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Literature summary extracted from

  • Liu, G.; Cheng, T.; Chu, J.; Li, S.; He, B.
    Efficient synthesis of purine nucleoside analogs by a new trimeric purine nucleoside phosphorylase from Aneurinibacillus migulanus AM007 (2019), Molecules, 25, 100 .
    View publication on PubMedView publication on EuropePMC

Application

EC Number Application Comment Organism
2.4.2.1 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)

EC Number Cloned (Comment) Organism
2.4.2.1 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

EC Number Protein Variants Comment Organism
2.4.2.1 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]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.4.2.1 1.16
-
 guanosine pH 8.0, 50°C, recombinant enzyme Aneurinibacillus migulanus
2.4.2.1 1.45
-
 adenosine pH 8.0, 50°C, recombinant enzyme Aneurinibacillus migulanus
2.4.2.1 5.3
-
 Inosine pH 8.0, 50°C, recombinant enzyme Aneurinibacillus migulanus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.4.2.1 87000
-
 recombinant His6-tagged enzyme, gel filtration Aneurinibacillus migulanus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.4.2.1 guanosine + phosphate Aneurinibacillus migulanus
-
 guanine + alpha-D-ribose 1-phosphate
-
 r
2.4.2.1 guanosine + phosphate Aneurinibacillus migulanus AM007
-
 guanine + alpha-D-ribose 1-phosphate
-
 r
2.4.2.1 inosine + phosphate Aneurinibacillus migulanus
-
 hypoxanthine + alpha-D-ribose 1-phosphate
-
 r
2.4.2.1 inosine + phosphate Aneurinibacillus migulanus AM007
-
 hypoxanthine + alpha-D-ribose 1-phosphate
-
 r

Organism

EC Number Organism UniProt Comment Textmining
2.4.2.1 Aneurinibacillus migulanus A0A3G5E8S8
-
-
2.4.2.1 Aneurinibacillus migulanus AM007 A0A3G5E8S8
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.4.2.1 2'-deoxyguanosine + phosphate moderate activity Aneurinibacillus migulanus guanine + 2-deoxy-alpha-D-ribose 1-phosphate
-
 r
2.4.2.1 2,6-diaminopurine + alpha-D-ribose 1-phosphate
-
 Aneurinibacillus migulanus 2,6-diaminopurine ribonucleoside + phosphate
-
 r
2.4.2.1 2-amino-6-chloropurine + alpha-D-ribose 1-phosphate
-
 Aneurinibacillus migulanus 2-amino-6-chloropurine ribonucleoside + phosphate
-
 r
2.4.2.1 6-thioguanine + alpha-D-ribose 1-phosphate
-
 Aneurinibacillus migulanus 6-thioguanine ribonucleoside + phosphate
-
 r
2.4.2.1 adenosine + phosphate low activity Aneurinibacillus migulanus adenine + alpha-D-ribose 1-phosphate
-
 r
2.4.2.1 guanosine + phosphate
-
 Aneurinibacillus migulanus guanine + alpha-D-ribose 1-phosphate
-
 r
2.4.2.1 guanosine + phosphate moderate activity Aneurinibacillus migulanus guanine + alpha-D-ribose 1-phosphate
-
 r
2.4.2.1 guanosine + phosphate
-
 Aneurinibacillus migulanus AM007 guanine + alpha-D-ribose 1-phosphate
-
 r
2.4.2.1 guanosine + phosphate moderate activity Aneurinibacillus migulanus AM007 guanine + alpha-D-ribose 1-phosphate
-
 r
2.4.2.1 inosine + phosphate
-
 Aneurinibacillus migulanus hypoxanthine + alpha-D-ribose 1-phosphate
-
 r
2.4.2.1 inosine + phosphate best substrate Aneurinibacillus migulanus hypoxanthine + alpha-D-ribose 1-phosphate
-
 r
2.4.2.1 inosine + phosphate
-
 Aneurinibacillus migulanus AM007 hypoxanthine + alpha-D-ribose 1-phosphate
-
 r
2.4.2.1 inosine + phosphate best substrate Aneurinibacillus migulanus AM007 hypoxanthine + alpha-D-ribose 1-phosphate
-
 r
2.4.2.1 additional information substrate specificity, detailed overview, no activity with 2'-deoxyadenosine Aneurinibacillus migulanus ?
-
-
2.4.2.1 additional information substrate specificity, detailed overview, no activity with 2'-deoxyadenosine Aneurinibacillus migulanus AM007 ?
-
-

Subunits

EC Number Subunits Comment Organism
2.4.2.1 homotrimer 3 * 29000, about, recombinant His6-tagged enzyme, SDS-PAGE Aneurinibacillus migulanus

Synonyms

EC Number Synonyms Comment Organism
2.4.2.1 AmPNP
-
 Aneurinibacillus migulanus
2.4.2.1 inosine-guanosine phosphorylase UniProt Aneurinibacillus migulanus
2.4.2.1 PNP
-
 Aneurinibacillus migulanus
2.4.2.1 purine nucleoside phosphorylase
-
 Aneurinibacillus migulanus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.4.2.1 70
-
 recombinant enzyme Aneurinibacillus migulanus

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
2.4.2.1 30 80 activity range, profile overview, recombinant enzyme Aneurinibacillus migulanus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
2.4.2.1 55
-
 and below, purified recombinant enzyme, 12 h, completely stable, half-life is 78.6 h Aneurinibacillus migulanus
2.4.2.1 60
-
 purified recombinant enzyme, 2 h, pH 7.0-9.0, completely stable, half-life is 37.3 h Aneurinibacillus migulanus
2.4.2.1 65
-
 purified recombinant enzyme, half-life is 6.9 h Aneurinibacillus migulanus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.4.2.1 14.3
-
 adenosine pH 8.0, 50°C, recombinant enzyme Aneurinibacillus migulanus
2.4.2.1 18400
-
 guanosine pH 8.0, 50°C, recombinant enzyme Aneurinibacillus migulanus
2.4.2.1 63800
-
 Inosine pH 8.0, 50°C, recombinant enzyme Aneurinibacillus migulanus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.4.2.1 7.5
-
 recombinant enzyme Aneurinibacillus migulanus

pH Range

EC Number pH Minimum pH Maximum Comment Organism
2.4.2.1 6 8 over 80% of maximal activity within this range, recombinant enzyme Aneurinibacillus migulanus

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
2.4.2.1 7 9 purified recombinant enzyme, 2 h, 60°C, completely stable Aneurinibacillus migulanus

General Information

EC Number General Information Comment Organism
2.4.2.1 evolution most PNPs are divided into trimeric PNPs and hexametric PNPs based on molecular mass, protein structure, or substrate specificity Aneurinibacillus migulanus
2.4.2.1 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]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.4.2.1 9.86
-
 adenosine pH 8.0, 50°C, recombinant enzyme Aneurinibacillus migulanus
2.4.2.1 12038
-
 Inosine pH 8.0, 50°C, recombinant enzyme Aneurinibacillus migulanus
2.4.2.1 15862
-
 guanosine pH 8.0, 50°C, recombinant enzyme Aneurinibacillus migulanus