Part:BBa_K1921017
LPP-OmpA
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Usage
The Lpp-OmpA consists of the first nine N-terminal amino acids of major E. coli lipoprotein (Lpp) bined with a transmembrane domain (46-159 aa) from outer membrane protein A (OmpA). With the help of Lpp-OmpA, we can direct different polypeptides to the cell surface of E. coli, such as β-lactamase, cellulases, etal. Now we hope to display our PETase on the surface of E. coli by using it.
Biology
Surface expression of recombinant proteins was first described more than 30 years ago.The Lpp-OmpA consists of the first nine N-terminal amino acids of major E. coli lipoprotein (Lpp) bined with a transmembrane domain (46-159 aa) from outer membrane protein A (OmpA). We got its sequence from the genome of Escherichia coli str. K-12 substr.
Reference
[1] Ali Karami, Ali Mohamad Latifi*, and Samaneh Khodi，etal. 2014. Comparison of the Organophosphorus Hydrolase Surface Display Using InaVN and Lpp-OmpA Systems in Escherichia coli. J. Microbiol. Biotechnol. (2014),24(3), 379–385 [2] Joseph A. Francisco*, etal. 1992. Transport and anchoring of β-lactamase to the external surface of Escherichia coli.
Pre-expression
Figure 1.This is the pre-expression using E.coli BL21 at 37 ℃.
Figure 2. This is the pre-expression using E.coli BL21 at 16 ℃.
Figure 3. This is the pre-expression using E.coli BL21 at 25 ℃.
Surface display HPLC results
Figure 4. Relative enzyme activity of engineering bacteria E.coli(BL21)/pET22b(+)LAP when induced at 16℃.
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