Part:BBa_K4427021
pSB1C3-stuffer-LP2
Our engineered bacteria were able to synthesise 2-PE in a yield of 84mg/L, but this is only 30% of the theoretical expression of the overexpressed photosensitive blue light system and is lower than the theoretical amount of 130mg/L for the KDCA and ADH1 co-expression system.
During the reflective research phase, we learned that the BUCT team was also synthesising linalool. After initial communication, it was found that our two teams differed in the design of the phenylethanol synthesis pathway. The synthetic pathways and genetic pathways of the two teams are as follows.
After further exploration, we conducted a comparison experiment. Meanwhile, in yet another literature research, I learned that tyrB encodes an aromatic amino acid transaminase that plays a key role in the synthesis of aromatic amino acids and 2-PE. deletion of the tyrB gene leads to cellular flow imbalance, massive accumulation of phenylethanol and inhibition of cell growth.
We conjectured that the addition of an exogenous tyrB gene to the engineered bacteria might enhance 2-PE expression. Thus, we carried out the third generation design of the blue light expression system. Based on the original blue light expression system, we used aromatic amino acid transaminase to reduce the inhibition of cell growth by acetaldehyde, thus appropriately increasing the expression of 2-PE.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal suffix found in sequence at 6035
Illegal EcoRI site found at 1
Illegal XbaI site found at 16
Illegal PstI site found at 2917 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 1
Illegal SpeI site found at 6036
Illegal PstI site found at 2917
Illegal PstI site found at 6050
Illegal NotI site found at 7
Illegal NotI site found at 6043 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 1
Illegal BamHI site found at 1165
Illegal BamHI site found at 5958
Illegal XhoI site found at 1743
Illegal XhoI site found at 5967
Illegal XhoI site found at 7067
Illegal XhoI site found at 7959 - 23INCOMPATIBLE WITH RFC[23]Illegal prefix found in sequence at 1
Illegal suffix found in sequence at 6036
Illegal PstI site found at 2917 - 25INCOMPATIBLE WITH RFC[25]Illegal prefix found in sequence at 1
Illegal XbaI site found at 16
Illegal SpeI site found at 6036
Illegal PstI site found at 2917
Illegal PstI site found at 6050
Illegal NgoMIV site found at 1313
Illegal AgeI site found at 1000
Illegal AgeI site found at 3084 - 1000COMPATIBLE WITH RFC[1000]
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