Cloned (Comment) | Organism |
---|---|
expression of exchange mutants in Escherichia coli | Saccharomyces cerevisiae |
overexpression in Escherichia coli | Saccharomyces cerevisiae |
Protein Variants | Comment | Organism |
---|---|---|
D65L | potentially responsible for substrate binding to leucine by site directed mutagenesis, unchanged activity | Saccharomyces cerevisiae |
D79L | potentially responsible for substrate binding to leucine by site directed mutagenesis, not decreased activity, 1 new activity to form 24-methylzymosterol from zymosterol | Saccharomyces cerevisiae |
E64L | potentially responsible for substrate binding to leucine by site directed mutagenesis, unchanged activity | Saccharomyces cerevisiae |
E68L | potentially responsible for substrate binding to leucine by site directed mutagenesis, no activity | Saccharomyces cerevisiae |
E82L | potentially responsible for substrate binding to leucine by site directed mutagenesis, not decreased activity, 1 new activity to form 24-methylzymosterol from zymosterol | Saccharomyces cerevisiae |
E98L | potentially responsible for substrate binding to leucine by site directed mutagenesis, unchanged activity | Saccharomyces cerevisiae |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
26,27-dehydrozymosterol | mechanism-based irreversible inhibition | Saccharomyces cerevisiae |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.013 | - |
26,27-dehydrozymosterol | mutant E64L | Saccharomyces cerevisiae | |
0.015 | - |
26,27-dehydrozymosterol | mutant D65L | Saccharomyces cerevisiae | |
0.015 | 0.017 | 26,27-dehydrozymosterol | wild-type | Saccharomyces cerevisiae | |
0.017 | - |
26,27-dehydrozymosterol | mutant D79L | Saccharomyces cerevisiae | |
0.022 | - |
26,27-dehydrozymosterol | mutant E82L | Saccharomyces cerevisiae | |
0.074 | - |
26,27-dehydrozymosterol | mutant E98L | Saccharomyces cerevisiae |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
zymosterol + S-adenosyl-L-methionine | Saccharomyces cerevisiae | - |
fecosterol + S-adenosyl-L-homocysteine | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Saccharomyces cerevisiae | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant mutant from E. coli | Saccharomyces cerevisiae |
recombinant wild-type from E. coli | Saccharomyces cerevisiae |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
S-adenosyl-L-methionine + 5alpha-cholesta-8,24-dien-3beta-ol = S-adenosyl-L-homocysteine + 24-methylene-5alpha-cholest-8-en-3beta-ol | substrate analogue inhibitor binding site structure | Saccharomyces cerevisiae |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
26,27-dehydrozymosterol + S-adenosyl-L-methionine | mutants | Saccharomyces cerevisiae | 26-homo-cholesta-8(9),23(24)E,26(26')-trienol + 26-homo-cholesta-8(9),26(26')-3beta,24beta-dienol + S-adenosyl-L-homocysteine | - |
? | |
zymosterol + S-adenosyl-L-methionine | - |
Saccharomyces cerevisiae | fecosterol + S-adenosyl-L-homocysteine | - |
? | |
zymosterol + S-adenosyl-L-methionine | wild-type and mutants | Saccharomyces cerevisiae | fecosterol + S-adenosyl-L-homocysteine | - |
? | |
zymosterol + S-adenosyl-L-methionine | mutants D79L and E82L | Saccharomyces cerevisiae | 24-methylzymosta-8,25(27)-dienol + S-adenosyl-L-homocysteine | - |
? |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0008 | - |
26,27-dehydrozymosterol | - |
Saccharomyces cerevisiae | |
0.001 | - |
Zymosterol | mutant E98L | Saccharomyces cerevisiae | |
0.005 | - |
Zymosterol | mutants E82L and E64L | Saccharomyces cerevisiae | |
0.007 | - |
Zymosterol | mutant D65L | Saccharomyces cerevisiae | |
0.01 | - |
Zymosterol | wild-type and mutant D79L | Saccharomyces cerevisiae |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0011 | - |
26,27-dehydrozymosterol | - |
Saccharomyces cerevisiae |