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Literature summary extracted from

  • Giersberg, M.; Degelmann, A.; Bode, R.; Piontek, M.; Kunze, G.
    Production of a thermostable alcohol dehydrogenase from Rhodococcus ruber in three different yeast species using the Xplor2 transformation/expression platform (2012), J. Ind. Microbiol. Biotechnol., 39, 1385-1396.
    View publication on PubMed

Application

EC Number Application Comment Organism
1.1.1.1 synthesis alcohol dehydrogenases represent an important group of biocatalysts due to their ability to stereospecifically reduce prochiral carbonyl compounds Rhodococcus ruber
1.1.1.1 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(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.1 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]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.1 0.14
-
 capronaldehyde pH 6.0, 37°C, recombinant enzyme expressed from Arxula adeninivorans Rhodococcus ruber
1.1.1.1 0.14
-
 4-chloroacetophenone pH 6.0, 37°C, recombinant enzyme expressed from Hansenula polymorpha Rhodococcus ruber
1.1.1.1 0.17
-
 Valeraldehyde pH 6.0, 37°C, recombinant enzyme expressed from Saccharomyces cerevisiae Rhodococcus ruber
1.1.1.1 0.19
-
 phenylacetaldehyde pH 6.0, 37°C, recombinant enzyme expressed from Saccharomyces cerevisiae Rhodococcus ruber
1.1.1.1 0.2
-
 acetophenone pH 6.0, 37°C, recombinant enzyme expressed from Hansenula polymorpha Rhodococcus ruber
1.1.1.1 0.22
-
 capronaldehyde pH 6.0, 37°C, recombinant enzyme expressed from Saccharomyces cerevisiae Rhodococcus ruber
1.1.1.1 0.25
-
 capronaldehyde pH 6.0, 37°C, recombinant enzyme expressed from Hansenula polymorpha Rhodococcus ruber
1.1.1.1 0.28
-
 Valeraldehyde pH 6.0, 37°C, recombinant enzyme expressed from Arxula adeninivorans Rhodococcus ruber
1.1.1.1 0.33
-
 phenylacetaldehyde pH 6.0, 37°C, recombinant enzyme expressed from Hansenula polymorpha Rhodococcus ruber
1.1.1.1 0.4
-
 acetophenone pH 6.0, 37°C, recombinant enzyme expressed from Arxula adeninivorans Rhodococcus ruber
1.1.1.1 0.53
-
 4-chloroacetophenone pH 6.0, 37°C, recombinant enzyme expressed from Saccharomyces cerevisiae Rhodococcus ruber
1.1.1.1 0.66
-
 acetophenone pH 6.0, 37°C, recombinant enzyme expressed from Saccharomyces cerevisiae Rhodococcus ruber
1.1.1.1 0.91
-
 phenylacetaldehyde pH 6.0, 37°C, recombinant enzyme expressed from Arxula adeninivorans Rhodococcus ruber
1.1.1.1 1.11
-
 4-chloroacetophenone pH 6.0, 37°C, recombinant enzyme expressed from Arxula adeninivorans Rhodococcus ruber
1.1.1.1 2
-
 Valeraldehyde pH 6.0, 37°C, recombinant enzyme expressed from Hansenula polymorpha Rhodococcus ruber

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.1 38000
-
 2 * 38000, recombinant enzyme, SDS-PAGE Rhodococcus ruber
1.1.1.1 66000
-
 recombinant enzyme expressed from Hansenula polymorpha, gel filtration Rhodococcus ruber
1.1.1.1 72000
-
 recombinant enzyme expressed from Arxula adeninivorans and Saccharomyces cerevisiae, gel filtration Rhodococcus ruber

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.1 Rhodococcus ruber
-
-
-
1.1.1.1 Rhodococcus ruber
-
 DSM44514
-
1.1.1.1 Rhodococcus ruber 219
-
 DSM44514
-
1.1.1.1 Rhodococcus ruber DSM 44514
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.1 recombinant His-tagged enzyme by nickel affinity chromatography and gel filtration Rhodococcus ruber

