Application | Comment | Organism |
---|---|---|
synthesis | alcohol dehydrogenases represent an important group of biocatalysts due to their ability to stereospecifically reduce prochiral carbonyl compounds | Rhodococcus ruber |
synthesis | expression of enzyme in auxotrophic Arxula adeninivorans, Hansenula polymorpha, and Saccharomyces cerevisiae strains using yeast ribosomal DNA integrative expression cassettes. Recombinant ADH accumulates intracellularly in all strains tested. The best yields of active enzyme are obtained from A. adeninivorans, with Saccharomyces cerevisiae producing intermediate amounts. Although Hansenula polymorpha is the least efficient producer overall, the product obtained is most similar to the enzyme synthesized by Rhodococcus ruber 219 with respect to its thermostability | Rhodococcus ruber |
Cloned (Comment) | Organism |
---|---|
DNA and amino acid sequence determination and analysis, expression of His-tagged enzyme under the control of the strong constitutive Arxula adeninivorans-derived TEF1 promoter in auxotrophic Arxula adeninivorans strain G1214, Hansenula polymorpha strain RB11, and Saccharomyces cerevisiae strainSEY6210, using different expression modules for transformation, evaluation of effeciency, overview. Expression in Arxula adeninivorans is most effective | Rhodococcus ruber |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.14 | - |
capronaldehyde | pH 6.0, 37°C, recombinant enzyme expressed from Arxula adeninivorans | Rhodococcus ruber | |
0.14 | - |
4-chloroacetophenone | pH 6.0, 37°C, recombinant enzyme expressed from Hansenula polymorpha | Rhodococcus ruber | |
0.17 | - |
Valeraldehyde | pH 6.0, 37°C, recombinant enzyme expressed from Saccharomyces cerevisiae | Rhodococcus ruber | |
0.19 | - |
phenylacetaldehyde | pH 6.0, 37°C, recombinant enzyme expressed from Saccharomyces cerevisiae | Rhodococcus ruber | |
0.2 | - |
acetophenone | pH 6.0, 37°C, recombinant enzyme expressed from Hansenula polymorpha | Rhodococcus ruber | |
0.22 | - |
capronaldehyde | pH 6.0, 37°C, recombinant enzyme expressed from Saccharomyces cerevisiae | Rhodococcus ruber | |
0.25 | - |
capronaldehyde | pH 6.0, 37°C, recombinant enzyme expressed from Hansenula polymorpha | Rhodococcus ruber | |
0.28 | - |
Valeraldehyde | pH 6.0, 37°C, recombinant enzyme expressed from Arxula adeninivorans | Rhodococcus ruber | |
0.33 | - |
phenylacetaldehyde | pH 6.0, 37°C, recombinant enzyme expressed from Hansenula polymorpha | Rhodococcus ruber | |
0.4 | - |
acetophenone | pH 6.0, 37°C, recombinant enzyme expressed from Arxula adeninivorans | Rhodococcus ruber | |
0.53 | - |
4-chloroacetophenone | pH 6.0, 37°C, recombinant enzyme expressed from Saccharomyces cerevisiae | Rhodococcus ruber | |
0.66 | - |
acetophenone | pH 6.0, 37°C, recombinant enzyme expressed from Saccharomyces cerevisiae | Rhodococcus ruber | |
0.91 | - |
phenylacetaldehyde | pH 6.0, 37°C, recombinant enzyme expressed from Arxula adeninivorans | Rhodococcus ruber | |
1.11 | - |
4-chloroacetophenone | pH 6.0, 37°C, recombinant enzyme expressed from Arxula adeninivorans | Rhodococcus ruber | |
2 | - |
Valeraldehyde | pH 6.0, 37°C, recombinant enzyme expressed from Hansenula polymorpha | Rhodococcus ruber |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
38000 | - |
2 * 38000, recombinant enzyme, SDS-PAGE | Rhodococcus ruber |
66000 | - |
recombinant enzyme expressed from Hansenula polymorpha, gel filtration | Rhodococcus ruber |
72000 | - |
recombinant enzyme expressed from Arxula adeninivorans and Saccharomyces cerevisiae, gel filtration | Rhodococcus ruber |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Rhodococcus ruber | - |
- |
- |
Rhodococcus ruber | - |
DSM44514 | - |
Rhodococcus ruber 219 | - |
DSM44514 | - |
Rhodococcus ruber DSM 44514 | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme by nickel affinity chromatography and gel filtration | Rhodococcus ruber |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
