Part:BBa_K2963009
capB*CA - encoding poly-γ-glutamic acid synthetase
The gene cluster capB*CA encode an multi-enzyme complex which can polymorize D/L glutamic acid to synthesize 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 γ-PGA. In Bacillus licheniformis, BCA are called capBCA. We used the mutant B* gene. 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 we have produce. The results as follow show that 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 is indicated that the capB*CA genes from Bacillus licheniformis heterologously express in Corynebacterium glutamicum, and the target product γ-PGA is 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.
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