Cloned (Comment) | Organism |
---|---|
DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic analysis, recombinant expression of N-terminally His-tagged isozyme PaDBR1 in Escherichia coli strain BL21(DE3) | Plagiochasma appendiculatum |
DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic analysis, recombinant expression of N-terminally His-tagged isozyme PaDBR2 in Escherichia coli strain BL21(DE3) | Plagiochasma appendiculatum |
Protein Variants | Comment | Organism |
---|---|---|
C56A | site-directed mutagenesis, the mutant shows altered substrate specificity with cinnamoyl aldehydes compared to wild-type enzyme | Plagiochasma appendiculatum |
C56F | site-directed mutagenesis, the mutant shows altered substrate specificity and increased activity with cinnamoyl aldehydes compared to wild-type enzyme | Plagiochasma appendiculatum |
C56S | site-directed mutagenesis, the mutant shows altered substrate specificity with cinnamoyl aldehydes compared to wild-type enzyme | Plagiochasma appendiculatum |
C56V | site-directed mutagenesis, the mutant shows altered substrate specificity with cinnamoyl aldehydes compared to wild-type enzyme | Plagiochasma appendiculatum |
C56Y | site-directed mutagenesis, the mutation turns the substrate selectivity and catalytic efficiency of isozyme PaDBR1 indistinguishable from those of isozyme PaDBR2, Docking arrangement of PaDBR1C56Y with NADP+/p-coumaryl aldehyde, overview. Increased activity compared to wild-type enzyme | Plagiochasma appendiculatum |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0943 | - |
4-coumaryl aldehyde | pH and temperature not specified in the publication, recombinant wild-type isozyme PaDBR2 | Plagiochasma appendiculatum | |
0.1183 | - |
4-coumaryl aldehyde | pH and temperature not specified in the publication, recombinant isozyme PaDBR1 mutant C56Y | Plagiochasma appendiculatum | |
0.1437 | - |
caffeoyl aldehyde | pH and temperature not specified in the publication, recombinant isozyme PaDBR1 mutant C56Y | Plagiochasma appendiculatum | |
0.1663 | - |
4-coumaryl aldehyde | pH and temperature not specified in the publication, recombinant isozyme PaDBR1 mutant C56V | Plagiochasma appendiculatum | |
0.1719 | - |
4-coumaryl aldehyde | pH and temperature not specified in the publication, recombinant wild-type isozyme PaDBR1 | Plagiochasma appendiculatum | |
0.1726 | - |
coniferyl aldehyde | pH and temperature not specified in the publication, recombinant isozyme PaDBR1 mutant C56Y | Plagiochasma appendiculatum | |
0.1812 | - |
caffeoyl aldehyde | pH and temperature not specified in the publication, recombinant wild-type isozyme PaDBR2 | Plagiochasma appendiculatum | |
0.1933 | - |
caffeoyl aldehyde | pH and temperature not specified in the publication, recombinant isozyme PaDBR1 mutant C56V | Plagiochasma appendiculatum | |
0.2026 | - |
5-hydroxyconiferyl aldehyde | pH and temperature not specified in the publication, recombinant isozyme PaDBR1 mutant C56Y | Plagiochasma appendiculatum | |
0.2099 | - |
coniferyl aldehyde | pH and temperature not specified in the publication, recombinant wild-type isozyme PaDBR2 | Plagiochasma appendiculatum | |
0.2214 | - |
caffeoyl aldehyde | pH and temperature not specified in the publication, recombinant wild-type isozyme PaDBR1 | Plagiochasma appendiculatum | |
0.3081 | - |
5-hydroxyconiferyl aldehyde | pH and temperature not specified in the publication, recombinant wild-type isozyme PaDBR2 | Plagiochasma appendiculatum |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | Plagiochasma appendiculatum | the recombinant isozyme PaDBR2 has a higher catalytic activity than isozyme PaDBR1 with respect to the reduction of the double bond present in hydroxycinnamyl aldehydes | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Plagiochasma appendiculatum | S5TMG0 | - |
- |
Plagiochasma appendiculatum | S5U4F4 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged isozyme PaDBR1 from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and ultrafiltration | Plagiochasma appendiculatum |
recombinant His-tagged isozyme PaDBR2 from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and ultrafiltration | Plagiochasma appendiculatum |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.0018 | - |
pH and temperature not specified in the publication, purified recombinant wild-type isozyme PaDBR1, substrate coniferyl aldeyde | Plagiochasma appendiculatum |
