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Literature summary for 3.1.26.4 extracted from

  • Chon, H.; Tadokoro, T.; Ohtani, N.; Koga, Y.; Takano, K.; Kanaya, S.
    Identification of RNase HII from psychrotrophic bacterium, Shewanella sp. SIB1 as a high-activity type RNase H (2006), FEBS J., 273, 2264-2275.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
gene rnhB, DNA and amino acid sequence determination and analysis, overexpression in Escherichia coli strain MIC2067(DE3), a rnhA/rnhB double mutant strain, as 60% soluble and 40% insoluble protein Shewanella sp.

General Stability

General Stability Organism
denaturation kinetics, midpoint at about 1.6 M urea and 5°C Shewanella sp.
denaturation kinetics, midpoint at about 1.8 M urea and 5°C Escherichia coli
metal ion and substrate stabilize the enzyme Escherichia coli
metal ion and substrate stabilize the enzyme Shewanella sp.

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.071
-
M13 DNA-RNA hybrid pH 8.5, 15°C, recombinant enzyme Shewanella sp.
0.075
-
M13 DNA-RNA hybrid pH 8.5, 30°C, recombinant enzyme Shewanella sp.
0.26
-
M13 DNA-RNA hybrid pH 8.5, 15°C or 30°C, recombinant enzyme Escherichia coli

Metals/Ions

Metals/Ions Comment Organism Structure
Co2+ activates Escherichia coli
Co2+ activates, best at 0.5 mM Shewanella sp.
KCl activates best at 110 mM, preferred to NaCl Shewanella sp.
KCl equally activating as NaCl Escherichia coli
Mg2+ activates Escherichia coli
Mg2+ activates, best at 5 mM Shewanella sp.
Mn2+ activates Escherichia coli
Mn2+ activates, best at 1 mM, strongly preferred divalent cation Shewanella sp.
additional information no activation by Ca2+, Zn2+, Ba2+, Ni2+, Cu2+, Fe2+, and Sr2+ Escherichia coli
additional information no activation by Ca2+, Zn2+, Ba2+, Ni2+, Cu2+, Fe2+, and Sr2+ Shewanella sp.
NaCl activates best at 60 mM Shewanella sp.
NaCl equally activating as KCl Escherichia coli

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
25000
-
recombinant enzyme, gel filtration Shewanella sp.

Organism

Organism UniProt Comment Textmining
Escherichia coli P10442 gene rnhB
-
Shewanella sp. Q25C12 psychrotrophic bacterium, gene rnhB
-
Shewanella sp. SIB1 Q25C12 psychrotrophic bacterium, gene rnhB
-

Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme from Escherichia coli strain MIC2067(DE3) to homogeneity by heparin affinity chromatography and gel filtration Shewanella sp.

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.31
-
15°C, pH 8.5, purified recombinant enzyme Escherichia coli
1.1
-
30°C, pH 8.5, purified recombinant enzyme Escherichia coli
7.6
-
15°C, pH 8.5, purified recombinant enzyme Shewanella sp.
22
-
30°C, pH 8.5, purified recombinant enzyme Shewanella sp.

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
DNA-RNA hybrid + H2O
-
Escherichia coli ?
-
?
DNA-RNA hybrid + H2O
-
Shewanella sp. ?
-
?
DNA-RNA hybrid + H2O
-
Shewanella sp. SIB1 ?
-
?
M13 DNA-RNA hybrid + H2O
-
Escherichia coli ?
-
?
M13 DNA-RNA hybrid + H2O
-
Shewanella sp. ?
-
?
M13 DNA-RNA hybrid + H2O
-
Shewanella sp. SIB1 ?
-
?
additional information substrate cleavage mode of the enzyme, cleavage site specificity Escherichia coli ?
-
?
additional information substrate cleavage mode of the enzyme, cleavage site specificity Shewanella sp. ?
-
?
additional information substrate cleavage mode of the enzyme, cleavage site specificity Shewanella sp. SIB1 ?
-
?

Subunits

Subunits Comment Organism
monomer 1 * 25000, recombinant enzyme, SDS-PAGE Shewanella sp.

Synonyms

Synonyms Comment Organism
RNase HII
-
Escherichia coli
RNase HII
-
Shewanella sp.
RnhB
-
Escherichia coli

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
40
-
-
Shewanella sp.
45
-
-
Escherichia coli

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
15 60
-
Escherichia coli
15 60
-
Shewanella sp.

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
30
-
half-life about 30 min Shewanella sp.
30
-
half-life about 45 min Escherichia coli

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
10
-
-
Escherichia coli
10
-
-
Shewanella sp.

pH Range

pH Minimum pH Maximum Comment Organism
7.1 12
-
Escherichia coli
7.1 12
-
Shewanella sp.