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show all sequences of 4.1.2.5

Improving thermal stability of thermophilic L-threonine aldolase from Thermotoga maritima

Wieteska, L.; Ionov, M.; Szemraj, J.; Feller, C.; Kolinski, A.; Gront, D.; J. Biotechnol. 199, 69-76 (2015)

Data extracted from this reference:

Application
Application
Commentary
Organism
synthesis
threonine aldolase is a very promising enzyme that can be used to prepare biologically active compounds or building blocks for pharmaceutical industry. Rational design is applied to thermophilic threonine aldolase from Thermotoga maritima to improve thermal stability by the incorporation of salt and disulfide bridges between subunits in the functional tetramer
Thermotoga maritima
Cloned(Commentary)
Commentary
Organism
the gene of L-threonine aldolase from Thermotoga maritima is cloned into c-LEctas expression vector pLE1A17 under control of the T7 promoter. Expression in Escherichia coli BL21(DE3)
Thermotoga maritima
Engineering
Amino acid exchange
Commentary
Organism
A21C
L-threonine cleavage activity is similar to the activity of the wild-type enzyme
Thermotoga maritima
F85Y
L-threonine cleavage activity is about 65% of the activity of the wild-type enzyme
Thermotoga maritima
I124D
L-threonine cleavage activity is about 65% of the activity of the wild-type enzyme
Thermotoga maritima
additional information
rational design is applied to the enzyme to improve thermal stability by the incorporation of salt and disulfide bridges between subunits in the functional tetramer
Thermotoga maritima
P56C
L-threonine cleavage activity is about 95% of the activity of the wild-type enzyme, the mutant enzyme displays significantly enhanced stability compared to the wild type enzyme
Thermotoga maritima
Q22K
L-threonine cleavage activity is about 75% of the activity of the wild-type enzyme
Thermotoga maritima
S198D
L-threonine cleavage activity is about 110% of the activity of the wild-type enzyme
Thermotoga maritima
T59D
L-threonine cleavage activity is about 10% of the activity of the wild-type enzyme
Thermotoga maritima
V235C
L-threonine cleavage activity is about 110% of the activity of the wild-type enzyme
Thermotoga maritima
V29D
L-threonine cleavage activity is about 90% of the activity of the wild-type enzyme
Thermotoga maritima
W86E
the mutant enzyme displays enhanced activity, with stability similar to the wild type enzyme
Thermotoga maritima
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
37000
-
SDS-PAGE
Thermotoga maritima
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Thermotoga maritima
Q9X266
-
-
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
glycine + acetaldehyde
-
748265
Thermotoga maritima
L-threonine
-
-
-
r
L-threonine
-
748265
Thermotoga maritima
glycine + acetaldehyde
-
-
-
r
Subunits
Subunits
Commentary
Organism
tetramer
4 * 37000, SDS-PAGE
Thermotoga maritima
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
75
-
-
Thermotoga maritima
Temperature Stability [°C]
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
75
-
15 h, 50% loss of activity
Thermotoga maritima
Cofactor
Cofactor
Commentary
Organism
Structure
pyridoxal 5'-phosphate
dependent on
Thermotoga maritima
Application (protein specific)
Application
Commentary
Organism
synthesis
threonine aldolase is a very promising enzyme that can be used to prepare biologically active compounds or building blocks for pharmaceutical industry. Rational design is applied to thermophilic threonine aldolase from Thermotoga maritima to improve thermal stability by the incorporation of salt and disulfide bridges between subunits in the functional tetramer
Thermotoga maritima
Cloned(Commentary) (protein specific)
Commentary
Organism
the gene of L-threonine aldolase from Thermotoga maritima is cloned into c-LEctas expression vector pLE1A17 under control of the T7 promoter. Expression in Escherichia coli BL21(DE3)
Thermotoga maritima
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
pyridoxal 5'-phosphate
dependent on
Thermotoga maritima
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
A21C
L-threonine cleavage activity is similar to the activity of the wild-type enzyme
Thermotoga maritima
F85Y
L-threonine cleavage activity is about 65% of the activity of the wild-type enzyme
Thermotoga maritima
I124D
L-threonine cleavage activity is about 65% of the activity of the wild-type enzyme
Thermotoga maritima
additional information
rational design is applied to the enzyme to improve thermal stability by the incorporation of salt and disulfide bridges between subunits in the functional tetramer
Thermotoga maritima
P56C
L-threonine cleavage activity is about 95% of the activity of the wild-type enzyme, the mutant enzyme displays significantly enhanced stability compared to the wild type enzyme
Thermotoga maritima
Q22K
L-threonine cleavage activity is about 75% of the activity of the wild-type enzyme
Thermotoga maritima
S198D
L-threonine cleavage activity is about 110% of the activity of the wild-type enzyme
Thermotoga maritima
T59D
L-threonine cleavage activity is about 10% of the activity of the wild-type enzyme
Thermotoga maritima
V235C
L-threonine cleavage activity is about 110% of the activity of the wild-type enzyme
Thermotoga maritima
V29D
L-threonine cleavage activity is about 90% of the activity of the wild-type enzyme
Thermotoga maritima
W86E
the mutant enzyme displays enhanced activity, with stability similar to the wild type enzyme
Thermotoga maritima
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
37000
-
SDS-PAGE
Thermotoga maritima
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
glycine + acetaldehyde
-
748265
Thermotoga maritima
L-threonine
-
-
-
r
L-threonine
-
748265
Thermotoga maritima
glycine + acetaldehyde
-
-
-
r
Subunits (protein specific)