0.0026 | - |
pH and temperature not specified in the publication, purified recombinant isozyme PaDBR1 mutant C56A, substrate coniferyl aldeyde | Plagiochasma appendiculatum |
0.0027 | - |
pH and temperature not specified in the publication, purified recombinant wild-type isozyme PaDBR1, substrate 5-hydroxyconiferyl aldeyde | Plagiochasma appendiculatum |
0.0041 | - |
pH and temperature not specified in the publication, purified recombinant isozyme PaDBR1 mutant C56A, substrate 4-coumaryl aldeyde | Plagiochasma appendiculatum |
0.0046 | - |
pH and temperature not specified in the publication, purified recombinant isozyme PaDBR1 mutant C56A, substrate 5-hydroxyconiferyl aldeyde | Plagiochasma appendiculatum |
0.0078 | - |
pH and temperature not specified in the publication, purified recombinant isozyme PaDBR1 mutant C56A, substrate caffeoyl aldeyde | Plagiochasma appendiculatum |
0.0079 | - |
pH and temperature not specified in the publication, purified recombinant isozyme PaDBR1 mutant C56S, substrate 4-coumaryl aldeyde | Plagiochasma appendiculatum |
0.008 | 4 | pH and temperature not specified in the publication, purified recombinant isozyme PaDBR1 mutant C56V, substrate 5-hydroxyconiferyl aldeyde | Plagiochasma appendiculatum |
0.008 | 4 | pH and temperature not specified in the publication, purified recombinant isozyme PaDBR1 mutant C56V, substrate coniferyl aldeyde | Plagiochasma appendiculatum |
0.0091 | - |
pH and temperature not specified in the publication, purified recombinant isozyme PaDBR1 mutant C56S, substrate caffeoyl aldeyde | Plagiochasma appendiculatum |
0.0115 | - |
pH and temperature not specified in the publication, purified recombinant isozyme PaDBR1 mutant C56S, substrate coniferyl aldeyde | Plagiochasma appendiculatum |
0.0116 | - |
pH and temperature not specified in the publication, purified recombinant isozyme PaDBR1 mutant C56S, substrate 5-hydroxyconiferyl aldeyde | Plagiochasma appendiculatum |
0.0124 | - |
pH and temperature not specified in the publication, purified recombinant isozyme PaDBR1 mutant C56V, substrate 4-coumaryl aldeyde | Plagiochasma appendiculatum |
0.015 | - |
pH and temperature not specified in the publication, purified recombinant isozyme PaDBR1 mutant C56V, substrate caffeoyl aldeyde | Plagiochasma appendiculatum |
0.0177 | - |
pH and temperature not specified in the publication, purified recombinant wild-type isozyme PaDBR1, substrate caffeoyl aldeyde | Plagiochasma appendiculatum |
0.022 | - |
pH and temperature not specified in the publication, purified recombinant isozyme PaDBR1 mutant C56F, substrate 5-hydroxyconiferyl aldeyde | Plagiochasma appendiculatum |
0.0234 | - |
pH and temperature not specified in the publication, purified recombinant isozyme PaDBR1 mutant C56Y, substrate 5-hydroxyconiferyl aldeyde | Plagiochasma appendiculatum |
0.0237 | - |
pH and temperature not specified in the publication, purified recombinant wild-type isozyme PaDBR1, substrate 4-coumaryl aldeyde | Plagiochasma appendiculatum |
0.0239 | - |
pH and temperature not specified in the publication, purified recombinant isozyme PaDBR1 mutant C56Y, substrate coniferyl aldeyde | Plagiochasma appendiculatum |
0.0271 | - |
pH and temperature not specified in the publication, purified recombinant isozyme PaDBR1 mutant C56F, substrate coniferyl aldeyde | Plagiochasma appendiculatum |
0.0454 | - |
pH and temperature not specified in the publication, purified recombinant isozyme PaDBR1 mutant C56F, substrate caffeoyl aldeyde | Plagiochasma appendiculatum |
0.0497 | - |
pH and temperature not specified in the publication, purified recombinant isozyme PaDBR1 mutant C56Y, substrate caffeoyl aldeyde | Plagiochasma appendiculatum |
0.066 | - |
pH and temperature not specified in the publication, purified recombinant isozyme PaDBR1 mutant C56F, substrate 4-coumaryl aldeyde | Plagiochasma appendiculatum |
0.0743 | - |
pH and temperature not specified in the publication, purified recombinant isozyme PaDBR1 mutant C56Y, substrate 4-coumaryl aldeyde | Plagiochasma appendiculatum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-coumaryl aldehyde + NADP+ | - |
Plagiochasma appendiculatum | dihydro-4-coumaryl aldehyde + NADPH + H+ | - |
? | |
5-hydroxyconiferyl aldehyde + NADP+ | - |
Plagiochasma appendiculatum | dihydro-5-hydroxyconiferyl aldehyde + NADPH + H+ | - |
? | |
