Our main focus is on exploration and exploitation of India`s
rich microbial diversity (both cultivable and uncultivable)
for novel bioactive molecules and biocatalysts with application
in health, agriculture and industry, and their overproduction
using conventional and molecular biology tools. The group
is also working on development of new fungal expression
systems and diagnostic Microbiology.
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Development of microbial repository
from extreme environments and endophytes of endemic
plants |
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Bioprospecting and characterization
of novel endophytes and extremophiles for bioactives
and biocatalysts |
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Development of metagenomic libraries
from unexplored ecosystems for new/novel bioactives/biocatalysts
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Production of microbial enzyme and
high value microbial products from wild type and
genetically modified strains |
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Metabolic pathway engineering for
production of novel/ desired metabolites |
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Hyperexpression and protein engineering
of biocatalysts for their application in biotransformation
of important drug auxillaries/intermediates |
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Repository of psychrotrophic, mesophilic
and thermo-tolerant microorganism including bacteria,
actinomycetes, fungi and endophytes |
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Metagenomic libraries of environmental
samples from unique niches |
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High altitude medicinal plant gene
bank |
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Indigenous in vitro propagation
methods for many high value medicinal plants |
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Indigenous methods for rapid molecular
detection of several microbial pathogens |
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Indigenous novel expression system
for heterologous protein production in fungi |
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Hyperexpression and protein engineering
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Strain development for production
of bioactive molecules and novel enzymes using traditional
and molecular tools (genomics and proteomics) |
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| Sr. NO |
Name |
Expertise |
E-mail |
| 1. |
Sarojini Johri |
Microbial (enzymes) Biotechnology
|
sjohri@iiim.res.in
|
| 2. |
Meenu Katoch |
Microbial Biotechnology, Molecular
Biology, virology |
mkatoch@iiim.res.in |
| 3. |
Qazi Parvaiz Hassan |
Genetic Engineering and Molecular
Biology |
qphassan@iiim.res.in |
| 4. |
Rajinder Kumar |
DNA sequencing, maintenance of instruments
|
rkumar@iiim.res.in
|
| 5. |
Syed Riyaz-Ul-Hassan |
Microbiology, Molecular Biology, Diagnostics |
srhassan@iiim.res.in
|
| 6. |
Ashok Mehra |
Sectional stores and lab maintenance |
|
|
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Molecular and biochemical characterization
of bioactive molecules and enzymes from cultivable
microorganisms with special properties from extremophiles
(thermophiles, Psychrophiles, alkalophiles etc)
and endophytes |
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Characterization of cultivable and
non-cultivable microbial populations from extreme
environments |
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Screening and characterization of
bioactive molecules and new/novel enzymes from metagenomic
libraries constructed from environmental samples
from unique niches |
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Heterologous protein production in
filamentous fungi and use of novel promoters in
expression system |
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Mechanism of drug resistance in Salmonella,
particularly with reference to role of different
efflux pumps |
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Development of molecular methods for
culture-independent and rapid detection of food-borne
and clinical pathogens |
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Well equipped Microbiology laboratories
at IIIM, Jammu and branch laboratory Srinagar to
carryout cutting edge research on isolation and
characterization of microorganisms and their bioactive
metabolites |
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State-of-the-art facility to undertake
nucleic acid amplification, biomarker development,
gene cloning, heterologous gene expression, protein
purification, DNA sequencing and differential expression
studies |
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Mycology unit at Branch Laboratory
Srinagar undertaking collection, culture and identification
of mushrooms for characterization of bioactive molecules
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Experimental farms at various locations
in Jammu and Srinagar with a large collection of
medicinal and aromatic plants suitable for studying
plant-microbe interactions, endophytic, ectophytic
and other symbiotic associations |
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Well equipped plant tissue culture
Lab at Srinagar to carryout in vitro propagation
studies for high value medicinal plants |
Bioprospecting for endophytes and their bioactive
molecules
The project seeks to establish a repository of endophytes
and isolation and characterization of novel bioactive
agents. These organisms are obtained from specific areas
of the state in which there are areas of endemism, ethnobotanical
applications of plants and strong botanical biodiversity.
Our rationale is based on the fact that certain microbes,
selected from their natural plant ecological settings,
may provide new and useful leads in industrial, medicinal
and agricultural product discovery. The microbes are screened
for antimicrobial, anti-cancer and immune suppressant
activities. The endophytes are taxonomically characterized
with conventional and molecular phylogeny. Preference
is given to microbes representing unique texa. The bioactive
compounds are purified and characterized. Potent bioactive
microorganisms will be selected for metabolic engineering
of the polyketide pathway to produce new and desired molecules.
The project will lead to establishment of a repository
of endophytic microorganisms, isolation of new bioactive
molecules against infectious agents, cancer and inflammation.
It will also generate intellectual property and publications.
Characterization of Lovastatin Gene Cluster of Aspergillus
niger
Filamentous fungi produce many secondary metabolites with
complex chemical structure via the polyketide pathway.
Lovastatin and its derivatives are the most commercially
important polyketide drugs, with a market value of over
US$12 billion in 2007 (Broad Institute). Lovastatin can
be produced by Aspergillus terreus, and A. flavipes.
The biosynthetic pathway of lovastatin in A. terreus
has been investigated. A. niger is an important
industrial strain and given ‘Generally Regarded As Safe’
status. Recently its genome has been sequenced. Annotation
of genomic sequences suggested that it also has lovastatin
biosynthetic gene cluster. Because of the importance of
this molecule, gene cluster of lovastatin biosynthesis
will be characterized from A. niger and recombinant clones
expressing these proteins will be utilized for generating
the new analogues of lovastatin which can be exploited
for various bioactivity other than cholesterol lowering.
