Application | Comment | Organism |
---|---|---|
synthesis | enzyme HSPHZZ can be used for the enzymatic production of 2,5-dihydroxypyridine in biotechnology applications. 85.3 mg/l 2,5-dihydroxypyridine is produced in 40 min with a conversion of 74.9% at 30°C, pH 8.5, 1.0 mM substrate concentration, and 0.001 mM enzyme concentration | Pseudomonas sp. ZZ-5 |
Cloned (Comment) | Organism |
---|---|
recombinant expression of His-tagged enzyme in Escherichia coli strain B21-Codon Plus(DE3)-RIL | Pseudomonas sp. ZZ-5 |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
acetone | inhibits at 20% v/v | Pseudomonas sp. ZZ-5 | |
chloroform | inhibits at 20% v/v | Pseudomonas sp. ZZ-5 | |
Co2+ | inhibits at 5 mM | Pseudomonas sp. ZZ-5 | |
Cu2+ | inhibits at 5 mM | Pseudomonas sp. ZZ-5 | |
dimethylformamide | inhibits at 20% v/v | Pseudomonas sp. ZZ-5 | |
DMSO | inhibits slightly by 10% at 20% v/v | Pseudomonas sp. ZZ-5 | |
EDTA | inhibits 15-20% at 0.5-5 mM | Pseudomonas sp. ZZ-5 | |
Fe2+ | inhibits at 5 mM | Pseudomonas sp. ZZ-5 | |
formaldehyde | inhibits at 20% v/v | Pseudomonas sp. ZZ-5 | |
methanol | inhibits at 20% v/v | Pseudomonas sp. ZZ-5 | |
Mg2+ | slight inhibition | Pseudomonas sp. ZZ-5 | |
Mn2+ | slight inhibition | Pseudomonas sp. ZZ-5 | |
additional information | no inhibition by Twen-20, Tween-40, and Tween-80 at 1% w/v | Pseudomonas sp. ZZ-5 | |
Span-20 | at 1% w/v by over 50% | Pseudomonas sp. ZZ-5 | |
SPAN-40 | at 1% w/v by over 50% | Pseudomonas sp. ZZ-5 | |
Toluene | inhibits at 20% v/v | Pseudomonas sp. ZZ-5 | |
Triton X-100 | at 1% w/v by over 50% | Pseudomonas sp. ZZ-5 | |
Zn2+ | inhibits at 5 mM | Pseudomonas sp. ZZ-5 |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.18 | - |
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate | pH 8.5, 30°C | Pseudomonas sp. ZZ-5 | |
0.23 | - |
NADH | pH 8.5, 30°C | Pseudomonas sp. ZZ-5 |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
additional information | the enzyme is not significantly affected by Ni2+, Ca2+, K+, and Na+ at 5 mM. The enzyme is no metalloenzyme | Pseudomonas sp. ZZ-5 |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate + 2 NADH + 2 H+ + O2 | Pseudomonas sp. ZZ-5 | - |
2,5-dihydroxypyridine + succinate semialdehyde + 2 NAD+ + H2O | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas sp. ZZ-5 | - |
isolated from soil samples obtained from a field under continuous tobacco cropping in Henan, P.R. China | - |
Purification (Comment) | Organism |
---|---|
native enzyme 17.6fold to homogeneity by ammonium sulfate fractionation, anion exchange and hydrophobic interaction chromatography, and gel filtration, recombinant His-tagged enzyme 6.2fold from Escherichia coli strain B21-Codon Plus(DE3)-RIL by nickel affinity chromatography and gel filtration | Pseudomonas sp. ZZ-5 |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
5.1 | - |
purified native enzyme, pH 8.5, 30°C | Pseudomonas sp. ZZ-5 |
31.8 | - |
purified recombinant His-tagged enzyme, pH 8.5, 30°C | Pseudomonas sp. ZZ-5 |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate + 2 NADH + 2 H+ + O2 | - |
Pseudomonas sp. ZZ-5 | 2,5-dihydroxypyridine + succinate semialdehyde + 2 NAD+ + H2O | - |
? | |
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate + 2 NADH + 2 H+ + O2 | the enzyme efficiently catalyzes the conversion of 6-hydroxy-3-succinoylpyridine (HSP) into 2,5-dihydroxypyridine (2,5-DHP) and succinic acid in the presence of NADH and FAD | Pseudomonas sp. ZZ-5 | 2,5-dihydroxypyridine + succinate semialdehyde + 2 NAD+ + H2O | - |
? | |
additional information | assay method optimization, overview. Product identification by LC-MS | Pseudomonas sp. ZZ-5 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 45000, SDS-PAGE, x * 44300 , about, sequence calculation | Pseudomonas sp. ZZ-5 |
Synonyms | Comment | Organism |
---|---|---|
HSP hydroxylase | - |
Pseudomonas sp. ZZ-5 |
HSPHZZ | - |
Pseudomonas sp. ZZ-5 |
nicotine hydroxylase | - |
Pseudomonas sp. ZZ-5 |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
- |
Pseudomonas sp. ZZ-5 |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
5 | 45 | activity range, profile overview | Pseudomonas sp. ZZ-5 |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
purified enzyme, pH 8.5, 120 min, only slight loss of activity | Pseudomonas sp. ZZ-5 |
40 | - |
purified enzyme, pH 8.5, 30 min, loss of 60% activity | Pseudomonas sp. ZZ-5 |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.3 | - |
NADH | pH 8.5, 30°C | Pseudomonas sp. ZZ-5 | |
2.1 | - |
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate | pH 8.5, 30°C | Pseudomonas sp. ZZ-5 |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.5 | - |
- |
Pseudomonas sp. ZZ-5 |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
6 | 9.7 | activity range, profile overview | Pseudomonas sp. ZZ-5 |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
7 | 9 | purified enzyme, 30°C, 30 min, over 50% activity remaining within this range | Pseudomonas sp. ZZ-5 |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | - |
Pseudomonas sp. ZZ-5 | |
NADH | - |
Pseudomonas sp. ZZ-5 |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
5.7 | - |
NADH | pH 8.5, 30°C | Pseudomonas sp. ZZ-5 | |
11.7 | - |
4-(6-hydroxypyridin-3-yl)-4-oxobutanoate | pH 8.5, 30°C | Pseudomonas sp. ZZ-5 |