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Literature summary for 5.3.1.14 extracted from

  • Lin, C.J.; Tseng, W.C.; Fang, T.Y.
    Characterization of a thermophilic L-rhamnose isomerase from Caldicellulosiruptor saccharolyticus ATCC 43494 (2011), J. Agric. Food Chem., 59, 8702-8708.
    View publication on PubMed
Search for more literature for 5.3.1.14

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21-CodonPlus (DE3)-RIL cells Caldicellulosiruptor saccharolyticus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.03
-
 L-rhamnose in 50 mM potassium phosphate buffer (pH 7.0) and 1 mM Co2+, at 65°C Caldicellulosiruptor saccharolyticus
3 3.5 D-ribose in 50 mM potassium phosphate buffer (pH 7.0) and 1 mM Co2+, at 65°C Caldicellulosiruptor saccharolyticus
3.64
-
 L-Mannose in 50 mM potassium phosphate buffer (pH 7.0) and 1 mM Co2+, at 65°C Caldicellulosiruptor saccharolyticus
3.65
-
 L-Lyxose in 50 mM potassium phosphate buffer (pH 7.0) and 1 mM Co2+, at 65°C Caldicellulosiruptor saccharolyticus
14.3
-
 D-allose in 50 mM potassium phosphate buffer (pH 7.0) and 1 mM Co2+, at 65°C Caldicellulosiruptor saccharolyticus

Metals/Ions

Metals/Ions Comment Organism Structure
Co2+ maximum activity in the presence of Co2+ Caldicellulosiruptor saccharolyticus
Mn2+ the enzyme shows 58% of the maximum activity in the presence of Mn2+ Caldicellulosiruptor saccharolyticus
additional information no activity in the presence of Na+, Mg2+, Ca2+, Cu2+, K+, and Zn2+ or in the absence of any ions Caldicellulosiruptor saccharolyticus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
48000
-
 4 * 48000, SDS-PAGE Caldicellulosiruptor saccharolyticus
48292
-
 4 * 48292, MALDI-TOF mass spectrometry Caldicellulosiruptor saccharolyticus
48294
-
 4 * 48294, calculated from amino acid sequence Caldicellulosiruptor saccharolyticus
193400
-
 gel filtration Caldicellulosiruptor saccharolyticus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-Rhamnose Caldicellulosiruptor saccharolyticus
-
 L-Rhamnulose
-
 r
L-Rhamnose Caldicellulosiruptor saccharolyticus ATCC 43494
-
 L-Rhamnulose
-
 r

Organism

Organism UniProt Comment Textmining
Caldicellulosiruptor saccharolyticus A4XHV7
-
-
Caldicellulosiruptor saccharolyticus ATCC 43494 A4XHV7
-
-

Purification (Commentary)

Purification (Comment) Organism
heat treatment, nucleic acid precipitation, and Q-Sepharose ion-exchange chromatography Caldicellulosiruptor saccharolyticus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
14.8
-
 crude extract, in 50 mM potassium phosphate buffer (pH 7.0) and 1 mM Co2+, at 90°C Caldicellulosiruptor saccharolyticus
379
-
 after 25.6fold purification, in 50 mM potassium phosphate buffer (pH 7.0) and 1 mM Co2+, at 90°C Caldicellulosiruptor saccharolyticus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
D-allose
-
 Caldicellulosiruptor saccharolyticus D-psicose
-
 ?
D-allose
-
 Caldicellulosiruptor saccharolyticus ATCC 43494 D-psicose
-
 ?
D-ribose very minimal activity toward D-ribose Caldicellulosiruptor saccharolyticus D-ribulose
-
 ?
D-ribose very minimal activity toward D-ribose Caldicellulosiruptor saccharolyticus ATCC 43494 D-ribulose
-
 ?
L-lyxose
-
 Caldicellulosiruptor saccharolyticus L-xylulose
-
 ?
L-lyxose
-
 Caldicellulosiruptor saccharolyticus ATCC 43494 L-xylulose
-
 ?
L-Mannose
-
 Caldicellulosiruptor saccharolyticus L-Fructose
-
 ?
L-Mannose
-
 Caldicellulosiruptor saccharolyticus ATCC 43494 L-Fructose
-
 ?
L-Rhamnose
-
 Caldicellulosiruptor saccharolyticus L-Rhamnulose
-
 r
L-Rhamnose
-
 Caldicellulosiruptor saccharolyticus ATCC 43494 L-Rhamnulose
-
 r

Subunits

Subunits Comment Organism
homotetramer 4 * 48000, SDS-PAGE Caldicellulosiruptor saccharolyticus
homotetramer 4 * 48292, MALDI-TOF mass spectrometry Caldicellulosiruptor saccharolyticus
homotetramer 4 * 48294, calculated from amino acid sequence Caldicellulosiruptor saccharolyticus

Synonyms

Synonyms Comment Organism
L-RhI
-
 Caldicellulosiruptor saccharolyticus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
90
-
-
 Caldicellulosiruptor saccharolyticus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
80 90 the enzyme shows good thermostability and retains more than 90% activity after a 6 h incubation at 80°C and pH 7.0-8.0. The enzyme is stable at 85°C for 4 h. However, the enzyme activity drops sharply to 30% after 2 h at 90°C, and the half-life at 90°C is around 65 min Caldicellulosiruptor saccharolyticus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
68.1
-
 D-allose in 50 mM potassium phosphate buffer (pH 7.0) and 1 mM Co2+, at 65°C Caldicellulosiruptor saccharolyticus
78.7
-
 L-Mannose in 50 mM potassium phosphate buffer (pH 7.0) and 1 mM Co2+, at 65°C Caldicellulosiruptor saccharolyticus
99.9
-
 L-rhamnose in 50 mM potassium phosphate buffer (pH 7.0) and 1 mM Co2+, at 65°C Caldicellulosiruptor saccharolyticus
118
-
 D-ribose in 50 mM potassium phosphate buffer (pH 7.0) and 1 mM Co2+, at 65°C Caldicellulosiruptor saccharolyticus
186
-
 L-Lyxose in 50 mM potassium phosphate buffer (pH 7.0) and 1 mM Co2+, at 65°C Caldicellulosiruptor saccharolyticus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
-
 Caldicellulosiruptor saccharolyticus

pH Stability

pH Stability pH Stability Maximum Comment Organism
4 11 the enzyme is stable at pH values ranging from 4.0 to 11.0 with more than 80% remaining activity and retains more than 90% activity after a 6 h incubation at 80°C and pH 7.0-8.0. The enzyme activity is fully retained at pH 7-8, 32% remaining at pH 9.0, but becomes inactivated in the pH range from 4.0 to 6.0 Caldicellulosiruptor saccharolyticus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.54
-
 D-ribose in 50 mM potassium phosphate buffer (pH 7.0) and 1 mM Co2+, at 65°C Caldicellulosiruptor saccharolyticus
4.77
-
 D-allose in 50 mM potassium phosphate buffer (pH 7.0) and 1 mM Co2+, at 65°C Caldicellulosiruptor saccharolyticus
21.6
-
 L-Mannose in 50 mM potassium phosphate buffer (pH 7.0) and 1 mM Co2+, at 65°C Caldicellulosiruptor saccharolyticus
50.8
-
 L-Lyxose in 50 mM potassium phosphate buffer (pH 7.0) and 1 mM Co2+, at 65°C Caldicellulosiruptor saccharolyticus
97.1
-
 L-rhamnose in 50 mM potassium phosphate buffer (pH 7.0) and 1 mM Co2+, at 65°C Caldicellulosiruptor saccharolyticus