sequence_variant_note: Resolved canonical sequence from UniProt Q38087 direct FASTA check plus local substitution reconstruction and applied mutation signature Y567A to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: DNA-directed DNA polymerase(Enzyme) mutant Y567A


enzyme_display_name: DNA-directed DNA polymerase

organism: Escherichia coli

substrate_name: deoxynucleoside triphosphate

source_db: brenda

kcat_display: Not reported

ki_display: Not reported

kcat_over_km_display: Not reported

protein_accession_database: UniProt

source_license: CC BY 4.0

km_unit: mM

sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature Y824A to reconstruct the assayed variant sequence.

mutation_type: substitution

proof_lines: mutant enzyme Y824A, in 50 mM Tris-HCl (pH 7.8), 1 mM dithiothreitol, at 25°C

enzyme: DNA-directed DNA polymerase

substrate: deoxynucleoside triphosphate

source: brenda


organism: Homo sapiens

substrate_name: DNA; dATP

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

sequence_variant_note: Mutation signature Y52E matches the mutant residue at offset +1 and the reference residue at offset +0; variant reconstruction was not applied because multiple plausible residue-numbering offsets are present.

proof_lines: SABIO-RK modifier species: DNA-directed DNA polymerase(Enzyme) mutant Y52E

substrate: DNA; dATP


substrate_name: dTTP; DNA

sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_mutation_ranked_match and applied mutation signature E895A to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: DNA-directed DNA polymerase(Enzyme) mutant E895A

substrate: dTTP; DNA


substrate_name: DNA; dTTP

sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_mutation_ranked_match and applied mutation signature Y951F to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: DNA-directed DNA polymerase(Enzyme) mutant Y951F

substrate: DNA; dTTP


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_mutation_ranked_match and applied mutation signature Y955A to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: DNA-directed DNA polymerase(Enzyme) mutant Y955A


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_mutation_ranked_match and applied mutation signature Y951A to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: DNA-directed DNA polymerase(Enzyme) mutant Y951A


organism: Bacteriophage T4

substrate_name: H2O; DNA

sequence_variant_note: Resolved canonical sequence from reconstructed_from_uniprot_accession and applied mutation signature D219A/D324A to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: DNA-directed DNA polymerase(Enzyme) mutant D219A/D324A

substrate: H2O; DNA


sequence_variant_note: Resolved canonical sequence from reconstructed_from_uniprot_accession and applied mutation signature R821A to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme R821A, in 50 mM Tris-HCl (pH 7.8), 1 mM dithiothreitol, at 25°C


sequence_variant_note: Resolved canonical sequence from reconstructed_from_uniprot_accession and applied mutation signature R822A to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme R822A, in 50 mM Tris-HCl (pH 7.8), 1 mM dithiothreitol, at 25°C


sequence_variant_note: Resolved canonical sequence from reconstructed_from_uniprot_accession and applied mutation signature R822A/Y824A to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme R822A/Y824A in 50 mM Tris-HCl (pH 7.8), 1 mM dithiothreitol, at 25°C


sequence_variant_note: Mutation signature R823A could not be applied safely because the resolved sequence does not match the expected reference residue(s).

proof_lines: mutant enzyme R823A, in 50 mM Tris-HCl (pH 7.8), 1 mM dithiothreitol, at 25°C


mutation_signature: Unknown

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

proof_lines: wild type enzyme, in 50 mM Tris-HCl (pH 7.8), 1 mM dithiothreitol, at 25°C


organism: Plasmodium falciparum (isolate 3D7)

substrate_name: dNTPs

source_db: uniprot

proof_lines: kcat is 22 sec(-1) for polymerase activity (PubMed:27487482). kcat is 10 min(-1) for polymerase activity (PubMed:39490679). kcat is 1.69 min(-1) for polymerase activity on dA template with dTTP (PubMed:35220686, PubMed:32636411). kcat is 1.

substrate: dNTPs

source: uniprot


organism: Saccharolobus solfataricus

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

proof_lines: pH 7.5, 60°C, incorporation opposite the 5-thymine of a cis-syn thymine dimer of the template

proof_lines: pH 7.4, 37°C, steady-state kinetics for single nucleotide primer extension with guanine as template

proof_lines: pH 7.5, 60°C, incorporation opposite the 5-thymine of an nondamaged template

source_databases_merged: brenda skid


substrate_name: dTTP

verification_status: corrected

assay_conditions_summary: pH 7.5; 310.15 K; buffer 50 mM Tris-HCl

column_label: TABLE 2 (Steady-state kinetics of dNTP incorporation opposite G by Dpo 1, Dpo2, Dpo3, and Dpo4); Polymerase column = Dpo2; dNTP column = T (dTTP); column 'Km (uM)' = 3700 +/- 1300; column 'kcat (min^-1)' = 0.0000072 +/- 0.0000009; column 'kcat/Km (min^-1 uM^-1)' = 1.9 x 10^-9; column 'fins' = 3.5 x 10^-4.

normalized_value_unit_evidence: Km: printed TABLE 2 cell 3700 uM; 3700 / 1000 = 3.7 mM. kcat: printed TABLE 2 cell 0.0000072 min^-1, under the column head 'kcat' whose unit line reads 'min' with the superscript -1; one min^-1 is 1/60 s^-1, so 0.0000072 / 60 = 0.00000012 s^-1. kcat/Km: printed TABLE 2 cell 1.9 x 10^-9 min^-1 uM^-1, under the column head 'kcat/Km' whose unit line reads 'min' with the superscript -1 followed by 'uM' with the superscript -1; one min^-1 uM^-1 is 1000/60 s^-1 mM^-1, so 0.0000000019 x 1000 / 60 = 0.00000003166666666666667 s^-1 mM^-1.

raw_value_unit_evidence: TABLE 2, Polymerase Dpo2, dNTP row T (dTTP): Km = 3700 uM; kcat = 0.0000072 min^-1; printed kcat/Km = 1.9 x 10^-9 min^-1 uM^-1.

row_label: TABLE 2, Polymerase block 'Dpo2', dNTP row 'T' (incoming nucleotide dTTP), insertion opposite template G

table_label: TABLE 2: Steady-state kinetics of dNTP incorporation opposite G by Dpo 1, Dpo2, Dpo3, and Dpo4 (J. Biol. Chem. 2011, 286(36):31180-31193)

data_origin: experimental

substrate: dTTP

status: corrected


substrate_name: dCTP

column_label: TABLE 2 (Steady-state kinetics of dNTP incorporation opposite G by Dpo 1, Dpo2, Dpo3, and Dpo4); Polymerase column = Dpo2; dNTP column = C (dCTP); column 'Km (uM)' = 9.8 +/- 2.1; column 'kcat (min^-1)' = 0.000053 +/- 0.000003; column 'kcat/Km (min^-1 uM^-1)' = 5.4 x 10^-6; column 'fins' = 1.

normalized_value_unit_evidence: Km: printed TABLE 2 cell 9.8 uM; 9.8 / 1000 = 0.0098 mM. kcat: printed TABLE 2 cell 0.000053 min^-1, under the column head 'kcat' whose unit line reads 'min' with the superscript -1; one min^-1 is 1/60 s^-1, so 0.000053 / 60 = 0.0000008833333333333334 s^-1. kcat/Km: printed TABLE 2 cell 5.4 x 10^-6 min^-1 uM^-1, under the column head 'kcat/Km' whose unit line reads 'min' with the superscript -1 followed by 'uM' with the superscript -1; one min^-1 uM^-1 is 1000/60 s^-1 mM^-1, so 0.0000054 x 1000 / 60 = 0.00009 s^-1 mM^-1.

raw_value_unit_evidence: TABLE 2, Polymerase Dpo2, dNTP row C (dCTP): Km = 9.8 uM; kcat = 0.000053 min^-1; printed kcat/Km = 5.4 x 10^-6 min^-1 uM^-1.

row_label: TABLE 2, Polymerase block 'Dpo2', dNTP row 'C' (incoming nucleotide dCTP), insertion opposite template G

substrate: dCTP


substrate_name: dATP

column_label: TABLE 2 (Steady-state kinetics of dNTP incorporation opposite G by Dpo 1, Dpo2, Dpo3, and Dpo4); Polymerase column = Dpo3; dNTP column = A (dATP); column 'Km (uM)' = 390 +/- 10; column 'kcat (min^-1)' = 0.000016 +/- 0.000001; column 'kcat/Km (min^-1 uM^-1)' = 4.1 x 10^-8; column 'fins' = 5.7 x 10^-3.

normalized_value_unit_evidence: Km: printed TABLE 2 cell 390 uM; 390 / 1000 = 0.39 mM. kcat: printed TABLE 2 cell 0.000016 min^-1, under the column head 'kcat' whose unit line reads 'min' with the superscript -1; one min^-1 is 1/60 s^-1, so 0.000016 / 60 = 0.00000026666666666666667 s^-1. kcat/Km: printed TABLE 2 cell 4.1 x 10^-8 min^-1 uM^-1, under the column head 'kcat/Km' whose unit line reads 'min' with the superscript -1 followed by 'uM' with the superscript -1; one min^-1 uM^-1 is 1000/60 s^-1 mM^-1, so 0.000000040999999999999997 x 1000 / 60 = 0.0000006833333333333333 s^-1 mM^-1.

raw_value_unit_evidence: TABLE 2, Polymerase Dpo3, dNTP row A (dATP): Km = 390 uM; kcat = 0.000016 min^-1; printed kcat/Km = 4.1 x 10^-8 min^-1 uM^-1.

row_label: TABLE 2, Polymerase block 'Dpo3', dNTP row 'A' (incoming nucleotide dATP), insertion opposite template G

substrate: dATP


substrate_name: dGTP

column_label: TABLE 2 (Steady-state kinetics of dNTP incorporation opposite G by Dpo 1, Dpo2, Dpo3, and Dpo4); Polymerase column = Dpo3; dNTP column = G (dGTP); column 'Km (uM)' = 190 +/- 40; column 'kcat (min^-1)' = 0.000013 +/- 0.000001; column 'kcat/Km (min^-1 uM^-1)' = 6.8 x 10^-8; column 'fins' = 9.5 x 10^-3.

normalized_value_unit_evidence: Km: printed TABLE 2 cell 190 uM; 190 / 1000 = 0.19 mM. kcat: printed TABLE 2 cell 0.000013 min^-1, under the column head 'kcat' whose unit line reads 'min' with the superscript -1; one min^-1 is 1/60 s^-1, so 0.000013 / 60 = 0.00000021666666666666665 s^-1. kcat/Km: printed TABLE 2 cell 6.8 x 10^-8 min^-1 uM^-1, under the column head 'kcat/Km' whose unit line reads 'min' with the superscript -1 followed by 'uM' with the superscript -1; one min^-1 uM^-1 is 1000/60 s^-1 mM^-1, so 0.000000068 x 1000 / 60 = 0.0000011333333333333334 s^-1 mM^-1.

raw_value_unit_evidence: TABLE 2, Polymerase Dpo3, dNTP row G (dGTP): Km = 190 uM; kcat = 0.000013 min^-1; printed kcat/Km = 6.8 x 10^-8 min^-1 uM^-1.

row_label: TABLE 2, Polymerase block 'Dpo3', dNTP row 'G' (incoming nucleotide dGTP), insertion opposite template G

substrate: dGTP


sequence_variant_note: Resolved canonical sequence from uniprot_ec_organism_mutation_ranked_match and applied mutation signature Y955F to reconstruct the assayed variant sequence.

proof_lines: SABIO-RK modifier species: DNA-directed DNA polymerase(Enzyme) mutant Y955F


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

proof_lines: SABIO-RK modifier species: DNA-directed DNA polymerase(Enzyme) mutant DNA polymerase lambda (lacking BRCT-like and Proline-rich domains)


substrate_name: DNA with an intact AP site

assay_conditions_summary: pH 7

proof_lines: kcat is 0.075 sec(-1) for 5'-deoxyribose-phosphate lyase activity (at pH 7.0 and 37 degrees Celsius) (PubMed:9614142). kcat is 0.004 sec(-1) for AP lyase activity (at pH 7.0 and 37 degrees Celsius) (PubMed:9614142).

substrate: DNA with an intact AP site


substrate_name: Unknown

km_display: Not reported

sequence_variant_note: Mutation context was captured, but multiple structural mutation events could not be translated into an exact protein sequence safely.

