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 | + | 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 | + | 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.
Figure 1. The synthesis of Xylulose from Glycolaldehyde and Dihydroxyacetone(DHA), catalyzed by TalB-F187Y.
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.
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.
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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 861
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]