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

 
 
(One intermediate revision by one other user not shown)
Line 5: Line 5:
  
 
===Applications of BBa_K2738104===
 
===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.
 +
https://static.igem.org/mediawiki/2018/5/50/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.
 +
 +
https://static.igem.org/mediawiki/2018/a/a9/T--Paris_Bettencourt--BB_104.png
 +
 +
Biocompatibility towards mammalian cell membrane
 +
https://static.igem.org/mediawiki/2018/2/2d/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.
 +
https://static.igem.org/mediawiki/2018/2/2d/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===
 
===User Reviews===

Latest revision as of 05:59, 9 November 2018


This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

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.

T--Paris_Bettencourt--BB_104.png

Biocompatibility towards mammalian cell membrane 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

UNIQ7538aafb4c4e10c8-partinfo-00000000-QINU UNIQ7538aafb4c4e10c8-partinfo-00000001-QINU