Difference between revisions of "Part:BBa K2963009"

Revision as of 12:37, 19 October 2019

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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 <partinfo>BBa_K2963009 short</partinfo>
-The gene cluster <i>capB*CA</i> 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&#65294;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.
+The gene cluster <i>capB*CA</i> 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 poly-γ-glutamic acid. In Bacillus licheniformis, BCA are called capBCA.This part is used for producing L-glutamate-rich γ-PGA.
+The <i>BCA</i> genes from <i>Bacillus sp.</i>, encoding a polyglutamate synthetase located on the cell membrane, is capable of polymerizing glutamic acid to form γ-PGA. In <i>Bacillus licheniformis</i>, <i>BCA</i> are called <i>capBCA</i>. We used the mutant <i>B* </i>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. The results showed we have successfully produced L-glutamate-rich γ-PGA.
+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.
 [[image:NMR.png|400px]]
-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 is indicated that the <i>capB*CA</i> genes from <i>Bacillus licheniformis</i> heterologously express in <i>Corynebacterium glutamicum</i>, and the target product γ-PGA is successfully produced.
 [[image:HPLC.png|400px]]