lyophilized cell | - |
Rhodococcus ruber | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-chloroacetophenone + NADH | - |
Rhodococcus ruber | 1-(4-chlorophenyl)ethanol + NAD+ | - |
r | |
4-chloroacetophenone + NADH | - |
Rhodococcus ruber 219 | 1-(4-chlorophenyl)ethanol + NAD+ | - |
r | |
4-chloroacetophenone + NADH | - |
Rhodococcus ruber DSM 44514 | 1-(4-chlorophenyl)ethanol + NAD+ | - |
r | |
acetophenone + NADH + H+ | - |
Rhodococcus ruber | 1-phenylethanol + NAD+ | - |
r | |
acetophenone + NADH + H+ | - |
Rhodococcus ruber 219 | 1-phenylethanol + NAD+ | - |
r | |
acetophenone + NADH + H+ | - |
Rhodococcus ruber DSM 44514 | 1-phenylethanol + NAD+ | - |
r | |
capronaldehyde + NADH | best substrate | Rhodococcus ruber | 1-hexanol + NAD+ | - |
r | |
capronaldehyde + NADH | best substrate | Rhodococcus ruber 219 | 1-hexanol + NAD+ | - |
r | |
capronaldehyde + NADH | best substrate | Rhodococcus ruber DSM 44514 | 1-hexanol + NAD+ | - |
r | |
phenylacetaldehyde + NADH | - |
Rhodococcus ruber | phenylethanol + NAD+ | - |
r | |
phenylacetaldehyde + NADH | - |
Rhodococcus ruber 219 | phenylethanol + NAD+ | - |
r | |
phenylacetaldehyde + NADH | - |
Rhodococcus ruber DSM 44514 | phenylethanol + NAD+ | - |
r | |
valeraldehyde + NADH | - |
Rhodococcus ruber | 1-pentanol + NAD+ | - |
r | |
valeraldehyde + NADH | - |
Rhodococcus ruber 219 | 1-pentanol + NAD+ | - |
r | |
valeraldehyde + NADH | - |
Rhodococcus ruber DSM 44514 | 1-pentanol + NAD+ | - |
r |
Subunits | Comment | Organism |
---|---|---|
dimer | 2 * 38000, recombinant enzyme, SDS-PAGE | Rhodococcus ruber |
Synonyms | Comment | Organism |
---|---|---|
ADH6Hp | - |
Rhodococcus ruber |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
35 | - |
reductive reaction, recombinant enzymes expressed from Saccharomyces cerevisiae and Hansenula polymorpha | Rhodococcus ruber |
35 | - |
recombinant enzyme expressed in Hansenula polymorpha | Rhodococcus ruber |
35 | - |
recombinant enzyme expressed in Saccharomyces cerevisiae | Rhodococcus ruber |
39 | - |
reductive reaction, recombinant enzyme expressed from Arxula adeninivorans | Rhodococcus ruber |
39 | - |
recombinant enzyme expressed in Arxula adeninivorans | Rhodococcus ruber |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
additional information | - |
temperature stability profiles of recombinantly expressed enzymes, overview | Rhodococcus ruber |
32 | 41 | purified recombinant enzyme expressed from Saccharomyces cerevisiae, at least 80% of the initial activity is retained after 4 min | Rhodococcus ruber |
32 | 41 | more than 80% of maximum activity, recombinant enzyme expressed in Saccharomyces cerevisiae | Rhodococcus ruber |
34 | 38 | purified recombinant enzyme expressed from Hansenula polymorpha, at least 80% of the initial activity is retained after 4 min | Rhodococcus ruber |
34 | 38 | more than 80% of maximum activity, recombinant enzyme expressed in Hansenula polymorpha | Rhodococcus ruber |
38.5 | 42.5 | purified recombinant enzyme expressed from Arxula adeninivorans, at least 80% of the initial activity is retained after 4 min | Rhodococcus ruber |
38.5 | 42.5 | more than 80% of maximum activity, recombinant enzyme expressed in Arxula adeninivorans | Rhodococcus ruber |
70 | - |
purified recombinant enzyme expressed from Hansenula polymorpha, over 80% of the initial activity is retained after 60 min | Rhodococcus ruber |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
145 | - |
4-chloroacetophenone | pH 6.0, 37°C, recombinant enzyme expressed from Hansenula polymorpha | Rhodococcus ruber | |
202 | - |
phenylacetaldehyde | pH 6.0, 37°C, recombinant enzyme expressed from Saccharomyces cerevisiae | Rhodococcus ruber | |
211 | - |
acetophenone | pH 6.0, 37°C, recombinant enzyme expressed from Hansenula polymorpha | Rhodococcus ruber | |
230 | - |
4-chloroacetophenone | pH 6.0, 37°C, recombinant enzyme expressed from Saccharomyces cerevisiae | Rhodococcus ruber | |
259 | - |
