Any feedback?
Literature summary for 2.4.2.1 extracted from
- Functional and structural characterization of purine nucleoside phosphorylase from Kluyveromyces lactis and its potential applications in reducing purine content in food (2016), PLoS ONE, 11, e0164279 .
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| sequence comparisons, recombinant expression of N- and C-terminally His6-tagged wild-type and mutant enzymes in Escherichia coli | Kluyveromyces lactis |
Crystallization (Commentary)
| Crystallization (Comment) | Organism |
|---|---|
| purified enzyme KlacPNP in complex with hypoxanthine, screening of recombinant N- and C-terminally His6-tagged wild-type enzyme, vapor diffusion sitting drop, mixing of 0.00125 ml of 8 mg/ml of protein in 25 mM Tris-HCl, 100 mM NaCl, pH 8.0, with 0.00125 ml of mother liquor consisting of 0.1 M sodium acetate trihydrate, pH 4.6, and 8% PEG 4000, X-ray diffraction structure determination and analysis at 1.97 A resolution, molecular replacement using the human PNP as a search template (PDB ID 3PHB) | Kluyveromyces lactis |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| N256D | site-directed mutagenesis, mutant KlacPNPN256D accepts both 6-oxopurines and 6-aminopurines as substrates, it shows the highest activity with adenosine | Kluyveromyces lactis |
| N256E | site-directed mutagenesis, the mutant is only active with inosine, no activity with guanosine, xanthosine, and adenosine | Kluyveromyces lactis |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.0103 | - |
adenosine | recombinant mutant N256D, pH 7.0, 25°C | Kluyveromyces lactis |  |
| 0.021 | - |
Inosine | recombinant wild-type enzyme, pH 7.0, 25°C | Kluyveromyces lactis | |
| 0.144 | - |
Inosine | recombinant mutant N256E, pH 7.0, 25°C | Kluyveromyces lactis | |
| 0.222 | - |
Inosine | recombinant mutant N256D, pH 7.0, 25°C | Kluyveromyces lactis | |
| 0.5 | - |
guanosine | recombinant wild-type enzyme, pH 7.0, 25°C | Kluyveromyces lactis | |
| 0.536 | - |
Xanthosine | recombinant mutant N256D, pH 7.0, 25°C | Kluyveromyces lactis | |
| 0.555 | - |
Xanthosine | recombinant wild-type enzyme, pH 7.0, 25°C | Kluyveromyces lactis | |
| 0.842 | - |
guanosine | recombinant mutant N256D, pH 7.0, 25°C | Kluyveromyces lactis | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 105000 | - |
recombinant His-tagged enzyme, gel filtration | Kluyveromyces lactis |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| guanosine + phosphate | Kluyveromyces lactis | - |
guanine + alpha-D-ribose 1-phosphate | - |
r | |
| guanosine + phosphate | Kluyveromyces lactis CBS 2359 | - |
guanine + alpha-D-ribose 1-phosphate | - |
r | |
| guanosine + phosphate | Kluyveromyces lactis NRRL Y-1140 | - |
guanine + alpha-D-ribose 1-phosphate | - |
r | |
| guanosine + phosphate | Kluyveromyces lactis DSM 70799 | - |
guanine + alpha-D-ribose 1-phosphate | - |
r | |
| guanosine + phosphate | Kluyveromyces lactis WM37 | - |
guanine + alpha-D-ribose 1-phosphate | - |
r | |
| guanosine + phosphate | Kluyveromyces lactis ATCC 8585 | - |
guanine + alpha-D-ribose 1-phosphate | - |
r | |
| guanosine + phosphate | Kluyveromyces lactis NBRC 1267 | - |
guanine + alpha-D-ribose 1-phosphate | - |
r | |
| inosine + phosphate | Kluyveromyces lactis | - |
hypoxanthine + alpha-D-ribose 1-phosphate | - |
r | |
| inosine + phosphate | Kluyveromyces lactis CBS 2359 | - |
hypoxanthine + alpha-D-ribose 1-phosphate | - |
r | |
| inosine + phosphate | Kluyveromyces lactis NRRL Y-1140 | - |
hypoxanthine + alpha-D-ribose 1-phosphate | - |
r | |
| inosine + phosphate | Kluyveromyces lactis DSM 70799 | - |
hypoxanthine + alpha-D-ribose 1-phosphate | - |
r | |
| inosine + phosphate | Kluyveromyces lactis WM37 | - |
hypoxanthine + alpha-D-ribose 1-phosphate | - |
r | |
| inosine + phosphate | Kluyveromyces lactis ATCC 8585 | - |
hypoxanthine + alpha-D-ribose 1-phosphate | - |
r | |
| inosine + phosphate | Kluyveromyces lactis NBRC 1267 | - |
hypoxanthine + alpha-D-ribose 1-phosphate | - |
r | |
| purine ribonucleoside + phosphate | Kluyveromyces lactis | - |
purine + alpha-D-ribose 1-phosphate | - |
r | |
| xanthosine + phosphate | Kluyveromyces lactis | - |
xanthine + alpha-D-ribose 1-phosphate | - |
r | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Kluyveromyces lactis | Q6CSZ6 | - |
- |
| Kluyveromyces lactis ATCC 8585 | Q6CSZ6 | - |
- |
| Kluyveromyces lactis CBS 2359 | Q6CSZ6 | - |
- |
| Kluyveromyces lactis DSM 70799 | Q6CSZ6 | - |
- |
| Kluyveromyces lactis NBRC 1267 | Q6CSZ6 | - |
- |
| Kluyveromyces lactis NRRL Y-1140 | Q6CSZ6 | - |
- |
| Kluyveromyces lactis WM37 | Q6CSZ6 | - |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant N- and C-terminally His6-tagged wild-type and mutant enzymes from Escherichia coli by nickel affinity chromatography, dialysis, ultrafiltration, and gel filtration | Kluyveromyces lactis |
Storage Stability
| Storage Stability | Organism |
|---|---|
| purified recombinant His-tagged enzyme at 25 mg/ml is stable for months when stored at 8°C in an elution buffer containing 25 mM Tris-HCl, 100 mM NaCl, 10% glycerol, pH 7.0, up to 65% activity is retained after storage under these conditions for over 30 days | Kluyveromyces lactis |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| adenosine + phosphate | - |
Kluyveromyces lactis | adenine + alpha-D-ribose 1-phosphate | - |
r | |
| adenosine + phosphate | - |
Kluyveromyces lactis CBS 2359 | adenine + alpha-D-ribose 1-phosphate | - |
r | |
| adenosine + phosphate | - |
Kluyveromyces lactis NRRL Y-1140 | adenine + alpha-D-ribose 1-phosphate | - |
r | |
| adenosine + phosphate | - |
Kluyveromyces lactis DSM 70799 | adenine + alpha-D-ribose 1-phosphate | - |
r | |
| adenosine + phosphate | - |
Kluyveromyces lactis WM37 | adenine + alpha-D-ribose 1-phosphate | - |
r | |
| adenosine + phosphate | - |
Kluyveromyces lactis ATCC 8585 | adenine + alpha-D-ribose 1-phosphate | - |
r | |
| adenosine + phosphate | - |
Kluyveromyces lactis NBRC 1267 | adenine + alpha-D-ribose 1-phosphate | - |
r | |
| guanosine + phosphate | - |
Kluyveromyces lactis | guanine + alpha-D-ribose 1-phosphate | - |
r | |
| guanosine + phosphate | - |
Kluyveromyces lactis CBS 2359 | guanine + alpha-D-ribose 1-phosphate | - |
r | |
| guanosine + phosphate | - |
Kluyveromyces lactis NRRL Y-1140 | guanine + alpha-D-ribose 1-phosphate | - |
r | |
| guanosine + phosphate | - |
Kluyveromyces lactis DSM 70799 | guanine + alpha-D-ribose 1-phosphate | - |
r | |
| guanosine + phosphate | - |
Kluyveromyces lactis WM37 | guanine + alpha-D-ribose 1-phosphate | - |
r | |
| guanosine + phosphate | - |
Kluyveromyces lactis ATCC 8585 | guanine + alpha-D-ribose 1-phosphate | - |
r | |
| guanosine + phosphate | - |
Kluyveromyces lactis NBRC 1267 | guanine + alpha-D-ribose 1-phosphate | - |
r | |
| inosine + phosphate | - |
Kluyveromyces lactis | hypoxanthine + alpha-D-ribose 1-phosphate | - |
r | |
| inosine + phosphate | preferred substrate | Kluyveromyces lactis | hypoxanthine + alpha-D-ribose 1-phosphate | - |
r | |
| inosine + phosphate | - |
Kluyveromyces lactis CBS 2359 | hypoxanthine + alpha-D-ribose 1-phosphate | - |
r | |
| inosine + phosphate | preferred substrate | Kluyveromyces lactis CBS 2359 | hypoxanthine + alpha-D-ribose 1-phosphate | - |
r | |
| inosine + phosphate | - |
Kluyveromyces lactis NRRL Y-1140 | hypoxanthine + alpha-D-ribose 1-phosphate | - |
r | |
| inosine + phosphate | preferred substrate | Kluyveromyces lactis NRRL Y-1140 | hypoxanthine + alpha-D-ribose 1-phosphate | - |
r | |
| inosine + phosphate | - |
Kluyveromyces lactis DSM 70799 | hypoxanthine + alpha-D-ribose 1-phosphate | - |
r | |
| inosine + phosphate | preferred substrate | Kluyveromyces lactis DSM 70799 | hypoxanthine + alpha-D-ribose 1-phosphate | - |
r | |
| inosine + phosphate | - |
Kluyveromyces lactis WM37 | hypoxanthine + alpha-D-ribose 1-phosphate | - |
r | |
| inosine + phosphate | preferred substrate | Kluyveromyces lactis WM37 | hypoxanthine + alpha-D-ribose 1-phosphate | - |
r | |
| inosine + phosphate | - |
Kluyveromyces lactis ATCC 8585 | hypoxanthine + alpha-D-ribose 1-phosphate | - |
r | |
| inosine + phosphate | preferred substrate | Kluyveromyces lactis ATCC 8585 | hypoxanthine + alpha-D-ribose 1-phosphate | - |
r | |
| inosine + phosphate | - |
Kluyveromyces lactis NBRC 1267 | hypoxanthine + alpha-D-ribose 1-phosphate | - |
r | |
| inosine + phosphate | preferred substrate | Kluyveromyces lactis NBRC 1267 | hypoxanthine + alpha-D-ribose 1-phosphate | - |
r | |
