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Literature summary for 1.1.1.B65 extracted from

  • Wang, J.; Feng, J.; Li, W.; Yang, C.; Chen, X.; Bao, B.; Yang, J.; Wang, P.; Li, D.; Shi, R.
    Characterization of a novel (R)-mandelate dehydrogenase from Pseudomonas putida NUST506 (2015), J. Mol. Catal. B, 120, 23-27 .
No PubMed abstract available

Activating Compound

Activating Compound Comment Organism Structure
DTT enhancement of enzyme activity of 26% and 33% in the concentration of 10 mM and 50 mM Pseudomonas putida
Triton X-100 0.25% of TritonX-100 has slight improvement on enzyme activity, whereas 0.75% of TritonX-100 slightly inhibits the enzyme activity Pseudomonas putida

Inhibitors

Inhibitors Comment Organism Structure
Ca2+ inhibits 4% aT 1 mM and 33% at 10 mM Pseudomonas putida
EDTA inhibits the enzyme activity about 68% and 89% in the concentration of 10 mM and 50 mM,respectively Pseudomonas putida
Hg2+ strong inhibition at 1 mM Pseudomonas putida
Li+ inhibits 17% at 10 mM Pseudomonas putida
Mg2+ inhibits 19% at 1 mM and 31% at 10 mM Pseudomonas putida
Mn2+ 21% activation at 1 mM, and 61% inhibition at 10 mM Pseudomonas putida
Ni2+ inhibits 39% at 1 mM and 63% at 10 mM Pseudomonas putida
SDS strongly inhibits the enzyme activity at 0.25-0.75% Pseudomonas putida
Triton X-100 0.25% of TritonX-100 has slight improvement on enzyme activity, whereas 0.75% of TritonX-100 slightly inhibits the enzyme activity Pseudomonas putida
Zn2+ strong inhibition at 10 mM Pseudomonas putida

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.015
-
NADP+ pH 8.5, 30°C Pseudomonas putida
0.018
-
NAD+ pH 8.5, 30°C Pseudomonas putida
0.02
-
(R)-mandelate pH 8.5, 30°C Pseudomonas putida

Metals/Ions

Metals/Ions Comment Organism Structure
K+ activates 44% at 1 mM and 69% at 10 mM Pseudomonas putida
Mn2+ 21% activation at 1 mM, and 61% inhibition at 10 mM Pseudomonas putida
Na+ activates 12% at 10 mM Pseudomonas putida

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
(R)-mandelate + NAD+ Pseudomonas putida
-
phenylglyoxylate + NADH + H+
-
r
(R)-mandelate + NAD+ Pseudomonas putida NUST506
-
phenylglyoxylate + NADH + H+
-
r

Organism

Organism UniProt Comment Textmining
Pseudomonas putida
-
isolated from soil around the schoolyard of Nanjing University of Science and Technology (Nanjing, China)
-
Pseudomonas putida NUST506
-
isolated from soil around the schoolyard of Nanjing University of Science and Technology (Nanjing, China)
-

Purification (Commentary)

Purification (Comment) Organism
native enzyme 11fold by ammonium sulfate fractionation and hydrophobic interaction chromatography Pseudomonas putida

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.03
-
crude enzyme extract, pH 8.5, 30°C Pseudomonas putida
0.33
-
purified native enzyme, pH 8.5, 30°C Pseudomonas putida

Storage Stability

Storage Stability Organism
4°C, purified enzyme, 96 h, 78% of enzyme activity is retained at pH 6.0, and 30% at pH 8.5 Pseudomonas putida

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(R)-mandelate + NAD+
-
Pseudomonas putida phenylglyoxylate + NADH + H+
-
r
(R)-mandelate + NAD+
-
Pseudomonas putida NUST506 phenylglyoxylate + NADH + H+
-
r
(R)-mandelate + NADP+
-
Pseudomonas putida phenylglyoxylate + NADPH + H+
-
r
(R)-mandelate + NADP+
-
Pseudomonas putida NUST506 phenylglyoxylate + NADPH + H+
-
r

Subunits

Subunits Comment Organism
? x * 61000, SDS-PAGE Pseudomonas putida

Synonyms

Synonyms Comment Organism
RMDH
-
Pseudomonas putida

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
-
Pseudomonas putida

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
20 55 activity range, profile overview Pseudomonas putida

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
30
-
purified enzyme, pH 8.5, 31% activity remaining after 0.5 h, and 20% after 1 h Pseudomonas putida

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.3
-
NADP+ pH 8.5, 30°C Pseudomonas putida
0.9
-
NAD+ pH 8.5, 30°C Pseudomonas putida
0.9
-
(R)-mandelate pH 8.5, 30°C Pseudomonas putida

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.5
-
-
Pseudomonas putida

pH Range

pH Minimum pH Maximum Comment Organism
5 10 activity range, profile overview Pseudomonas putida

pH Stability

pH Stability pH Stability Maximum Comment Organism
6 9 purified enzyme, the stability drops significantly at pH 8.5 and pH 9.0, the enzyme loses 85% and 94% of the activity when incubated for 3.5 h at 30°C, and it loses 11% of activity at pH 6.0 after 3.5 h at 30°C. After 96 h at 4°C, 78% of enzyme activity is retained at pH 6.0, and 30% at pH 8.5 Pseudomonas putida

Cofactor

Cofactor Comment Organism Structure
NAD+
-
Pseudomonas putida
NADH
-
Pseudomonas putida
NADP+
-
Pseudomonas putida
NADPH
-
Pseudomonas putida