Difference between revisions of "Part:BBa K2599000"

Latest revision as of 12:14, 1 October 2018

Antimicrobial peptide - Subtilosin

This biobrick is the basic part of the Subtilosin, more information please look for the composite part (BBa_K2599008).

Figure 1. Basic part of Subtilosin

Introduction

Bacillus subtilis produces a bacteriocin called Subtilosin that targets the surface receptors and electrostatically binds to the bacterial cell membrane. Bacteriocins are a group of anitmicrobial peptides that are often distinguished from traditioinal antibiotics by their narrow range of avtivity against closely related bacteria.

Subtilosin was evaluated as a quorum sensing (QS) inhibitor in gram-positive bacteria using Fe(III) reduction assay. In gram-positive bacteria, subtilosin was evaluated as a QS inhibitor utilizing Chromobacterium voilaceum as a microbial reporter.

Mechanism of Subtilosin

The bacteriocins inhibit their target organisms through pore formation. Though the mechanism of each inhibition is vary from species to species, the general process is conserved. Details are on our project page.

According to the reference, Subtilosin acts by fully depleting the transmembrane pH gradient (ΔpH) and causing an immediate efflux of intracellular ATP, but has no effect on the transmembrane electric potential. In addition, the paper also shows that membrane permeabilization occurred at concentrations of subtilosin that were significantly higher than the MIC level of E. coli tested strains.

Features of Subtilosin

1. Species Specific

Bacteriocin target strains or closely related species. Subtilosin has specificity to Bacillus subtilis , Listeria monocytogenes , Enterococcus faecalis , etc. More target organisms can be found on [http://bactibase.hammamilab.org/BAC098 bactibase].

2. Eco-friendly

Since subtilosin is a polypeptide naturally produced by bacteria itself and can inhibit other bacteria without much environment impact. It don't pose threat to other organisms like farm animals or humans. Therefore, this toxin will not cause safety problem.

3. Biodegradable

Subtilosin is a short peptide that will degrade in a short time. After degradation, this antibacterial peptide is harmless to our environment.

Experiment Result

Cloning

We conbined our toxic gene to pSB1C3 backbone and conducted PCR to check the size of our part. The Subtilosin sequence length is around 147 b.p.

Figure 2 PCR

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

Reference

@@ Line 8: / Line 8: @@
-{{#tag:html|<img style="max-width: 20%; padding-left: 30%;" src="https://static.igem.org/mediawiki/2018/2/2c/T--NCTU_Formosa--Sub.png" alt="" />}}
+{{#tag:html|<img style="width: 20%; padding-left: 30%;" src="https://static.igem.org/mediawiki/2018/2/2c/T--NCTU_Formosa--Sub.png" alt="" />}}
+<div style="width:40%; padding-left: 20%;"><p style="padding-top: 10px; font-size: 10px; text-align: center;"><b>Figure 1.</b> Basic part of Subtilosin</p></div>
-'''''Figure 1:SDS-PAGE gels of TDPs after protein purification.'''
 <p style="padding-top:20px;font-size:20px"><b>Introduction</b></p>
-<i> Bacillus subtilis </i> produces a bacteriocin called Subtilosin that possesses antibacterial activity against certain gram-positive bacteria. The bacteriocins are a group of anitmicrobial peptides that are often distinguished from traditioinal antibiotics by their narrow range of avtivity against closely related bacteria.
+<i> Bacillus subtilis </i> produces a bacteriocin called Subtilosin that targets the surface receptors and electrostatically binds to the bacterial cell membrane. Bacteriocins are a group of anitmicrobial peptides that are often distinguished from traditioinal antibiotics by their narrow range of avtivity against closely related bacteria.
+Subtilosin was evaluated as a quorum sensing (QS) inhibitor in gram-positive bacteria using Fe(III) reduction assay. In gram-positive bacteria, subtilosin was evaluated as a QS inhibitor utilizing Chromobacterium voilaceum as a microbial reporter.
+<p style="padding-top:20px;font-size:20px"><b>Mechanism of Subtilosin</b></p>
+The bacteriocins inhibit their target organisms through pore formation. Though the mechanism of each inhibition is vary from species to species, the general process is conserved. Details are on our project page.
+According to the reference, Subtilosin acts by fully depleting the transmembrane pH gradient (ΔpH) and causing an immediate efflux of intracellular ATP, but has no effect on the transmembrane electric potential. In addition, the paper also shows that membrane permeabilization occurred at concentrations of subtilosin that were significantly higher than the MIC level of <i>E. coli</i> tested strains.
+<p style="padding-top:10px;font-size:20px;"><b>Features of Subtilosin</b></p>
+<p style="padding-top:16px;font-size:16px"><b>1. Species Specific</b></p>
+Bacteriocin target strains or closely related species. Subtilosin has specificity to <i> Bacillus subtilis </i>, <i> Listeria monocytogenes </i>, <i> Enterococcus faecalis </i>, etc. More target organisms can be found on [http://bactibase.hammamilab.org/BAC098 bactibase].
+<p style="padding-top:16px;font-size:16px"><b>2. Eco-friendly</b></p>
+Since subtilosin is a polypeptide naturally produced by bacteria itself and can inhibit other bacteria without much environment impact. It don't pose threat to other organisms like farm animals or humans. Therefore, this toxin will not cause safety problem.
+<p style="padding-top:16px;font-size:16px"><b>3. Biodegradable</b></p>
+Subtilosin is a short peptide that will degrade in a short time. After degradation, this antibacterial peptide is harmless to our environment.