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Literature summary extracted from

  • Raj, S.; Rathnasingh, C.; Jung, W.; Selvakumar, E.; Park, S.
    A novel NAD+-dependent aldehyde dehydrogenase encoded by the puuC gene of Klebsiella pneumoniae DSM 2026 that utilizes 3-hydroxypropionaldehyde as a substrate (2010), Biotechnol. Bioprocess Eng., 15, 131-138.
No PubMed abstract available

Activating Compound

EC Number Activating Compound Comment Organism Structure
1.2.1.3 beta-mercaptoethanol 161.09% activity at 1 mM Klebsiella pneumoniae
1.2.1.3 dithiothreitol 147.48% activity at 1 mM Klebsiella pneumoniae
1.2.1.3 Semicarbazide hydrochloride 118.98% activity at 1 mM Klebsiella pneumoniae
1.2.1.99 2-mercaptoethanol 1 mM, activity increases by 61% Klebsiella pneumoniae
1.2.1.99 dithiothreitol 1 mM, activity increases by 47.5% Klebsiella pneumoniae
1.2.1.99 Semicarbazide hydrochloride 1 mM, activity increases by 19% Klebsiella pneumoniae

Application

EC Number Application Comment Organism
1.2.1.99 synthesis PuuC is an active enzyme that can be utilized effectively for several purposes, including biological production of 3-hydroxypropionate Klebsiella pneumoniae

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.2.1.3 expressed in Escherichia coli BL21(DE3) cells (strain SH-KAD2) Klebsiella pneumoniae
1.2.1.99 expressed in Escherichia coli Klebsiella pneumoniae

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.2.1.3 Ag+ complete inhibition at 1 mM Klebsiella pneumoniae
1.2.1.3 Co2+ 89.01% residual activity at 1 mM Klebsiella pneumoniae
1.2.1.3 Cu2+ complete inhibition at 1 mM Klebsiella pneumoniae
1.2.1.3 Fe2+ 26.34% residual activity at 1 mM Klebsiella pneumoniae
1.2.1.3 Hg2+ complete inhibition at 1 mM Klebsiella pneumoniae
1.2.1.3 Li+ 90.64% residual activity at 1 mM Klebsiella pneumoniae
1.2.1.3 Mg2+ 93.64% residual activity at 1 mM Klebsiella pneumoniae
1.2.1.3 Na+ 89.17% residual activity at 1 mM Klebsiella pneumoniae
1.2.1.3 NH3 92.06% residual activity at 1 mM Klebsiella pneumoniae
1.2.1.3 Ni2+ 89.54% residual activity at 1 mM Klebsiella pneumoniae
1.2.1.3 Zn2+ 35.91% residual activity at 1 mM Klebsiella pneumoniae
1.2.1.99 Ag+ 1 mM, complete inhibition Klebsiella pneumoniae
1.2.1.99 Co2+ 1 mM, 11% inhibition Klebsiella pneumoniae
1.2.1.99 Cu2+ 1 mM, complete inhibition Klebsiella pneumoniae
1.2.1.99 EDTA 1 mM, 8.3% inhibition Klebsiella pneumoniae
1.2.1.99 Fe2+ 1 mM, 73.6% inhibition Klebsiella pneumoniae
1.2.1.99 Hg2+ 1 mM, complete inhibition Klebsiella pneumoniae
1.2.1.99 Li+ 1 mM, 9.4% inhibition Klebsiella pneumoniae
1.2.1.99 Mg2+ 1 mM, 6.3% inhibition Klebsiella pneumoniae
1.2.1.99 NaCl mM, 11% inhibition Klebsiella pneumoniae
1.2.1.99 NH4Cl 8% inhibition Klebsiella pneumoniae
1.2.1.99 Ni2+ 1 mM, 10.5% inhibition Klebsiella pneumoniae
1.2.1.99 Zn2+ 1 mM, 64% inhibition Klebsiella pneumoniae

