Literature summary for 1.1.1.1 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.

Application

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(Commentary)

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

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 [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
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 [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

Organism	UniProt	Comment	Textmining
Rhodococcus ruber	-	-	-
Rhodococcus ruber	-	DSM44514	-
Rhodococcus ruber 219	-	DSM44514	-
Rhodococcus ruber DSM 44514	-	-	-

Purification (Commentary)

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

Source Tissue

Source Tissue	Comment	Organism	Textmining
lyophilized cell	-	Rhodococcus ruber	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
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

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

Synonyms

Synonyms	Comment	Organism
ADH6Hp	-	Rhodococcus ruber

Temperature Optimum [°C]

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 [°C]

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 [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
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

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

pH Stability

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

Cofactor	Comment	Organism	Structure
NAD+	-	Rhodococcus ruber
NADH	-	Rhodococcus ruber

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism
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