Difference between revisions of "Part:BBa K2963009"
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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. | 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. | ||
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| − | + | 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. | |
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Revision as of 15:08, 18 October 2019
capB*CA - encoding poly-γ-glutamic acid synthetase
CapBCA complex, consisting of three subunits, is composed of CapB、CapC and CapA. CapB catalyzes poly-gamma-Glutamic acid synthesis．And CapC links CapB and CapA in the membrane. While CapA transports poly-gamma-Glutamic acid outside the cell.In our project, we use the CapBCA complex to biosynthesize poly-gamma-Glutamic acid.
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.