acetophenone | pH 6.0, 37°C, recombinant enzyme expressed from Saccharomyces cerevisiae | Rhodococcus ruber | |
264 | - |
phenylacetaldehyde | pH 6.0, 37°C, recombinant enzyme expressed from Hansenula polymorpha | Rhodococcus ruber | |
288 | - |
acetophenone | pH 6.0, 37°C, recombinant enzyme expressed from Arxula adeninivorans | Rhodococcus ruber | |
303 | - |
phenylacetaldehyde | pH 6.0, 37°C, recombinant enzyme expressed from Arxula adeninivorans | Rhodococcus ruber | |
317 | - |
capronaldehyde | pH 6.0, 37°C, recombinant enzyme expressed from Arxula adeninivorans | Rhodococcus ruber | |
410 | - |
capronaldehyde | pH 6.0, 37°C, recombinant enzyme expressed from Hansenula polymorpha | Rhodococcus ruber | |
418 | - |
Valeraldehyde | pH 6.0, 37°C, recombinant enzyme expressed from Saccharomyces cerevisiae | Rhodococcus ruber | |
432 | - |
Valeraldehyde | pH 6.0, 37°C, recombinant enzyme expressed from Arxula adeninivorans | Rhodococcus ruber | |
432 | - |
4-chloroacetophenone | pH 6.0, 37°C, recombinant enzyme expressed from Arxula adeninivorans | Rhodococcus ruber | |
528 | - |
Valeraldehyde | pH 6.0, 37°C, recombinant enzyme expressed from Hansenula polymorpha | Rhodococcus ruber | |
576 | - |
capronaldehyde | pH 6.0, 37°C, recombinant enzyme expressed from Saccharomyces cerevisiae | Rhodococcus ruber |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6 | - |
reductive reaction, all recombinant enzymes | Rhodococcus ruber |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
5.2 | 6.3 | purified recombinant enzyme expressed from Saccharomyces cerevisiae, at least 80% of the initial activity is retained after 4 min, 30°C | Rhodococcus ruber |
5.5 | 6.3 | purified recombinant enzyme expressed from Hansenula polymorpha, at least 80% of the initial activity is retained after 4 min, 30°C | Rhodococcus ruber |
5.8 | 6.6 | purified recombinant enzyme expressed from Arxula adeninivorans, at least 80% of the initial activity is retained after 4 min, 30°C | Rhodococcus ruber |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | - |
Rhodococcus ruber | |
NADH | - |
Rhodococcus ruber |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
264 | - |
Valeraldehyde | pH 6.0, 37°C, recombinant enzyme expressed from Hansenula polymorpha | Rhodococcus ruber | |
333 | - |
phenylacetaldehyde | pH 6.0, 37°C, recombinant enzyme expressed from Arxula adeninivorans | Rhodococcus ruber | |
389 | - |
4-chloroacetophenone | pH 6.0, 37°C, recombinant enzyme expressed from Arxula adeninivorans | Rhodococcus ruber | |
392 | - |
acetophenone | pH 6.0, 37°C, recombinant enzyme expressed from Saccharomyces cerevisiae | Rhodococcus ruber | |
434 | - |
4-chloroacetophenone | pH 6.0, 37°C, recombinant enzyme expressed from Saccharomyces cerevisiae | Rhodococcus ruber | |
589 | - |
Valeraldehyde | pH 6.0, 37°C, recombinant enzyme expressed from Saccharomyces cerevisiae | Rhodococcus ruber | |
720 | - |
acetophenone | pH 6.0, 37°C, recombinant enzyme expressed from Arxula adeninivorans | Rhodococcus ruber | |
800 | - |
phenylacetaldehyde | pH 6.0, 37°C, recombinant enzyme expressed from Hansenula polymorpha | Rhodococcus ruber | |
1036 | - |
4-chloroacetophenone | pH 6.0, 37°C, recombinant enzyme expressed from Hansenula polymorpha | Rhodococcus ruber | |
1055 | - |
acetophenone | pH 6.0, 37°C, recombinant enzyme expressed from Hansenula polymorpha | Rhodococcus ruber | |
1063 | - |
phenylacetaldehyde | pH 6.0, 37°C, recombinant enzyme expressed from Saccharomyces cerevisiae | Rhodococcus ruber | |
1543 | - |
Valeraldehyde | pH 6.0, 37°C, recombinant enzyme expressed from Arxula adeninivorans | Rhodococcus ruber | |
1640 | - |
capronaldehyde | pH 6.0, 37°C, recombinant enzyme expressed from Hansenula polymorpha | Rhodococcus ruber | |
2264 | - |
capronaldehyde | pH 6.0, 37°C, recombinant enzyme expressed from Arxula adeninivorans | Rhodococcus ruber | |
2618 | - |
capronaldehyde | pH 6.0, 37°C, recombinant enzyme expressed from Saccharomyces cerevisiae | Rhodococcus ruber |