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.2.1.3 0.09
-
 NAD+ recombinant enzyme, in 50 mM potassium phosphate buffer, at pH 8.0 and 45°C Klebsiella pneumoniae
1.2.1.3 0.14
-
 Valeraldehyde recombinant enzyme, in 50 mM potassium phosphate buffer, at pH 8.0 and 45°C Klebsiella pneumoniae
1.2.1.3 0.21
-
 Isovaleraldehyde recombinant enzyme, in 50 mM potassium phosphate buffer, at pH 8.0 and 45°C Klebsiella pneumoniae
1.2.1.3 0.26
-
 Butyraldehyde recombinant enzyme, in 50 mM potassium phosphate buffer, at pH 8.0 and 45°C Klebsiella pneumoniae
1.2.1.3 0.48
-
 3-hydroxypropionaldehyde recombinant enzyme, in 50 mM potassium phosphate buffer, at pH 8.0 and 45°C Klebsiella pneumoniae
1.2.1.99 0.14
-
 Valeraldehyde pH 8.0, 45°C Klebsiella pneumoniae
1.2.1.99 0.21
-
 Isovaleraldehyde pH 8.0, 45°C Klebsiella pneumoniae
1.2.1.99 0.26
-
 Butyraldehyde pH 8.0, 45°C Klebsiella pneumoniae
1.2.1.99 0.48
-
 3-hydroxypropionaldehyde pH 8.0, 45°C Klebsiella pneumoniae

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.2.1.3 53000
-
 recombinant enzyme, SDS-PAGE Klebsiella pneumoniae
1.2.1.99 53800
-
 x * 53800, SDS-PAGE Klebsiella pneumoniae

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.2.1.99 3-hydroxypropionaldehyde + NAD+ + H2O Klebsiella pneumoniae the enzyme might play a role in putresine and propionate metabolism 3-hydroxypropanoate + NADH + 2 H+
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.2.1.3 Klebsiella pneumoniae
-
-
-
1.2.1.3 Klebsiella pneumoniae DSM 2026
-
-
-
1.2.1.99 Klebsiella pneumoniae
-
-
-
1.2.1.99 Klebsiella pneumoniae DSM 2026
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.2.1.3 Ni-NTA column chromatography Klebsiella pneumoniae
1.2.1.99
-
 Klebsiella pneumoniae

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.2.1.3 3.13
-
 crude extract, using 3-hydroxypropionaldehyde as substrate, in 50 mM, potassium phosphate buffer, at pH 8.0 and 45°C Klebsiella pneumoniae
1.2.1.3 27.73
-
 after 8.86fold purification, using 3-hydroxypropionaldehyde as substrate, in 50 mM, potassium phosphate buffer, at pH 8.0 and 45°C Klebsiella pneumoniae
1.2.1.99 22.25
-
 Vmax: 3-hydroxypropionaldehyde, pH 8.0, 45°C Klebsiella pneumoniae
1.2.1.99 28.61
-
 Vmax: isovaleraldehyde, pH 8.0, 45°C Klebsiella pneumoniae
1.2.1.99 33.26
-
 Vmax: valeraldehyde, pH 8.0, 45°C Klebsiella pneumoniae
1.2.1.99 34.08
-
 Vmax: butyraldehyde, pH 8.0, 45°C Klebsiella pneumoniae

