Good morning everyone.

Batın, Deniz and I will present

the properties of bacteria
Clostridium botulinum, the difference between the close
relative Clostridium tetani, cellular mechanism of
botulism, symptoms, treatments, and biosecurity.
C. botulinum was discovered by Emile van Ermengem
in 1895 while there is an outbreak caused by pork. It is
named after sausage poisoning.
It is a rod shaped, gram-positive, and motile bacterium
that forms anaerobic spores. C. botulinum releases
botulinum toxin which damages body’s nerves, deprives
breathing, causes muscle paralysis and leads to death.
Most importantly, it is the most dangerous bacterial
toxin.
There are four different groups based on physiological
differences and 16s rRNA genome sequencing. Group 1
and group 2 members causes botulism in humans. Also,
there are seven types of botulinum neurotoxin that
cleaves SNARE proteins enzymatically to prevent
acetylcholine release at the neuromuscular junctions.
BoNT is synthesized as 150kDa single polypeptide
which then post translationally cleaved into 100 kda
heavy chain and 50 kda light chain which are connected
to each other via disulphide bond.
Accessory proteins help infection to human by forming
complexes with the botulinum neurotoxin. Spore
staining or gram staining methods can be utilized to
observe and differentiate the components and the
structures of the bacteria.
C. botulinum causes flaccid paralysis in muscles. Its
prevalence is rare in developed countries and seldom in
developing countries. Close relative of C. botulinum, C.
tetani produces a neurotoxin, as well. It is called tetanus
neurotoxin (TeNT). It is gram positive, spore forming,
anaerobic, as well. But, more prevalent than BoNT.

Botulinum Neurotoxin (BoNT) synthesised as a 150 k Dalton

single polypeptide chain. Then it is cleaved as 50 kDa Light
chain (Lc) or catalytic subunit that is zinc dependent
endopeptidase, 100 kDa Heavy chain (Hc) that is consist
of to equal part; N terminal (Hn) translocation domain and C
terminal (Hc) receptor binding domain respectively (Lacy et
al. ,1998 and Swaminathan and
Eswaramoorthy,2000). BoNT arranged in linear fashion,
Lc, Hcn and Hcc respectively. Lc noncovalently bound to
translocation domain. Also, there are a disulphide bridge
between catalytic and translocation domain and
translocation domain also cover Lc with belt loop.
Furthermore, receptor binding domain consist of two super
secondary structure ß-trefoil and jelly
roll fold(Tighe and Schiavo,2013). Pathogenesis
of BoNT consists of several step: receptor binding-
internalization-endosome formation-translocation-catalysis
(Montal, 2010). In brief Receptor binding domains recognize
ganglioside oligosaccharides and synaptotagmin ectodomain
peptide (Chai et al., 2006 and Stenmark,2008). Then
neurotoxin internalize and form endosome. Translocation
domain chaperones catalytic domain translocation across
endosome by unfold and stabilize unfolded state during
translocation by pH gradient. After catalytic domain fully
reach cytosol disulphide bridge is reduced and catalytic
domain refold (Kumaran et al., 2009 and Montal,2010).
Catalytic domain binds to SNARE and hydrolase amide bonds
by Zn- dependent catalytic mechanism (Hanson and
Stevens, 2000 and Breidenbach and Brunger,2004). As a
result, SNARE proteins degraded and membrane fusion
events at synapse terminal is blocked. Thus, acetylcholine
release is prevented, this causes flaccid paralysis at
motoneurons and ultimately leads to dead due to paralysis
in respiratory system (Turton, Chaddock & Acharya,2002
and Shukla and Sharma, 2005).