Difference between revisions of "Part:BBa K3209002"

 
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This transaldolase mutant (TalB-F178Y) is derived from E.coli, which is different from the part BBa_K2155003, containing mutation F178Y. It catalyzes xylulose synthesis from glycolaldehyde and dihydroxyacetone (DHA). Along with the BFD-M7, TalB-F178Y catalyzes the xylulose production which is a precursor of rare sugar.
 
This transaldolase mutant (TalB-F178Y) is derived from E.coli, which is different from the part BBa_K2155003, containing mutation F178Y. It catalyzes xylulose synthesis from glycolaldehyde and dihydroxyacetone (DHA). Along with the BFD-M7, TalB-F178Y catalyzes the xylulose production which is a precursor of rare sugar.
 
[[File:K3209002-1.jpg|center]]
 
[[File:K3209002-1.jpg|center]]
Figure 1. The synthesis of xylulose from Glycolaldehyde and Dihydroxyacetone(DHA), catalyzed by TalB-F187Y.
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Figure 1. The synthesis of Xylulose from Glycolaldehyde and Dihydroxyacetone(DHA), catalyzed by TalB-F187Y.
  
 
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[[File:K3209000-2-1.jpg|center]]
 
[[File:K3209000-2-1.jpg|center]]
Figure 2. Identification of BDD-M7 and TalB-F187Y expression vectors construction.  
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Figure 2. Identification of BFD-M7 (BBa_K3209000) and TalB-F187Y (BBa_K3209002) expression vectors construction.  
 
1:BFD-M7 plasmid; 2:Digestion of BFD-M7 plasmid by EcoRI and PstI; 3:TalB-F187Y plasmid; 4:Digestion of TalB-F187Y plasmid by EcoRI and PstI; M: Marker.
 
1:BFD-M7 plasmid; 2:Digestion of BFD-M7 plasmid by EcoRI and PstI; 3:TalB-F187Y plasmid; 4:Digestion of TalB-F187Y plasmid by EcoRI and PstI; M: Marker.
  

Latest revision as of 00:27, 21 October 2019


Transaldolase mutant (TalB-F187Y)

This transaldolase mutant (TalB-F178Y) is derived from E.coli, which is different from the part BBa_K2155003, containing mutation F178Y. It catalyzes xylulose synthesis from glycolaldehyde and dihydroxyacetone (DHA). Along with the BFD-M7, TalB-F178Y catalyzes the xylulose production which is a precursor of rare sugar.

K3209002-1.jpg

Figure 1. The synthesis of Xylulose from Glycolaldehyde and Dihydroxyacetone(DHA), catalyzed by TalB-F187Y.



K3209000-2-1.jpg

Figure 2. Identification of BFD-M7 (BBa_K3209000) and TalB-F187Y (BBa_K3209002) expression vectors construction. 1:BFD-M7 plasmid; 2:Digestion of BFD-M7 plasmid by EcoRI and PstI; 3:TalB-F187Y plasmid; 4:Digestion of TalB-F187Y plasmid by EcoRI and PstI; M: Marker.



K3209002-3.jpg

Figure 3. Expression of TalB-F187Y and purification by Ni-NTA affinity chromatography. M: protein marker; 1: precipitation samples in the cell lysates; 2: supernatant samples in the cell lysates; 3: 50 mM imidazole eluent; 4: 100 mM imidazole eluent; 5: 200 mM imidazole. The results showed that 200 mM imidazole eluent is the best concentration for elution expressed TalB-F187Y.



K3209002-4.jpg

Figure 4. HPLC analysis of the xylulose product catalyzed by TalB-F187Y. A: Standard Xylulose; B: The xylulose product catalyzed by TalB-F187Y in vitro. The results showed that our expression vector expressed active TalB-F187Y, which catalyzed the synthesis of xylulose from glycolaldehyde and DHA.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 861
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]