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.2.1.3 2-furaldehyde + NAD+ + H2O the activity on 2-furaldehyde is 22% of that on 3-hydroxypropionaldehyde Klebsiella pneumoniae 2-furanoic acid + NADH + H+
-
 ?
1.2.1.3 3-hydroxypropionaldehyde + NAD+ + H2O the activity on 3-hydroxypropionaldehyde is about 70% of that on valeraldehyde Klebsiella pneumoniae 3-hydroxypropanoate + NADH + 2 H+
-
 ?
1.2.1.3 acetaldehyde + NAD+ + H2O the activity on acetaldehyde is about 50% of that on valeraldehyde Klebsiella pneumoniae acetate + NADH + H+
-
 ?
1.2.1.3 acetaldehyde + NAD+ + H2O the activity on acetaldehyde is about 50% of that on valeraldehyde Klebsiella pneumoniae DSM 2026 acetate + NADH + H+
-
 ?
1.2.1.3 benzaldehyde + NAD+ + H2O the activity on benzaldehyde is 35% of that on 3-hydroxypropionaldehyde Klebsiella pneumoniae benzoate + NADH + H+
-
 ?
1.2.1.3 benzaldehyde + NAD+ + H2O the activity on benzaldehyde is 35% of that on 3-hydroxypropionaldehyde Klebsiella pneumoniae DSM 2026 benzoate + NADH + H+
-
 ?
1.2.1.3 butyraldehyde + NAD+ + H2O the activity on butyraldehyde is about 95% of that on valeraldehyde Klebsiella pneumoniae butanoate + NADH + H+
-
 ?
1.2.1.3 isovaleraldehyde + NAD+ + H2O the activity on isovaleraldehyde is about 80% of that on valeraldehyde Klebsiella pneumoniae isovalerate + NADH + H+
-
 ?
1.2.1.3 additional information no appreciable activity is observed on formaldehyde Klebsiella pneumoniae ?
-
 ?
1.2.1.3 additional information no appreciable activity is observed on formaldehyde Klebsiella pneumoniae DSM 2026 ?
-
 ?
1.2.1.3 propionaldehyde + NAD+ + H2O the activity on propionaldehyde is about 55% of that on valeraldehyde Klebsiella pneumoniae propionate + NADH + H+
-
 ?
1.2.1.3 propionaldehyde + NAD+ + H2O the activity on propionaldehyde is about 55% of that on valeraldehyde Klebsiella pneumoniae DSM 2026 propionate + NADH + H+
-
 ?
1.2.1.3 valeraldehyde + NAD+ + H2O most preferred substrate, 100% activity Klebsiella pneumoniae valerate + NADH + H+
-
 ?
1.2.1.3 valeraldehyde + NAD+ + H2O most preferred substrate, 100% activity Klebsiella pneumoniae DSM 2026 valerate + NADH + H+
-
 ?
1.2.1.99 2-furaldehyde + NAD+ + H2O 22% of the activity with 3-hydroxypropionaldehyde Klebsiella pneumoniae furan-2-carboxylate + NADH + 2 H+
-
 ?
1.2.1.99 3-hydroxypropionaldehyde + NAD+ + H2O
-
 Klebsiella pneumoniae 3-hydroxypropanoate + NADH + 2 H+
-
 ?
1.2.1.99 3-hydroxypropionaldehyde + NAD+ + H2O the enzyme might play a role in putresine and propionate metabolism Klebsiella pneumoniae 3-hydroxypropanoate + NADH + 2 H+
-
 ?
1.2.1.99 acetaldehyde + NAD+ + H2O
-
 Klebsiella pneumoniae acetate + NADH + H+
-
 ?
1.2.1.99 benzaldehyde + NAD+ + H2O 35% of the activity with 3-hydroxypropionaldehyde Klebsiella pneumoniae benzoate + NADH + H+
-
 ?
1.2.1.99 butyraldehyde + NAD+ + H2O
-
 Klebsiella pneumoniae butyrate + NADH + 2 H+
-
 ?
1.2.1.99 isovaleraldehyde + NAD+ + H2O
-
 Klebsiella pneumoniae 3-methylbutanoate + NADH + 2 H+
-
 ?
1.2.1.99 additional information no activity with formaldehyde Klebsiella pneumoniae ?
-
 ?
1.2.1.99 propionaldehyde + NAD+ + H2O
-
 Klebsiella pneumoniae propionate + NADH + 2 H+
-
 ?
1.2.1.99 valeraldehyde + NAD+ + H2O most preferred substrate Klebsiella pneumoniae pentanoate + NADH + H+
-
 ?