mutation_type: deletion

proof_lines: kcat is 87 sec(-1) for DNA synthesis by Pol-delta4 and 19 sec(-1) for Pol-delta3. kcat for exonuclease activity determined using a 26mer/40mer duplex DNA gives a value of 0.003 sec(-1) for Pol-delta4 and 0.026 sec(-1) for Pol-delta3. When u

substrate: Unknown


organism: Human alphaherpesvirus 1

ki_unit: mM

assay_conditions_summary: pH 8; 310.15 K; buffer context in 50 mM Tris-HCl

proof_lines: in 50 mM Tris-HCl (pH 8.0), 8 mM MgCl2, and 0.5 mM dithiohtreitol, at 37°C


kcat_over_km_origin: calculated


organism: Thermococcus waiotapuensis

substrate_name: DNAn

sequence_variant_note: Mutation signature H633R could not be applied safely because the resolved sequence does not match the expected reference residue(s).

proof_lines: pH 8.6, 75°C, mM of template, in the presence of an excess of annealed primer, wild-type enzyme

proof_lines: pH 8.6, 75°C, mM of each nucleotide in an equimolar mixture of the four nucleotides, wild-type enzyme

proof_lines: pH 8.6, 75°C, mM of template, in the presence of an excess of annealed prime, wild-type enzyme

substrate: DNAn


proof_lines: pH 8.6, 75°C, mM of template, in the presence of an excess of annealed primer, mutant enzyme H633R

proof_lines: pH 8.6, 75°C, mM of each nucleotide in an equimolar mixture of the four nucleotides, mutant enzyme H633R


organism: Bacteriophage RB69

source_db: oed

sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature D222A/D327A/K560A to reconstruct the assayed variant sequence.

source: oed


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature D222A/D327A/N564A to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature D222A/D327A/R482A to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature D222A/D327A/Y567A to reconstruct the assayed variant sequence.


substrate_name: dGDP

sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature D222A/D327A/K486A to reconstruct the assayed variant sequence.

substrate: dGDP


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature D222A/D327A to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature D222A/D327A/K483A to reconstruct the assayed variant sequence.


verification_status: unverified

status: unverified


enzyme_display_name: hexokinase

organism: Entamoeba histolytica

substrate_name: D-Glucose

verification_status: verified

column_label: PMID 10613700 Results 3.2 same prose segment for HXK2 glucose; kcat and glucose Km are reported together and kcat/Km is locked from those values.

raw_value_unit_evidence: Results section 3.2: kcat 237 s^-1 and apparent Km for glucose 47 uM for HXK2.

row_label: HXK2 glucose, optimal purified preparation prose segment

table_label: Results section 3.2 kinetic-properties prose

enzyme: hexokinase

substrate: D-Glucose

status: verified


substrate_name: D-Mannose

substrate: D-Mannose


organism: Rattus norvegicus

substrate_name: D-glucose

assay_conditions_summary: cofactor ATP

substrate: D-glucose


substrate_name: D-fructose

substrate: D-fructose


substrate_name: ATP

substrate: ATP


organism: Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)

substrate_name: fructose 6-phosphate

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

smiles: C(C(C(C(C(=O)CO)O)O)O)OP(=O)(O)O

substrate: fructose 6-phosphate


substrate_name: fructose 1,6-bisphosphate (for gluconeogenic reaction)


substrate_name: ADP (for gluconeogenic reaction)

assay_conditions_summary: cofactor ADP

substrate: ADP (for gluconeogenic reaction)


substrate_name: IDP (for gluconeogenic reaction)

substrate: IDP (for gluconeogenic reaction)


substrate_name: GDP (for gluconeogenic reaction)

substrate: GDP (for gluconeogenic reaction)


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature I270S to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature R265T/Y266F to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature S144T to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature S144T/R265T/Y266F to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature S144T/V183T/I270S to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature S144T/V183T/I270S/R265T/Y266F to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature V183T to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature V183T/I270S to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature V183T/I270S/R265T/Y266F to reconstruct the assayed variant sequence.


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


organism: Solanum chacoense

assay_conditions_summary: pH 8; 303.15 K; cofactor ATP

column_label: Table 3. Kinetic constants of pure ScHK2; row D-Glucose; column Kmapp (mM): raw value 0.023 +/- 0.003 mM, normalized value 0.023 mM; column kcat (s^-1): raw value 5.01 +/- 0.75 s^-1, normalized value 5.01 s^-1; column kcat/Kmapp (mM^-1 s^-1): the table prints only 218, a rounded two-significant-figure value, so the absolute ratio recorded here is the quotient of this same row's own two printed cells, 5.01 / 0.023 = 217.82608695652172 s^-1 mM^-1.

normalized_value_unit_evidence: metric: kcat; raw value: 5.01 +/- 0.75; raw unit: s^-1; normalized value: 5.01; normalized unit: s^-1; metric: Km; raw value: 0.023 +/- 0.003; raw unit: mM; normalized value: 0.023; normalized unit: mM; metric: kcat/Km; the table prints only the rounded cell 218 mM^-1 s^-1, so the absolute ratio recorded here is the quotient of this row's own two printed cells; normalized value: 217.82608695652172; normalized unit: s^-1 mM^-1; assay condition: Purified hexahistidine-tagged recombinant ScHK2 hexokinase from Solanum chacoense expressed in Escherichia coli; HK reaction coupled to the glucose-6-phosphate dehydrogenase (EC 1.1.1.49) reaction and assayed at 30 C by monitoring NAD+ reduction at 340 nm; 200 uL reaction mixture containing 50 mM Tris-HCl, pH 8.0, 50 mM KCl, 5 mM MgCl2, 5 mM DTT and 0.3 mM NAD+; apparent Km values calculated from the Michaelis-Menten equation by non-linear least-squares regression and kcat values calculated from the Vmax of Michaelis-Menten plots using a subunit molecular mass of 56,776 Da.

raw_value_unit_evidence: Table 3. Kinetic constants of pure ScHK2 (journal page 336), row D-Glucose: the Kmapp (mM) column prints 0.023 +/- 0.003; the kcat (s^-1) column prints 5.01 +/- 0.75; the kcat/Kmapp (mM^-1 s^-1) column prints 218. Footnote a: For hexoses and ATP, kinetic parameters are given as means +/- SE of values from three independent enzyme preparations.

row_label: D-Glucose


substrate_name: GTP

column_label: Table 3. Kinetic constants of pure ScHK2; row GTP; column Kmapp (mM): raw value 0.231 mM, normalized value 0.231 mM; column kcat (s^-1): raw value 0.85 s^-1, normalized value 0.85 s^-1; column kcat/Kmapp (mM^-1 s^-1): the table prints only 3.7, a rounded two-significant-figure value, so the absolute ratio recorded here is the quotient of this same row's own two printed cells, 0.85 / 0.231 = 3.679653679653679 s^-1 mM^-1.

normalized_value_unit_evidence: metric: kcat; raw value: 0.85; raw unit: s^-1; normalized value: 0.85; normalized unit: s^-1; metric: Km; raw value: 0.231; raw unit: mM; normalized value: 0.231; normalized unit: mM; metric: kcat/Km; the table prints only the rounded cell 3.7 mM^-1 s^-1, so the absolute ratio recorded here is the quotient of this row's own two printed cells; normalized value: 3.679653679653679; normalized unit: s^-1 mM^-1; assay condition: Purified hexahistidine-tagged recombinant ScHK2 hexokinase from Solanum chacoense expressed in Escherichia coli; HK reaction coupled to the glucose-6-phosphate dehydrogenase (EC 1.1.1.49) reaction and assayed at 30 C by monitoring NAD+ reduction at 340 nm; 200 uL reaction mixture containing 50 mM Tris-HCl, pH 8.0, 50 mM KCl, 5 mM MgCl2, 5 mM DTT and 0.3 mM NAD+; apparent Km values calculated from the Michaelis-Menten equation by non-linear least-squares regression and kcat values calculated from the Vmax of Michaelis-Menten plots using a subunit molecular mass of 56,776 Da.

raw_value_unit_evidence: Table 3. Kinetic constants of pure ScHK2 (journal page 336), row GTP: the Kmapp (mM) column prints 0.231; the kcat (s^-1) column prints 0.85; the kcat/Kmapp (mM^-1 s^-1) column prints 3.7. Footnote a: Values for TTP, GTP, UTP, and CTP, are representative of two independent enzyme preparations.

row_label: GTP

substrate: GTP


substrate_name: UTP

column_label: Table 3. Kinetic constants of pure ScHK2; row UTP; column Kmapp (mM): raw value 2.2 mM, normalized value 2.2 mM; column kcat (s^-1): raw value 3.02 s^-1, normalized value 3.02 s^-1; column kcat/Kmapp (mM^-1 s^-1): the table prints only 1.4, a rounded two-significant-figure value, so the absolute ratio recorded here is the quotient of this same row's own two printed cells, 3.02 / 2.2 = 1.3727272727272726 s^-1 mM^-1.

normalized_value_unit_evidence: metric: kcat; raw value: 3.02; raw unit: s^-1; normalized value: 3.02; normalized unit: s^-1; metric: Km; raw value: 2.2; raw unit: mM; normalized value: 2.2; normalized unit: mM; metric: kcat/Km; the table prints only the rounded cell 1.4 mM^-1 s^-1, so the absolute ratio recorded here is the quotient of this row's own two printed cells; normalized value: 1.3727272727272726; normalized unit: s^-1 mM^-1; assay condition: Purified hexahistidine-tagged recombinant ScHK2 hexokinase from Solanum chacoense expressed in Escherichia coli; HK reaction coupled to the glucose-6-phosphate dehydrogenase (EC 1.1.1.49) reaction and assayed at 30 C by monitoring NAD+ reduction at 340 nm; 200 uL reaction mixture containing 50 mM Tris-HCl, pH 8.0, 50 mM KCl, 5 mM MgCl2, 5 mM DTT and 0.3 mM NAD+; apparent Km values calculated from the Michaelis-Menten equation by non-linear least-squares regression and kcat values calculated from the Vmax of Michaelis-Menten plots using a subunit molecular mass of 56,776 Da.

raw_value_unit_evidence: Table 3. Kinetic constants of pure ScHK2 (journal page 336), row UTP: the Kmapp (mM) column prints 2.2; the kcat (s^-1) column prints 3.02; the kcat/Kmapp (mM^-1 s^-1) column prints 1.4. Footnote a: Values for TTP, GTP, UTP, and CTP, are representative of two independent enzyme preparations.

row_label: UTP

substrate: UTP


substrate_name: dTTP

substrate: dTTP


substrate_name: dUTP

substrate: dUTP


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature D222A/D327A/K560A to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature D222A/D327A/N564A to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature D222A/D327A/R482A to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from uniprot_accession and applied mutation signature D222A/D327A/Y567A to reconstruct the assayed variant sequence.


assay_conditions_summary: pH 8; 303.15 K; cofactor NAD+

table_label: Table 3. Kinetic constants of pure ScHK2


column_label: Table 3. Kinetic constants of pure ScHK2; row ATP; column Kmapp (mM): raw value 0.061 +/- 0.01 mM, normalized value 0.061 mM; column kcat (s^-1): raw value 3.50 +/- 0.37 s^-1, normalized value 3.5 s^-1; column kcat/Kmapp (mM^-1 s^-1): the table prints only 57, a rounded two-significant-figure value, so the absolute ratio recorded here is the quotient of this same row's own two printed cells, 3.5 / 0.061 = 57.37704918032787 s^-1 mM^-1.

normalized_value_unit_evidence: metric: kcat; raw value: 3.50 +/- 0.37; raw unit: s^-1; normalized value: 3.5; normalized unit: s^-1; metric: Km; raw value: 0.061 +/- 0.01; raw unit: mM; normalized value: 0.061; normalized unit: mM; metric: kcat/Km; the table prints only the rounded cell 57 mM^-1 s^-1, so the absolute ratio recorded here is the quotient of this row's own two printed cells; normalized value: 57.37704918032787; normalized unit: s^-1 mM^-1; assay condition: Purified hexahistidine-tagged recombinant ScHK2 hexokinase from Solanum chacoense expressed in Escherichia coli; HK reaction coupled to the glucose-6-phosphate dehydrogenase (EC 1.1.1.49) reaction and assayed at 30 C by monitoring NAD+ reduction at 340 nm; 200 uL reaction mixture containing 50 mM Tris-HCl, pH 8.0, 50 mM KCl, 5 mM MgCl2, 5 mM DTT and 0.3 mM NAD+; apparent Km values calculated from the Michaelis-Menten equation by non-linear least-squares regression and kcat values calculated from the Vmax of Michaelis-Menten plots using a subunit molecular mass of 56,776 Da.

