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Part:BBa_K2738104:Experience

Designed by: Darshak Bhatt   Group: iGEM18_Paris_Bettencourt   (2018-10-10)
Revision as of 00:41, 18 October 2018 by Aleksa (Talk | contribs)


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Applications of BBa_K2738104

Results Expression data The lyase-cage-enterocinA fusion protein was produced via cell-free expression system. The expression was verified by SDS gel as shown in the figure below. T--Paris_Bettencourt--BBa_2738104.png 2. MIC and growth curve We determined the MIC of lyase-cage-enterocinA StarCore to inhibit the growth of E coli and B subtilis (figure A and B). We performed growth curve assay for E coli and B subtilis with and without antimicrobial peptides as shown in figure C and D.

The results indicate that the lyase-cage-enterocinA StarCore has the MIC activity at 150 ug/ml for B subtilis but not for E coli. Moreover, at 7.3 uM it also affects the growth kinetics B. subtilis to a significant extent indicating bactericidal activity. The antibacterial activity of lyase-cage-enterocinA StarCore is comparable to ovispirin at its MIC concentration (5 uM) alone as shown in the figure.

<img scr="T--Paris_Bettencourt--BB_104.png">

Biocompatibility towards mammalian cell membrane <img scr="T--Paris_Bettencourt--liposome_c104.png">

Figure 2. Mammalian liposome leakage induced by lyase cage-Enterocin A StarCore (A) and the kinetics of liposome leakage (B). Ovispirin (5 uM) and lyase-ovispirin (3.9 uM) are used at their MIC concentration obtained from previous results. T--Paris_Bettencourt--liposome_c104.png Ovispirin alone lyses the liposome indicating damage to mammalian cell membrane. Lyase-cage-enterocin has a reduced effect on the liposome leakage indicating an improvement in biocompatibility towards mammalian cell membrane. The Starcore ferritin-ovispirin has a safer profile in terms of membrane activity as compared to ovispirin alone at their respective MIC concentration.

References: Protein lyase cage obtained from RCSB: 4QCC PDB Enterocin reference from Bactibase: http://bactibase.hammamilab.org/BAC088 Star shaped antimicrobial peptides : 10.1038/NMICROBIOL.2016.162 Liposome leakage assay: https://www.sciencedirect.com/science/article/pii/S007668799778025X



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