Part:BBa_K5292348:Design
MnfaA-12
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
During the design of the MnfaA signal peptide, we considered the following factors: H-region optimization: We modified the H-region of the nfaA signal peptide to enhance its secretion efficiency in Escherichia coli. By utilizing a Markov transfer frequency matrix to screen and substitute key amino acids, we aimed to optimize its hydrophobicity and transmembrane capability.
Secretion efficiency of the signal peptide: Ensuring that the signal peptide effectively guides the secretion of the ICCG protein to the periplasmic space was a key objective. Therefore, we focused on the length, amino acid composition, and fusion expression effects of the signal peptide in the design.
Compatibility: Since the MnfaA signal peptide will be expressed in Escherichia coli, we ensured its compatibility with the E. coli secretion system, ensuring that it can function effectively in the E. coli environment. Protein stability: We also considered the potential impact of the fusion of the signal peptide with the ICCG protein on protein stability, aiming to avoid expression level reduction due to misfolding or insufficient secretion efficiency.
Source
The MnfaA signal peptide is derived from the nfaA signal peptide, which was obtained by modifying and optimizing the H region of the nfaA signal peptide. The nfaA signal peptide originates from the genome of Thermobifida fusca, a species of actinobacteria capable of degrading various complex organic compounds. Thermobifida fusca is widely used in research on the degradation of cellulose and polymers. The nfaA signal peptide can be used for the secretion of recombinant proteins in Escherichia coli.