Source Tissue

EC Number Source Tissue Comment Organism Textmining
1.1.1.1 lyophilized cell
-
 Rhodococcus ruber
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.1 4-chloroacetophenone + NADH
-
 Rhodococcus ruber 1-(4-chlorophenyl)ethanol + NAD+
-
 r
1.1.1.1 4-chloroacetophenone + NADH
-
 Rhodococcus ruber 219 1-(4-chlorophenyl)ethanol + NAD+
-
 r
1.1.1.1 4-chloroacetophenone + NADH
-
 Rhodococcus ruber DSM 44514 1-(4-chlorophenyl)ethanol + NAD+
-
 r
1.1.1.1 acetophenone + NADH + H+
-
 Rhodococcus ruber 1-phenylethanol + NAD+
-
 r
1.1.1.1 acetophenone + NADH + H+
-
 Rhodococcus ruber 219 1-phenylethanol + NAD+
-
 r
1.1.1.1 acetophenone + NADH + H+
-
 Rhodococcus ruber DSM 44514 1-phenylethanol + NAD+
-
 r
1.1.1.1 capronaldehyde + NADH best substrate Rhodococcus ruber 1-hexanol + NAD+
-
 r
1.1.1.1 capronaldehyde + NADH best substrate Rhodococcus ruber 219 1-hexanol + NAD+
-
 r
1.1.1.1 capronaldehyde + NADH best substrate Rhodococcus ruber DSM 44514 1-hexanol + NAD+
-
 r
1.1.1.1 phenylacetaldehyde + NADH
-
 Rhodococcus ruber phenylethanol + NAD+
-
 r
1.1.1.1 phenylacetaldehyde + NADH
-
 Rhodococcus ruber 219 phenylethanol + NAD+
-
 r
1.1.1.1 phenylacetaldehyde + NADH
-
 Rhodococcus ruber DSM 44514 phenylethanol + NAD+
-
 r
1.1.1.1 valeraldehyde + NADH
-
 Rhodococcus ruber 1-pentanol + NAD+
-
 r
1.1.1.1 valeraldehyde + NADH
-
 Rhodococcus ruber 219 1-pentanol + NAD+
-
 r
1.1.1.1 valeraldehyde + NADH
-
 Rhodococcus ruber DSM 44514 1-pentanol + NAD+
-
 r

Subunits

EC Number Subunits Comment Organism
1.1.1.1 dimer 2 * 38000, recombinant enzyme, SDS-PAGE Rhodococcus ruber

Synonyms

EC Number Synonyms Comment Organism
1.1.1.1 ADH6Hp
-
 Rhodococcus ruber

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.1 35
-
 reductive reaction, recombinant enzymes expressed from Saccharomyces cerevisiae and Hansenula polymorpha Rhodococcus ruber
1.1.1.1 35
-
 recombinant enzyme expressed in Hansenula polymorpha Rhodococcus ruber
1.1.1.1 35
-
 recombinant enzyme expressed in Saccharomyces cerevisiae Rhodococcus ruber
1.1.1.1 39
-
 reductive reaction, recombinant enzyme expressed from Arxula adeninivorans Rhodococcus ruber
1.1.1.1 39
-
 recombinant enzyme expressed in Arxula adeninivorans Rhodococcus ruber

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.1.1.1 additional information
-
 temperature stability profiles of recombinantly expressed enzymes, overview Rhodococcus ruber
1.1.1.1 32 41 purified recombinant enzyme expressed from Saccharomyces cerevisiae, at least 80% of the initial activity is retained after 4 min Rhodococcus ruber
1.1.1.1 32 41 more than 80% of maximum activity, recombinant enzyme expressed in Saccharomyces cerevisiae Rhodococcus ruber
1.1.1.1 34 38 purified recombinant enzyme expressed from Hansenula polymorpha, at least 80% of the initial activity is retained after 4 min Rhodococcus ruber
1.1.1.1 34 38 more than 80% of maximum activity, recombinant enzyme expressed in Hansenula polymorpha Rhodococcus ruber
1.1.1.1 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
1.1.1.1 38.5 42.5 more than 80% of maximum activity, recombinant enzyme expressed in Arxula adeninivorans Rhodococcus ruber
1.1.1.1 70
-
 purified recombinant enzyme expressed from Hansenula polymorpha, over 80% of the initial activity is retained after 60 min Rhodococcus ruber

