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Literature summary for 3.5.4.3 extracted from

  • Trautwein-Schult, A.; Jankowska, D.; Cordes, A.; Hoferichter, P.; Klein, C.; Matros, A.; Mock, H.P.; Baronian, K.; Bode, R.; Kunze, G.
    Arxula adeninivorans recombinant guanine deaminase and its application in the production of food with low purine content (2014), J. Mol. Microbiol. Biotechnol., 24, 67-81.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Arxula adeninivorans strain G1212 Blastobotrys adeninivorans

Inhibitors

Inhibitors Comment Organism Structure
additional information not inhibited by EDTA, 1,10-phenanthroline, dithiothreitol, L-cysteine, and phenylmethylsulfonyl fluoride Blastobotrys adeninivorans

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.026
-
guanine recombinant enzyme, at pH 6.0 and 37°C Blastobotrys adeninivorans
0.056
-
guanine native enzyme, at pH 6.0 and 37°C Blastobotrys adeninivorans

Localization

Localization Comment Organism GeneOntology No. Textmining
cytoplasm
-
Blastobotrys adeninivorans 5737
-
vacuole
-
Blastobotrys adeninivorans 5773
-

Metals/Ions

Metals/Ions Comment Organism Structure
Cu2+ at 1 mM Cu2+, the native enzyme shows about 80% inhibition and the recombinant enzyme almost complete (99%) inhibition Blastobotrys adeninivorans
Hg2+ complete inhibition at 1 mM Blastobotrys adeninivorans
additional information not influenced by Ca2+, Fe3+, K+, Mg2+, Ni2+, and Zn2+ Blastobotrys adeninivorans

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
53000
-
2 * 53000, calculated from amino acid sequence Blastobotrys adeninivorans
55000
-
2 * 55000, SDS-PAGE and MALDI-TOF mass spectrometry Blastobotrys adeninivorans
100000 110000 recombinant and native enzyme, gel filtration Blastobotrys adeninivorans

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
guanine + H2O Blastobotrys adeninivorans 100% activity xanthine + NH3
-
r
guanine + H2O Blastobotrys adeninivorans LS3 100% activity xanthine + NH3
-
r

Organism

Organism UniProt Comment Textmining
Blastobotrys adeninivorans
-
-
-
Blastobotrys adeninivorans LS3
-
-
-

Purification (Commentary)

Purification (Comment) Organism
heat precipitation, DEAE Sepharose column chromatography, and Superdex 200 gel filtration Blastobotrys adeninivorans

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.1
-
crude extract, at pH 6.0 and 37°C Blastobotrys adeninivorans
4.7
-
after 51.5fold purification, at pH 6.0 and 37°C Blastobotrys adeninivorans

Storage Stability

Storage Stability Organism
-20°C, 28 days, 123% activity Blastobotrys adeninivorans
-20°C, with 20% (v/v) glycine, 28 days, 65% residual activity Blastobotrys adeninivorans
22°C, 28 days, 34% residual activity Blastobotrys adeninivorans
4°C, 28 days, 44% residual activity Blastobotrys adeninivorans

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2,6-diaminopurine + H2O the native enzyme shows 107% activity and the recombinant enzyme 98% activity compared to guanine Blastobotrys adeninivorans ?
-
?
2-amino-6-chloropurine + H2O the native enzyme shows 93% activity and the recombinant enzyme 88% activity compared to guanine Blastobotrys adeninivorans 6-chloro-9H-purin-2-ol + NH3
-
?
8-azaguanine + H2O the native enzyme shows 99% activity and the recombinant enzyme 95% activity compared to guanine Blastobotrys adeninivorans 8-azaxanthine + NH3
-
?
8-azaguanine + H2O the native enzyme shows 99% activity and the recombinant enzyme 95% activity compared to guanine Blastobotrys adeninivorans LS3 8-azaxanthine + NH3
-
?
8-azaxanthine + H2O the native enzyme shows 95% activity and the recombinant enzyme 88% activity compared to guanine Blastobotrys adeninivorans ?
-
?
adenine + H2O the native enzyme shows 102% activity and the recombinant enzyme 85% activity compared to guanine Blastobotrys adeninivorans allopurinol + NH3
-
?
adenine + H2O the native enzyme shows 102% activity and the recombinant enzyme 85% activity compared to guanine Blastobotrys adeninivorans LS3 allopurinol + NH3
-
?
adenosine + H2O the native enzyme shows 98% activity and the recombinant enzyme 91% activity compared to guanine Blastobotrys adeninivorans inosine + NH3
-
?
adenosine + H2O the native enzyme shows 98% activity and the recombinant enzyme 91% activity compared to guanine Blastobotrys adeninivorans LS3 inosine + NH3
-
?
guanine + H2O 100% activity Blastobotrys adeninivorans xanthine + NH3
-
r
guanine + H2O 100% activity Blastobotrys adeninivorans LS3 xanthine + NH3
-
r
guanosine + H2O the native enzyme shows 48% activity and the recombinant enzyme 70% activity compared to guanine Blastobotrys adeninivorans ?
-
?
hypoxanthine + H2O the native enzyme shows 100% activity and the recombinant enzyme 87% activity compared to guanine Blastobotrys adeninivorans ?
-
?
uric acid + H2O the native enzyme shows 89% activity and the recombinant enzyme 86% activity compared to guanine Blastobotrys adeninivorans ?
-
?
uric acid + H2O the native enzyme shows 89% activity and the recombinant enzyme 86% activity compared to guanine Blastobotrys adeninivorans LS3 ?
-
?
xanthine + H2O the native enzyme shows 01% activity and the recombinant enzyme 87% activity compared to guanine Blastobotrys adeninivorans ?
-
?

Subunits

Subunits Comment Organism
homodimer 2 * 53000, calculated from amino acid sequence Blastobotrys adeninivorans
homodimer 2 * 55000, SDS-PAGE and MALDI-TOF mass spectrometry Blastobotrys adeninivorans

Synonyms

Synonyms Comment Organism
Agdap
-
Blastobotrys adeninivorans
GDA
-
Blastobotrys adeninivorans
Guanine aminohydrolase
-
Blastobotrys adeninivorans

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
55
-
-
Blastobotrys adeninivorans

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
30
-
the native enzyme remains stable for 1 h at up to 30°C Blastobotrys adeninivorans
60
-
the recombinant enzyme remains stable for 1 h at up to 60°C Blastobotrys adeninivorans

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.5
-
guanine recombinant enzyme, at pH 6.0 and 37°C Blastobotrys adeninivorans
3.9
-
guanine native enzyme, at pH 6.0 and 37°C Blastobotrys adeninivorans

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.5
-
the native enzyme shows optimal pH values of 6.5 and 8.0 while the recombinant enzyme has a single pH optimum at pH 6.5 Blastobotrys adeninivorans
8
-
the native enzyme shows optimal pH values of 6.5 and 8.0 Blastobotrys adeninivorans

pH Stability

pH Stability pH Stability Maximum Comment Organism
6.5 8 native and recombinant enzymes are stable during incubation for 1 h at 37°C with pH in the range of 6.5-8.0 Blastobotrys adeninivorans

pI Value

Organism Comment pI Value Maximum pI Value
Blastobotrys adeninivorans calculated from amino acid sequence
-
5.7

Expression

Organism Comment Expression
Blastobotrys adeninivorans immediately after the shift of cells into media with purine as nitrogen source, the enzyme transcript level increases and reached maximum level at 2 h with adenine and 4 h with hypoxanthine, after which transcription fell to the basal level up