Subunits

EC Number Subunits Comment Organism
1.2.1.99 ? x * 53800, SDS-PAGE Klebsiella pneumoniae

Synonyms

EC Number Synonyms Comment Organism
1.2.1.3 ALDH
-
 Klebsiella pneumoniae
1.2.1.3 NAD+-dependent aldehyde dehydrogenase
-
 Klebsiella pneumoniae
1.2.1.3 NAD+-specific ALDH
-
 Klebsiella pneumoniae
1.2.1.3 PuuC
-
 Klebsiella pneumoniae
1.2.1.99 PuuC
-
 Klebsiella pneumoniae

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.2.1.3 45
-
-
 Klebsiella pneumoniae
1.2.1.99 45
-
-
 Klebsiella pneumoniae

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
1.2.1.99 20 60 20°C: about 30% of maximal activity, 60°C: about 35% of maximal activity Klebsiella pneumoniae

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.2.1.3 30 60 the enzyme is stable between 30°C and 50°C, but becomes unstable at 60°C Klebsiella pneumoniae
1.2.1.99 60
-
 becomes unstable at 60°C Klebsiella pneumoniae

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.2.1.99 19.81
-
 3-hydroxypropionaldehyde pH 8.0, 45°C Klebsiella pneumoniae
1.2.1.99 25.47
-
 Isovaleraldehyde pH 8.0, 45°C Klebsiella pneumoniae
1.2.1.99 29.61
-
 Valeraldehyde pH 8.0, 45°C Klebsiella pneumoniae
1.2.1.99 30.34
-
 Butyraldehyde pH 8.0, 45°C Klebsiella pneumoniae

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.2.1.3 8
-
-
 Klebsiella pneumoniae
1.2.1.99 8
-
-
 Klebsiella pneumoniae

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.2.1.3 7 9
-
 Klebsiella pneumoniae
1.2.1.99 7 9 pH 7.0: about 50% of maximal activity, pH 9.0: about 80% of maximal activity Klebsiella pneumoniae

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
1.2.1.3 6
-
 the enzyme activity is completely abolished at pH 6.0 Klebsiella pneumoniae

Cofactor

EC Number Cofactor Comment Organism Structure
1.2.1.3 NAD+ NAD+ is the preferred coenzyme over NADP+ Klebsiella pneumoniae
1.2.1.99 NAD+ preferred coenzyme over NADP+ Klebsiella pneumoniae

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.2.1.3 41.44
-
 3-hydroxypropionaldehyde recombinant enzyme, in 50 mM potassium phosphate buffer, at pH 8.0 and 45°C Klebsiella pneumoniae
1.2.1.3 117.2
-
 Butyraldehyde recombinant enzyme, in 50 mM potassium phosphate buffer, at pH 8.0 and 45°C Klebsiella pneumoniae
1.2.1.3 121.9
-
 Isovaleraldehyde recombinant enzyme, in 50 mM potassium phosphate buffer, at pH 8.0 and 45°C Klebsiella pneumoniae
1.2.1.3 207.1
-
 Valeraldehyde recombinant enzyme, in 50 mM potassium phosphate buffer, at pH 8.0 and 45°C Klebsiella pneumoniae
1.2.1.3 254.7
-
 NAD+ recombinant enzyme, in 50 mM potassium phosphate buffer, at pH 8.0 and 45°C Klebsiella pneumoniae
1.2.1.99 41.44
-
 3-hydroxypropionaldehyde pH 8.0, 45°C Klebsiella pneumoniae
1.2.1.99 117.2
-
 Butyraldehyde pH 8.0, 45°C Klebsiella pneumoniae
1.2.1.99 121.9
-
 Isovaleraldehyde pH 8.0, 45°C Klebsiella pneumoniae
1.2.1.99 207.1
-
 Valeraldehyde pH 8.0, 45°C Klebsiella pneumoniae