Cloned (Comment) | Organism |
---|---|
isozyme COR-B, DNA and amino acid sequence determination and analysis, sequence comparison with codeinone reductase isoforms COR1.2, COR1.3, COR1.4, and COR1.1, quantitative real-time PCR expression analysis, recombinant coexpression of isozyme COR-B with thebaine 6-O-demethylase (T6ODM) in Saccharomyces cerevisiae strain CEN.PK fed with thebaine | Papaver somniferum |
isozyme COR1.1, DNA and amino acid sequence determination and analysis, sequence comparison with codeinone reductase isoforms COR1.2, COR1.3, COR1.4, and COR-B, quantitative real-time PCR expression analysis | Papaver somniferum |
isozyme COR1.2, DNA and amino acid sequence determination and analysis, sequence comparison with codeinone reductase isoforms COR1.1, COR1.3, COR1.4, and COR-B, quantitative real-time PCR expression analysis. Isozyme COR1.2 exhibits lower expression levels compared to the other isozymes | Papaver somniferum |
isozyme COR1.3, DNA and amino acid sequence determination and analysis, sequence comparison with codeinone reductase isoforms COR1.2, COR1.1, COR1.4, and COR-B, quantitative real-time PCR expression analysis, recombinant coexpression of isozyme COR1.3 with thebaine 6-O-demethylase (T6ODM) in Saccharomyces cerevisiae strain CEN.PK fed with thebaine | Papaver somniferum |
isozyme COR1.4, DNA and amino acid sequence determination and analysis, sequence comparison with codeinone reductase isoforms COR1.2, COR1.3, COR1.1, and COR-B, quantitative real-time PCR expression analysis | Papaver somniferum |
Protein Variants | Comment | Organism |
---|---|---|
A25V | site-directed mutagenesis, the mutant shows a marginal increase in neopine formation compared to wild-type | Papaver somniferum |
A25V/E111D/T145S/S165C | site-directed mutagenesis, the mutant shows a reduction in neopine formation compared to wild-type | Papaver somniferum |
E111D/T145S/S165C | site-directed mutagenesis, the mutant shows a reduction in neopine formation compared to wild-type | Papaver somniferum |
F129L | site-directed mutagenesis, the mutant of COR-B shows unaltered neopine formation compared to wild-type | Papaver somniferum |
K41N | site-directed mutagenesis, the mutant of COR-B shows unaltered neopine formation compared to wild-type | Papaver somniferum |
L129F | site-directed mutagenesis, the mutant shows a higher neopine yield compared to wild-type | Papaver somniferum |
additional information | combinations and permutations of the three substitutions N41K, L129F and S279W do not produce a synergistic effect | Papaver somniferum |
additional information | recombinant coexpression of isozyme COR-B with thebaine 6-O-demethylase (T6ODM) in Saccharomyces cerevisiae strain CEN.PK fed with thebaine, the strain expressing T6ODM and COR-B accumulates approximately 2fold more codeine and 10fold more neopine compared to wild-type | Papaver somniferum |
additional information | recombinant coexpression of isozyme COR1.3 with thebaine 6-O-demethylase (T6ODM) in Saccharomyces cerevisiae strain CEN.PK fed with thebaine. In the recombinant strain, codeine and neopine accumulate to similar levels | Papaver somniferum |
additional information | the COR1.4 single-substitution mutant A (A25V) results in a marginal increase in neopine formation, whereas the triple C (E111D), E (T145S) and F (S165C), and the quadruple A (A25V), C (E111D), E (T145S) and F (S165C), mutants result in higher reductions in neopine levels | Papaver somniferum |
N41K | site-directed mutagenesis, the mutant shows a higher neopine yield compared to wild-type | Papaver somniferum |