raw_value_unit_evidence: Table 3. Kinetic constants of pure ScHK2 (journal page 336), row ATP: the Kmapp (mM) column prints 0.061 +/- 0.01; the kcat (s^-1) column prints 3.50 +/- 0.37; the kcat/Kmapp (mM^-1 s^-1) column prints 57. Footnote a: For hexoses and ATP, kinetic parameters are given as means +/- SE of values from three independent enzyme preparations.

row_label: ATP


substrate_name: TTP

column_label: Table 3. Kinetic constants of pure ScHK2; row TTP; column Kmapp (mM): raw value 0.231 mM, normalized value 0.231 mM; column kcat (s^-1): raw value 1.42 s^-1, normalized value 1.42 s^-1; column kcat/Kmapp (mM^-1 s^-1): the table prints only 6.1, a rounded two-significant-figure value, so the absolute ratio recorded here is the quotient of this same row's own two printed cells, 1.42 / 0.231 = 6.147186147186146 s^-1 mM^-1.

normalized_value_unit_evidence: metric: kcat; raw value: 1.42; raw unit: s^-1; normalized value: 1.42; normalized unit: s^-1; metric: Km; raw value: 0.231; raw unit: mM; normalized value: 0.231; normalized unit: mM; metric: kcat/Km; the table prints only the rounded cell 6.1 mM^-1 s^-1, so the absolute ratio recorded here is the quotient of this row's own two printed cells; normalized value: 6.147186147186146; normalized unit: s^-1 mM^-1; assay condition: Purified hexahistidine-tagged recombinant ScHK2 hexokinase from Solanum chacoense expressed in Escherichia coli; HK reaction coupled to the glucose-6-phosphate dehydrogenase (EC 1.1.1.49) reaction and assayed at 30 C by monitoring NAD+ reduction at 340 nm; 200 uL reaction mixture containing 50 mM Tris-HCl, pH 8.0, 50 mM KCl, 5 mM MgCl2, 5 mM DTT and 0.3 mM NAD+; apparent Km values calculated from the Michaelis-Menten equation by non-linear least-squares regression and kcat values calculated from the Vmax of Michaelis-Menten plots using a subunit molecular mass of 56,776 Da.

raw_value_unit_evidence: Table 3. Kinetic constants of pure ScHK2 (journal page 336), row TTP: the Kmapp (mM) column prints 0.231; the kcat (s^-1) column prints 1.42; the kcat/Kmapp (mM^-1 s^-1) column prints 6.1. Footnote a: Values for TTP, GTP, UTP, and CTP, are representative of two independent enzyme preparations.

row_label: TTP

substrate: TTP


substrate_name: CTP

column_label: Table 3. Kinetic constants of pure ScHK2; row CTP; column Kmapp (mM): raw value 2.0 mM, normalized value 2.0 mM; column kcat (s^-1): raw value 1.61 s^-1, normalized value 1.61 s^-1; column kcat/Kmapp (mM^-1 s^-1): the table prints only 0.8, a rounded two-significant-figure value, so the absolute ratio recorded here is the quotient of this same row's own two printed cells, 1.61 / 2.0 = 0.805 s^-1 mM^-1.

normalized_value_unit_evidence: metric: kcat; raw value: 1.61; raw unit: s^-1; normalized value: 1.61; normalized unit: s^-1; metric: Km; raw value: 2.0; raw unit: mM; normalized value: 2.0; normalized unit: mM; metric: kcat/Km; the table prints only the rounded cell 0.8 mM^-1 s^-1, so the absolute ratio recorded here is the quotient of this row's own two printed cells; normalized value: 0.805; normalized unit: s^-1 mM^-1; assay condition: Purified hexahistidine-tagged recombinant ScHK2 hexokinase from Solanum chacoense expressed in Escherichia coli; HK reaction coupled to the glucose-6-phosphate dehydrogenase (EC 1.1.1.49) reaction and assayed at 30 C by monitoring NAD+ reduction at 340 nm; 200 uL reaction mixture containing 50 mM Tris-HCl, pH 8.0, 50 mM KCl, 5 mM MgCl2, 5 mM DTT and 0.3 mM NAD+; apparent Km values calculated from the Michaelis-Menten equation by non-linear least-squares regression and kcat values calculated from the Vmax of Michaelis-Menten plots using a subunit molecular mass of 56,776 Da.

raw_value_unit_evidence: Table 3. Kinetic constants of pure ScHK2 (journal page 336), row CTP: the Kmapp (mM) column prints 2.0; the kcat (s^-1) column prints 1.61; the kcat/Kmapp (mM^-1 s^-1) column prints 0.8. Footnote a: Values for TTP, GTP, UTP, and CTP, are representative of two independent enzyme preparations.

row_label: CTP

substrate: CTP


column_label: Table 3 row ATP; columns Kmapp (mM), kcat (s^-1), and kcat/Kmapp (mM^-1 s^-1).

normalized_value_unit_evidence: Central values are Kmapp 0.061 mM, kcat 3.5 s^-1, and independently printed kcat/Kmapp 57.0 s^-1 mM^-1.

table_label: Table 3, Kinetic constants of pure ScHK2


column_label: Table 3 row CTP; columns Kmapp (mM), kcat (s^-1), and kcat/Kmapp (mM^-1 s^-1).

normalized_value_unit_evidence: Central values are Kmapp 2.0 mM, kcat 1.61 s^-1, and independently printed kcat/Kmapp 0.8 s^-1 mM^-1.

raw_value_unit_evidence: CTP: Kmapp 2.0 mM; kcat 1.61 s^-1; kcat/Kmapp 0.8 mM^-1 s^-1


substrate_name: D-Fructose

column_label: Table 3 row D-Fructose; columns Kmapp (mM), kcat (s^-1), and kcat/Kmapp (mM^-1 s^-1).

normalized_value_unit_evidence: Central values are Kmapp 5.2 mM, kcat 3.78 s^-1, and independently printed kcat/Kmapp 0.7 s^-1 mM^-1.

row_label: D-Fructose

substrate: D-Fructose


organism: Helianthus annuus


assay_conditions_summary: pH 7; 295.15 K; cofactor ATP


enzyme_display_name: exo-alpha-sialidase

organism: Bacteroides thetaiotaomicron

enzyme: exo-alpha-sialidase


organism: Clostridium perfringens

substrate_name: Unknown

substrate: Unknown


organism: Corynebacterium diphtheriae


organism: Cutibacterium acnes


organism: Danio rerio


organism: Homo sapiens


sequence_source: UniProtKB reviewed accession Q9Y3R4; direct REST lookup on 2026-05-25


sequence_source: UniProtKB reviewed accession Q9UQ49; direct REST lookup on 2026-05-25; variant reconstructed from mutation signature A160G/H277F

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

proof_lines: recombinant mutants A160G and H277F, pH 5.0, 37°C


sequence_source: UniProtKB reviewed accession Q9UQ49; direct REST lookup on 2026-05-25; variant reconstructed from mutation signature R48S


sequence_source: UniProtKB reviewed accession Q9UQ49; direct REST lookup on 2026-05-25; variant reconstructed from mutation signature H277F


sequence_source: UniProtKB reviewed accession Q9UQ49; direct REST lookup on 2026-05-25; variant reconstructed from mutation signature R48N


sequence_source: UniProtKB reviewed accession Q9UQ49; direct REST lookup on 2026-05-25; variant reconstructed from mutation signature A160G

sequence_variant_note: Mutation signature A160G matches the mutant residue at offset +0 and the reference residue at offset -3; variant reconstruction was not applied because multiple plausible residue-numbering offsets are present.


sequence_source: UniProtKB reviewed accession Q9UQ49; direct REST lookup on 2026-05-25

proof_lines: recombinant wild-type enzyme, pH 5.0, 37°C


sequence_source: UniProtKB reviewed accession Q9UQ49; direct REST lookup on 2026-05-25; variant reconstructed from mutation signature E51S

sequence_variant_note: Mutation signature E51S matches the mutant residue at offset +0 and the reference residue at offset -8; variant reconstruction was not applied because multiple plausible residue-numbering offsets are present.


organism: Aspergillus fumigatus

sequence_variant_note: Resolved canonical sequence from UniProtKB:Q4WQS0 direct REST/FASTA lookup; Y358H reconstructed from canonical sequence and applied mutation signature Y358H to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme Y358H, pH 5.2, 16 mM sodium tartrate, at 37°C


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

proof_lines: mutant enzyme R71L, pH 5.2, 16 mM sodium tartrate, at 37°C


sequence_variant_note: Resolved canonical sequence from UniProtKB:Q4WQS0 direct REST/FASTA lookup; D84A reconstructed from canonical sequence and applied mutation signature D84A to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme D84A, pH 5.2, 16 mM sodium tartrate, at 37°C


sequence_source: UniProtKB:Q4WQS0 direct REST/FASTA lookup

proof_lines: wild type enzyme, pH 5.2, 16 mM sodium tartrate, at 37°C


source_db: skid

source_license: CC BY-NC-ND 4.0

sequence_source: UniProtKB:Q4WQS0 direct REST/FASTA lookup from source-provided SKiD accession

source: skid


organism: Aspergillus terreus

sequence_source: UniProtKB:Q0CMX0 direct REST/FASTA lookup

proof_lines: in 100 mM sodium acetate, at pH 4.0 and 37°C


organism: Avian orthoavulavirus 1

sequence_source: UniProtKB:Q9Q2W5 direct REST/FASTA lookup from source-provided SKiD accession


organism: avian paramyxovirus 1

sequence_variant_note: Resolved canonical sequence from UniProtKB:Q9Q2W5 direct REST/FASTA lookup; S418A reconstructed from source-provided accession but paper strain needs arbitration and applied mutation signature S418A to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from UniProt Q9UQ49 canonical sequence plus selected mutation M87G; residue check on 2026-05-25 confirmed canonical M87 before reconstruction. and applied mutation signature M87G to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from UniProt Q9UQ49 canonical sequence plus selected mutation H277F; residue check on 2026-05-25 confirmed canonical H277 before reconstruction. and applied mutation signature H277F to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from UniProt Q9UQ49 canonical sequence plus selected mutation A160G; residue check on 2026-05-25 confirmed canonical A160 before reconstruction. and applied mutation signature A160G to reconstruct the assayed variant sequence.


organism: Influenza A virus

sequence_source: UniProt REST direct check on 2026-05-25: queried P03469, primary accession P03469.


sequence_source: UniProt REST direct check on 2026-05-25: queried P03468, primary accession P03468.


sequence_source: UniProt REST direct check on 2026-05-25: queried secondary accession Q288X1, live primary accession Q1K9J1.


sequence_source: UniProt REST direct check on 2026-05-25: queried Q8JSD9, primary accession Q8JSD9.


sequence_source: UniProt REST direct check on 2026-05-25: queried Q20N24, primary accession Q20N24.


organism: influenza A virus H1N1

sequence_variant_note: Resolved canonical sequence from UniProtKB:C3W6G3 REST lookup on 2026-05-25; applied H275Y and applied mutation signature H275Y to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme H275Y, at 37°C, pH not specified in the publication


sequence_variant_note: Resolved canonical sequence from UniProtKB:C3W6G3 REST lookup on 2026-05-25; applied H275Y/S247N and applied mutation signature H275Y/S247N to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme H275Y/S247N, at 37°C, pH not specified in the publication


sequence_source: UniProtKB:C3W6G3 REST lookup on 2026-05-25

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


organism: influenza A virus H5N1

sequence_source: UniProtKB:A5Z227 REST lookup on 2026-05-25


organism: Influenza B virus

sequence_source: UniProtKB:P03474 REST lookup on 2026-05-25


sequence_variant_note: Resolved canonical sequence from UniProtKB:P03474 REST lookup on 2026-05-25; applied D149E and applied mutation signature D149E to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from UniProtKB:P03474 REST lookup on 2026-05-25; applied R150K and applied mutation signature R150K to reconstruct the assayed variant sequence.


