Difference between revisions of "Part:BBa K2963009"
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<partinfo>BBa_K2963009 short</partinfo> | <partinfo>BBa_K2963009 short</partinfo> | ||
| − | The gene cluster <i>capB*CA | + | The gene cluster <i>capB*CA encode an multi-enzyme complex on cell membrane. This multi-enzyme complex is composed of capB*;capC and capA. capB catalyzes poly-γ-glutamic acid（γ-PGA） synthesis．And capC links CapB and CapA on the membrane. While CapA transports γ-PGA outside the cell.In our project, we use the this part to biosynthesize γ-PGA. |
===Usage and Biology=== | ===Usage and Biology=== | ||
Revision as of 12:07, 19 October 2019
capB*CA - encoding poly-γ-glutamic acid synthetase
The gene cluster capB*CA encode an multi-enzyme complex on cell membrane. This multi-enzyme complex is composed of capB*;capC and capA. capB catalyzes poly-γ-glutamic acid（γ-PGA） synthesis．And capC links CapB and CapA on the membrane. While CapA transports γ-PGA outside the cell.In our project, we use the this part to biosynthesize γ-PGA.
Usage and Biology
The BCA genes from Bacillus sp., encoding a polyglutamate synthetase located on the cell membrane, is capable of polymerizing glutamic acid to form poly-γ-glutamic acid. In Bacillus licheniformis, BCA are called capBCA.This part is used for producing L-glutamate-rich γ-PGA.
Characterization
We used NMR to detect γ-PGA and HPLC to analyze L- glutamate ratio of γ-PGA. The results showed we have successfully produced L-glutamate-rich γ-PGA.
By the NMR detection of the fermentation product, the specific hydrogen peaks a, b, and c on the γ-amide bond on the γ-polyglutamic acid could be detected at the corresponding time points. It was indicated that the synthetase capB*CA genes of Bacillus licheniformis heterologously expressed in Corynebacterium glutamicum, and the target product γ-polyglutamic acid was successfully produced.
We used HPLC to detect the L-glutamate monomer ratio of the γ-PGA we have produced. The results show that the L-glutamic acid monomer ratio reaches more than 90%. This part is working and we have produced L-glutamate-rich γ-PGA.