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Literature summary for 5.1.3.B12 extracted from

  • Choi, J.G.; Ju, Y.H.; Yeom, S.J.; Oh, D.K.
    Improvement in the thermostability of D-psicose 3-epimerase from Agrobacterium tumefaciens by random and site-directed mutagenesis (2011), Appl. Environ. Microbiol., 77, 7316-7320.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 5.1.3.B12

Protein Variants

Protein Variants Comment Organism
G67C the variant shows remarkably decreased specific activity Agrobacterium tumefaciens
I33E the enzyme variant shows no activity Agrobacterium tumefaciens
I33K the enzyme variant shows no activity Agrobacterium tumefaciens
I33L increase of 5°C in the temperature for maximal enzyme activity, increase of 7.2-fold in the half-life at 50°C, and increase of 4.3°C in apparent melting temperature, respectively, compared with the wild-type enzyme Agrobacterium tumefaciens
I33L/S213C increase of 7.5°C in the temperature for maximal enzyme activity, increase of 29.9-fold in the half-life at 50°C, and increase of 7.6°C in apparent melting temperature, respectively, compared with the wild-type enzyme. After 8 or 16 days, the enzyme activity gradually decreases, and the conversion yields with and without borate are reduced to 22 and 9.6%, respectively, at 30 days. In contrast, the activity of the immobilized I33L/S213C variant with and without borate does not decrease during the operation time of 30 days Agrobacterium tumefaciens
I33P the enzyme variant shows no activity Agrobacterium tumefaciens
S213C increase of 2.5°C in the temperature for maximal enzyme activity, increase of 3.3-fold in the half-life at 50°C, and increase of 3.1°C in apparent melting temperature, respectively, compared with the wild-type enzyme. Immobilized wild-type enzyme with and without borate maintains activity for 8 days at a conversion yield of 70% (350 g/l psicose) and for 16 days at a conversion yield of 30% (150 g/l psicose), respectively Agrobacterium tumefaciens
S213E decrease of 7.2°C in half-life at 55°C Agrobacterium tumefaciens
S213K the variant shows no activity Agrobacterium tumefaciens
S213M decrease of 6.8°C in half-life at 55°C Agrobacterium tumefaciens
S213P decrease of 6°C in half-life at 55°C Agrobacterium tumefaciens
S213T decrease of 5°C in half-life at 55°C Agrobacterium tumefaciens
S8T the variant displays 29% of the wild-type activity Agrobacterium tumefaciens
V96A the variant displays 51% of the wild-type activity Agrobacterium tumefaciens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
31
-
 D-fructose pH 8.0, 50°C, mutant enzyme I33L/S213C Agrobacterium tumefaciens
40
-
 D-fructose pH 8.0, 50°C, mutant enzyme I33L Agrobacterium tumefaciens
42
-
 D-fructose pH 8.0, 50°C, mutant enzyme S213C Agrobacterium tumefaciens
44
-
 D-fructose pH 8.0, 50°C, wild-type enzyme Agrobacterium tumefaciens

Organism

Organism UniProt Comment Textmining
Agrobacterium tumefaciens A9CH28
-
-
Agrobacterium tumefaciens ATCC 33970 A9CH28
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Agrobacterium tumefaciens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
D-fructose
-
 Agrobacterium tumefaciens D-psicose
-
 ?
D-fructose
-
 Agrobacterium tumefaciens ATCC 33970 D-psicose
-
 ?

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
50
-
 assay at Agrobacterium tumefaciens

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
50
-
 the wild-type, S213C, I33L, and I33L S213C variant enzymes have half-lives of 62, 202, 445, and 1,853 min at 50°C, respectively Agrobacterium tumefaciens
55
-
 half-life of wild-type enzyme: 10 min, half-lives of mutant enzymes: G67C (132 min), I33L (64 min), S213C (28 min), V96A (18 min), S8T (15 min), S213P (6 min), S213T (5 min), S213M (3.2 min), S213E (2.8 min) Agrobacterium tumefaciens
60.5
-
 melting temperature of wild-type enzyme Agrobacterium tumefaciens
63.6
-
 melting temperature of S213C enzyme Agrobacterium tumefaciens
64.8
-
 melting temperature of I33L enzyme Agrobacterium tumefaciens
68.1
-
 melting temperature of S213C enzyme Agrobacterium tumefaciens

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
68.3
-
 D-fructose pH 8.0, 50°C, mutant enzyme I33L/S213C Agrobacterium tumefaciens
70
-
 D-fructose pH 8.0, 50°C, mutant enzyme I33L Agrobacterium tumefaciens
70
-
 D-fructose pH 8.0, 50°C, mutant enzyme S213C Agrobacterium tumefaciens
71.6
-
 D-fructose pH 8.0, 50°C, wild-type enzyme Agrobacterium tumefaciens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
 assay at Agrobacterium tumefaciens

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.65
-
 D-fructose pH 8.0, 50°C, wild-type enzyme Agrobacterium tumefaciens
1.68
-
 D-fructose pH 8.0, 50°C, mutant enzyme S213C Agrobacterium tumefaciens
1.75
-
 D-fructose pH 8.0, 50°C, mutant enzyme I33L Agrobacterium tumefaciens
2.23
-
 D-fructose pH 8.0, 50°C, mutant enzyme I33L/S213C Agrobacterium tumefaciens