sequence_source: UniProtKB:Q9Q2W5 canonical sequence fetched directly on 2026-05-25


sequence_source: UniProtKB:Q9Q2W5 canonical sequence fetched directly on 2026-05-25, with Y262F applied after confirming residue 262 is Y


organism: Bacteroides fragilis

sequence_source: UniProtKB:Q5L868 direct lookup redirects to UniProtKB:A0A380YS77 canonical sequence, fetched on 2026-05-25


substrate_name: sialic acid

substrate: sialic acid


sequence_source: UniProtKB:Q8AAK9 canonical sequence fetched directly on 2026-05-25


proof_lines:
pH 7.0 and 37°C
Raw SKiD Env: Temp=37°C, pH=7


sequence_source: UniProtKB:Q1K9L1 canonical sequence


organism: influenza A virus

substrate_name: fetuin

sequence_source: UniProtKB:A0A076VGS2 canonical sequence

substrate: fetuin


sequence_source: UniProtKB:C7FH46 canonical sequence

proof_lines:
at pH 5.5 and 37°C
Raw SKiD Env: Temp=37°C, pH=5.5


sequence_source: UniProtKB:C3W6G3 canonical sequence

proof_lines: wild type enzyme, at 37°C, pH not specified in the publication


sequence_variant_note: Resolved canonical sequence from UniProtKB:C3W6G3 canonical sequence with direct 1-based substitution reconstruction H275Y/I223V and applied mutation signature H275Y/I223V to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme H275Y/I223V, at 37°C, pH not specified in the publication


sequence_variant_note: Resolved canonical sequence from UniProtKB:C3W6G3 canonical sequence with direct 1-based substitution reconstruction I223V and applied mutation signature I223V to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme I223V, at 37°C, pH not specified in the publication


sequence_variant_note: Resolved canonical sequence from UniProtKB:C3W6G3 canonical sequence with direct 1-based substitution reconstruction S247N and applied mutation signature S247N to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme S247N, at 37°C, pH not specified in the publication


organism: Glutamicibacter nicotianae

substrate_name: glucomacropeptide

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

proof_lines:
37°C, sialidase isoenzyme NA2
37°C, sialidase isoenzyme NA1

substrate: glucomacropeptide


organism: Aeropyrum pernix

substrate_name: D-glucose 6-phosphate

smiles: C(C(C(C(C(C=O)O)O)O)O)OP(=O)(O)O

assay_conditions_summary: pH 6.7; 358.15 K; cofactor ATP

substrate: D-glucose 6-phosphate


substrate_name: adenosine

substrate: adenosine


substrate_name: fructose

substrate: fructose


substrate_name: D-ribose 5-phosphate

substrate: D-ribose 5-phosphate


substrate_name: D-ribose

substrate: D-ribose


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

substrate_name: 4-methylumbelliferyl alpha-D-N-acetylneuraminic acid (MUN)


substrate_name: ganglioside GD1a

substrate: ganglioside GD1a


substrate_name: ganglioside GM3 (in the presence of Triton X-100)


sequence_source: UniProtKB:P03474 canonical FASTA (sp|P03474|NRAM_INBLE Neuraminidase OS=Influenza B virus (strain B/Lee/1940) OX=518987 GN=NA PE=1 SV=1) with selected R374N substitution applied after coordinate check on 2026-05-25

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


sequence_source: UniProtKB:P03474 canonical FASTA (sp|P03474|NRAM_INBLE Neuraminidase OS=Influenza B virus (strain B/Lee/1940) OX=518987 GN=NA PE=1 SV=1) with selected R374K substitution applied after coordinate check on 2026-05-25


sequence_source: UniProtKB:P03474 canonical FASTA (sp|P03474|NRAM_INBLE Neuraminidase OS=Influenza B virus (strain B/Lee/1940) OX=518987 GN=NA PE=1 SV=1) with selected E275D substitution applied after coordinate check on 2026-05-25


sequence_source: UniProtKB:P03474 canonical FASTA (sp|P03474|NRAM_INBLE Neuraminidase OS=Influenza B virus (strain B/Lee/1940) OX=518987 GN=NA PE=1 SV=1) with selected E117G substitution applied after coordinate check on 2026-05-25


sequence_source: UniProtKB:P03474 canonical FASTA (sp|P03474|NRAM_INBLE Neuraminidase OS=Influenza B virus (strain B/Lee/1940) OX=518987 GN=NA PE=1 SV=1) with selected R223K substitution applied after coordinate check on 2026-05-25


organism: Micromonospora viridifaciens

substrate_name: phenyl alpha-sialoside

sequence_source: UniProtKB:Q02834 canonical FASTA (sp|Q02834|NANH_MICVI Sialidase OS=Micromonospora viridifaciens OX=1881 GN=nedA PE=1 SV=1) with selected Y370G substitution applied after coordinate check on 2026-05-25

substrate: phenyl alpha-sialoside


sequence_source: UniProtKB:Q02834 canonical FASTA (sp|Q02834|NANH_MICVI Sialidase OS=Micromonospora viridifaciens OX=1881 GN=nedA PE=1 SV=1) with selected E260D substitution applied after coordinate check on 2026-05-25


sequence_source: UniProtKB:Q02834 canonical FASTA (sp|Q02834|NANH_MICVI Sialidase OS=Micromonospora viridifaciens OX=1881 GN=nedA PE=1 SV=1) with selected E260C substitution applied after coordinate check on 2026-05-25


sequence_source: UniProtKB:Q02834 reviewed Swiss-Prot; REST check 2026-05-25


organism: Mumps orthorubulavirus

sequence_source: UniProtKB:P19762 reviewed Swiss-Prot; PMID 34778199 methods cite P19762-1

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


organism: Porphyromonas gingivalis

sequence_source: GenBank:AUR47485.1 and RCSB PDB:8GN6; source UniProt A0A1R4DV85 is deleted/inactive


organism: Tannerella forsythia

sequence_variant_note: Resolved canonical sequence from UniProtKB:G8UIQ1 canonical sequence with PMID 35916484/PDB author-numbering offset -13 applied and applied mutation signature R306A with inferred residue-numbering offset -13 to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from UniProtKB:G8UIQ1 canonical sequence with PMID 35916484/PDB author-numbering offset -13 applied and applied mutation signature V236Y with inferred residue-numbering offset -13 to reconstruct the assayed variant sequence.


sequence_source: UniProtKB:G8UIQ1 unreviewed TrEMBL; REST check 2026-05-25; RCSB 7QYP maps to G8UIQ1


sequence_variant_note: Resolved canonical sequence from UniProtKB:G8UIQ1 canonical sequence with PMID 35916484/PDB author-numbering offset -13 applied and applied mutation signature V236Q with inferred residue-numbering offset -13 to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from UniProtKB:G8UIQ1 canonical sequence with PMID 35916484/PDB author-numbering offset -13 applied and applied mutation signature I456Y with inferred residue-numbering offset -13 to reconstruct the assayed variant sequence.


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

proof_lines: mutant enzyme N234Y/S235Y, at pH 7.4 and 25°C

proof_lines: mutant enzyme N234YS235Y, at pH 7.4 and 25°C


organism: Treponema denticola

sequence_source: UniProtKB REST/FASTA accession Q73QH2 checked 2026-05-25

proof_lines: in 70 mM sodium citrate, at pH 7.0 and 37°C


organism: Trichophyton rubrum

sequence_source: UniProtKB REST/FASTA accession A0A178EUH2 checked 2026-05-25

proof_lines: in 100 mM sodium acetate,at pH 4.0 and 37°C


organism: Geobacillus stearothermophilus

substrate_name: ATP

assay_conditions_summary: pH 8.2; 303.15 K; cofactor ATP

column_label: Table 2: Alternative Nucleoside Triphosphate Substrates (Biochemistry 1994, 33(11):3424-3431); row ATP; the Michaelis constant 0.07 mM comes from the column Km(NTP) (mM) of that row, whose footnote a states that Fru-6P concentration was kept fixed at 1.0 mM (saturating) for these determinations; the turnover number 112 s-1 comes from the Results text eq 3 fit of the forward reaction, which is the 100 percent reference of the Table 2 column % of kcat(ATP) and is the only forward-reaction turnover number the article prints. There is no kcat/Km column.

normalized_value_unit_evidence: kcat: printed value 112 s-1; already in s^-1, so no unit conversion applies; the paper value is 112.0 s^(-1). Km: printed cell 0.07 mM; already in mM, so no unit conversion applies; the paper value is 0.07 mM. kcat/Km: this article prints no kcat/Km cell for this measurement, so the catalog value 1599.9999999999998 s^(-1)*mM^(-1) is not a printed value; it is the quotient 112 / 0.07 of this measurement's own two printed values, typed derived_same_measurement.

raw_value_unit_evidence: Table 2, row ATP: Km = 0.07 mM printed in the column Km(NTP) (mM). Results text, forward reaction fit to eq 3 with A as MgATP and B as Fru-6P: kcat = 112 s-1. No kcat/Km value is printed for this measurement.

row_label: Table 2, row ATP, alternative nucleoside triphosphate substrates, read with the forward-reaction turnover number of the Results text

substrate: ATP


substrate_name: D-Fructose 1,6-bisphosphate

assay_conditions_summary: pH 8.2; 303.15 K; cofactor ADP

column_label: Results text, reverse reaction initial velocity analysis (Biochemistry 1994, 33(11):3424-3431); the paragraph states that the initial velocity data were fit to the equation for a rapid-equilibrium random or ordered bireactant mechanism (eq 3), assuming that A is MgADP and B is Fru-1,6BP, and prints kcat = 3.2 s-1 together with Kb = 0.44 mM, the Michaelis constant of substrate B, which that fit declares to be Fru-1,6BP. The same paragraph prints Ka = 0.11 mM for MgADP and the two dissociation constants Kia and Kib. There is no kcat/Km value.

normalized_value_unit_evidence: kcat: printed value 3.2 s-1; already in s^-1, so no unit conversion applies; the paper value is 3.2 s^(-1). Km: printed value 0.44 mM; already in mM, so no unit conversion applies; the paper value is 0.44 mM. kcat/Km: this article prints no kcat/Km cell for this measurement, so the catalog value 7.272727272727273 s^(-1)*mM^(-1) is not a printed value; it is the quotient 3.2 / 0.44 of this measurement's own two printed values, typed derived_same_measurement.

raw_value_unit_evidence: Results text, reverse reaction fit to eq 3 with A as MgADP and B as Fru-1,6BP: kcat = 3.2 s-1 and Kb = 0.44 mM, the Michaelis constant of substrate B, Fru-1,6BP. No kcat/Km value is printed for this measurement.

row_label: Results text, reverse reaction initial velocity fit to eq 3 with A as MgADP and B as Fru-1,6BP

substrate: D-Fructose 1,6-bisphosphate


substrate_name: GTP

substrate: GTP


substrate_name: UTP

substrate: UTP


organism: Homo sapiens

substrate_name: dATP (at 30 degrees Celsius)

substrate: dATP (at 30 degrees Celsius)


substrate_name: dGTP (at 30 degrees Celsius)

substrate: dGTP (at 30 degrees Celsius)


substrate_name: dCTP (at 30 degrees Celsius)

substrate: dCTP (at 30 degrees Celsius)


substrate_name: dTTP (at 30 degrees Celsius)

substrate: dTTP (at 30 degrees Celsius)


enzyme_display_name: mandelate racemase

organism: Pseudomonas putida

sequence_source: UniProtKB P11444 canonical FASTA direct check plus selected source mutation_signature V22L; V22L checked against canonical residue V22.

enzyme: mandelate racemase


sequence_source: UniProtKB P11444 canonical FASTA direct check plus selected source mutation_signature Y54A; Y54A checked against canonical residue Y54.

sequence_variant_note: Mutation signature Y54A matches the mutant residue at offset +0 and the reference residue at offset -4; variant reconstruction was not applied because multiple plausible residue-numbering offsets are present.


sequence_source: UniProtKB P11444 canonical FASTA direct check plus selected source mutation_signature V26I; V26I checked against canonical residue V26.


sequence_source: UniProtKB P11444 canonical FASTA direct check plus selected source mutation_signature Y54I; Y54I checked against canonical residue Y54.

sequence_variant_note: Mutation signature Y54I matches the mutant residue at offset +0 and the reference residue at offset -4; variant reconstruction was not applied because multiple plausible residue-numbering offsets are present.


sequence_source: UniProtKB P11444 canonical FASTA direct check plus selected source mutation_signature Y54F; Y54F checked against canonical residue Y54.

sequence_variant_note: Mutation signature Y54F matches the mutant residue at offset +0 and the reference residue at offset -4; variant reconstruction was not applied because multiple plausible residue-numbering offsets are present.


sequence_source: UniProtKB P11444 FASTA direct check; reviewed Swiss-Prot mandelate racemase from Pseudomonas putida.


sequence_source: UniProtKB P11444 canonical sequence reconstructed with selected mutation signature


sequence_variant_note: Mutation signature Y54V matches the mutant residue at offset +0 and the reference residue at offset -4; variant reconstruction was not applied because multiple plausible residue-numbering offsets are present.


sequence_variant_note: Mutation signature Y54L matches the mutant residue at offset +0 and the reference residue at offset -4; variant reconstruction was not applied because multiple plausible residue-numbering offsets are present.


substrate_name: (R)-mandelamide

substrate: (R)-mandelamide


sequence_variant_note: Mutation signature V26I/Y54I matches the mutant residue at offset +0 and the reference residue at offset -4; variant reconstruction was not applied because multiple plausible residue-numbering offsets are present.


sequence_source: UniProtKB REST FASTA P11444 canonical sequence; reconstructed V29F after confirming canonical residue 29 is V.

proof_lines: pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V29F


substrate_name: (R)-mandelate

sequence_source: UniProtKB REST FASTA P11444 canonical sequence.