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.1 145
-
 4-chloroacetophenone pH 6.0, 37°C, recombinant enzyme expressed from Hansenula polymorpha Rhodococcus ruber
1.1.1.1 202
-
 phenylacetaldehyde pH 6.0, 37°C, recombinant enzyme expressed from Saccharomyces cerevisiae Rhodococcus ruber
1.1.1.1 211
-
 acetophenone pH 6.0, 37°C, recombinant enzyme expressed from Hansenula polymorpha Rhodococcus ruber
1.1.1.1 230
-
 4-chloroacetophenone pH 6.0, 37°C, recombinant enzyme expressed from Saccharomyces cerevisiae Rhodococcus ruber
1.1.1.1 259
-
 acetophenone pH 6.0, 37°C, recombinant enzyme expressed from Saccharomyces cerevisiae Rhodococcus ruber
1.1.1.1 264
-
 phenylacetaldehyde pH 6.0, 37°C, recombinant enzyme expressed from Hansenula polymorpha Rhodococcus ruber
1.1.1.1 288
-
 acetophenone pH 6.0, 37°C, recombinant enzyme expressed from Arxula adeninivorans Rhodococcus ruber
1.1.1.1 303
-
 phenylacetaldehyde pH 6.0, 37°C, recombinant enzyme expressed from Arxula adeninivorans Rhodococcus ruber
1.1.1.1 317
-
 capronaldehyde pH 6.0, 37°C, recombinant enzyme expressed from Arxula adeninivorans Rhodococcus ruber
1.1.1.1 410
-
 capronaldehyde pH 6.0, 37°C, recombinant enzyme expressed from Hansenula polymorpha Rhodococcus ruber
1.1.1.1 418
-
 Valeraldehyde pH 6.0, 37°C, recombinant enzyme expressed from Saccharomyces cerevisiae Rhodococcus ruber
1.1.1.1 432
-
 Valeraldehyde pH 6.0, 37°C, recombinant enzyme expressed from Arxula adeninivorans Rhodococcus ruber
1.1.1.1 432
-
 4-chloroacetophenone pH 6.0, 37°C, recombinant enzyme expressed from Arxula adeninivorans Rhodococcus ruber
1.1.1.1 528
-
 Valeraldehyde pH 6.0, 37°C, recombinant enzyme expressed from Hansenula polymorpha Rhodococcus ruber
1.1.1.1 576
-
 capronaldehyde pH 6.0, 37°C, recombinant enzyme expressed from Saccharomyces cerevisiae Rhodococcus ruber

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.1 6
-
 reductive reaction, all recombinant enzymes Rhodococcus ruber

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
1.1.1.1 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
1.1.1.1 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
1.1.1.1 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

EC Number Cofactor Comment Organism Structure
1.1.1.1 NAD+
-
 Rhodococcus ruber
1.1.1.1 NADH
-
 Rhodococcus ruber

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.1.1.1 264
-
 Valeraldehyde pH 6.0, 37°C, recombinant enzyme expressed from Hansenula polymorpha Rhodococcus ruber
1.1.1.1 333
-
 phenylacetaldehyde pH 6.0, 37°C, recombinant enzyme expressed from Arxula adeninivorans Rhodococcus ruber
1.1.1.1 389
-
 4-chloroacetophenone pH 6.0, 37°C, recombinant enzyme expressed from Arxula adeninivorans Rhodococcus ruber
1.1.1.1 392
-
 acetophenone pH 6.0, 37°C, recombinant enzyme expressed from Saccharomyces cerevisiae Rhodococcus ruber
1.1.1.1 434
-
 4-chloroacetophenone pH 6.0, 37°C, recombinant enzyme expressed from Saccharomyces cerevisiae Rhodococcus ruber
1.1.1.1 589
-
 Valeraldehyde pH 6.0, 37°C, recombinant enzyme expressed from Saccharomyces cerevisiae Rhodococcus ruber
1.1.1.1 720
-
 acetophenone pH 6.0, 37°C, recombinant enzyme expressed from Arxula adeninivorans Rhodococcus ruber
1.1.1.1 800
-
 phenylacetaldehyde pH 6.0, 37°C, recombinant enzyme expressed from Hansenula polymorpha Rhodococcus ruber
1.1.1.1 1036
-
 4-chloroacetophenone pH 6.0, 37°C, recombinant enzyme expressed from Hansenula polymorpha Rhodococcus ruber
1.1.1.1 1055
-
 acetophenone pH 6.0, 37°C, recombinant enzyme expressed from Hansenula polymorpha Rhodococcus ruber
1.1.1.1 1063
-
 phenylacetaldehyde pH 6.0, 37°C, recombinant enzyme expressed from Saccharomyces cerevisiae Rhodococcus ruber
1.1.1.1 1543
-
 Valeraldehyde pH 6.0, 37°C, recombinant enzyme expressed from Arxula adeninivorans Rhodococcus ruber
1.1.1.1 1640
-
 capronaldehyde pH 6.0, 37°C, recombinant enzyme expressed from Hansenula polymorpha Rhodococcus ruber
1.1.1.1 2264
-
 capronaldehyde pH 6.0, 37°C, recombinant enzyme expressed from Arxula adeninivorans Rhodococcus ruber
1.1.1.1 2618
-
 capronaldehyde pH 6.0, 37°C, recombinant enzyme expressed from Saccharomyces cerevisiae Rhodococcus ruber