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</i> encode an multi-enzyme complex | + | 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=== | ===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 | + | 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 produce γ-PGA. In <i>Bacillus licheniformis</i>, <i>BCA</i> are called <i>capBCA</i>. We used the mutant <i>B* </i>gene of <i>B</i> gene. This part is used for producing L-glutamate-rich γ-PGA. |
===Characterization=== | ===Characterization=== | ||
| − | We used NMR to detect γ-PGA and HPLC to analyze L- glutamate ratio of γ-PGA. The results | + | 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]] | [[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 | + | 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. |
| − | |||
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. | 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. | ||
| + | [[image:HPLC.png|400px]] | ||
| + | |||
| + | ===References=== | ||
| + | 1. Xu P, Vansiri A, Bhan N, et al. ePathBrick: a synthetic biology platform for engineering metabolic pathways in E. coli[J]. ACS Synthetic Biology, 2012, 1(7): 256-266. | ||
| + | |||
| + | 2. Peng Yingyun. Study on the production, synthesis mechanism and antifreeze of γ-polyglutamic acid. Diss. Jiangnan University, 2015. | ||
| + | |||
| + | 3. Sung M H, Park C, Kim C J, et al. Natural and edible biopolymer poly-gamma-glutamic acid: synthesis, production, and applications [J]. Chemical Record, 2005, 5(6): 352-366. | ||
<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here | ||
Latest revision as of 14:54, 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 produce γ-PGA. In Bacillus licheniformis, BCA are called capBCA. We used the mutant B* gene of 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.
References
1. Xu P, Vansiri A, Bhan N, et al. ePathBrick: a synthetic biology platform for engineering metabolic pathways in E. coli[J]. ACS Synthetic Biology, 2012, 1(7): 256-266.
2. Peng Yingyun. Study on the production, synthesis mechanism and antifreeze of γ-polyglutamic acid. Diss. Jiangnan University, 2015.
3. Sung M H, Park C, Kim C J, et al. Natural and edible biopolymer poly-gamma-glutamic acid: synthesis, production, and applications [J]. Chemical Record, 2005, 5(6): 352-366.