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

substrate: (R)-mandelate


sequence_source: UniProtKB REST FASTA P11444 canonical sequence; reconstructed V22A after confirming canonical residue 22 is V.

proof_lines:
mutant V22A, pH 7.5, 25°C
pH 7.5, 3.3 mM Mg2+, mutant V22A
pH 7.5, 3.3 mM Mg2+ mutant V22A


sequence_source: UniProtKB REST FASTA P11444 canonical sequence; reconstructed V29L after confirming canonical residue 29 is V.

proof_lines:
mutant V29L, pH 7.5, 25°C
pH 7.5, 3.3 mM Mg2+, mutant V29L
pH 7.5, 25°C, mutant V29L


substrate_name: (S)-trifluorolactate

substrate: (S)-trifluorolactate


substrate_name: (R)-trifluorolactate

substrate: (R)-trifluorolactate


sequence_source: UniProtKB REST FASTA P11444 canonical sequence; reconstructed V26I after confirming canonical residue 26 is V.


sequence_source: UniProtKB REST FASTA P11444 canonical sequence; reconstructed L319F after confirming canonical residue 319 is L.


sequence_source: UniProtKB:P11444 canonical sequence with V26L reconstructed after direct residue check


sequence_source: UniProtKB:P11444 canonical sequence with Y54F reconstructed after direct residue check


sequence_source: UniProtKB:P11444 canonical sequence with Y54L reconstructed after direct residue check


sequence_source: UniProtKB:P11444 canonical sequence with V26I/Y54I reconstructed after direct residue checks


sequence_source: UniProtKB:P11444 canonical sequence


sequence_source: UniProtKB:P11444 canonical sequence with V26I/Y54V reconstructed after direct residue checks

sequence_variant_note: Mutation signature V26I/Y54V matches the mutant residue at offset +0 and the reference residue at offset -4; variant reconstruction was not applied because multiple plausible residue-numbering offsets are present.


sequence_source: UniProtKB:P11444 canonical sequence with V26S reconstructed after direct residue check


sequence_source: UniProt REST P11444 canonical sequence checked on 2026-05-25; mutation V26A applied after validating the canonical residue coordinate.

proof_lines: pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V26A


sequence_source: UniProt REST P11444 canonical sequence checked on 2026-05-25; mutation V22I/V29L applied after validating the canonical residue coordinate.

proof_lines: mutant V22I/V29L, pH 7.5, 25°C, presence of 20 mM Mg2+

proof_lines: pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V22I/V29L


substrate_name: (S)-Mandelate

sequence_source: UniProt REST P11444 canonical sequence checked on 2026-05-25; mutation V26A/V29L applied after validating the canonical residue coordinate.

proof_lines: pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V26A/V29L

substrate: (S)-Mandelate


sequence_source: UniProt REST P11444 canonical sequence checked on 2026-05-25; mutation V22F applied after validating the canonical residue coordinate.

proof_lines: pH 7.5, mutant V22F, addition of either 10 mM (R)- or (S)-mandelate


sequence_source: UniProt REST P11444 canonical sequence checked on 2026-05-25; mutation T24S applied after validating the canonical residue coordinate.

proof_lines: pH 7.5, mutant T24S, addition of either 10 mM (R)- or (S)-mandelate


sequence_source: UniProt REST P11444 canonical sequence checked on 2026-05-25; mutation V26F applied after validating the canonical residue coordinate.

proof_lines: pH 7.5, mutant V26F, addition of either 10 mM (R)- or (S)-mandelate


sequence_source: UniProt REST P11444 canonical sequence checked on 2026-05-25; mutation V22I applied after validating the canonical residue coordinate.

proof_lines: mutant V22I, pH 7.5, 25°C, presence of 20 mM Mg2+


sequence_source: UniProt REST P11444 canonical sequence checked on 2026-05-25; mutation A25V applied after validating the canonical residue coordinate.

proof_lines: pH 7.5 at 25°C, 3.3 mM Mg2+, mutant A25V


sequence_source: UniProt P11444 FASTA retrieved directly; V26I and Y54V applied after confirming P11444 has V at 26 and Y at 54.


sequence_source: UniProt P11444 FASTA retrieved directly; V29L applied after confirming P11444 has V at 29.


sequence_source: UniProt P11444 FASTA retrieved directly; Y54A applied after confirming P11444 has Y at 54.


sequence_source: UniProt P11444 FASTA retrieved directly; V26S applied after confirming P11444 has V at 26.


sequence_source: UniProt P11444 FASTA retrieved directly; V22I applied after confirming P11444 has V at 22.


sequence_source: UniProt P11444 FASTA checked directly; variant sequence not reconstructed because Y45F conflicts with the P11444 reference residue at position 45.


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:P11444 and applied mutation signature V22I/V29I to reconstruct the assayed variant sequence.

assay_conditions_summary: pH 7.5; 298.15 K; buffer Tris-HCl

column_label: Table 2: Kinetic analysis of the WT and mutants of MR toward (S)-o-chloromandelic acid and (R)-o-chloromandelic acid; group header column '(R)-2-CMA' covering the three kinetic columns 'Km (mM)', 'kcat (s-1)' and 'kcat/Km (s-1 M-1)'; line for enzyme form 'V22I/V29I': Km = 15.10 mM, kcat = 2.60 s^(-1), kcat/Km = 172.25 M^-1 s^-1; the trailing 'Keq' column of that line is not a kinetic metric of the export schema.

normalized_value_unit_evidence: Km: printed Table 2 cell 15.10 mM; the printed unit is already the export unit, so no unit conversion applies and the normalized value is 15.1 mM. kcat: printed Table 2 cell 2.60 s^(-1); the printed unit is already the export unit, so no unit conversion applies and the normalized value is 2.6 s^(-1). kcat/Km: printed Table 2 cell 172.25 M^-1 s^-1; 1 M^-1 s^-1 is 0.001 s^-1 mM^-1, so 172.25 / 1000 = 0.17225 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 2, (R)-2-CMA half of the printed line for enzyme form V22I/V29I: Km = 15.10 mM; kcat = 2.60 s^(-1); printed kcat/Km = 172.25 M^-1 s^-1.


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:P11444 and applied mutation signature V22I/V29I/Y54F to reconstruct the assayed variant sequence.

column_label: Table 2: Kinetic analysis of the WT and mutants of MR toward (S)-o-chloromandelic acid and (R)-o-chloromandelic acid; group header column '(R)-2-CMA' covering the three kinetic columns 'Km (mM)', 'kcat (s-1)' and 'kcat/Km (s-1 M-1)'; line for enzyme form 'V22I/V29I/Y54F': Km = 14.52 mM, kcat = 2.57 s^(-1), kcat/Km = 176.70 M^-1 s^-1; the trailing 'Keq' column of that line is not a kinetic metric of the export schema.

normalized_value_unit_evidence: Km: printed Table 2 cell 14.52 mM; the printed unit is already the export unit, so no unit conversion applies and the normalized value is 14.52 mM. kcat: printed Table 2 cell 2.57 s^(-1); the printed unit is already the export unit, so no unit conversion applies and the normalized value is 2.57 s^(-1). kcat/Km: printed Table 2 cell 176.70 M^-1 s^-1; 1 M^-1 s^-1 is 0.001 s^-1 mM^-1, so 176.70 / 1000 = 0.1767 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 2, (R)-2-CMA half of the printed line for enzyme form V22I/V29I/Y54F: Km = 14.52 mM; kcat = 2.57 s^(-1); printed kcat/Km = 176.70 M^-1 s^-1.


sequence_source: UniProtKB:P11444 canonical FASTA, direct lookup on 2026-05-25

assay_conditions_summary: pH 7.5; 298.15 K; buffer Na-HEPES

proof_lines: recombinant mandelate racemase, putative misfolded form with N-terminal hexahistidine tag, Na-HEPES buffer (0.1 M, pH 7.5), 3.3 mM MgCl2

proof_lines: recombinant mandelate racemase represents correctly folded enzyme, with N-terminal hexahistidine tag, in Na-HEPES buffer (0.1 M, pH 7.5), 3.3 mM MgCl2


sequence_source: UniProtKB:P11444 canonical FASTA with validated F52W substitution, reconstructed on 2026-05-25


sequence_source: UniProtKB:P11444 canonical FASTA with validated Y54W substitution, reconstructed on 2026-05-25

sequence_variant_note: Mutation signature Y54W matches the mutant residue at offset +0 and the reference residue at offset -4; variant reconstruction was not applied because multiple plausible residue-numbering offsets are present.


sequence_source: UniProtKB:P11444 canonical FASTA with validated F52W/Y54W substitutions, reconstructed on 2026-05-25

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


sequence_source: UniProtKB:P11444 canonical FASTA with validated V26L substitution, reconstructed on 2026-05-25

proof_lines:
mutant V26L, pH 7.5, 25°C
pH 7.5, 3.3 mM Mg2+, mutant V26L
pH 7.5, 25°C, mutant V26L


sequence_source: UniProtKB:P11444 canonical FASTA with validated V29A substitution, reconstructed on 2026-05-25

proof_lines: pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V29A


organism: Unknown

substrate_name: Unknown

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

substrate: Unknown


sequence_source: UniProt P11444 FASTA direct check, canonical wild-type sequence

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

proof_lines: wild-type, pH 7.5, 25°C, presence of 20 mM Mg2+


column_label: Table 2: Kinetic analysis of the WT and mutants of MR toward (S)-o-chloromandelic acid and (R)-o-chloromandelic acid; group header column '(S)-2-CMA' covering the three kinetic columns 'Km (mM)', 'kcat (s-1)' and 'kcat/Km (s-1 M-1)'; line for enzyme form 'V22I/V29I/Y54F': Km = 17.94 mM, kcat = 3.49 s^(-1), kcat/Km = 194.69 M^-1 s^-1; the trailing 'Keq' column of that line is not a kinetic metric of the export schema.

normalized_value_unit_evidence: Km: printed Table 2 cell 17.94 mM; the printed unit is already the export unit, so no unit conversion applies and the normalized value is 17.94 mM. kcat: printed Table 2 cell 3.49 s^(-1); the printed unit is already the export unit, so no unit conversion applies and the normalized value is 3.49 s^(-1). kcat/Km: printed Table 2 cell 194.69 M^-1 s^-1; 1 M^-1 s^-1 is 0.001 s^-1 mM^-1, so 194.69 / 1000 = 0.19469 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 2, (S)-2-CMA half of the printed line for enzyme form V22I/V29I/Y54F: Km = 17.94 mM; kcat = 3.49 s^(-1); printed kcat/Km = 194.69 M^-1 s^-1.


row_label: Table 2, (R)-2-CMA half of the printed line for enzyme form V22I/V29I

table_label: Table 2: Kinetic analysis of the WT and mutants of MR toward (S)-o-chloromandelic acid and (R)-o-chloromandelic acid


column_label: Table 2: Kinetic analysis of the WT and mutants of MR toward (S)-o-chloromandelic acid and (R)-o-chloromandelic acid; group header column '(R)-2-CMA' covering the three kinetic columns 'Km (mM)', 'kcat (s-1)' and 'kcat/Km (s-1 M-1)'; line for enzyme form 'WT': Km = 14.82 mM, kcat = 0.90 s^(-1), kcat/Km = 60.74 M^-1 s^-1; the trailing 'Keq' column of that line is not a kinetic metric of the export schema.

normalized_value_unit_evidence: Km: printed Table 2 cell 14.82 mM; the printed unit is already the export unit, so no unit conversion applies and the normalized value is 14.82 mM. kcat: printed Table 2 cell 0.90 s^(-1); the printed unit is already the export unit, so no unit conversion applies and the normalized value is 0.9 s^(-1). kcat/Km: printed Table 2 cell 60.74 M^-1 s^-1; 1 M^-1 s^-1 is 0.001 s^-1 mM^-1, so 60.74 / 1000 = 0.06074 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 2, (R)-2-CMA half of the printed line for enzyme form WT: Km = 14.82 mM; kcat = 0.90 s^(-1); printed kcat/Km = 60.74 M^-1 s^-1.


