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Literature summary for 5.3.1.6 extracted from

  • Yeom, S.J.; Seo, E.S.; Kim, Y.S.; Oh, D.K.
    Increased D-allose production by the R132E mutant of ribose-5-phosphate isomerase from Clostridium thermocellum (2011), Appl. Microbiol. Biotechnol., 89, 1859-1866.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli ER2566 cells Acetivibrio thermocellus

Protein Variants

Protein Variants Comment Organism
C65A the mutant shows no activity for D-psicose Acetivibrio thermocellus
D8A the mutant shows no activity for D-psicose Acetivibrio thermocellus
H133A the mutant shows 15% activity compared to the wild type enzyme Acetivibrio thermocellus
H98A the mutant shows 13% activity compared to the wild type enzyme Acetivibrio thermocellus
H9A the mutant shows 63% activity compared to the wild type enzyme Acetivibrio thermocellus
N99A the mutant shows 35% activity compared to the wild type enzyme Acetivibrio thermocellus
R132A the mutant exhibits an increase in D-psicose isomerization activity (116% activity compared to the wild type enzyme) Acetivibrio thermocellus
R132D the mutant shows increased catalytic efficiency toward D-psicose compared to the wild type enzyme Acetivibrio thermocellus
R132E the mutant shows increased catalytic efficiency toward D-psicose compared to the wild type enzyme Acetivibrio thermocellus
R132E the specific activity and catalytic efficiency (kcat/Km) of the R132E mutant for D-psicose are 1.4 and 1.5fold higher than those of the wild type enzyme, respectively Acetivibrio thermocellus
R132I the mutant shows decreased catalytic efficiency toward D-psicose compared to the wild type enzyme Acetivibrio thermocellus
R132K the mutant shows decreased catalytic efficiency toward D-psicose compared to the wild type enzyme Acetivibrio thermocellus
R132Q the mutant shows decreased catalytic efficiency toward D-psicose compared to the wild type enzyme Acetivibrio thermocellus
R136A the mutant shows 15% activity compared to the wild type enzyme Acetivibrio thermocellus
T135A the mutant shows 91% activity compared to the wild type enzyme Acetivibrio thermocellus
T67A the mutant shows 46% activity compared to the wild type enzyme Acetivibrio thermocellus
Y42A the mutant shows no activity for D-psicose Acetivibrio thermocellus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
43
-
D-psicose mutant enzyme R132E, pH 7.5, 80°C Acetivibrio thermocellus
46
-
D-psicose mutant enzyme R132D, pH 7.5, 80°C Acetivibrio thermocellus
53
-
D-psicose mutant enzyme R132K, pH 7.5, 80°C Acetivibrio thermocellus
53
-
D-psicose wild type enzyme, pH 7.5, 80°C Acetivibrio thermocellus
54
-
D-psicose mutant enzyme R132Q, pH 7.5, 80°C Acetivibrio thermocellus
56
-
D-psicose mutant enzyme R132A, pH 7.5, 80°C Acetivibrio thermocellus
78
-
D-psicose mutant enzyme R132I, pH 7.5, 80°C Acetivibrio thermocellus

Organism

Organism UniProt Comment Textmining
Acetivibrio thermocellus A3DIL8
-
-
Acetivibrio thermocellus ATCC 27405 A3DIL8
-
-

Purification (Commentary)

Purification (Comment) Organism
HisTrap HP column chromatography Acetivibrio thermocellus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.259
-
mutant enzyme H98A, using D-psicose as substrate, pH 7.5, 80°C Acetivibrio thermocellus
0.299
-
mutant enzyme H133A, using D-psicose as substrate, pH 7.5, 80°C Acetivibrio thermocellus
0.299
-
mutant enzyme R136A, using D-psicose as substrate, pH 7.5, 80°C Acetivibrio thermocellus
0.698
-
mutant enzyme N99A, using D-psicose as substrate, pH 7.5, 80°C Acetivibrio thermocellus
0.917
-
mutant enzyme T67A, using D-psicose as substrate, pH 7.5, 80°C Acetivibrio thermocellus
1.256
-
mutant enzyme H9A, using D-psicose as substrate, pH 7.5, 80°C Acetivibrio thermocellus
1.815
-
mutant enzyme T135A, using D-psicose as substrate, pH 7.5, 80°C Acetivibrio thermocellus
1.994
-
wild type enzyme, using D-psicose as substrate, pH 7.5, 80°C Acetivibrio thermocellus
2.313
-
mutant enzyme R132A, using D-psicose as substrate, pH 7.5, 80°C Acetivibrio thermocellus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
D-psicose the conversion yield of D-psicose to D-allose is 32% for the R132E mutant enzyme and 25% for the wild type enzyme after 80 min Acetivibrio thermocellus D-allose
-
r
D-psicose the conversion yield of D-psicose to D-allose is 32% for the R132E mutant enzyme and 25% for the wild type enzyme after 80 min Acetivibrio thermocellus ATCC 27405 D-allose
-
r
D-ribulose 5-phosphate
-
Acetivibrio thermocellus D-ribose 5-phosphate
-
r
D-ribulose 5-phosphate
-
Acetivibrio thermocellus ATCC 27405 D-ribose 5-phosphate
-
r

Synonyms

Synonyms Comment Organism
CTRpiB
-
Acetivibrio thermocellus
ribose-5-phosphate isomerase
-
Acetivibrio thermocellus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
80
-
-
Acetivibrio thermocellus

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
60 90
-
Acetivibrio thermocellus

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
60 75 the half-lives of the wild type enzyme at 60°C, 65°C, 70°C, 75°C, and 80°C are 11, 7.0, 4.2, 1.5, and 0.6 h, respectively Acetivibrio thermocellus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1962
-
D-psicose mutant enzyme R132K, pH 7.5, 80°C Acetivibrio thermocellus
2211
-
D-psicose mutant enzyme R132I, pH 7.5, 80°C Acetivibrio thermocellus
2335
-
D-psicose mutant enzyme R132Q, pH 7.5, 80°C Acetivibrio thermocellus
2347
-
D-psicose wild type enzyme, pH 7.5, 80°C Acetivibrio thermocellus
2580
-
D-psicose mutant enzyme R132A, pH 7.5, 80°C Acetivibrio thermocellus
2674
-
D-psicose mutant enzyme R132D, pH 7.5, 80°C Acetivibrio thermocellus
2743
-
D-psicose mutant enzyme R132E, pH 7.5, 80°C Acetivibrio thermocellus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
-
Acetivibrio thermocellus

pH Range

pH Minimum pH Maximum Comment Organism
6 9
-
Acetivibrio thermocellus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
28
-
D-psicose mutant enzyme R132I, pH 7.5, 80°C Acetivibrio thermocellus
37
-
D-psicose mutant enzyme R132K, pH 7.5, 80°C Acetivibrio thermocellus
43
-
D-psicose mutant enzyme R132Q, pH 7.5, 80°C Acetivibrio thermocellus
44
-
D-psicose wild type enzyme, pH 7.5, 80°C Acetivibrio thermocellus
46
-
D-psicose mutant enzyme R132A, pH 7.5, 80°C Acetivibrio thermocellus
58
-
D-psicose mutant enzyme R132D, pH 7.5, 80°C Acetivibrio thermocellus
64
-
D-psicose mutant enzyme R132E, pH 7.5, 80°C Acetivibrio thermocellus