Composite

Part:BBa_K2963009

Designed by: Experiment group members of JNU-China   Group: iGEM19_JNU-China   (2019-09-08)
Revision as of 15:04, 18 October 2019 by GHX (Talk | contribs)


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.

NMR.png 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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