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:P11444 and applied mutation signature V29I to reconstruct the assayed variant sequence.

column_label: Table 2: Kinetic analysis of the WT and mutants of MR toward (S)-o-chloromandelic acid and (R)-o-chloromandelic acid; group header column '(S)-2-CMA' covering the three kinetic columns 'Km (mM)', 'kcat (s-1)' and 'kcat/Km (s-1 M-1)'; line for enzyme form 'V29I': Km = 14.35 mM, kcat = 1.83 s^(-1), kcat/Km = 127.66 M^-1 s^-1; the trailing 'Keq' column of that line is not a kinetic metric of the export schema.

normalized_value_unit_evidence: Km: printed Table 2 cell 14.35 mM; the printed unit is already the export unit, so no unit conversion applies and the normalized value is 14.35 mM. kcat: printed Table 2 cell 1.83 s^(-1); the printed unit is already the export unit, so no unit conversion applies and the normalized value is 1.83 s^(-1). kcat/Km: printed Table 2 cell 127.66 M^-1 s^-1; 1 M^-1 s^-1 is 0.001 s^-1 mM^-1, so 127.66 / 1000 = 0.12766 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 2, (S)-2-CMA half of the printed line for enzyme form V29I: Km = 14.35 mM; kcat = 1.83 s^(-1); printed kcat/Km = 127.66 M^-1 s^-1.


column_label: Table 2: Kinetic analysis of the WT and mutants of MR toward (S)-o-chloromandelic acid and (R)-o-chloromandelic acid; group header column '(S)-2-CMA' covering the three kinetic columns 'Km (mM)', 'kcat (s-1)' and 'kcat/Km (s-1 M-1)'; line for enzyme form 'V22I/V29I': Km = 13.74 mM, kcat = 2.35 s^(-1), kcat/Km = 159.31 M^-1 s^-1; the trailing 'Keq' column of that line is not a kinetic metric of the export schema.

normalized_value_unit_evidence: Km: printed Table 2 cell 13.74 mM; the printed unit is already the export unit, so no unit conversion applies and the normalized value is 13.74 mM. kcat: printed Table 2 cell 2.35 s^(-1); the printed unit is already the export unit, so no unit conversion applies and the normalized value is 2.35 s^(-1). kcat/Km: printed Table 2 cell 159.31 M^-1 s^-1; 1 M^-1 s^-1 is 0.001 s^-1 mM^-1, so 159.31 / 1000 = 0.15931 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 2, (S)-2-CMA half of the printed line for enzyme form V22I/V29I: Km = 13.74 mM; kcat = 2.35 s^(-1); printed kcat/Km = 159.31 M^-1 s^-1.


organism: Thermoproteus tenax

assay_conditions_summary: pH 7.5; 323.15 K; cofactor ATP


substrate_name: D-glucose

substrate: D-glucose


smiles: C(C=O)C(C(C(CO)O)O)O


substrate_name: D-fructose

substrate: D-fructose


sequence_source: UniProtKB P11444 REST/FASTA direct check; single-substitution reconstruction Y137L from canonical sequence.


sequence_source: UniProtKB P11444 REST/FASTA direct check; single-substitution reconstruction Y137S from canonical sequence.


sequence_source: UniProtKB P11444 REST/FASTA direct check; single-substitution reconstruction K166R from canonical sequence.

sequence_variant_note: Mutation signature K166R matches the mutant residue at offset +0 and the reference residue at offset -2; variant reconstruction was not applied because multiple plausible residue-numbering offsets are present.

proof_lines:
pH 7.5, 25°C, mutant enzyme K166R
mutant K166R
25°C, pH 7.5, mutant enzyme K166R


sequence_source: UniProtKB P11444 REST/FASTA direct check; single-substitution reconstruction K164R from canonical sequence.

sequence_variant_note: Mutation signature K164R matches the mutant residue at offset +0 and the reference residue at offset +2; variant reconstruction was not applied because multiple plausible residue-numbering offsets are present.

proof_lines:
pH 7.5, 25°C, mutant enzyme K164R
25°C, pH 7.5, mutant enzyme K164R


sequence_source: UniProtKB P11444 REST/FASTA direct check; single-substitution reconstruction L93R from canonical sequence.


sequence_source: UniProtKB P11444 REST/FASTA direct check; single-substitution reconstruction L93N from canonical sequence.


sequence_source: UniProtKB P11444 REST/FASTA direct check; single-substitution reconstruction L93A from canonical sequence.


sequence_source: UniProtKB P11444 REST/FASTA direct check; single-substitution reconstruction L93F from canonical sequence.


sequence_variant_note: Resolved canonical sequence from uniprot_rest_accession:P11444 and applied mutation signature V22I to reconstruct the assayed variant sequence.

column_label: Table 2: Kinetic analysis of the WT and mutants of MR toward (S)-o-chloromandelic acid and (R)-o-chloromandelic acid; group header column '(R)-2-CMA' covering the three kinetic columns 'Km (mM)', 'kcat (s-1)' and 'kcat/Km (s-1 M-1)'; line for enzyme form 'V22I': Km = 15.51 mM, kcat = 1.82 s^(-1), kcat/Km = 117.82 M^-1 s^-1; the trailing 'Keq' column of that line is not a kinetic metric of the export schema.

normalized_value_unit_evidence: Km: printed Table 2 cell 15.51 mM; the printed unit is already the export unit, so no unit conversion applies and the normalized value is 15.51 mM. kcat: printed Table 2 cell 1.82 s^(-1); the printed unit is already the export unit, so no unit conversion applies and the normalized value is 1.82 s^(-1). kcat/Km: printed Table 2 cell 117.82 M^-1 s^-1; 1 M^-1 s^-1 is 0.001 s^-1 mM^-1, so 117.82 / 1000 = 0.11782 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 2, (R)-2-CMA half of the printed line for enzyme form V22I: Km = 15.51 mM; kcat = 1.82 s^(-1); printed kcat/Km = 117.82 M^-1 s^-1.


column_label: Table 2: Kinetic analysis of the WT and mutants of MR toward (S)-o-chloromandelic acid and (R)-o-chloromandelic acid; group header column '(R)-2-CMA' covering the three kinetic columns 'Km (mM)', 'kcat (s-1)' and 'kcat/Km (s-1 M-1)'; line for enzyme form 'V29I': Km = 11.99 mM, kcat = 1.45 s^(-1), kcat/Km = 121.37 M^-1 s^-1; the trailing 'Keq' column of that line is not a kinetic metric of the export schema.

normalized_value_unit_evidence: Km: printed Table 2 cell 11.99 mM; the printed unit is already the export unit, so no unit conversion applies and the normalized value is 11.99 mM. kcat: printed Table 2 cell 1.45 s^(-1); the printed unit is already the export unit, so no unit conversion applies and the normalized value is 1.45 s^(-1). kcat/Km: printed Table 2 cell 121.37 M^-1 s^-1; 1 M^-1 s^-1 is 0.001 s^-1 mM^-1, so 121.37 / 1000 = 0.12137 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 2, (R)-2-CMA half of the printed line for enzyme form V29I: Km = 11.99 mM; kcat = 1.45 s^(-1); printed kcat/Km = 121.37 M^-1 s^-1.


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


proof_lines: 25°C, pH 7.5, mutant enzyme tmMR-thiaLys


proof_lines: 25°C, pH 7.5, mutant enzyme C92S/C264S/166thiaK


sequence_source: UniProtKB:P11444 canonical FASTA with validated Y137F substitution, checked 2026-05-25


sequence_source: UniProtKB:P11444 canonical FASTA with validated Y137T substitution, checked 2026-05-25


row_label: Table 2, (R)-2-CMA half of the printed line for enzyme form V22I/V29I/Y54F


column_label: Table 2: Kinetic analysis of the WT and mutants of MR toward (S)-o-chloromandelic acid and (R)-o-chloromandelic acid; group header column '(S)-2-CMA' covering the three kinetic columns 'Km (mM)', 'kcat (s-1)' and 'kcat/Km (s-1 M-1)'; line for enzyme form 'V22I': Km = 19.91 mM, kcat = 2.47 s^(-1), kcat/Km = 123.97 M^-1 s^-1; the trailing 'Keq' column of that line is not a kinetic metric of the export schema.

normalized_value_unit_evidence: Km: printed Table 2 cell 19.91 mM; the printed unit is already the export unit, so no unit conversion applies and the normalized value is 19.91 mM. kcat: printed Table 2 cell 2.47 s^(-1); the printed unit is already the export unit, so no unit conversion applies and the normalized value is 2.47 s^(-1). kcat/Km: printed Table 2 cell 123.97 M^-1 s^-1; 1 M^-1 s^-1 is 0.001 s^-1 mM^-1, so 123.97 / 1000 = 0.12397 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 2, (S)-2-CMA half of the printed line for enzyme form V22I: Km = 19.91 mM; kcat = 2.47 s^(-1); printed kcat/Km = 123.97 M^-1 s^-1.


row_label: Table 2, (R)-2-CMA half of the printed line for enzyme form V29I


sequence_source: UniProtKB P11444 canonical FASTA fetched 2026-05-25.

column_label: Table 2: Kinetic analysis of the WT and mutants of MR toward (S)-o-chloromandelic acid and (R)-o-chloromandelic acid; group header column '(S)-2-CMA' covering the three kinetic columns 'Km (mM)', 'kcat (s-1)' and 'kcat/Km (s-1 M-1)'; line for enzyme form 'WT': Km = 17.74 mM, kcat = 0.99 s^(-1), kcat/Km = 56.34 M^-1 s^-1; the trailing 'Keq' column of that line is not a kinetic metric of the export schema.

normalized_value_unit_evidence: Km: printed Table 2 cell 17.74 mM; the printed unit is already the export unit, so no unit conversion applies and the normalized value is 17.74 mM. kcat: printed Table 2 cell 0.99 s^(-1); the printed unit is already the export unit, so no unit conversion applies and the normalized value is 0.99 s^(-1). kcat/Km: printed Table 2 cell 56.34 M^-1 s^-1; 1 M^-1 s^-1 is 0.001 s^-1 mM^-1, so 56.34 / 1000 = 0.05634 s^(-1)*mM^(-1).

raw_value_unit_evidence: Table 2, (S)-2-CMA half of the printed line for enzyme form WT: Km = 17.74 mM; kcat = 0.99 s^(-1); printed kcat/Km = 56.34 M^-1 s^-1.


organism: Escherichia coli

substrate_name: ITP

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

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

substrate: ITP


assay_conditions_summary: pH 7.2; 303.15 K; cofactor ATP


substrate_name: CTP

substrate: CTP


sequence_variant_note: Resolved canonical sequence from UniProtKB:P11444 FASTA; row-specific L93K reconstruction after confirming canonical residue L at position 93. and applied mutation signature L93K to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from UniProtKB:P11444 FASTA; row-specific L93W reconstruction after confirming canonical residue L at position 93. and applied mutation signature L93W to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from UniProtKB:P11444 FASTA; row-specific L93D reconstruction after confirming canonical residue L at position 93. and applied mutation signature L93D to reconstruct the assayed variant sequence.


sequence_variant_note: Resolved canonical sequence from UniProtKB:P11444 FASTA; row-specific D270N reconstruction after confirming canonical residue D at position 270. and applied mutation signature D270N to reconstruct the assayed variant sequence.


organism: Saccharolobus solfataricus

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


substrate_name: dCTP

substrate: dCTP


substrate_name: dPTP

substrate: dPTP


organism: Saccharomyces cerevisiae (strain ATCC 204508 / S288c)

substrate_name: ATP

substrate: ATP


substrate_name: fructose 6-phosphate

substrate: fructose 6-phosphate


organism: African swine fever virus (strain Badajoz 1971 Vero-adapted)

substrate_name: dGTP

substrate: dGTP


substrate_name: dATP

substrate: dATP


substrate_name: dTTP

substrate: dTTP


substrate_name: dGTP (insertion opposite abasic site)

substrate: dGTP (insertion opposite abasic site)


substrate_name: dATP (insertion opposite abasic site)

substrate: dATP (insertion opposite abasic site)


substrate_name: dTTP (insertion opposite abasic site)

substrate: dTTP (insertion opposite abasic site)


substrate_name: dCTP (insertion opposite abasic site)

substrate: dCTP (insertion opposite abasic site)


organism: Brugia malayi

substrate_name: glucose

substrate: glucose


substrate_name: fructose

substrate: fructose


organism: Toxoplasma gondii

assay_conditions_summary: pH 7; 310.15 K; cofactor ATP


substrate_name: D-glucose

substrate: D-glucose


organism: Schizosaccharomyces pombe

source_databases_merged: brenda uniprot


substrate_name: D-mannose

substrate: D-mannose


substrate_name: D-fructose

substrate: D-fructose


organism: Macrobrachium nipponense


substrate_name: sedoheptulose 7-phosphate

smiles: C(C(C(C(C(C(=O)CO)O)O)O)O)OP(=O)(O)O

proof_lines:
pH 7.0, 55°C
Raw SKiD Env: Temp=55°C, pH=7

substrate: sedoheptulose 7-phosphate


assay_conditions_summary: pH 7; 328.15 K; cofactor NADH

column_label: Table 1; row PfkA E.co PFK; cofactor ATP; substrate S7P; Km cell 2.5 mM; kcat cell 0.25 s^-1; kcat/Km cell 0.15 x 10^3 s^-1 M^-1; assay at 55 C and pH 7.0.

