Difference between revisions of "Part:BBa K2963021"

Revision as of 10:47, 20 October 2019

pgsBCA- encoding a poly-γ-glutamic acid synthetase

PgsBCA complex, consisting of three subunits, is composed of PgsB、PgsC and PgsA. PgsB catalyzes poly-γ-glutamic acid synthesis． And PgsC links PgsB and PgsA in the membrane. While PgsA transports poly-γ-glutamic acid outside the cell. In our project, we use the PgsBCA complex to biosynthesize poly-γ-glutamic acid.

Usage and Biology

The BCA genes from Bacillus sp. encode a γ-PGA synthetase located on the cell membrane which is capable of polymerizing glutamic acid to form poly-γ-glutamic acid. In Bacillus subtilis, BCA are called pgsBCA. This part is used 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 show 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 synthetase pgsBCA genes of Bacillus subtilis heterologously expressed in Corynebacterium glutamicum, and the target product γ-polyglutamic acid 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.

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal XbaI site found at 2207
Illegal PstI site found at 2188
Illegal PstI site found at 3467
Illegal PstI site found at 3753
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 2365
Illegal NheI site found at 2963
Illegal NheI site found at 4254
Illegal PstI site found at 2188
Illegal PstI site found at 3467
Illegal PstI site found at 3753
21
COMPATIBLE WITH RFC[21]
23
INCOMPATIBLE WITH RFC[23]
Illegal XbaI site found at 2207
Illegal PstI site found at 2188
Illegal PstI site found at 3467
Illegal PstI site found at 3753
25
INCOMPATIBLE WITH RFC[25]
Illegal XbaI site found at 2207
Illegal PstI site found at 2188
Illegal PstI site found at 3467
Illegal PstI site found at 3753
Illegal NgoMIV site found at 2586
Illegal AgeI site found at 4111
1000
COMPATIBLE WITH RFC[1000]

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K2963021 short</partinfo>
-PgsBCA complex, consisting of three subunits, is composed of PgsB&#12289;PgsC and PgsA. PgsB catalyzes poly-gamma-Glutamic acid synthesis&#65294;And PgsC links PgsB and PgsA in the membrane. While PgsA transports poly-gamma-Glutamic acid outside the cell.In our project, we use the PgsBCA complex to biosynthesize poly-gamma-Glutamic acid.
+PgsBCA complex, consisting of three subunits, is composed of PgsB&#12289;PgsC and PgsA. PgsB catalyzes poly-γ-glutamic acid synthesis&#65294; And PgsC links PgsB and PgsA in the membrane. While PgsA transports poly-γ-glutamic acid outside the cell. In our project, we use the PgsBCA complex to biosynthesize poly-γ-glutamic acid.
 ===Usage and Biology===
-The BCA genes from Bacillus sp. encode a γ-PGA synthetase located on the cell membrane which is capable of polymerizing glutamic acid to form poly-γ-glutamic acid. In Bacillus subtilis, BCA are called pgsBCA.This part is used producing L-glutamate-rich γ-PGA.
+The <i>BCA</i> genes from <i>Bacillus sp.</i> encode a γ-PGA synthetase located on the cell membrane which is capable of polymerizing glutamic acid to form poly-γ-glutamic acid. In <i>Bacillus subtilis</i>, <i>BCA</i> are called <i>pgsBCA</i>. This part is used 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. The results show 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 pgsB*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 synthetase <i>pgsBCA</i> genes of <i>Bacillus subtilis</i> heterologously expressed in <i>Corynebacterium glutamicum</i>, and the target product γ-polyglutamic acid is successfully produced.
 [[image:HPLC.png|400px]]