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Literature summary 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)

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

Protein Variants

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

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.6.1.19 additional information
-
 additional information Km values for mutant enzymes with diverse dinucleotide substrates, overview Rhizopus niveus
4.6.1.19 0.044
-
 GpG wild-type enzyme, pH 5.0, 23°C Rhizopus niveus
4.6.1.19 0.047
-
 CpC wild-type enzyme, pH 5.0, 23°C Rhizopus niveus
4.6.1.19 0.052
-
 ApG wild-type enzyme, pH 5.0, 23°C Rhizopus niveus
4.6.1.19 0.06
-
 UpG wild-type enzyme, pH 5.0, 23°C Rhizopus niveus
4.6.1.19 0.065
-
 ApA wild-type enzyme, pH 5.0, 23°C Rhizopus niveus
4.6.1.19 0.066
-
 UpC wild-type enzyme, pH 5.0, 23°C Rhizopus niveus
4.6.1.19 0.069
-
 GpA wild-type enzyme, pH 5.0, 23°C Rhizopus niveus
4.6.1.19 0.072
-
 ApU wild-type enzyme, pH 5.0, 23°C Rhizopus niveus
4.6.1.19 0.075
-
 GpU wild-type enzyme, pH 5.0, 23°C Rhizopus niveus
4.6.1.19 0.081
-
 CpU wild-type enzyme, pH 5.0, 23°C Rhizopus niveus
4.6.1.19 0.107
-
 GpC wild-type enzyme, pH 5.0, 23°C Rhizopus niveus
4.6.1.19 0.113
-
 UpU wild-type enzyme, pH 5.0, 23°C Rhizopus niveus
4.6.1.19 0.117
-
 CpG wild-type enzyme, pH 5.0, 23°C Rhizopus niveus
4.6.1.19 0.12
-
 ApC wild-type enzyme, pH 5.0, 23°C Rhizopus niveus
4.6.1.19 0.13
-
 CpA wild-type enzyme, pH 5.0, 23°C Rhizopus niveus
4.6.1.19 0.144
-
 UpA wild-type enzyme, pH 5.0, 23°C Rhizopus niveus

Organism

EC Number Organism UniProt Comment Textmining
4.6.1.19 Rhizopus niveus
-
-
-

Purification (Commentary)

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

Reaction

EC Number Reaction Comment Organism Reaction ID
4.6.1.19 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
a

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
4.6.1.19 150
-
 purified recombinant mutant Q32F Rhizopus niveus
4.6.1.19 330
-
 purified recombinant mutant Q32N Rhizopus niveus
4.6.1.19 420
-
 purified recombinant mutant Q32D Rhizopus niveus
4.6.1.19 620
-
 purified recombinant mutant Q32V Rhizopus niveus
4.6.1.19 1250
-
 purified recombinant mutant Q32E Rhizopus niveus
4.6.1.19 1600
-
 purified recombinant wild-type enzyme Rhizopus niveus
4.6.1.19 1800
-
 purified recombinant mutant Q32L Rhizopus niveus
4.6.1.19 2000
-
 purified recombinant mutant Q32T Rhizopus niveus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.6.1.19 ApA + H2O
-
 Rhizopus niveus AMP + adenosine
-
 ?
4.6.1.19 ApC + H2O
-
 Rhizopus niveus AMP + cytosine
-
 ?
4.6.1.19 ApG + H2O
-
 Rhizopus niveus AMP + guanosine
-
 ?
4.6.1.19 ApU + H2O
-
 Rhizopus niveus AMP + uridine
-
 ?
4.6.1.19 CpA + H2O
-
 Rhizopus niveus CMP + adenosine
-
 ?
4.6.1.19 CpC + H2O
-
 Rhizopus niveus CMP + cytidine
-
 ?
4.6.1.19 CpG + H2O
-
 Rhizopus niveus CMP + guanosine
-
 ?
4.6.1.19 CpU + H2O
-
 Rhizopus niveus 3'-CMP + uridine
-
 ?
4.6.1.19 GpA + H2O
-
 Rhizopus niveus GMP + adenosine
-
 ?
4.6.1.19 GpC + H2O
-
 Rhizopus niveus GMP + cytosine
-
 ?
4.6.1.19 GpG + H2O best substrate Rhizopus niveus GMP + guanosine
-
 ?
4.6.1.19 GpU + H2O
-
 Rhizopus niveus GMP + uridine
-
 ?
4.6.1.19 additional information subsite base specificities at the B1 and B2 site of wild-type and mutant enzymes, overview Rhizopus niveus ?
-
 ?
4.6.1.19 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 ?
4.6.1.19 UpA + H2O
-
 Rhizopus niveus UMP + adenosine
-
 ?
4.6.1.19 UpC + H2O
-
 Rhizopus niveus UMP + cytosine
-
 ?
4.6.1.19 UpG + H2O
-
 Rhizopus niveus UMP + guanosine
-
 ?
4.6.1.19 UpU + H2O
-
 Rhizopus niveus UMP + uridine
-
 ?

Synonyms

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

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
4.6.1.19 37
-
 assay at Rhizopus niveus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
4.6.1.19 5
-
 assay at Rhizopus niveus