raw_value_unit_evidence: Table 1 PfkA E.co PFK S7P cells: Km 2.5 mM; kcat 0.25 s^-1; kcat/Km 0.15 x 10^3 s^-1 M^-1.

row_label: PfkA E.co PFK | sedoheptulose 7-phosphate

table_label: Table 1. Kinetics of the PFKs tested


substrate_name: D-Fructose 6-phosphate

column_label: Table 1; row PfkA E.co PFK; cofactor ATP; substrate F6P; Km cell 0.065 mM; kcat cell 1.2 s^-1; kcat/Km cell 19 x 10^3 s^-1 M^-1; assay at 55 C and pH 7.0.

raw_value_unit_evidence: Table 1 PfkA E.co PFK F6P cells: Km 0.065 mM; kcat 1.2 s^-1; kcat/Km 19 x 10^3 s^-1 M^-1.

row_label: PfkA E.co PFK | F6P

substrate: D-Fructose 6-phosphate


organism: Thermotoga maritima

sequence_source: multiple

assay_conditions_summary: pH 7.8; 323.15 K; cofactor ATP


substrate_name: D-fructose 6-phosphate

substrate: D-fructose 6-phosphate


substrate_name: GTP

substrate: GTP


substrate_name: ATP

substrate: ATP


substrate_name: dGTP

substrate: dGTP


organism: Botrytis cinerea


organism: Candida albicans (strain SC5314 / ATCC MYA-2876)

substrate_name: N-acetylglucosamine (GlcNAc)

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

assay_conditions_summary: pH 7.5; cofactor ATP

substrate: N-acetylglucosamine (GlcNAc)


organism: Ogataea angusta

assay_conditions_summary: pH 7.5; 310.15 K; cofactor ATP


organism: Thermus thermophilus


organism: Amycolatopsis methanolica


normalized_value_unit_evidence: Km 0.6 mM.

raw_value_unit_evidence: Table 4 row ATP: Kapp 0.6 mM with table uncertainty 0.1 mM; prose identifies this as the apparent Km for ATP.

row_label: ATP; PPi absent; F-6-P fixed at 10 mM

table_label: Table 4. Kinetics of the ATP-PFK enzyme from A. methanolica WVI and effects of PPi on affinities for substrates F-6-P and ATP


organism: Bacillus licheniformis


assay_conditions_summary: pH 6.8; 303.15 K; cofactor ATP


organism: Schistosoma mansoni

assay_conditions_summary: pH 8.5; cofactor ATP


organism: Lactococcus lactis subsp. lactis

assay_conditions_summary: cofactor ATP


substrate_name: fructose 6-phosphate

substrate: fructose 6-phosphate


organism: Saccharomyces cerevisiae

substrate_name: D-Glucose

assay_conditions_summary: pH 6.5; 303.15 K; cofactor NADPH

column_label: Table 2; isoenzyme block HXK / Hxk1p; row "HXK | Hxk1p | kcat | 10.2 |  |  | s−1" column Value raw value 10.2 with column Unit s−1; row "HXK | Hxk1p | Kglc | 0.15 |  | 0.08 | mM" column Value raw value 0.15 with column Unit mM. Both cells sit in the same Hxk1p isoenzyme block of Table 2, so the kcat and Km cells are one tuple.

normalized_value_unit_evidence: kcat 10.2 s-1 normalizes to 10.2 s^-1. Kglc 0.15 mM normalizes to 0.15 mM. kcat/Km is not printed in Table 2, so paper kcat/Km = paper kcat / paper Km = 10.2 / 0.15 = 68.0 s^-1 mM^-1.

raw_value_unit_evidence: Table 2 line "HXK | Hxk1p | kcat | 10.2 |  |  | s−1" gives kcat 10.2 s−1. Table 2 line "HXK | Hxk1p | Kglc | 0.15 |  | 0.08 | mM" gives Kglc 0.15 mM. Table 2 prints no kcat/Km or catalytic-efficiency column.

row_label: HXK / Hxk1p

table_label: Table 2. Enzyme kinetics parameters determined in this project and contrasted with those of Ref. [23]

substrate: D-Glucose


column_label: Table 2; isoenzyme block HXK / Hxk2p; row "HXK | Hxk2p | kcat | 63.1 |  |  | s−1" column Value raw value 63.1 with column Unit s−1; row "HXK | Hxk2p | Kglc | 0.2 |  | 0.08 | mM" column Value raw value 0.2 with column Unit mM. Both cells sit in the same Hxk2p isoenzyme block of Table 2, so the kcat and Km cells are one tuple.

normalized_value_unit_evidence: kcat 63.1 s-1 normalizes to 63.1 s^-1. Kglc 0.2 mM normalizes to 0.2 mM. kcat/Km is not printed in Table 2, so paper kcat/Km = paper kcat / paper Km = 63.1 / 0.2 = 315.5 s^-1 mM^-1.

raw_value_unit_evidence: Table 2 line "HXK | Hxk2p | kcat | 63.1 |  |  | s−1" gives kcat 63.1 s−1. Table 2 line "HXK | Hxk2p | Kglc | 0.2 |  | 0.08 | mM" gives Kglc 0.2 mM. Table 2 prints no kcat/Km or catalytic-efficiency column.

row_label: HXK / Hxk2p


organism: Lysinibacillus sphaericus


organism: Escherichia coli (strain K12)


assay_conditions_summary: pH 6.9; cofactor ATP


column_label: Results 3.3, journal page 174, and Figure 4/caption on page 175: recombinant Schistosoma mansoni PFK Fru-6-P saturation under 1 mM AMP reports apparent Km 0.8 mM; the figure caption specifies pH 6.9 and 1 mM ATP.

normalized_value_unit_evidence: Apparent Fru-6-P Km is already canonical at 0.8 mM.

raw_value_unit_evidence: Apparent Km for Fru-6-P under 1 mM AMP: raw 0.8 mM.

row_label: recombinant Schistosoma mansoni PFK; Fru-6-P saturation with 1 mM AMP

table_label: Results 3.3 and Figure 4


organism: Leishmania donovani


organism: Spinacia oleracea

assay_conditions_summary: pH 7.8; 303.15 K; cofactor ATP


organism: Saccharolobus solfataricus

substrate_name: North-methanocarba-dATP

substrate: North-methanocarba-dATP


substrate_name: South-methanocarba-dATP

substrate: South-methanocarba-dATP


organism: Schizosaccharomyces pombe

assay_conditions_summary: pH 7.2; 298.15 K; cofactor ATP


organism: Streptomyces coelicolor (strain ATCC BAA-471 / A3(2) / M145)


organism: Schizosaccharomyces pombe (strain 972 / ATCC 24843)

substrate_name: fructose 6-phosphate (without effector)

substrate: fructose 6-phosphate (without effector)


organism: Homo sapiens

substrate_name: dTTP (POLG polymerase activity at matched G:C primer-template)


organism: Drosophila melanogaster

substrate_name: deoxythymidine triphosphate

substrate: deoxythymidine triphosphate


organism: Danio rerio

substrate_name: dTTP

substrate: dTTP


organism: Thermococcus thioreducens

substrate_name: dNTP

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

substrate: dNTP


organism: Trypanosoma cruzi (strain CL Brener)


organism: Dictyostelium discoideum


organism: Thermococcus kodakarensis

substrate_name: ADP

assay_conditions_summary: pH 7.5; 323.15 K; cofactor ADP

substrate: ADP


substrate_name: AMP

substrate: AMP


substrate_name: D-fructose 1,6-bisphosphate

substrate: D-fructose 1,6-bisphosphate


substrate_name: D-glucose

substrate: D-glucose


substrate_name: D-Fructose 6-phosphate

substrate: D-Fructose 6-phosphate


organism: Komagataella pastoris

substrate_name: D-fructose 6-phosphate

substrate: D-fructose 6-phosphate


substrate_name: dCTP

sequence_variant_note: Resolved canonical sequence from uniprot_Q97W02_reconstructed_variant_from_PMID_18984592 and applied mutation signature N188W to reconstruct the assayed variant sequence.

substrate: dCTP


sequence_variant_note: Resolved canonical sequence from uniprot_Q97W02_reconstructed_variant_from_PMID_18984592 and applied mutation signature T239W to reconstruct the assayed variant sequence.


organism: Lactobacillus delbrueckii subsp. bulgaricus

substrate_name: ATP

assay_conditions_summary: pH 8.2; cofactor ATP

substrate: ATP


substrate_name: fructose 6-phosphate

assay_conditions_summary: pH 6; cofactor ATP


organism: Trypanosoma brucei

assay_conditions_summary: pH 7.6; 298.15 K; cofactor ATP

proof_lines:
Trypanosoma brucei
phosphofructokinase
ATP


substrate_name: Fructose 6-phosphate

proof_lines:
Trypanosoma brucei
phosphofructokinase
fructose 6-phosphate

substrate: Fructose 6-phosphate


organism: Thermotoga maritima

assay_conditions_summary: pH 7; 348.15 K; cofactor ATP


organism: Geobacillus stearothermophilus

assay_conditions_summary: pH 7.5; 298.15 K; cofactor ATP


organism: Escherichia coli

assay_conditions_summary: pH 8.2; 301.15 K; cofactor ATP

source_databases_merged: skid uniprot


assay_conditions_summary: pH 8.2; 298.15 K; cofactor ATP


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


substrate: fructose 6-phosphate


organism: Oceanimonas doudoroffii

assay_conditions_summary: pH 8; 301.15 K; cofactor ATP


substrate_name: dATP

sequence_source: UniProtKB P52026 canonical FASTA retrieved by direct UniProt REST check on 2026-05-25

proof_lines: pH and temperature not specified in the publication

substrate: dATP


substrate_name: dGTP

substrate: dGTP


substrate_name: TTP

substrate: TTP


substrate_name: D-mannose

sequence_source: UniProtKB:P35557 canonical sequence with row-specific substitution reconstruction

substrate: D-mannose


substrate_name: D-fructose

substrate: D-fructose


assay_conditions_summary: pH 7.5; 303.15 K; cofactor ATP


substrate_name: fructose 1,6-bisphosphate

substrate: fructose 1,6-bisphosphate


assay_conditions_summary: pH 8.2; 303.15 K; cofactor ATP

proof_lines: pH 8.2, 30°C, at fructose 6-phosphate concentrations that are less than approx. one-third the fixed MgATP concentration


substrate_name: GTP

substrate: GTP


substrate_name: UTP

substrate: UTP


substrate_name: CTP

substrate: CTP


assay_conditions_summary: pH 8; 303.15 K; cofactor ATP

proof_lines: pH 8.0, mutant enzyme Q466_H467insHMNLAEQ


sequence_source: UniProt P35557 canonical sequence reconstructed with A208G after direct residue check

sequence_variant_note: Mutation signature A208G matches the mutant residue at offset +0 and the reference residue at offset -7; variant reconstruction was not applied because multiple plausible residue-numbering offsets are present.

assay_conditions_summary: 303.15 K; cofactor ATP


sequence_source: UniProt P35557 canonical sequence reconstructed with K458R after direct residue check

sequence_variant_note: Mutation signature K458R matches the mutant residue at offset +0 and the reference residue at offset +1; variant reconstruction was not applied because multiple plausible residue-numbering offsets are present.


sequence_source: UniProt P35557 canonical sequence reconstructed with C457V after direct residue check

sequence_variant_note: Mutation signature C457V matches the mutant residue at offset +0 and the reference residue at offset +4; variant reconstruction was not applied because multiple plausible residue-numbering offsets are present.


sequence_source: UniProt P35557 canonical sequence reconstructed with Y215A after direct residue check

sequence_variant_note: Mutation signature Y215A matches the mutant residue at offset +0 and the reference residue at offset -1; variant reconstruction was not applied because multiple plausible residue-numbering offsets are present.


sequence_source: UniProt P35557 canonical sequence reconstructed with A460R after direct residue check

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


sequence_source: UniProt P35557 canonical sequence reconstructed with K459L after direct residue check

sequence_variant_note: Mutation signature K459L matches the mutant residue at offset +0 and the reference residue at offset -1; variant reconstruction was not applied because multiple plausible residue-numbering offsets are present.


