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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 caused by
Mycobacteriumtuberculosis
. Due
to the emergence of multiple drug resistance organisms,
research in this area is focused to identify new drug targets.
In TB database, several genes have been annotated as
hypothetical and needs characterization for assigning a role.
Rv1075c
has been annotated GDSL lipase. GDSL esterases/
lipases possess multi-functional properties due to broad
substrate specificity, so some of them have thioesterase,
protease, arylesterase, and lysophospholipase activity. In
this study, we have cloned
Rv1075c
gene in pET28a vector,
expressed in E. coli BL21 (DE3) pGro7 strain and protein
was purified by Ni-NTA chromatography. Also, the active
site mutants were created using site-directed mutagenesis
technique. Based on biochemical characterization, it
was found to posses’ lipase activity toward mid-carbon
chain length having pNP-laurate as its optimal substrate.
Its optimum temperature and pH were 37˚C and 9˚C,
respectively; and stable up to 60˚C and long pH range, pH5 to
11. It was also confirmed to be belonged to SGNH hydrolase
subgroup of GDSL category of lipases by the activity analysis
of active site mutants. The active site mutant’s activity was
found to be tampered as compared to wild-type protein.
Speaker Biography
Jashandeep Kaur is a PhD Scholar in the Department of Biotechnology, Panjab
University, Chandigarh, India. She has been working on deciphering the role of lipases
in the life-cycle of
Mycobacterium tuberculosis
under the supervision of Prof. Jagdeep
Kaur. Her expertise is in Molecular Biology, Protein Biochemistry and Animal Tissue
Culture.
e:
jashandhanoa89@gmail.comCharacterization of a probable GDSL lipase,
Rv1075c of Mycobacterium tuberculosis
Jashandeep Kaur
and
Jagdeep Kaur
Panjab University, India