrate: Phosphate; Uridine


organism: Mus musculus

sequence_source: UniProtKB Q99N42 / TYPH_MOUSE, Mus musculus thymidine phosphorylase, EC 2.4.2.4; direct UniProt REST accession check on 2026-06-16


substrate_name: n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n RNA (poly(A)); ATP

assay_conditions_summary: cofactor ATP

proof_lines: SABIO-RK modifier species: polynucleotide adenylyltransferase(Enzyme) mutant bPAP-513 F220Y


mutation_type: deletion

proof_lines: SABIO-RK modifier species: polynucleotide adenylyltransferase(Enzyme) wildtype bPAP-513 (C-terminal deletion)


substrate_name: ATP

substrate: ATP


proof_lines: SABIO-RK modifier species: polynucleotide adenylyltransferase(Enzyme) mutant bPAP-513 F243A

substrate: n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n RNA (poly(A)); ATP


proof_lines: SABIO-RK modifier species: polynucleotide adenylyltransferase(Enzyme) mutant bPAP-513 L193M


substrate_name: ATP; n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n RNA (poly(A))

proof_lines: SABIO-RK modifier species: polynucleotide adenylyltransferase(Enzyme) mutant bPAP-513 R221A


proof_lines: SABIO-RK modifier species: polynucleotide adenylyltransferase(Enzyme) mutant bPAP-513 R104A

substrate: ATP; n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n n+1 n RNA (poly(A))


proof_lines: SABIO-RK modifier species: polynucleotide adenylyltransferase(Enzyme) mutant bPAP-513 K191A


proof_lines: SABIO-RK modifier species: polynucleotide adenylyltransferase(Enzyme) mutant bPAP-513 G203H

proof_lines: SABIO-RK modifier species: polynucleotide adenylyltransferase(Enzyme) mutant G203H


sequence_variant_note: Resolved canonical sequence from UniProtKB P25500 displayed canonical sequence; variant reconstructed by one-residue substitution from selected mutation_signature. and applied mutation signature F220R to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from UniProtKB P25500 displayed canonical sequence; variant reconstructed by one-residue substitution from selected mutation_signature. and applied mutation signature L229R to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from UniProtKB P25500 displayed canonical sequence; variant reconstructed by one-residue substitution from selected mutation_signature. and applied mutation signature R225N to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: polynucleotide adenylyltransferase(Enzyme) mutant bPAP-513 R225N


sequence_variant_note: Resolved canonical sequence from UniProtKB P25500 displayed canonical sequence; variant reconstructed by one-residue substitution from selected mutation_signature. and applied mutation signature F243A to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from UniProtKB P25500 displayed canonical sequence; variant reconstructed by one-residue substitution from selected mutation_signature. and applied mutation signature L211K to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: polynucleotide adenylyltransferase(Enzyme) mutant bPAP-513 L211K