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Literature summary for 4.6.1.19 extracted from

  • Ohgi, K.; Iwama, M.; Inokuchi, N.; Irie, M.
    Enzymatic properties of glutamine 32 mutants of RNase Rh from Rhizopus niveus, a trial to alter the most preferential inter-nucleotidic linkages of RNase Rh (2003), Biosci. Biotechnol. Biochem., 67, 570-576.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type and mutant enzymes in Saccharomyces cerevisiae strain R27-7C-1C Rhizopus niveus

Protein Variants

Protein Variants Comment Organism
additional information construction of enzyme mutants which possess designed substrate specificity, conversion of the base-nonspecific wild-type to base-specific mutants Rhizopus niveus
Q32D site-directed mutagenesis, 26% activity compared to the wild-type enzyme Rhizopus niveus
Q32E site-directed mutagenesis, 78% activity compared to the wild-type enzyme Rhizopus niveus
Q32F site-directed mutagenesis, 9.3% activity compared to the wild-type enzyme Rhizopus niveus
Q32L site-directed mutagenesis, 112% activity compared to the wild-type enzyme Rhizopus niveus
Q32N site-directed mutagenesis, 21% activity compared to the wild-type enzyme Rhizopus niveus
Q32T site-directed mutagenesis, 125% activity compared to the wild-type enzyme Rhizopus niveus
Q32V site-directed mutagenesis, 39% activity compared to the wild-type enzyme Rhizopus niveus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information Km values for mutant enzymes with diverse dinucleotide substrates, overview Rhizopus niveus
0.044
-
GpG wild-type enzyme, pH 5.0, 23°C Rhizopus niveus
0.047
-
CpC wild-type enzyme, pH 5.0, 23°C Rhizopus niveus
0.052
-
ApG wild-type enzyme, pH 5.0, 23°C Rhizopus niveus
0.06
-
UpG wild-type enzyme, pH 5.0, 23°C Rhizopus niveus
0.065
-
ApA wild-type enzyme, pH 5.0, 23°C Rhizopus niveus
0.066
-
UpC wild-type enzyme, pH 5.0, 23°C Rhizopus niveus
0.069
-
GpA wild-type enzyme, pH 5.0, 23°C Rhizopus niveus
0.072
-
ApU wild-type enzyme, pH 5.0, 23°C Rhizopus niveus
0.075
-
GpU wild-type enzyme, pH 5.0, 23°C Rhizopus niveus
0.081
-
CpU wild-type enzyme, pH 5.0, 23°C Rhizopus niveus
0.107
-
GpC wild-type enzyme, pH 5.0, 23°C Rhizopus niveus
0.113
-
UpU wild-type enzyme, pH 5.0, 23°C Rhizopus niveus
0.117
-
CpG wild-type enzyme, pH 5.0, 23°C Rhizopus niveus
0.12
-
ApC wild-type enzyme, pH 5.0, 23°C Rhizopus niveus
0.13
-
CpA wild-type enzyme, pH 5.0, 23°C Rhizopus niveus
0.144
-
UpA wild-type enzyme, pH 5.0, 23°C Rhizopus niveus

Organism

Organism UniProt Comment Textmining
Rhizopus niveus
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant enzymes from Saccharomyces cerevisiae Rhizopus niveus

Reaction

Reaction Comment Organism Reaction ID
RNA + H2O = an [RNA fragment]-3'-nucleoside-3'-phosphate + a 5'-hydroxy-ribonucleotide-3'-[RNA fragment] active site structure, Glu32 is an essential part of the B2-substrate recognition and binding site Rhizopus niveus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
150
-
purified recombinant mutant Q32F Rhizopus niveus
330
-
purified recombinant mutant Q32N Rhizopus niveus
420
-
purified recombinant mutant Q32D Rhizopus niveus
620
-
purified recombinant mutant Q32V Rhizopus niveus
1250
-
purified recombinant mutant Q32E Rhizopus niveus
1600
-
purified recombinant wild-type enzyme Rhizopus niveus
1800
-
purified recombinant mutant Q32L Rhizopus niveus
2000
-
purified recombinant mutant Q32T Rhizopus niveus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ApA + H2O
-
Rhizopus niveus AMP + adenosine
-
?
ApC + H2O
-
Rhizopus niveus AMP + cytosine
-
?
ApG + H2O
-
Rhizopus niveus AMP + guanosine
-
?
ApU + H2O
-
Rhizopus niveus AMP + uridine
-
?
CpA + H2O
-
Rhizopus niveus CMP + adenosine
-
?
CpC + H2O
-
Rhizopus niveus CMP + cytidine
-
?
CpG + H2O
-
Rhizopus niveus CMP + guanosine
-
?
CpU + H2O
-
Rhizopus niveus 3'-CMP + uridine
-
?
GpA + H2O
-
Rhizopus niveus GMP + adenosine
-
?
GpC + H2O
-
Rhizopus niveus GMP + cytosine
-
?
GpG + H2O best substrate Rhizopus niveus GMP + guanosine
-
?
GpU + H2O
-
Rhizopus niveus GMP + uridine
-
?
additional information subsite base specificities at the B1 and B2 site of wild-type and mutant enzymes, overview Rhizopus niveus ?
-
?
RNA + H2O from yeast Rhizopus niveus nucleoside 3'-phosphates and 3'-phosphooligonucleotides via oligonucleotides or mononucleotides with 2',3'-cyclic phosphate at the 3'-side ?
UpA + H2O
-
Rhizopus niveus UMP + adenosine
-
?
UpC + H2O
-
Rhizopus niveus UMP + cytosine
-
?
UpG + H2O
-
Rhizopus niveus UMP + guanosine
-
?
UpU + H2O
-
Rhizopus niveus UMP + uridine
-
?

Synonyms

Synonyms Comment Organism
base-nonspecific RNase
-
Rhizopus niveus
More enzyme belongs to the RNase T2 family Rhizopus niveus
RNase Rh
-
Rhizopus niveus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Rhizopus niveus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
5
-
assay at Rhizopus niveus