Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 4.3.1.19 extracted from

  • Wang, Y.; Li, G.S.; Qiao, P.; Lin, L.; Xue, H.L.; Zhu, L.; Wu, M.B.; Lin, J.P.; Yang, L.R.
    Increased productivity of L-2-aminobutyric acid and total turnover number of NAD+/NADH in a one-pot system through enhanced thermostability of L-threonine deaminase (2018), Biotechnol. Lett., 40, 1551-1559 .
    View publication on PubMed

Application

Application Comment Organism
synthesis the mutant L-threonine deaminase G323D/F510L/T344A together with thermostable L-leucine dehydrogenase from Bacillus sphaericus DSM730 and formate dehydrogenase from Candida boidinii constitute a one-pot system for biosynthesis of L-2-aminobutyric acid Escherichia coli

Cloned(Commentary)

Cloned (Comment) Organism
expression of the mutant enzyme G323D/F510L/T344A in Escherichia coli BL21(DE3) Escherichia coli

Protein Variants

Protein Variants Comment Organism
A14T the mutant enzyme shows higher thermostability than wild-type enzyme without considerable loss in activity Escherichia coli
F510L mutant enzyme with greater thermostability compared to wild-type enzyme Escherichia coli
G323D mutant enzyme with greater thermostability compared to wild-type enzyme Escherichia coli
G323D/F510L/T344A the half-life of mutant enzyme at 42°C increases from 10 to 210 min, a 20fold increase compared to the wild-type enzyme, and the temperature at which the activity of the enzyme decreases by 50% in 15 min increases from 39 to 53°C Escherichia coli
R449C the mutant enzyme shows higher thermostability than wild-type enzyme without considerable loss in activity Escherichia coli
T344A the mutant enzyme shows higher thermostability than wild-type enzyme without considerable loss in activity Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3 3.5 L-threonine pH 7.5, 30°C, mutant enzyme G323D/R449C Escherichia coli
14.5
-
L-threonine pH 7.5, 30°C, wild-type enzyme Escherichia coli
26.2
-
L-threonine pH 7.5, 30°C, mutant enzyme G323D/F510L/T344A Escherichia coli
26.4
-
L-threonine pH 7.5, 30°C, mutant enzyme G323D/F510L Escherichia coli
26.9
-
L-threonine pH 7.5, 30°C, mutant enzyme G323D/F510L/T344A/R449C Escherichia coli
27.1
-
L-threonine pH 7.5, 30°C, mutant enzyme G323D/F510L/R449C Escherichia coli
28.3
-
L-threonine pH 7.5, 30°C, mutant enzyme F510L Escherichia coli
30.1
-
L-threonine pH 7.5, 30°C, mutant enzyme A14T/G323D Escherichia coli
32.5
-
L-threonine pH 7.5, 30°C, mutant enzyme A14T Escherichia coli
32.8
-
L-threonine pH 7.5, 30°C, mutant enzyme R449C Escherichia coli
33.3
-
L-threonine pH 7.5, 30°C, mutant enzyme T344A Escherichia coli
33.6
-
L-threonine pH 7.5, 30°C, mutant enzyme G323D Escherichia coli
33.7
-
L-threonine pH 7.5, 30°C, mutant enzyme G323D/T344A Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-threonine Escherichia coli
-
2-oxobutanoate + NH3
-
?

Organism

Organism UniProt Comment Textmining
Escherichia coli P04968
-
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
68.5
-
pH 7.5, 30°C, mutant enzyme A14T Escherichia coli
77.4
-
pH 7.5, 30°C, mutant enzyme T344A Escherichia coli
78.8
-
pH 7.5, 30°C, mutant enzyme F510L Escherichia coli
89.5
-
pH 7.5, 30°C, mutant enzyme R449C Escherichia coli
99.4
-
pH 7.5, 30°C, mutant enzyme G323D/F510L Escherichia coli
99.5
-
pH 7.5, 30°C, mutant enzyme G323D/F510L/T344A Escherichia coli
99.5
-
pH 7.5, 30°C, mutant enzyme G323D/F510L/T344A/R449C Escherichia coli
99.5
-
pH 7.5, 30°C, mutant enzyme G323D/T344A Escherichia coli
100.2
-
pH 7.5, 30°C, mutant enzyme G323D/R449C Escherichia coli
100.4
-
pH 7.5, 30°C, mutant enzyme A14T/G323D Escherichia coli
101
-
pH 7.5, 30°C, mutant enzyme G323D/F510L/R449C Escherichia coli
104.6
-
pH 7.5, 30°C, mutant enzyme G323D Escherichia coli
108
-
pH 7.5, 30°C, wild-type enzyme Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-threonine
-
Escherichia coli 2-oxobutanoate + NH3
-
?