S279W | site-directed mutagenesis, the mutant shows a higher neopine yield compared to wild-type | Papaver somniferum |
V25A | site-directed mutagenesis, the mutant of COR-B shows 2fold reduced neopine formation compared to wild-type | Papaver somniferum |
W279S | site-directed mutagenesis, the mutant of COR-B shows 8fold reduced neopine formation compared to wild-type | Papaver somniferum |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | steady-state Michaelis-Menten kinetics | Papaver somniferum | |
0.0117 | - |
codeinone | pH 6.8, 30°C | Papaver somniferum | |
0.0183 | - |
codeinone | pH 6.8, 30°C | Papaver somniferum | |
0.0298 | - |
codeine | pH 9.0, 30°C | Papaver somniferum | |
0.0312 | - |
codeine | pH 9.0, 30°C | Papaver somniferum | |
0.036 | - |
NADPH | pH 6.8, 30°C | Papaver somniferum | |
0.0462 | - |
NADP+ | pH 9.0, 30°C | Papaver somniferum | |
0.0648 | - |
NADPH | pH 6.8, 30°C | Papaver somniferum | |
0.0697 | - |
NADP+ | pH 9.0, 30°C | Papaver somniferum |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
codeine + NADP+ | Papaver somniferum | - |
codeinone + NADPH + H+ | - |
r | |
additional information | Papaver somniferum | codeinone and morphinone are spontaneously converted to neopinone and neomorphinone, respectively. Latex protein extracts assayed for COR activity show the highest turnover of codeinone and morphinone to codeine and morphine, respectively. Codeine and morphine are also converted at reduced levels, whereas no conversion of either neopine or neomorphine is detected | ? | - |
- |
|
morphine + NADP+ | Papaver somniferum | - |
morphinone + NADPH + H+ | - |
r | |
neomorphinone + NADPH + H+ | Papaver somniferum | - |
neomorphine + NADP+ | - |
ir | |
neopinone + NADPH + H+ | Papaver somniferum | - |
neopine + NADP+ | - |
ir |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Papaver somniferum | A0A4P2SMY1 | isozyme COR-B; chemotype Bea's Choice | - |
Papaver somniferum | Q9SQ67 | isozyme COR1.4; chemotype Bea's Choice | - |
Papaver somniferum | Q9SQ68 | isozyme COR1.3; chemotype Bea's Choice | - |
Papaver somniferum | Q9SQ69 | isozyme COR1.2; chemotype Bea's Choice | - |
Papaver somniferum | Q9SQ70 | isozyme COR1.1; chemotype Bea's Choice | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
codeine + NADP+ | - |
Papaver somniferum | codeinone + NADPH + H+ | - |
r | |
additional information | codeinone and morphinone are spontaneously converted to neopinone and neomorphinone, respectively. Latex protein extracts assayed for COR activity show the highest turnover of codeinone and morphinone to codeine and morphine, respectively. Codeine and morphine are also converted at reduced levels, whereas no conversion of either neopine or neomorphine is detected | Papaver somniferum | ? | - |
- |
|
additional information | codeinone and morphinone are spontaneously converted to neopinone and neomorphinone, respectively | Papaver somniferum | ? | - |
- |
|
morphine + NADP+ | - |
Papaver somniferum | morphinone + NADPH + H+ | - |
r | |
neomorphinone + NADPH + H+ | - |
Papaver somniferum | neomorphine + NADP+ | - |
ir | |
neopinone + NADPH + H+ | - |
Papaver somniferum | neopine + NADP+ | - |
ir |
Synonyms | Comment | Organism |
---|---|---|
codeinone reductase | - |
Papaver somniferum |
COR | - |
Papaver somniferum |
COR-B | - |
Papaver somniferum |
COR1.1 | - |
Papaver somniferum |
COR1.2 | - |
Papaver somniferum |
COR1.3 | - |
Papaver somniferum |
COR1.4 | - |
Papaver somniferum |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
- |
Papaver somniferum |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
additional information | - |
addition of NADP+ and NADPH cofactors only a marginal effect on COR1.3 thermostability | Papaver somniferum |
