Protein Variants | Comment | Organism |
---|---|---|
E43K/S97C/T148S/A155H/P210C/L227V/Y244F | site-directed mutagenesis, mutant ADHA-0381, the mutant shows increased thermostability and altered pH conditions compared to the wild-type enzyme | Starmerella magnoliae |
E43K/S97C/T148S/T194V/M206D/P210C/L227V | site-directed mutagenesis, mutant ADHA-0398, the mutant shows increased thermostability and altered pH conditions compared to the wild-type enzyme | Starmerella magnoliae |
additional information | development of improved enzyme variants ADHA-0398 and -0381 shows +26°C improved melting temperature and 17fold higher oxidative activity at pH 6.5 towards (4R,6S)-6-(chloromethyl)oxane-2,4-diol but only 6fold at pH 7.5, method development, overview | Starmerella magnoliae |
S97C/M206D | site-directed mutagenesis, mutant ADHA-0279, the mutant shows increased thermostability and altered pH conditions compared to the wild-type enzyme | Starmerella magnoliae |
S97C/M206D/P210C | site-directed mutagenesis, mutant ADHA-0278, the mutant shows increased thermostability and altered pH conditions compared to the wild-type enzyme | Starmerella magnoliae |
S97C/T148S/A155H/P210C/L227V | site-directed mutagenesis, mutant ADHA-0391, the mutant shows increased thermostability and altered pH conditions compared to the wild-type enzyme | Starmerella magnoliae |
S97C/T148S/A155H/P210C/L227V/Y244F | site-directed mutagenesis, mutant ADHA-0384, the mutant shows increased thermostability and altered pH conditions compared to the wild-type enzyme | Starmerella magnoliae |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Starmerella magnoliae | - |
- |
- |
Starmerella magnoliae DSMZ 70638 | - |
- |
- |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
specific activities of enzyme mutants with the different substrates, overview | Starmerella magnoliae |
0.33 | - |
wild-type enzyme, pH 6.5, 28°C, substrate (4R,6S)-6-(chloromethyl)oxane-2,4-diol | Starmerella magnoliae |
4.11 | - |
wild-type enzyme, pH 7.0, 28°C, substrate ethyl acetoacetate | Starmerella magnoliae |
4.11 | - |
wild-type enzyme, pH 7.5, 28°C, substrate (4R,6S)-6-(chloromethyl)oxane-2,4-diol | Starmerella magnoliae |
12.9 | - |
wild-type enzyme, pH 7.0, 28°C, substrate (4R,6R)-6-butyloxane-2,4-diol | Starmerella magnoliae |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(4R,6R)-6-butyloxane-2,4-diol + NAD+ | - |
Starmerella magnoliae | (4R,6R)-6-butyl-4-hydroxyoxan-2-one + NADH + H+ | - |
? | |
(4R,6R)-6-butyloxane-2,4-diol + NAD+ | - |
Starmerella magnoliae DSMZ 70638 | (4R,6R)-6-butyl-4-hydroxyoxan-2-one + NADH + H+ | - |
? | |
(4R,6R)-6-butyloxane-2,4-diol + NADP+ | - |
Starmerella magnoliae | (4R,6R)-6-butyl-4-hydroxyoxan-2-one + NADPH + H+ | - |
? | |
(4R,6R)-6-butyloxane-2,4-diol + NADP+ | - |
Starmerella magnoliae DSMZ 70638 | (4R,6R)-6-butyl-4-hydroxyoxan-2-one + NADPH + H+ | - |
? | |
(4R,6R)-6-pentyloxane-2,4-diol + NAD+ | - |
Starmerella magnoliae | (4R,6R)-4-hydroxy-6-pentyloxan-2-one + NADH + H+ | - |
? | |
(4R,6R)-6-pentyloxane-2,4-diol + NAD+ | - |
Starmerella magnoliae DSMZ 70638 | (4R,6R)-4-hydroxy-6-pentyloxan-2-one + NADH + H+ | - |
? | |
(4R,6R)-6-pentyloxane-2,4-diol + NADP+ | - |
Starmerella magnoliae | (4R,6R)-4-hydroxy-6-pentyloxan-2-one + NADPH + H+ | - |
? | |
(4R,6R)-6-pentyloxane-2,4-diol + NADP+ | - |
Starmerella magnoliae DSMZ 70638 | (4R,6R)-4-hydroxy-6-pentyloxan-2-one + NADPH + H+ | - |
? | |
(4R,6S)-6-(chloromethyl)oxane-2,4-diol + NAD+ | - |
Starmerella magnoliae | (4R,6S)-6-(chloromethyl)-4-hydroxyoxan-2-one + NADH + H+ | - |
