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Literature summary for 5.3.1.14 extracted from

  • Prabhu, P.; Doan, T.T.; Jeya, M.; Kang, L.W.; Lee, J.K.
    Cloning and characterization of a rhamnose isomerase from Bacillus halodurans (2011), Appl. Microbiol. Biotechnol., 89, 635-644.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Halalkalibacterium halodurans

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
119
-
L-Mannose in 50 mM malate buffer, at pH 7.0 and 70°C Halalkalibacterium halodurans
528
-
L-rhamnose in 50 mM malate buffer, at pH 7.0 and 70°C Halalkalibacterium halodurans
771
-
L-Lyxose in 50 mM malate buffer, at pH 7.0 and 70°C Halalkalibacterium halodurans

Metals/Ions

Metals/Ions Comment Organism Structure
Co2+ required for catalytic activity, 93% activity at 1 mM compared to Mn2+ Halalkalibacterium halodurans
Mg2+ required for catalytic activity, 57% activity at 1 mM compared to Mn2+ Halalkalibacterium halodurans
Mn2+ required for catalytic activity, 100% activity at 1 mM Halalkalibacterium halodurans
additional information enzyme without any metal ions shows a negligible activity. The enzyme activity is not stimulated by Ca2+, Ba2+, Na2+, Fe2+, or Cu2+ Halalkalibacterium halodurans
Zn2+ required for catalytic activity, 20% activity at 1 mM compared to Mn2+ Halalkalibacterium halodurans

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
48000
-
2 * 48000, SDS-PAGE Halalkalibacterium halodurans
48178
-
2 * 48178, calculated from amino acid sequence Halalkalibacterium halodurans
121000
-
gel filtration Halalkalibacterium halodurans

Organism

Organism UniProt Comment Textmining
Halalkalibacterium halodurans
-
-
-
Halalkalibacterium halodurans ATCC BAA-125
-
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA column chromatography Halalkalibacterium halodurans

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
D-gulose 5% activity compared to L-rhamnose Halalkalibacterium halodurans D-sorbose
-
?
D-gulose 5% activity compared to L-rhamnose Halalkalibacterium halodurans ATCC BAA-125 D-sorbose
-
?
L-lyxose 144% activity compared to L-rhamnose Halalkalibacterium halodurans L-xylulose
-
?
L-lyxose 144% activity compared to L-rhamnose Halalkalibacterium halodurans ATCC BAA-125 L-xylulose
-
?
L-Mannose 59% activity compared to L-rhamnose Halalkalibacterium halodurans L-Fructose
-
?
L-Mannose 59% activity compared to L-rhamnose Halalkalibacterium halodurans ATCC BAA-125 L-Fructose
-
?
L-Rhamnose 100% activity Halalkalibacterium halodurans L-Rhamnulose
-
r
L-Rhamnose 100% activity Halalkalibacterium halodurans ATCC BAA-125 L-Rhamnulose
-
r
L-talose 3% activity compared to L-rhamnose Halalkalibacterium halodurans ?
-
?
L-talose 3% activity compared to L-rhamnose Halalkalibacterium halodurans ATCC BAA-125 ?
-
?

Subunits

Subunits Comment Organism
homodimer 2 * 48000, SDS-PAGE Halalkalibacterium halodurans
homodimer 2 * 48178, calculated from amino acid sequence Halalkalibacterium halodurans

Synonyms

Synonyms Comment Organism
L-RhI
-
Halalkalibacterium halodurans

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
70
-
-
Halalkalibacterium halodurans

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
60 80 the enzyme shows 100% activity for 10 h at 60°C, the half-life of the enzyme is more than 900 min, about 25 min, and 5 min at 60°C, 70°C, and 80°C, respectively Halalkalibacterium halodurans

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
88.88
-
L-Mannose in 50 mM malate buffer, at pH 7.0 and 70°C Halalkalibacterium halodurans
149.5
-
L-rhamnose in 50 mM malate buffer, at pH 7.0 and 70°C Halalkalibacterium halodurans
216
-
L-Lyxose in 50 mM malate buffer, at pH 7.0 and 70°C Halalkalibacterium halodurans

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
-
Halalkalibacterium halodurans

pH Range

pH Minimum pH Maximum Comment Organism
6 10 the optimum pH for isomerization is pH 7.0, with 75%, 85%, 92%, and 90% of maximum activity at pH 6.0, 8.0, 9.0 and 10.0, respectively Halalkalibacterium halodurans

pI Value

Organism Comment pI Value Maximum pI Value
Halalkalibacterium halodurans isoelectric focusing
-
5.5

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.283
-
L-rhamnose in 50 mM malate buffer, at pH 7.0 and 70°C Halalkalibacterium halodurans