Difference between revisions of "Part:BBa K2738104:Experience"

 
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===Applications of BBa_K2738104===
 
===Applications of BBa_K2738104===
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Results
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Expression data
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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.
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https://static.igem.org/mediawiki/2018/5/50/T--Paris_Bettencourt--BBa_2738104.png
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2. MIC and growth curve
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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. 
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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.
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<img scr="https://static.igem.org/mediawiki/2018/a/a9/T--Paris_Bettencourt--BB_104.png">
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Biocompatibility towards mammalian cell membrane
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<img scr="https://static.igem.org/mediawiki/2018/2/2d/T--Paris_Bettencourt--liposome_c104.png">
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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.
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https://static.igem.org/mediawiki/2018/2/2d/T--Paris_Bettencourt--liposome_c104.png
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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.
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References:
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Protein lyase cage obtained from RCSB: 4QCC PDB 
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Enterocin reference from Bactibase: http://bactibase.hammamilab.org/BAC088
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Star shaped antimicrobial peptides : 10.1038/NMICROBIOL.2016.162
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Liposome leakage assay: https://www.sciencedirect.com/science/article/pii/S007668799778025X
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===User Reviews===
 
===User Reviews===

Revision as of 00:41, 18 October 2018


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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



User Reviews

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