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Literature summary for 1.2.1.44 extracted from
- Characterization of the cinnamoyl-CoA reductase (CCR) gene family in Populus tomentosa reveals the enzymatic active sites and evolution of CCR (2017), Planta, 245, 61-75 .
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| gene CCR1, DNA and amino acid sequence determination and analysis of diverse isozymes from Populus tomentosa, phylogenetic analysis, recombinant expression of His-tagged isozyme CCR1 in Escherichia coli strain M15, subcloning in Escherichia coli strain DH5alpha | Populus tomentosa |
| gene CCR7, DNA and amino acid sequence determination and analysis of diverse isozymes from Populus tomentosa, phylogenetic analysis, recombinant expression of His-tagged wild-type and mutant isozymes CCR7 in Escherichia coli strain M15, subcloning in Escherichia coli strain DH5alpha | Populus tomentosa |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| A132S | site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme | Populus tomentosa |
| A132T | site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme | Populus tomentosa |
| A43V | site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme | Populus tomentosa |
| F155H | site-directed mutagenesis, the mutant is only active with feruloyl-CoA as substrate in contrast to the wild-type enzyme | Populus tomentosa |
| F155Y | site-directed mutagenesis, the mutant exhibits greater catalytic efficiency for sinapoyl-CoA compared to the wild-type PtoCCR7 | Populus tomentosa |
| H208M | site-directed mutagenesis, inactive mutant | Populus tomentosa |
| H208V | site-directed mutagenesis, inactive mutant | Populus tomentosa |
| H208Y | site-directed mutagenesis, inactive mutant | Populus tomentosa |
| L192M | site-directed mutagenesis, the mutant shows altered substrate specificity compared to the wild-type enzyme | Populus tomentosa |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | Michaelis-Menten kinetics | Populus tomentosa | |
| 0.00986 | - |
feruloyl-CoA | pH 6.0, 40°C, recombinant His-tagged enzyme | Populus tomentosa |  |
| 0.01431 | - |
feruloyl-CoA | pH 6.0, 40°C, recombinant His-tagged wild-type enzyme | Populus tomentosa | |
| 0.01595 | - |
4-coumaroyl-CoA | pH 6.0, 40°C, recombinant His-tagged wild-type enzyme | Populus tomentosa | |
| 0.01643 | - |
feruloyl-CoA | pH 6.0, 40°C, recombinant His-tagged mutant A132T | Populus tomentosa | |
| 0.01731 | - |
feruloyl-CoA | pH 6.0, 40°C, recombinant His-tagged mutant A132S | Populus tomentosa | |
| 0.01924 | - |
sinapoyl-CoA | pH 6.0, 40°C, recombinant His-tagged mutant F155Y | Populus tomentosa | |
| 0.01973 | - |
4-coumaroyl-CoA | pH 6.0, 40°C, recombinant His-tagged mutant F155Y | Populus tomentosa | |
| 0.02376 | - |
caffeoyl-CoA | pH 6.0, 40°C, recombinant His-tagged wild-type enzyme | Populus tomentosa | |
| 0.03123 | - |
feruloyl-CoA | pH 6.0, 40°C, recombinant His-tagged mutant F155H | Populus tomentosa | |
| 0.03494 | - |
sinapoyl-CoA | pH 6.0, 40°C, recombinant His-tagged mutant A43V | Populus tomentosa | |
| 0.04102 | - |
sinapoyl-CoA | pH 6.0, 40°C, recombinant His-tagged wild-type enzyme | Populus tomentosa | |
| 0.04295 | - |
caffeoyl-CoA | pH 6.0, 40°C, recombinant His-tagged mutant F155Y | Populus tomentosa | |
| 0.04887 | - |
feruloyl-CoA | pH 6.0, 40°C, recombinant His-tagged mutant A43V | Populus tomentosa | |
| 0.07474 | - |
feruloyl-CoA | pH 6.0, 40°C, recombinant His-tagged mutant L192M | Populus tomentosa | |
| 0.0933 | - |
