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academies
September 20-21, 2017 | Philadelphia, USA
Global summit on
TUBERCULOSIS AND LUNG DISEASE
Int J Respir Med 2017 Volume 2 Issue 2
T
uberculosis is still a global threatened disease caused
by intracellular pathogen
Mycobacterium tuberculosis
(Mtb)
. India accounts for 60% new cases worldwide. The
bacterium can outperform the host defense mechanism and
can persist inside the body for decades in dormant stage.
There are many factors that are known to play role during
this stage of bacterium; storage of triacylglycerol (TAG) is
one of them. The literature on
Mtb
is replete with strong
evidence that TAGs play a critical role in the intracellular/
intraphagosomal survival of this pathogen in the host.
Mtb
genome contains about 4000genes and26 (LipA- lipZ) of them
are annotated as putative lipases/esterases.
Rv1900c
of
Mtb
Lip family annotated as LipJ; conserved in all
Mycobacterium
species. It contains two domains, N- terminal α/β hydrolase
domain and C- terminal cyclase homology domain. In
this study we have cloned LipJ full length and N- Terminal
lipolytic domain in pET28a vector, expressed in
E. coli
BL21
(DE3) host cells. Recombinant protein was purified using Ni-
NTA affinity chromatography. Biochemical characterization
of holoenzyme and its N-terminal revealed esterase activity
towards pNP- caprate as their optimal substrate. Both the
holoenzyme and N-terminal has 40°C as optimal temperature
and is stable up to 60 °C, and their pH optima was found to
be pH9 and stable from pH6 to 9. Hence, it is confirmed that
the esterase activity of holoenzyme was just because of its
N- terminal domain. Biophysical characterization confirms
that it belongs to α/β hydrolase family.
Speaker Biography
Bandana Kumari is a PhD Scholar in Department of Biotechnology, Chandigarh,
India. She has been working on mycobacterial lipases since last two years under
the supervision of Prof. Jagdeep Kaur. Her expertise is in Molecular Biology, Protein
Chemistry and Animal Tissue Culture.
e:
vandanadhiman21@gmail.comFunctional analysis of
Rv1900c
gene product from
Mycobacterium tuberculosis
Bandana Kumari
and
Jagdeep Kaur
Panjab University, India