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Literature summary for 2.4.2.1 extracted from
- Efficient synthesis of purine nucleoside analogs by a new trimeric purine nucleoside phosphorylase from Aneurinibacillus migulanus AM007 (2019), Molecules, 25, 100 .
Application
| Application | Comment | Organism |
|---|---|---|
| synthesis | enzyme AmPNP is an excellent biocatalyst for the industrial production of purine nucleoside analogues, biosynthesis of purine nucleoside analogues, overview | Aneurinibacillus migulanus |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic analysis, recombinant expression of His6-tagged enzyme in Escherichia coli strain BL21(DE3) | Aneurinibacillus migulanus |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| additional information | a one-pot, two-enzyme mode of transglycosylation reaction is successfully constructed by combining pyrimidine nucleoside phosphorylase (BbPyNP, EC 2.4.2.2) derived from Brevibacillus borstelensis strain LK01 and Aneurinibacillus migulanus strain AM007 AmPNP for the production of purine nucleoside analogues. Conversions of 2,6-diaminopurine ribonucleoside, 2-amino-6-chloropurine ribonucleoside, and 6-thioguanine ribonucleoside synthesized still reaches over 90% on the higher concentrations of substrates (pentofuranosyl donor: purine base; 20:10 mM) with a low enzyme ratio of BbPyNP:AmPNP (2:20 microg/ml). Thus, the trimeric AmPNP is a promising biocatalyst for industrial production of purine nucleoside analogues. Method optimization and evaluation, overview | Aneurinibacillus migulanus |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 1.16 | - |
guanosine | pH 8.0, 50°C, recombinant enzyme | Aneurinibacillus migulanus |  |
| 1.45 | - |
adenosine | pH 8.0, 50°C, recombinant enzyme | Aneurinibacillus migulanus | |
| 5.3 | - |
Inosine | pH 8.0, 50°C, recombinant enzyme | Aneurinibacillus migulanus | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 87000 | - |
recombinant His6-tagged enzyme, gel filtration | Aneurinibacillus migulanus |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| guanosine + phosphate | Aneurinibacillus migulanus | - |
guanine + alpha-D-ribose 1-phosphate | - |
r | |
| guanosine + phosphate | Aneurinibacillus migulanus AM007 | - |
guanine + alpha-D-ribose 1-phosphate | - |
r | |
| inosine + phosphate | Aneurinibacillus migulanus | - |
hypoxanthine + alpha-D-ribose 1-phosphate | - |
r | |
| inosine + phosphate | Aneurinibacillus migulanus AM007 | - |
hypoxanthine + alpha-D-ribose 1-phosphate | - |
r | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Aneurinibacillus migulanus | A0A3G5E8S8 | - |
- |
| Aneurinibacillus migulanus AM007 | A0A3G5E8S8 | - |
- |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 2'-deoxyguanosine + phosphate | moderate activity | Aneurinibacillus migulanus | guanine + 2-deoxy-alpha-D-ribose 1-phosphate | - |
r | |
| 2,6-diaminopurine + alpha-D-ribose 1-phosphate | - |
Aneurinibacillus migulanus | 2,6-diaminopurine ribonucleoside + phosphate | - |
r | |
| 2-amino-6-chloropurine + alpha-D-ribose 1-phosphate | - |
Aneurinibacillus migulanus | 2-amino-6-chloropurine ribonucleoside + phosphate | - |
r | |
| 6-thioguanine + alpha-D-ribose 1-phosphate | - |
Aneurinibacillus migulanus | 6-thioguanine ribonucleoside + phosphate | - |
r | |
| adenosine + phosphate | low activity | Aneurinibacillus migulanus | adenine + alpha-D-ribose 1-phosphate | - |
r | |
| guanosine + phosphate | - |
Aneurinibacillus migulanus | guanine + alpha-D-ribose 1-phosphate | - |
r | |
| guanosine + phosphate | moderate activity | Aneurinibacillus migulanus | guanine + alpha-D-ribose 1-phosphate | - |
r | |
| guanosine + phosphate | - |
Aneurinibacillus migulanus AM007 | guanine + alpha-D-ribose 1-phosphate | - |
r | |
| guanosine + phosphate | moderate activity | Aneurinibacillus migulanus AM007 | guanine + alpha-D-ribose 1-phosphate | - |