? | |
(4R,6S)-6-(chloromethyl)oxane-2,4-diol + NADP+ | - |
Starmerella magnoliae | (4R,6S)-6-(chloromethyl)-4-hydroxyoxan-2-one + NADPH + H+ | - |
? | |
ethyl acetoacetate + NADPH + H+ | - |
Starmerella magnoliae | ? + NADP+ | - |
r | |
additional information | lactol substrates (4R,6S)-6-(chloromethyl)oxane-2,4-diol and (4R,6R)-6-butyloxane-2,4-diol for the biocatalytic oxidations are obtained from 2-deoxy-D-ribose 5-phosphate aldolase (DERA) catalyzed reactions of the corresponding acceptor aldehydes chloroacetaldehyde and pentanal, respectively and at least two equivalents of acetaldehyde as described previously. Development of an efficient biocatalytic process on pilot plant scale to oxidize the aliphatic hydroxylactol substrate (4R,6R)-6-butyloxane-2,4-diol to the corresponding lactone product (4R,6R)-6-butyl-4-hydroxyoxan-2-one employing the wild-type alcohol dehydrogenase ADHA | Starmerella magnoliae | ? | - |
- |
|
additional information | lactol substrates (4R,6S)-6-(chloromethyl)oxane-2,4-diol and (4R,6R)-6-butyloxane-2,4-diol for the biocatalytic oxidations are obtained from 2-deoxy-D-ribose 5-phosphate aldolase (DERA) catalyzed reactions of the corresponding acceptor aldehydes chloroacetaldehyde and pentanal, respectively and at least two equivalents of acetaldehyde as described previously. Development of an efficient biocatalytic process on pilot plant scale to oxidize the aliphatic hydroxylactol substrate (4R,6R)-6-butyloxane-2,4-diol to the corresponding lactone product (4R,6R)-6-butyl-4-hydroxyoxan-2-one employing the wild-type alcohol dehydrogenase ADHA | Starmerella magnoliae DSMZ 70638 | ? | - |
- |
Synonyms | Comment | Organism |
---|---|---|
ADH | - |
Starmerella magnoliae |
AdhA | - |
Starmerella magnoliae |
alcohol dehydrogenase | - |
Starmerella magnoliae |
NADPH-dependent ADHA | - |
Starmerella magnoliae |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
28 | 42 | assay at | Starmerella magnoliae |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
41 | - |
melting temperature T1/2 of wild-type enzyme ADHA | Starmerella magnoliae |
48 | - |
melting temperature T1/2 of mutant ADHA-0278 | Starmerella magnoliae |
53 | - |
melting temperature T1/2 of mutant ADHA-0279 | Starmerella magnoliae |
65 | - |
melting temperature T1/2 of mutant ADHA-0391 | Starmerella magnoliae |
66 | - |
melting temperature T1/2 of mutant ADHA-0384 | Starmerella magnoliae |
67 | - |
melting temperature T1/2 of mutant ADHA-0398 and of mutant ADHA-0381 | Starmerella magnoliae |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6.5 | 7.5 | dependent on the substrate | Starmerella magnoliae |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Starmerella magnoliae | |
NADH | - |
Starmerella magnoliae | |
NADP+ | - |
Starmerella magnoliae | |
NADPH | - |
Starmerella magnoliae |
General Information | Comment | Organism |
---|---|---|
malfunction | the optimized ADHA variant shows a 7fold higher oxidative activity, a 26°C increased stability, and is applied to develop an efficient chlorolactol oxidation process | Starmerella magnoliae |
physiological function | enzyme ADHA from Candida magnoliae strain DSMZ 70638 is identified to efficiently catalyze the regioselective hydroxy-lactone oxidations to hydroxy-lactones. Hydroxy-lactones are common intermediates in industrial processes to cholesterol lowering (va)statin drugs. ADH catalyzed oxidations can be developed to efficient and safe processes, but the ADHA wild-type enzyme is not productive enough in chlorolactol oxidation | Starmerella magnoliae |