Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes in Escherichia coli strain BL21 (DE3), sequence comparison of caffeine synthetic enzymes from coffee | Coffea sp. |
Protein Variants | Comment | Organism |
---|---|---|
A23S | site-directed mutagenesis, the mutant exhibits no 3-N methylation activity like the wild-type, the mutant shows over 80% of wild-type 7-methylation activity | Coffea sp. |
A23S/L191P/H219R | site-directed mutagenesis, the mutant exhibits no 3-N methylation activity like the wild-type, the mutant shows over 80% of wild-type 7-methylation activity | Coffea sp. |
A23S/P104Q/H219R | site-directed mutagenesis, the mutant exhibits no 3-N methylation activity like the wild-type, the mutant shows over 80% of wild-type 7-methylation activity | Coffea sp. |
A23S/P104Q/Q161H/H219R | site-directed mutagenesis, the mutant exhibits 3-N methylation activity in contrast to the wild-type enzyme, the mutant shows 60% of wild-type 7-methylation activity | Coffea sp. |
A23S/P104Q/Q161H/L191P/H219R | site-directed mutagenesis, the mutant exhibits 3-N methylation activity in contrast to the wild-type enzyme, the mutant shows over 80% of wild-type 7-methylation activity | Coffea sp. |
A23S/Q161H/H219R | site-directed mutagenesis, the mutant exhibits 3-N methylation activity in contrast to the wild-type enzyme, the mutant shows over 80% of wild-type 7-methylation activity | Coffea sp. |
A23S/Q161H/L191P/H219R | site-directed mutagenesis, the mutant exhibits 3-N methylation activity in contrast to the wild-type enzyme, the mutant shows over 80% of wild-type 7-methylation activity | Coffea sp. |
H219R | site-directed mutagenesis, the mutant exhibits no 3-N methylation activity like the wild-type, the mutant shows over 80% of wild-type 7-methylation activity | Coffea sp. |
L191P | site-directed mutagenesis, the mutant exhibits no 3-N methylation activity like the wild-type, the mutant shows over 80% of wild-type 7-methylation activity | Coffea sp. |
additional information | construction of several enzyme mutants, overview. The mutants of CmXRS1, that have 3-N methylation activity and produce caffeine from paraxanthine as a substrate, need to have replacement of the glutamine residue by histidine at position 161 in the CmXRS1 sequence, i.e. a Q161H mutation, overview | Coffea sp. |
P104Q | site-directed mutagenesis, the mutant exhibits no 3-N methylation activity like the wild-type, the mutant shows over 80% of wild-type 7-methylation activity | Coffea sp. |
P104Q/L191P | site-directed mutagenesis, the mutant exhibits no 3-N methylation activity like the wild-type, the mutant shows over 80% of wild-type 7-methylation activity | Coffea sp. |
P104Q/Q161H | site-directed mutagenesis, the mutant exhibits 3-N methylation activity in contrast to the wild-type enzyme, the mutant shows over 80% of wild-type 7-methylation activity | Coffea sp. |
P104Q/Q161H/L191P | site-directed mutagenesis, the mutant exhibits 3-N methylation activity in contrast to the wild-type enzyme, the mutant shows over 80% of wild-type 7-methylation activity | Coffea sp. |
Q161H | site-directed mutagenesis, the mutant exhibits 3-N methylation activity in contrast to the wild-type enzyme, the mutant shows over 80% of wild-type 7-methylation activity | Coffea sp. |
Q161H/L191P | site-directed mutagenesis, the mutant exhibits 3-N methylation activity in contrast to the wild-type enzyme, the mutant shows over 80% of wild-type 7-methylation activity | Coffea sp. |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Coffea sp. |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
S-adenosyl-L-methionine + xanthosine | Coffea sp. | - |
S-adenosyl-L-homocysteine + 7-methylxanthosine | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Coffea sp. | - |
- |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | 7-methylxanthosine synthase shows the most critical substrate specifi city of the three types of N-methyltransferases in Coffea species | Coffea sp. | ? | - |
? | |
S-adenosyl-L-methionine + xanthosine | - |
Coffea sp. | S-adenosyl-L-homocysteine + 7-methylxanthosine | - |
? |
Synonyms | Comment | Organism |
---|---|---|
CmXRS1 | - |
Coffea sp. |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
27 | - |
assay at | Coffea sp. |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Coffea sp. |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
S-adenosyl-L-methionine | - |
Coffea sp. |
General Information | Comment | Organism |
---|---|---|
metabolism | the enzyme catalyzes the first step in caffeine biosynthesis, pathway overview | Coffea sp. |