organism: Thermotoga maritima (strain ATCC 43589 / DSM 3109 / JCM 10099 / NBRC 100826 / MSB8)

substrate_name: fructose 1,6-bisphosphate (at 75 degrees Celsius)

assay_conditions_summary: cofactor ATP


sequence_variant_note: Resolved canonical sequence from UniProt P08237 canonical sequence plus PMID 22474333 human mutant muscle Pfk evidence and applied mutation signature N341A/R246A/K386A to reconstruct the assayed variant sequence.

assay_conditions_summary: pH 7; 298.15 K; cofactor ATP

proof_lines: mutant enzyme N341A/R246A/K386A, in the presence of 1 mM AMP, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C

proof_lines: mutant enzyme N341A/R246A/K386A, in the presence of 0.82 mM ADP, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C

proof_lines: mutant enzyme N341A/R246A/K386A, without effectors, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C


sequence_variant_note: Resolved canonical sequence from UniProt P08237 canonical sequence plus PMID 22474333 human mutant muscle Pfk evidence and applied mutation signature R246A to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme R246A, in the presence of 0.82 mM ADP, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C

proof_lines: mutant enzyme R246A, without effectors, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C

proof_lines: mutant enzyme R246A, in the presence of 1 mM AMP, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C


sequence_variant_note: Resolved canonical sequence from UniProt P08237 canonical sequence plus PMID 22474333 human mutant muscle Pfk evidence and applied mutation signature K678A to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme K678A, without effectors, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C

proof_lines: mutant enzyme K678A, in the presence of 1 mM AMP, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C

proof_lines: mutant enzyme K678A, in the presence of 0.82 mM ADP, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C


sequence_variant_note: Resolved canonical sequence from UniProt P08237 canonical sequence plus PMID 22474333 human mutant muscle Pfk evidence and applied mutation signature K386A to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme K386A, in the presence of 0.82 mM ADP, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C

proof_lines: mutant enzyme K386A, in the presence of 1 mM AMP, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C

proof_lines: mutant enzyme K386A, without effectors, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C


sequence_variant_note: Resolved canonical sequence from UniProt P08237 canonical sequence plus PMID 22474333 human mutant muscle Pfk evidence and applied mutation signature S377A to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme S377A, in the presence of 0.82 mM ADP, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C

proof_lines: mutant enzyme S377A, in the presence of 1 mM AMP, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C

proof_lines: mutant enzyme S377A, without effectors, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C


sequence_variant_note: Resolved canonical sequence from UniProt P08237 canonical sequence plus PMID 22474333 human mutant muscle Pfk evidence and applied mutation signature S377A/K678A to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme S377A/K678A, in the presence of 0.82 mM ADP, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C

proof_lines: mutant enzyme S377A/K678A, without effectors, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C

proof_lines: mutant enzyme S377A/K678A, in the presence of 1 mM AMP, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C


sequence_variant_note: Resolved canonical sequence from UniProt P08237 canonical sequence plus PMID 22474333 human mutant muscle Pfk evidence and applied mutation signature R246A/K386A to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme R246A/K386A, in the presence of 0.82 mM ADP, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C

proof_lines: mutant enzyme R246A/K386A, in the presence of 1 mM AMP, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C

proof_lines: mutant enzyme R246A/K386A, without effectors, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C


substrate_name: Unknown

sequence_source: UniProt P08237 canonical sequence plus PMID 22474333 human mutant muscle Pfk evidence

proof_lines: mutant enzyme S377A, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C

substrate: Unknown


proof_lines: mutant enzyme N341A/R246A/K386A, without effectors, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C

proof_lines: mutant enzyme N341A/R246A/K386A, in the presence of 0.82 mM ADP, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C

proof_lines: mutant enzyme N341A/R246A/K386A, in the presence of 1 mM AMP, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C


proof_lines: mutant enzyme K678A, without effectors, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C

proof_lines: mutant enzyme K678A, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C

proof_lines: mutant enzyme K678A, in the presence of 0.82 mM ADP, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C


proof_lines: Ki above 10 mM, mutant enzyme K386A, in the presence of 1 mM AMP, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C

proof_lines: mutant enzyme K386A, without effectors, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C

proof_lines: mutant enzyme K386A, in the presence of 0.82 mM ADP, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C


proof_lines: Ki above 10 mM, mutant enzyme R246A, in the presence of 0.82 mM ADP, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C

proof_lines: Ki above 10 mM, mutant enzyme R246A, in the presence of 1 mM AMP, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C

proof_lines: mutant enzyme R246A, without effectors, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C


proof_lines: Ki above 10 mM, mutant enzyme R246A/K386A, in the presence of 0.82 mM ADP, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C

proof_lines: Ki above 10 mM, mutant enzyme R246A/K386A, in the presence of 1 mM AMP, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C

proof_lines: Ki above 10 mM, mutant enzyme R246A/K386A, without effectors, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C


sequence_variant_note: Resolved canonical sequence from UniProt P08237 canonical sequence plus PMID 22474333 human mutant muscle Pfk evidence and applied mutation signature N341A to reconstruct the assayed variant sequence.

proof_lines: mutant enzyme N341A, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C

proof_lines: mutant enzyme N341A, in the presence of 0.82 mM ADP, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C

proof_lines: mutant enzyme N341A, in the presence of 1 mM AMP, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C


proof_lines: mutant enzyme S377A/K678A, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C

proof_lines: mutant enzyme S377A/K678A, in the presence of 0.82 mM ADP, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C

proof_lines: mutant enzyme S377A/K678A, in the presence of 1 mM AMP, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C


organism: Cucumis sativus

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

proof_lines:
pH 8.0, 25°C, cytosolic PFK
pH 7.2, 25°C, plastid PFK
pH 8.0, 25°C, plastid PFK


substrate_name: ITP

proof_lines: pH 8.0, 25°C, pH 7.2, 25°C, cytosolic PFK

substrate: ITP


proof_lines:
pH 7.2, 25°C, cytosolic PFK
pH 8.0, 25°C, plastid PFK
pH 8.0, 25°C, cytosolic PFK


proof_lines:
pH 8.0, 25°C, cytosolic PFK
pH 7.2, 25°C, cytosolic PFK
pH 8.0, 25°C, plastid PFK


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


substrate_name: D-fructose 6-phosphate


proof_lines: pH 8.5, 30°C, D127/R252Q double mutant PFK


assay_conditions_summary: pH 8.5; 303.15 K; cofactor ATP


organism: Homo sapiens

substrate_name: D-glucosamine

proof_lines:
hexokinase I, at 37°C, pH 8.1
hexokinases Ib and Ic, at 37°C, pH 7.2
hexokinase Ia, at 37°C, pH 7.2

substrate: D-glucosamine


proof_lines:
hexokinase I, at 37°C, pH 8.1
hexokinase Ib, at 37°C, pH 7.2
hexokinase Ia, at 37°C, pH 7.2


proof_lines: hexokinases Ia, Ib and Ic, at 37°C, pH 7.2


organism: Saccharomyces cerevisiae


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


substrate_name: D-mannosamine

substrate: D-mannosamine


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


smiles: C(C(C(C(C(C=O)F)O)O)O)O


organism: Pyrococcus furiosus

proof_lines: 55°C, pH not specified in the publication, mutant enzyme Y410I


proof_lines: 55°C, pH not specified in the publication, mutant enzyme Y410L


proof_lines: 55°C, pH not specified in the publication, mutant enzyme Y410V


proof_lines: 55°C, pH not specified in the publication, wild-type enzyme


proof_lines: 55°C, pH not specified in the publication, mutant enzyme L409V


proof_lines: 55°C, pH not specified in the publication, mutant enzyme L409M


proof_lines: 55°C, pH not specified in the publication, mutant enzyme L409I


substrate_name: D-glucose

sequence_variant_note: Resolved canonical sequence from uniprot_accession:P19367 and applied mutation signature A464P/E465G to reconstruct the assayed variant sequence.

assay_conditions_summary: pH 8

substrate: D-glucose


substrate_name: ATP

assay_conditions_summary: pH 8; cofactor ATP

substrate: ATP


sequence_variant_note: Resolved canonical sequence from uniprot_accession:P19367 and applied mutation signature Q466insHMNLAEQ to reconstruct the assayed variant sequence.

mutation_type: insertion

proof_lines: pH 8.0, mutant enzyme Q466_H467insHMNLAEQ


substrate_name: dATP

sequence_source: UniProt P00582 Klenow-fragment region; paper identity from PMID 29036327

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


substrate_name: dCTP

substrate: dCTP


substrate_name: dGTP


substrate_name: dTTP

substrate: dTTP


sequence_variant_note: Resolved canonical sequence from UniProtKB:P28340 plus PMID:20628184 Leu-606 paper context and applied mutation signature L606G to reconstruct the assayed variant sequence.

proof_lines: mutant L606G, pH 7.5, 37°C, misincorporation of dTTP with G at first position


sequence_source: UniProtKB:P28340 plus PMID:20628184 Leu-606 paper context

proof_lines: wild-type enzyme, pH 7.5, 37°C, misincorporation of dTTP with G at first position


sequence_variant_note: Resolved canonical sequence from UniProtKB:P28340 plus PMID:20628184 Leu-606 paper context and applied mutation signature L606K to reconstruct the assayed variant sequence.

proof_lines: mutant L606K, pH 7.5, 37°C, misincorporation of dTTP with G at first position


organism: Musa acuminata

assay_conditions_summary: pH 7.3; 298.15 K; cofactor ATP

proof_lines:
pH 7.3, 25°C, PFK II
pH 8.0, 25°C, PFK I


organism: Aspergillus niger


substrate_name: D-fructose


substrate_name: fructose 6-phosphate

proof_lines:
pH 8.0, 8.5°C, wild-type PFK
pH 8.5, 30°C, wild-type PFK

substrate: fructose 6-phosphate


organism: Leishmania mexicana


proof_lines:
inhibition with respect to ATP
inhibition with respect to glucose


organism: Oryctolagus cuniculus

proof_lines:
at 37°C, pH 8.1
hexokinase Ib
hexokinase Ia


proof_lines:
MgITP2-, at 37°C, pH 8.1
MgITP2-, erythrocyte enzyme
MgITP2-, hexokinase Ia


assay_conditions_summary: pH 8.1; 310.15 K; cofactor ATP

proof_lines: MgATP2-, bound mitochondrial HK I, at pH 7.5


proof_lines: mutant enzyme N341A, without effectors, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C

proof_lines: mutant enzyme N341A, in the presence of 0.82 mM ADP, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C

proof_lines: mutant enzyme N341A, in the presence of 1 mM AMP, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C


proof_lines: mutant enzyme D543A, in the presence of 0.1 mM 5-phospho-alpha-D ribose 1-diphosphate, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C

proof_lines: mutant enzyme D543A, in the presence of 0.82 mM ADP, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C

proof_lines: mutant enzyme D543A, without effectors, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C


assay_conditions_summary: pH 7; 298.15 K; cofactor ADP

proof_lines: mutant enzyme D543A, in the presence of 1 mM ADP, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C

proof_lines: mutant enzyme D543A, in the presence of 1 mM AMP, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C


proof_lines: wild type enzyme, in the presence of 0.82 mM ADP, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C

proof_lines: wild type enzyme, at pH 7.0 in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2 and 25°C

proof_lines: wild type enzyme, without effectors, in 50 mM HEPES, 100 mM KCl, 5 mM MgCl2, pH 7.0, at 25°C


substrate_name: ATP; D-Glucose

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

proof_lines: uncomplexed ATP, erythrocyte enzyme, competitive to MgATP2-

proof_lines: erythrocyte enzyme, competitive to glucose

proof_lines: erythrocyte enzyme, mixed inhibitor versus MgATP2-

substrate: ATP; D-Glucose


proof_lines:
hexokinase Ib
at 37°C, pH 8.1
erythrocyte enzyme


substrate_name: D-mannose

proof_lines:
hexokinase Ib
at 37°C, pH 8.1
reticulocyte hexokinase Ib


proof_lines:
hexokinase Ia
at 37°C, pH 8.1
hexokinase Ib


assay_conditions_summary: pH 8; 310.15 K; cofactor ATP

proof_lines: wild-type, 37°C, pH not specified in the puclication. Kinetic constants at high and low concentrations of the fixed substrate: 0.5 mM D-glucose


sequence_source: reconstructed_from_uniprot:P35557; L146R applied after direct residue check

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

proof_lines: mutant L146R, 37°C, pH not specified in the puclication


sequence_source: reconstructed_from_uniprot:P35557; T228M applied after direct residue check

proof_lines: mutant T228M, 37°C, pH not specified in the puclication


assay_conditions_summary: 310.15 K; cofactor ATP