r | |
| inosine + phosphate | - |
Aneurinibacillus migulanus | hypoxanthine + alpha-D-ribose 1-phosphate | - |
r | |
| inosine + phosphate | best substrate | Aneurinibacillus migulanus | hypoxanthine + alpha-D-ribose 1-phosphate | - |
r | |
| inosine + phosphate | - |
Aneurinibacillus migulanus AM007 | hypoxanthine + alpha-D-ribose 1-phosphate | - |
r | |
| inosine + phosphate | best substrate | Aneurinibacillus migulanus AM007 | hypoxanthine + alpha-D-ribose 1-phosphate | - |
r | |
| additional information | substrate specificity, detailed overview, no activity with 2'-deoxyadenosine | Aneurinibacillus migulanus | ? | - |
- |
|
| additional information | substrate specificity, detailed overview, no activity with 2'-deoxyadenosine | Aneurinibacillus migulanus AM007 | ? | - |
- |
|
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| homotrimer | 3 * 29000, about, recombinant His6-tagged enzyme, SDS-PAGE | Aneurinibacillus migulanus |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| AmPNP | - |
Aneurinibacillus migulanus |
| inosine-guanosine phosphorylase | UniProt | Aneurinibacillus migulanus |
| PNP | - |
Aneurinibacillus migulanus |
| purine nucleoside phosphorylase | - |
Aneurinibacillus migulanus |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 70 | - |
recombinant enzyme | Aneurinibacillus migulanus |
Temperature Range [°C]
| Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 30 | 80 | activity range, profile overview, recombinant enzyme | Aneurinibacillus migulanus |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 55 | - |
and below, purified recombinant enzyme, 12 h, completely stable, half-life is 78.6 h | Aneurinibacillus migulanus |
| 60 | - |
purified recombinant enzyme, 2 h, pH 7.0-9.0, completely stable, half-life is 37.3 h | Aneurinibacillus migulanus |
| 65 | - |
purified recombinant enzyme, half-life is 6.9 h | Aneurinibacillus migulanus |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 14.3 | - |
adenosine | pH 8.0, 50°C, recombinant enzyme | Aneurinibacillus migulanus | |
| 18400 | - |
guanosine | pH 8.0, 50°C, recombinant enzyme | Aneurinibacillus migulanus | |
| 63800 | - |
Inosine | pH 8.0, 50°C, recombinant enzyme | Aneurinibacillus migulanus | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.5 | - |
recombinant enzyme | Aneurinibacillus migulanus |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 6 | 8 | over 80% of maximal activity within this range, recombinant enzyme | Aneurinibacillus migulanus |
pH Stability
| pH Stability | pH Stability Maximum | Comment | Organism |
|---|---|---|---|
| 7 | 9 | purified recombinant enzyme, 2 h, 60°C, completely stable | Aneurinibacillus migulanus |
General Information
| General Information | Comment | Organism |
|---|---|---|
| evolution | most PNPs are divided into trimeric PNPs and hexametric PNPs based on molecular mass, protein structure, or substrate specificity | Aneurinibacillus migulanus |
| metabolism | nucleoside phosphorylases (NPs) catalyze the reversible cleavage of the glycosidic bond of (deoxy)ribonucleosides in the presence of phosphate to generate a nucleobase and alpha-D-(deoxy)ribose-1-phosphate. NPs include pyrimidine NPs (PyNPs, EC 2.4.2.2) and purine NPs (PNPs, EC 2.4.2.1). PyNPs show substrate specificity for the phosphorolysis of pyrimidine nucleosides, while PNPs are specific for purine nucleoside substrates | Aneurinibacillus migulanus |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 9.86 | - |
adenosine | pH 8.0, 50°C, recombinant enzyme | Aneurinibacillus migulanus | |
| 12038 | - |
Inosine | pH 8.0, 50°C, recombinant enzyme | Aneurinibacillus migulanus | |
| 15862 | - |
guanosine | pH 8.0, 50°C, recombinant enzyme | Aneurinibacillus migulanus | |
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