Difference between revisions of "Part:BBa K2963039"

 
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<partinfo>BBa_K2963039 short</partinfo>
 
<partinfo>BBa_K2963039 short</partinfo>
  
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===Usage and Biology===
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The BCA genes from Bacillus sp. encode a γ-PGA synthetase located on the cell membrane which is capable of polymerizing glutamic acid to form poly-γ-glutamic acid. In Bacillus licheniformis BCA are called capBCA. We mutated the B gene into B*.
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The racE gene is derived from Bacillus subtilis and it encodes a racemase which can converts L-glutamate to D-glutamate.
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We used this part to produce different D/L monomer ratios of γ-PGA. We assembled the capB*CA genes and racE gene (BBa_K2963032) together to construct this part using plasmid PZM1.
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===Characterization===
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We transferred this part into our chassis microorganism Corynebacterium glutamicum using plasmid PZM1. And we using HPLC to detect the D/L monomer ratio of γ-PGA.The result shows as below.
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[[image:HPLC.png|400px]]
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Picture1 is the L-glutamate monomer ratio in γ-PGA we have produced using part BBa_K2963009 and the result reaches about over 90%.
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[[image:HPLC2.png|400px]]
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This part is working. All the results show that we have achieved the goal of producing different D/L glutamate monomer ratio of γ-PGA preliminary.
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Picture2 is the result of BBa_K2963039. This part contains capBCA genes and racE gene which is under the control of tac promoter with one lacO. The L-glutamate monomer ratio reaches about 32%.
  
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===Usage and Biology===
 
  
 
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Revision as of 15:49, 18 October 2019


Producing γ-PGA with different D/L glutamate monomer ratio(R).

Usage and Biology

The BCA genes from Bacillus sp. encode a γ-PGA synthetase located on the cell membrane which is capable of polymerizing glutamic acid to form poly-γ-glutamic acid. In Bacillus licheniformis BCA are called capBCA. We mutated the B gene into B*.

The racE gene is derived from Bacillus subtilis and it encodes a racemase which can converts L-glutamate to D-glutamate.

We used this part to produce different D/L monomer ratios of γ-PGA. We assembled the capB*CA genes and racE gene (BBa_K2963032) together to construct this part using plasmid PZM1.

Characterization

We transferred this part into our chassis microorganism Corynebacterium glutamicum using plasmid PZM1. And we using HPLC to detect the D/L monomer ratio of γ-PGA.The result shows as below.

HPLC.png

Picture1 is the L-glutamate monomer ratio in γ-PGA we have produced using part BBa_K2963009 and the result reaches about over 90%.

HPLC2.png

This part is working. All the results show that we have achieved the goal of producing different D/L glutamate monomer ratio of γ-PGA preliminary.

Picture2 is the result of BBa_K2963039. This part contains capBCA genes and racE gene which is under the control of tac promoter with one lacO. The L-glutamate monomer ratio reaches about 32%.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1912
    Illegal EcoRI site found at 2756
    Illegal EcoRI site found at 2815
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1912
    Illegal EcoRI site found at 2756
    Illegal EcoRI site found at 2815
    Illegal NheI site found at 2365
    Illegal NheI site found at 2963
    Illegal NheI site found at 4281
    Illegal NheI site found at 5248
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1912
    Illegal EcoRI site found at 2756
    Illegal EcoRI site found at 2815
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1912
    Illegal EcoRI site found at 2756
    Illegal EcoRI site found at 2815
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1912
    Illegal EcoRI site found at 2756
    Illegal EcoRI site found at 2815
    Illegal NgoMIV site found at 2590
  • 1000
    COMPATIBLE WITH RFC[1000]