Synonyms

Synonyms Comment Organism
L-TD
-
Escherichia coli
L-threonine deaminase
-
Escherichia coli

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
39
-
15 min, 50% loss of activity, wild-type enzyme Escherichia coli
41
-
15 min, 50% loss of activity, mutant enzyme A14T Escherichia coli
41.2
-
15 min, 50% loss of activity, mutant enzyme R449C Escherichia coli
42
-
15 min, 50% loss of activity, mutant enzyme T344A Escherichia coli
42
-
half-life: 10 min (wild-type enzyme), 210 min (mutant enzyme G323D/F510L/T344A), 14 min (mutant enzyme A14T), 19 min (mutant enzyme T344A), 15 min (mutant enzyme R449C), 57 min (mutant enzyme G323D), 44 min (mutant enzyme F510L), 54 min (mutant enzyme A14T/G323D and mutant enzyme G323D/R449C), 61 min (mutant enzyme G323D/T344A), 201 min (mutant enzyme G323D/F510L), 200 min (mutant enzyme G323D/F510L/R449C), 211 min (mutant enzyme G323D/F510L/T344A/R449C) Escherichia coli
44.8
-
15 min, 50% loss of activity, mutant enzyme F510L Escherichia coli
46
-
15 min, 50% loss of activity, mutant enzyme A14T/G323D Escherichia coli
46
-
15 min, 50% loss of activity, mutant enzyme G323D/R449C Escherichia coli
46.2
-
15 min, 50% loss of activity, mutant enzyme G323D Escherichia coli
47.1
-
15 min, 50% loss of activity, mutant enzyme G323D/T344A Escherichia coli
52.4
-
15 min, 50% loss of activity, mutant enzyme G323D/F510L/R449C Escherichia coli
52.5
-
15 min, 50% loss of activity, mutant enzyme G323D/F510L Escherichia coli
53
-
15 min, 50% loss of activity, mutant enzyme G323D/F510L/T344A Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1230
-
L-threonine pH 7.5, 30°C, mutant enzyme F510L Escherichia coli
1256
-
L-threonine pH 7.5, 30°C, mutant enzyme T344A Escherichia coli
1261
-
L-threonine pH 7.5, 30°C, mutant enzyme A14T Escherichia coli
1273
-
L-threonine pH 7.5, 30°C, mutant enzyme R449C Escherichia coli
1283
-
L-threonine pH 7.5, 30°C, mutant enzyme G323D/F510L Escherichia coli
1298
-
L-threonine pH 7.5, 30°C, mutant enzyme G323D/F510L/R449C Escherichia coli
1301
-
L-threonine pH 7.5, 30°C, mutant enzyme G323D/F510L/T344A/R449C Escherichia coli
1309
-
L-threonine pH 7.5, 30°C, mutant enzyme G323D/F510L/T344A Escherichia coli
1312
-
L-threonine pH 7.5, 30°C, mutant enzyme G323D/R449C Escherichia coli
1369
-
L-threonine pH 7.5, 30°C, mutant enzyme A14T/G323D Escherichia coli
1394
-
L-threonine pH 7.5, 30°C, mutant enzyme G323D/T344A Escherichia coli
1398
-
L-threonine pH 7.5, 30°C, mutant enzyme G323D Escherichia coli
1414
-
L-threonine pH 7.5, 30°C, wild-type enzyme Escherichia coli