additional information | - |
addition of NADP+ and NADPH cofactors substantially increases the thermostability of COR-B | Papaver somniferum |
30 | 42 | COR1.3 activity is maintained at 30-42°C, sharply reduced above 42°C and is not detected at 62°C | Papaver somniferum |
30 | 62 | COR-B maintains activity from 30°C to 42°C and displays low activity at 62°C | Papaver somniferum |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.015 | - |
codeine | pH 9.0, 30°C | Papaver somniferum | |
0.023 | - |
codeinone | pH 6.8, 30°C | Papaver somniferum | |
0.027 | - |
codeine | pH 9.0, 30°C | Papaver somniferum | |
0.04 | - |
codeinone | pH 6.8, 30°C | Papaver somniferum | |
0.044 | - |
NADP+ | pH 9.0, 30°C | Papaver somniferum | |
0.045 | - |
NADPH | pH 6.8, 30°C | Papaver somniferum | |
0.06 | - |
NADPH | pH 6.8, 30°C | Papaver somniferum | |
0.067 | - |
NADP+ | pH 9.0, 30°C | Papaver somniferum |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6.8 | - |
reduction of codeinone to codeine | Papaver somniferum |
9 | - |
oxidation of codeine to codeinone | Papaver somniferum |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADP+ | - |
Papaver somniferum | |
NADPH | - |
Papaver somniferum |
General Information | Comment | Organism |
---|---|---|
evolution | deduction and calculation of sequences of codeinone reductase isoforms COR1.1, COR1.2, COR1.3, COR1.4, and COR-B, overview. In 3-h assays using codeine and NADP+ as substrates, isozyme COR-B yields the most neopine, followed by COR1.4, COR1.2, COR1.1, and COR1.3. The four residues (V25, K41, F129 and W279) responsible for the differential function of COR-B, compared with COR1.1-1.4, are predominant among COR isoforms in various opium poppy chemotypes | Papaver somniferum |
metabolism | codeinone reductase (COR) catalyzes the reversible NADPH-dependent reduction of codeinone to codeine as the penultimate step of morphine biosynthesis in opium poppy (Papaver somniferum). In a parallel pathway involving 3-O-demethylated analogs, COR converts morphinone to morphine, and neomorphinone to neomorphine. Similar to neopine, the formation of neomorphine by COR is irreversible | Papaver somniferum |
additional information | COR structure homology modeling using chain A of the crystal structure of CHR complexed with NADPH (PDB ID 1ZGD) as a template | Papaver somniferum |
additional information | COR structure homology modeling using chain A of the crystal structure of CHR complexed with NADPH (PDB ID 1ZGD) as a template. Structure-function relationships of isozymes COR1.3 and COR-B, comparisons, overview | Papaver somniferum |
physiological function | codeinone reductase (COR) catalyzes the reversible NADPH-dependent reduction of codeinone to codeine as the penultimate step of morphine biosynthesis in opium poppy (Papaver somniferum). It also irreversibly reduces neopinone, which forms by spontaneous isomerization in aqueous solution from codeinone, to neopine. In a parallel pathway involving 3-O-demethylated analogs, COR converts morphinone to morphine, and neomorphinone to neomorphine | Papaver somniferum |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.197 | - |
codeinone | pH 6.8, 30°C | Papaver somniferum | |
0.503 | - |
codeine | pH 9.0, 30°C | Papaver somniferum | |
0.631 | - |
NADP+ | pH 9.0, 30°C | Papaver somniferum | |
0.865 | - |
codeine | pH 9.0, 30°C | Papaver somniferum | |
0.925 | - |
NADPH | pH 6.8, 30°C | Papaver somniferum | |
1.25 | - |
NADPH | pH 6.8, 30°C | Papaver somniferum | |
1.45 | - |
NADP+ | pH 9.0, 30°C | Papaver somniferum | |
2.19 | - |
codeinone | pH 6.8, 30°C | Papaver somniferum |