5-hydroxyconiferyl aldehyde + NADP+ | only enzyme mutant PaDBR1C56Y, no activity with the wild-type enzyme | Plagiochasma appendiculatum | dihydro-5-hydroxyconiferyl aldehyde + NADPH + H+ | - |
? | |
caffeoyl aldehyde + NADP+ | - |
Plagiochasma appendiculatum | dihydrocaffeoyl aldehyde + NADPH + H+ | - |
? | |
coniferyl aldehyde + NADP+ | - |
Plagiochasma appendiculatum | dihydroconiferyl aldehyde + NADPH + H+ | - |
? | |
coniferyl aldehyde + NADP+ | only enzyme mutant PaDBR1C56Y, no activity with the wild-type enzyme | Plagiochasma appendiculatum | dihydroconiferyl aldehyde + NADPH + H+ | - |
? | |
additional information | the recombinant isozyme PaDBR2 has a higher catalytic activity than isozyme PaDBR1 with respect to the reduction of the double bond present in hydroxycinnamyl aldehydes | Plagiochasma appendiculatum | ? | - |
? | |
additional information | docking of the ligand and substrate into the active cavity, overview. The catalytic efficiency of isozyme PaDBR1 is substantially less than that of PaDBR2, especially towards cinnamyl aldehydes carrying a methoxy group (coniferyl, 5-hydroxyconiferyl and sinapyl aldehydes). No activity with sinapyl aldehyde, cinnamyl aldehyde, 4-coumaric acid, caffeic acid, 4-coumaryl alcohol, caffeyl alcohol, coniferyl alcohol, 5-hydroxyconiferyl alcohol, and sinapyl alcohol | Plagiochasma appendiculatum | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 55000, about, recombinant His-tagged enzyme, SDS-PAGE | Plagiochasma appendiculatum |
Synonyms | Comment | Organism |
---|---|---|
alkenal double bond reductase | - |
Plagiochasma appendiculatum |
double-bond reductase 1 | UniProt | Plagiochasma appendiculatum |
double-bond reductase 2 | UniProt | Plagiochasma appendiculatum |
PaDBR1 | - |
Plagiochasma appendiculatum |
PaDBR2 | - |
Plagiochasma appendiculatum |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.037 | - |
4-coumaryl aldehyde | pH and temperature not specified in the publication, recombinant isozyme PaDBR1 mutant C56V | Plagiochasma appendiculatum | |
0.068 | - |
caffeoyl aldehyde | pH and temperature not specified in the publication, recombinant isozyme PaDBR1 mutant C56V | Plagiochasma appendiculatum | |
0.098 | - |
4-coumaryl aldehyde | pH and temperature not specified in the publication, recombinant wild-type isozyme PaDBR1 | Plagiochasma appendiculatum | |
0.118 | - |
coniferyl aldehyde | pH and temperature not specified in the publication, recombinant wild-type isozyme PaDBR2 | Plagiochasma appendiculatum | |
0.132 | - |
caffeoyl aldehyde | pH and temperature not specified in the publication, recombinant wild-type isozyme PaDBR1 | Plagiochasma appendiculatum | |
0.134 | - |
coniferyl aldehyde | pH and temperature not specified in the publication, recombinant isozyme PaDBR1 mutant C56Y | Plagiochasma appendiculatum | |
0.161 | - |
5-hydroxyconiferyl aldehyde | pH and temperature not specified in the publication, recombinant isozyme PaDBR1 mutant C56Y | Plagiochasma appendiculatum | |
0.169 | - |
5-hydroxyconiferyl aldehyde | pH and temperature not specified in the publication, recombinant wild-type isozyme PaDBR2 | Plagiochasma appendiculatum | |
0.238 | - |
caffeoyl aldehyde | pH and temperature not specified in the publication, recombinant wild-type isozyme PaDBR2 | Plagiochasma appendiculatum | |
0.249 | - |
4-coumaryl aldehyde | pH and temperature not specified in the publication, recombinant isozyme PaDBR1 mutant C56Y | Plagiochasma appendiculatum | |
0.277 | - |
caffeoyl aldehyde | pH and temperature not specified in the publication, recombinant isozyme PaDBR1 mutant C56Y | Plagiochasma appendiculatum | |
0.278 | - |
4-coumaryl aldehyde | pH and temperature not specified in the publication, recombinant wild-type isozyme PaDBR2 | Plagiochasma appendiculatum |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADP+ | - |
Plagiochasma appendiculatum |
General Information | Comment | Organism |
---|---|---|
evolution | tyrosine at position 56 of PaDBR2 is highly conserved among oxidoreductases, and is proposed to be involved in binding with NADPH. In isozyme PaDBR1, this position is occupied by a Cystein residue | Plagiochasma appendiculatum |
evolution | tyrosine at position 56 of PaDBR2 is highly conserved among oxidoreductases, and is proposed to be involved in binding with NADPH. In isozyme PaDBR1, this position is occupied by a cysteine residue | Plagiochasma appendiculatum |