| additional information | no significant activity towards adenosine. Substrate binding structure analysis, active site structure comparisons, overview | Kluyveromyces lactis | ? | - |
- |
|
| additional information | no significant activity towards adenosine. Substrate binding structure analysis, active site structure comparisons, overview | Kluyveromyces lactis CBS 2359 | ? | - |
- |
|
| additional information | no significant activity towards adenosine. Substrate binding structure analysis, active site structure comparisons, overview | Kluyveromyces lactis NRRL Y-1140 | ? | - |
- |
|
| additional information | no significant activity towards adenosine. Substrate binding structure analysis, active site structure comparisons, overview | Kluyveromyces lactis DSM 70799 | ? | - |
- |
|
| additional information | no significant activity towards adenosine. Substrate binding structure analysis, active site structure comparisons, overview | Kluyveromyces lactis WM37 | ? | - |
- |
|
| additional information | no significant activity towards adenosine. Substrate binding structure analysis, active site structure comparisons, overview | Kluyveromyces lactis ATCC 8585 | ? | - |
- |
|
| additional information | no significant activity towards adenosine. Substrate binding structure analysis, active site structure comparisons, overview | Kluyveromyces lactis NBRC 1267 | ? | - |
- |
|
| purine ribonucleoside + phosphate | - |
Kluyveromyces lactis | purine + alpha-D-ribose 1-phosphate | - |
r | |
| xanthosine + phosphate | - |
Kluyveromyces lactis | xanthine + alpha-D-ribose 1-phosphate | - |
r | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| homotrimer | - |
Kluyveromyces lactis |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| KlacPNP | - |
Kluyveromyces lactis |
| KLLA0_C16621g | locus name | Kluyveromyces lactis |
| PNP | - |
Kluyveromyces lactis |
| purine nucleoside phosphorylase | - |
Kluyveromyces lactis |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 25 | - |
assay at | Kluyveromyces lactis |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 10.4 | - |
guanosine | recombinant mutant N256D, pH 7.0, 25°C | Kluyveromyces lactis | |
| 16.8 | - |
Xanthosine | recombinant wild-type enzyme, pH 7.0, 25°C | Kluyveromyces lactis | |
| 22.8 | - |
adenosine | recombinant mutant N256D, pH 7.0, 25°C | Kluyveromyces lactis | |
| 23.3 | - |
Xanthosine | recombinant mutant N256D, pH 7.0, 25°C | Kluyveromyces lactis | |
| 25.1 | - |
Inosine | recombinant mutant N256E, pH 7.0, 25°C | Kluyveromyces lactis | |
| 29.7 | - |
guanosine | recombinant wild-type enzyme, pH 7.0, 25°C | Kluyveromyces lactis | |
| 34.9 | - |
Inosine | recombinant mutant N256D, pH 7.0, 25°C | Kluyveromyces lactis | |
| 43.9 | - |
Inosine | recombinant wild-type enzyme, pH 7.0, 25°C | Kluyveromyces lactis | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7 | - |
assay at | Kluyveromyces lactis |
General Information
| General Information | Comment | Organism |
|---|---|---|
| metabolism | purine nucleoside phosphorylase (PNP) from Kluyveromyces lactis (KlacPNP) is a key enzyme involved in the purine degradation pathway, overview | Kluyveromyces lactis |
| additional information | KlacPNP and KlacPNPN256D reduce purine content in a beer sample | Kluyveromyces lactis |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 12.4 | - |
guanosine | recombinant mutant N256D, pH 7.0, 25°C | Kluyveromyces lactis | |
| 30.3 | - |
Xanthosine | recombinant wild-type enzyme, pH 7.0, 25°C | Kluyveromyces lactis | |
| 43.5 | - |
Xanthosine | recombinant mutant N256D, pH 7.0, 25°C | Kluyveromyces lactis | |
| 59.4 | - |
guanosine | recombinant wild-type enzyme, pH 7.0, 25°C | Kluyveromyces lactis | |
| 157.2 | - |
Inosine | recombinant mutant N256D, pH 7.0, 25°C | Kluyveromyces lactis | |
| 174.3 | - |
Inosine | recombinant mutant N256E, pH 7.0, 25°C | Kluyveromyces lactis | |
| 2090.5 | - |
Inosine | recombinant wild-type enzyme, pH 7.0, 25°C | Kluyveromyces lactis | |
| 2213.6 | - |
adenosine | recombinant mutant N256D, pH 7.0, 25°C | Kluyveromyces lactis | |
Use of this online version of BRENDA is free under the CC BY 4.0 license. See terms of use for full details.
Release 2023.1 (January 2023)
Legal
Curated at
Member of