feruloyl-CoA | pH 6.0, 40°C, recombinant His-tagged mutant F155Y | Populus tomentosa | |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 4-coumaroyl-CoA + NADPH + H+ | Populus tomentosa | - |
4-coumaraldehyde + CoA + NADP+ | - |
? | |
| caffeoyl-CoA + NADPH + H+ | Populus tomentosa | - |
caffealdehyde + CoA + NADP+ | - |
? | |
| feruloyl-CoA + NADPH + H+ | Populus tomentosa | preferred substrate | coniferaldehyde + CoA + NADP+ | - |
? | |
| feruloyl-CoA + NADPH + H+ | Populus tomentosa | highly specific for | coniferaldehyde + CoA + NADP+ | - |
? | |
| sinapoyl-CoA + NADPH + H+ | Populus tomentosa | - |
sinapaldehyde + CoA + NADP+ | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Populus tomentosa | A0A0F7EVN0 | - |
- |
| Populus tomentosa | T1WWB5 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His-tagged enzyme CCR1 from Escherichia coli strain M15 by nickel affinity chromatography | Populus tomentosa |
| recombinant His-tagged wild-type and mutant enzymes CCR7 from Escherichia coli strain M15 by nickel affinity chromatography | Populus tomentosa |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 4-coumaroyl-CoA + NADPH + H+ | - |
Populus tomentosa | 4-coumaraldehyde + CoA + NADP+ | - |
? | |
| caffeoyl-CoA + NADPH + H+ | - |
Populus tomentosa | caffealdehyde + CoA + NADP+ | - |
? | |
| feruloyl-CoA + NADPH + H+ | preferred substrate | Populus tomentosa | coniferaldehyde + CoA + NADP+ | - |
? | |
| feruloyl-CoA + NADPH + H+ | highly specific for | Populus tomentosa | coniferaldehyde + CoA + NADP+ | - |
? | |
| additional information | isozyme PtoCCR1 exhibits specificity for feruloyl-CoA, with no detectable activity for any other hydroxycinnamoyl-CoA esters. Substrate specificity, active site, molecular docking and modeling, overview | Populus tomentosa | ? | - |
? | |
| additional information | substrate specificity, active site, molecular docking and modeling, overview. A132 in CCR7 combined with the catalytic triad might comprisethe catalytic center. In CCR7, L192, F155, and H208 are the substrate-binding sites | Populus tomentosa | ? | - |
? | |
| sinapoyl-CoA + NADPH + H+ | - |
Populus tomentosa | sinapaldehyde + CoA + NADP+ | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| ? | x * 37080, CCR7, sequence calculation | Populus tomentosa |
| ? | x * 37540, CCR1, sequence calculation | Populus tomentosa |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| CCR | - |
Populus tomentosa |
| cinnamoyl-coenzyme A reductase | - |
Populus tomentosa |
| PtoCCR1 | - |
Populus tomentosa |
| PtoCCR7 | - |
Populus tomentosa |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 40 | - |
- |
Populus tomentosa |
Temperature Range [°C]
| Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 10 | 60 | activity range, profile overview, the enzyme is active even below 10°C | Populus tomentosa |
| 20 | 60 | activity range, profile overview | Populus tomentosa |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.0101 | - |
feruloyl-CoA | pH 6.0, 40°C, recombinant His-tagged mutant A132T | Populus tomentosa | |
| 0.0142 | - |
4-coumaroyl-CoA | pH 6.0, 40°C, recombinant His-tagged mutant F155Y | Populus tomentosa | |
| 0.0183 | - |
4-coumaroyl-CoA | pH 6.0, 40°C, recombinant His-tagged enzyme | Populus tomentosa | |
| 0.0192 | - |
feruloyl-CoA | pH 6.0, 40°C, recombinant His-tagged mutant A132S | Populus tomentosa | |
| 0.0327 | - |
caffeoyl-CoA | pH 6.0, 40°C, recombinant His-tagged mutant F155Y | Populus tomentosa | |
| 0.0362 | - |
