Part:BBa_K4075005
Engineered Antimicrobial Peptide CAM-W
Enginereed Cecropin A-Melittin hybrid antimicrobial peptides with leishmanicidal activity.
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]
Profile
Name: CAM-W
Base Pairs: 81 bp
Origin: Designed
Properties: Antimicrobial peptide with leishmanicidal activity.
Design considerations: Codon optimization for E. coli
Source: [1]
Usage and Biology
The CAM-W is a cecropin A-melittin hybrid peptide comprising the cationic N-terminal sequence of cecropin A and hydrophobic N-terminal sequence of melittin, which was modified through four-tryptophan-substitution (KWKLWKKIEKWGQGIGAVLKWLTTWL-NH2) from Cecropin A-melittin (CA(1-8)M(1-18), KWKLFKKIEKVGQGIGAVLKVLTTGL). Also, this peptide was already efficiently expressed in B. subtilis WB700 [1], which can make it easier to fine-tune for gram-positive expression.
Although only CAM was positively evaluated against Leishmania spp. promastigotes [2], CAM-W only was evaluated with increasing effects against a wide range of other pathogens and proteolytic stability under a series of gastrointestinal proteases and pHs (i.e. trypsin, pepsin). However, regarding safety aspects, these tryptophan-substitutions cooperate for moderate cytotoxicity (IC50 > 300mg/L) in erythrocyte lysis tests, and more analyses are needed [3].
The UNILA_LatAm 2021 team selected pro-peptides to act as an inactivator for leishmanicidal candidates such as CAM-W (BBa_K4075005), DRS-N1 (BBa_K4075004), DRS-S1 (BBa_K4075006) e DRS-H3 (BBa_K4075007). The composite structure contains a trypsin cleavage site between pro-peptide and AMP-activated form, this site allows the proAMPs to be split after trypsin digestion and the AMP-activated form restores the antimicrobial capacity.
| Pro-peptide | AMP | Final Composite |
|---|---|---|
| DRS-H3 pro-peptide | DRS-N1 | BBa_K4075008 |
| E10 pro-peptide | CAM-W | BBa_K4075009 |
| DRS-S1 pro-peptide | DRS-S1 | BBa_K4075010 |
| DRS-H3 pro-peptide | DRS-H3 | BBa_K4075011 |
| E10 pro-peptide | DRS-N1 | BBa_K4075012 |
[1] Ji, S. et al. (2017)"Efficient biosynthesis of a Cecropin A-melittin mutant in Bacillus subtilis WB700." Nature Scientific Reports. (7):40587.
[2] DIAZ-ACHIRICA, P. et al.(1998)"The plasma membrane of Leishmania donovani promastigotes is the main target for CA(1-8)M(1-18), a synthetic cecropin A-melittin hybrid peptide."Biochemical Journal. 330(1):453-460.
[3] Ji, S. et al.(2014) "Cecropin A-melittin mutant with improved proteolytic stability and enhanced antimicrobial activity against bacteria and fungi associated with gastroenteritis in vitro." Biochemical and Biophysical Research Communications. 451(4):650-655.
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