Subunits
Commentary
Organism
tetramer
4 * 37000, SDS-PAGE
Thermotoga maritima
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
75
-
-
Thermotoga maritima
Temperature Stability [°C] (protein specific)
Temperature Stability Minimum [°C]
Temperature Stability Maximum [°C]
Commentary
Organism
75
-
15 h, 50% loss of activity
Thermotoga maritima
Other publictions for EC 4.1.2.5
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
746894
Bulut
Development of a growth-depen ...
Pseudomonas putida
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1
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1
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1
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1
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1
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-
-
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-
-
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746900
Fesko
Expanding the threonine aldol ...
Aeromonas veronii
Appl. Microbiol. Biotechnol.
99
9651-9661
2015
-
-
1
-
-
-
-
3
-
-
-
-
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1
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1
-
-
-
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8
-
-
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3
-
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1
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1
1
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3
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1
-
-
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-
8
-
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-
3
-
-
-
-
-
-
-
-
3
3
748265
Wieteska
Improving thermal stability o ...
Thermotoga maritima
J. Biotechnol.
199
69-76
2015
-
1
1
-
11
-
-
-
-
-
1
-
-
2
-
-
-
-
-
-
-
-
2
1
1
-
1
-
-
-
-
1
-
-
-
-
1
1
1
-
11
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
2
1
1
-
1
-
-
-
-
-
-
-
-
-
-
-
713884
Dueckers
Threonine aldolases-screening, ...
Clostridium pasteurianum, Escherichia coli, Pseudomonas aeruginosa, Pseudomonas sp., Pseudomonas sp. NCIMB10558
Appl. Microbiol. Biotechnol.
88
409-424
2010
-
-
-
-
-
-
-
4
-
-
-
-
-
5
-
-
2
-
-
-
2
-
8
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
4
-
-
-
-
-
-
4
-
-
-
-
-
-
-
2
-
-
2
-
8
-
-
-
-
-
-
-
-
-
-
-
-
-
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5203
Pagani
DL-Allothreonine and L-threoni ...
Rattus norvegicus
Biochem. Soc. Trans.
19
346S
1991
-
-
-
-
-
-
-
-
-
-
-
-
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2
-
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-
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2
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1
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2
-
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1
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-
-
-
-
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-
-
-
-
-
-
-
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-
5191
Stoecklein
Evidence for L-threonine cleav ...
Clostridium pasteurianum
Biochem. J.
232
621-622
1985
-
-
-
-
-
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2
-
-
-
-
-
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1
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-
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1
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1
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1
2
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1
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2
2
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-
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1
-
-
-
-
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1
-
-
-
-
-
-
-
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-
5199
Dainty
Purification and properties of ...
Clostridium pasteurianum
Biochem. J.
117
585-592
1970
-
-
-
-
-
-
4
1
-
-
-
-
-
1
-
-
1
-
-
-
-
1
1
-
-
-
-
-
1
1
-
1
-
-
-
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-
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1
-
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4
-
1
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-
-
-
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1
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-
1
1
-
-
-
-
-
1
1
-
-
-
-
-
-
-
-
5200
Riario-Sforza
Threonine aldolase and allothr ...
Rattus norvegicus
Eur. J. Biochem.
8
88-92
1969
-
-
-
-
-
-
-
-
-
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1
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1
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1
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1
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1
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715411
Karasek
Greenberg, D.M.: Studies on th ...
Ovis aries
J. Biol. Chem.
227
191-205
1957
-
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3
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1
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1
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1
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1
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1
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1
1
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1
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1
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3
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1
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1
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1
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1
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1
1
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