Team:Bioriidl Somaiya/Design
Design
The biological design of our project was conducted using various assembly to optimize the functionality of the organism.
Initially we were in searching for the proteins which will act as an antimicrobial peptide, having potential to kill the contaminating bacteria present in the non-sterile media. Thus, this landed us with three main protein that can potentially affect the viability of the contaminants. The following are the description of the three desired proteins.
Nisin A
Normally this Lanthionine-containing peptide antibiotic
(lantibiotic) active on Gram-positive bacteria. The bactericidal
activity of lantibiotics is based on the depolarization of energized
bacterial cytoplasmic membranes, initiated by the formation of
aqueous transmembrane pores. In our project we have used this as an
anti-microbial peptide to kill all the bacterial contaminants
present in the media.
Mode of action: Lanthionine-containing peptide antibiotic
(lantibiotic). The bactericidal activity of lantibiotics is based on
depolarization of energized bacterial cytoplasmic membranes,
initiated by the formation of aqueous transmembrane pores.
Subtilosin-A
There are two genes used in combination which will help in releasing
the bactericidal protein Subtilosin-A. Subtilosin-A has bactericidal
activity against some gram-positive bacteria and some gram-negative
bacteria. Sbo-A gene is used in combination with the alb-A gene to
help in proper maturation and activation of Subtilosin-A protein. As
the gene alb-A catalyzes the formation of 3 thioether bonds during
the production of the sactipeptide subtilosin from SboA. In vitro
the thioether bonds cannot be made in the absence of the SboA
propeptide, suggesting this is the first reaction in subtilosin
maturation. This has garnered much attention for their use as safe,
natural food preservatives, as well as their potential in medical
applications. Subtilosin A (commonly referred to as subtilosin) has
a cyclical, cross-linked structure unique among characterized
bacteriocins.
Mode of Action- This synthesized peptides produced by bacteria that
have antimicrobial activity against organisms closely related to the
producer species. Due to its ability to be permeabilized into target
cell’s cytoplasmic membrane. Electrostatic interactions between the
membrane and protein provide a temporary linkage that allows the
hydrophobic peptide to insert itself into the membrane. In many
cases, these transient pores produce leakage of intracellular ions,
amino acids, and other low molecular weight molecules and eventually
cause a total depletion of the cell’s transmembrane ion potential.
Sporulation Killing Factor (SKF)
.The gene skfA produces a 26-residue extracellular sporulation
killing factor (skfA). The mature SKF is cyclic 26-amino acid
peptide that is posttranslationally modified with one disulfide and
one cysteine thioether bridged to the alpha-position of a
methionine, the thioether bond.
Mode of action- (SKF) that induces the lysis of other B.subtilis
cells that have not entered the sporulation pathway. Used in our
system to inhibit the spore bearer and to denature them