Novel Expression System for filamentous fungi
Filamentous fungi have ability to produce proteins
in large quantity (30g/l). To exploit this quality of
filamentous fungi and non-availability of commercial expression
system, an expression system was developed. Two promoters
(of glucose oxidase/catalase gene) from Aspergillus
niger strain were isolated and used in the development
of expression vector for expressing the heterologous proteins.
Under these promoters, expression of reporter proteins
was upregulated suggesting the efficient working of system.
Characterization of bioactive molecules and microbial
biocatalysts by metagenomic approach
Metagenomic libraries were constructed from soil sample
collected from Apharwat glacier (13,000 ft above sea level)
from North-west Himalayas and forest soil (Kupwara) of
Kashmir for characterization of new antifungal and antibacterial
compounds and novel enzymes. Identification of multifunctional
protease, phospholipase C and amylase from unculturables
was done by function driven approaches. Analysis of Microbial
(bacterial and archaeal) diversity in the soil samples
collected from Kupwara and Apharwat from Kashmir and saltpan
sediment of Mumbai was done using 16s ribosomal DNA sequencing.
Hyperexpression of genes encoding novel enzymes like carboxylesterase/lipases
(with broad substrate specificity and high enantioselectivity)
from culturable microbes was carried out. Protein engineering
(directed evolution/site directed mutagenesis) of these
enzymes for thermal stabilization and better catalytic
has been undertaken. Work is also being done on cloning
and hyperexpression of genes encoding highly thermostable
endocellulase from a salt tolerant mesophillic bacterium
and genes encoding cellobiohydrolase1 and glucose tolerant
ß-glucosidase from a biomass degrading fungal endophyte.
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Exploration of extremophiles/high altitude
medicinal plants from Kashmir valley for novel secondary
metabolites and biocatalysts
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Samples were collected from different geographical
high/low altitude locations of Kashmir valley and subjected
to isolation of extremophiles (psychrophiles, psychotrophs,
thermophiles, alkalophiles and acidophiles). Characterization
of one cold active and one thermostable stable lipase/cutinase
from two psychrotrophic bacterial strains IIIM3 and IIIM5
has been completed. These two enzymes are optimally active
both above and below the optimal growth temperature of
the microorganisms from which they were isolated. The
cloning & hyperexpression of genes encoding these enzymes
is underway.
We are also exploring high altitude medicinal plants
and their endophytes for bioactive molecules. Icariin
and Epimedin B two principal bioactive molecules from
Epimedium elatum have been isolated and characterized
so far. Our focus is study the pathways of these pharmaceutically
active secondary metabolites in extremophilic bacteria
/ fungi and high altitude medicinal plants and use of
genetic tools for their applications in human health,
agriculture etc.
Mechanism of Drug resistance in Salmonella
Salmonella spp. are facultative, intracellular
parasites that invade the mucous membrane, and are transmitted
to humans mainly through water and food. Salmonella infection
is the most frequent food-borne gastrointestinal disease
transmitted from animals to humans. It is estimated to
be responsible for the death of more than 500 people each
year, with costs of $1.1 billion to $1.5 billion annually
in the United States alone. The occurrence of resistance
to multiple antibiotics presents a serious problem in
the treatment of bacterial infections. Resistance can
be caused by a variety of mechanisms, active efflux of
the antimicrobial agents being one of the most important.
We are studying the role of known and putative efflux
pumps in conferring drug resistance to Salmonella
using molecular biology tools.
| |
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| Relative expression (9.5 fold) of AcrD in drug
resistant mutant of Salmonella typhimurium |
Molecular Diagnostics
Culture-independent Real-Time PCR assays were developed
for the detection of important pathogens in clinical and
food samples. The protocols were validated on the micro-PCR
system devised by the industrial partner, M/S Bigtec Pvt.
Ltd., Bangalore. The reaction mix was immobilized on the
PCR chip and real-time PCR tubes to simplify the whole
process.
Real-time PCR assays based on SYBR Green or Taqman chemistry
were developed for the detection of
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Salmonella spp. |
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Shigella spp. |
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E. coli |
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Shiga-toxic E. coli |
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Aspergillus flavus/A. parasiticus
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Hepatitis B |
A protocol was developed for the extraction of metagenomic
DNA from whole milk and subsequent detection of pathogens
by real-time PCR. 150 samples of raw milk were collected
from the local market and analyzed. All samples tested
positive for E. coli, nine samples for shiga toxin
producing E. coli (STEC) and 30 for Salmonella.
The validation and field application of these assays are
being carried out.
| |
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| Detection of Salmonella by real-time PCR |
PCR based Diagnosis of Cryptosporidium parvum
Cryptosporidium is a protozoan parasite responsible
for diarrhea to humans and animals. Cryptosporidiosis
is a zoonotic disease, transmits from animals to human
through food and water. In AIDS patients, diarrhea is
mainly because of this disease. It even becomes a cause
of death due to excessive dehydration. Diagnostics of
this parasite is a paramount issue. Different human and
animal samples were diagnosed for this disease using ELISA
and PCR. Identification of the species which would be
the main causative agent of human infection will lead
us to control the disease accordingly.
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Bio prospecting microbial strains
from North Western Himalayas (NWP006 CSIR Netwoking,
2007-2012) |
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Fractionation characterization and
molecular identification of SK-II Pitera (SSP0406
PNG-CSIR Project, 2007-2010) |
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Plant and microbial molecular biology
(Measure Lab Fund, 2007-2010) |
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Development of biocatalytic tool box
(2007-2010) |
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Bioprospecting of Epimedium elatum,
GAP-1108 (DBT, India, 2007-2010) |
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Bioprospecting of Melissa officinalis
(DST, 2008-2011) |
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