caffeoyl-CoA | pH 6.0, 40°C, recombinant His-tagged enzyme | Populus tomentosa | |
| 0.064 | - |
sinapoyl-CoA | pH 6.0, 40°C, recombinant His-tagged enzyme | Populus tomentosa | |
| 0.098 | - |
sinapoyl-CoA | pH 6.0, 40°C, recombinant His-tagged mutant A43V | Populus tomentosa | |
| 0.135 | - |
sinapoyl-CoA | pH 6.0, 40°C, recombinant His-tagged mutant F155Y | Populus tomentosa | |
| 0.1409 | - |
feruloyl-CoA | pH 6.0, 40°C, recombinant His-tagged mutant F155H | Populus tomentosa | |
| 0.18 | - |
feruloyl-CoA | pH 6.0, 40°C, recombinant His-tagged enzyme | Populus tomentosa | |
| 0.8375 | - |
feruloyl-CoA | pH 6.0, 40°C, recombinant His-tagged mutant A43V | Populus tomentosa | |
| 1.033 | - |
feruloyl-CoA | pH 6.0, 40°C, recombinant His-tagged enzyme | Populus tomentosa | |
| 1.905 | - |
feruloyl-CoA | pH 6.0, 40°C, recombinant His-tagged mutant L192M | Populus tomentosa | |
| 2.1 | - |
feruloyl-CoA | pH 6.0, 40°C, recombinant His-tagged mutant F155Y | Populus tomentosa | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 6 | - |
- |
Populus tomentosa |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 4 | 9 | activity range, profile overview | Populus tomentosa |
| 5 | 8 | activity range, profile overview | Populus tomentosa |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| NADPH | the conserved motifs G-X-X-G-X-X-A and D-X-X-D are reported to be involved in NAD(P) binding and adenine binding pocket stabilization. In addition, the NADP specificity motif R(X)5K is identified, which is a key structure that distinguishes CCR from other NAD(H)-dependent SDRs | Populus tomentosa | |
pI Value
| Organism | Comment | pI Value Maximum | pI Value |
|---|---|---|---|
| Populus tomentosa | sequence calculation | - |
6.04 |
| Populus tomentosa | sequence calculation | - |
8.22 |
General Information
| General Information | Comment | Organism |
|---|---|---|
| evolution | the conserved motifs G-X-X-G-X-X-A and D-X-X-D are reported to be involved in NAD(P) binding and adenine binding pocket stabilization. In addition, the NADP specificity motif R(X)5K is identified, which is a key structure that distinguishes CCR from other NAD(H)-dependent SDRs. Sequence comparisons and phylogenetic analysis of Populus tometosa CCRs, overview. Of the 11 PtoCCR and PtoCCR-like proteins, PtoCCR1 and 7 each catalyze at least one substrate. PtoCCR1 is active only with feruloyl-CoA as a substrate, while PtoCCR7 accepts all hydroxycinnamoyl-CoA esters | Populus tomentosa |
| malfunction | the positively charged extended structure of His208 was important in stabilizing CoA, while the mutants (M208, V208, and Y208) were unable to assume this function | Populus tomentosa |
| additional information | three-dimensional structure analysis of wild-type and mutant enzymes, molecular docking and modeling, overview | Populus tomentosa |
| additional information | three-dimensional structure analysis, molecular docking and modeling, overview | Populus tomentosa |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 1.15 | - |
caffeoyl-CoA | pH 6.0, 40°C, recombinant His-tagged enzyme | Populus tomentosa | |
| 1.28 | - |
4-coumaroyl-CoA | pH 6.0, 40°C, recombinant His-tagged enzyme | Populus tomentosa | |
| 1.56 | - |
sinapoyl-CoA | pH 6.0, 40°C, recombinant His-tagged enzyme | Populus tomentosa | |
| 18.26 | - |
feruloyl-CoA | pH 6.0, 40°C, recombinant His-tagged enzyme | Populus tomentosa | |
| 72.19 | - |
feruloyl-CoA | pH 6.0, 40°C, recombinant His-tagged enzyme | Populus tomentosa | |
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