proforma i proforma for submission of...
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PROFORMA – I
PROFORMA FOR SUBMISSION OF PROJECT PROPOSALS ON RESEARCH
AND
DEVELOPMENT, PROGRAMME SUPPORT
PART I: GENERAL INFORMATION
1. Name of the Institute/University/Organisation submitting the Project Proposal:
Vivekananda Institute of Tropical Mycology (VINSTROM), Ramakrishna Mission
Vidyapith, Chennai-600004.
2. State: Tamil Nadu
3. Status of the Institute: Non-profit organization of the Ramakrishna Mission
4. Name and designation of the Executive Authority of the Institute/University
forwarding the application : Swami Shukadevananda, Chairman, Vivekananda
Institute of Tropical Mycology, Ramakrishna Mission Vidyapith, 45, Oliver Road,
Mylapore, Chennai 600 004.
5. Project Title : A study of the biodiversity and bioactive natural products of non-
sporulating fungi associated with mangroves and sponges of
Andaman Islands 6. Category of the Project : R&D/ Programme Support
7. Specific Area : 6.2 Marine Biology
8. Duration : 3 Years
9. Total Cost (Rs.) : Rs. 92,49,960-
10. Is the project Single Institutional or Multiple-Institutional (S/M) ? : M
11. If the project is multi-institutional, please furnish the following:
Name of Project Coordinator : Dr. T.S. Suryanarayanan
Affiliation : Director, VINSTROM
Address : Ramakrishna Mission Vidyapith
45, Oliver Road
Mylapore, Chennai 600 004.
12. Scope of application indicating anticipated product and processes
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For the first time, non-sporulating endophytes of marine habitat would be
explored for their diversity and production of novel antibiotics, antialgal, antifungal, ,
antioxidant and antimalarial compounds.
13. Project Summary
Non-sporulating endophytic fungi associated with mangrove trees (leaf endophytes,
bark phellophytes and root endophytes) and sponges of the Andaman Islands (Rifleman,
Manjeri, Wandoor, and Pluto Islands) will be isolated using suitable growth media and
culture conditions. Emphasis will be laid on isolation and culturing of non-sporulating
fungi since these have not received enough attention for their diversity and metabolic
potential. The fungi will be identified based on DNA sequences and maintained in our
culture collection. The species diversity, tissue and host specificity of mangrove
endophytes and sponge fungi will be worked out. The extracts of conditioned media from
these fungi will be screened for production of antibiotic, antialgal, antifungal,
antioxidant and antimalarial metabolites. Those extracts which appear to be promising
will be purified and chemically characterized. A collection of rare and bioactive
metabolites producing fungi will be maintained. Likewise a library of extracts of
candidate fungi will be maintained. Voucher specimens of fungi which produce novel
and interesting metabolites will be deposited in a culture collection centre.
The secondary metabolites extracted from the culture filtrates of the fungi would
be tested for antialgal, antibacterial and antifungal activities by bio-autogram and agar
diffusion methods. The antioxidants will be assayed by DPPH radical scavenging assay,
hydroxyl radical scavenging assay and total reducing power assay.
Antibiotic metabolites active against Gram positive and negative microorganism
will be assayed by Muellar-Hinton agar plates. Metabolites active against Plasmodium
will be assayed by SYBR Green fluorescence based assay of P. falciparum growth in
microtiter plate wells.
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Those extracts that appear promising would be subjected to activity guided
RPHPLC purification in fraction collection mode and their structure determined using
FTIR, MS, and NMR
PART II: PARTICULARS OF INVESTIGATORS
Principal Investigator:
14. Name : Dr. T.S. SURYANARAYANAN
Date of Birth : 25.07.1951 Sex: Male.
Designation : Director
Department : -
Institute/University : Vivekananda Institute of Tropical Mycology
Address : Ramakrishna Mission Vidyapith
45, Oliver Road, Mylapore,
Chennai PIN: 600 004.
Telephone: 9791186036 E-mail: [email protected]
Number of research projects being handled at present: 2
Co-Investigator – 1
15. Name : Dr. MUKESH DOBLE
Date of Birth : 16.09.1950 Sex: Male
Designation : Professor
Department : Department of Biotechnology
Institute/University : Indian Institute of Technology Madras
Address : Indian Institute of Technology Madras, Chennai
PIN : 600 036
Telephone : 91-44- 22574107 Fax 91-44-22574102 E-mail : [email protected]
Number of Research projects being handled at present: 2
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Co-Investigator – 2
Name : Dr. DINKAR SAHAL
Date of Birth : 18TH
JANUARY 1956 Sex: Male
Designation : STAFF RESEARCH SCIENTIST
Department : MALARIA
Institute/University : International Centre For Genetic Engineering And
Biotechnology,
Address : Aruna Asaf Ali Road, New Delhi 110067
Telephone : : 9871980190
E-mail : : [email protected], [email protected]
Number of Research projects being handled at present: 1
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PART III: TECHNICAL DETAILS OF PROJECT
16. Introduction
16.1 Origin of the proposal
Fungi associated with sponges are the single most prolific producers of novel
metabolites (Bugni and Ireland, 2004; Rateb and Ebel, 2011). Similarly, endophytic fungi
of mangroves are also recognized as abundant producers of bioactive compounds (Bugni
and Ireland, 2004; Raghukumar, 2008; Rateb and Ebel, 2011). Furthermore, sterile fungi
isolated as endophytes also produce novel bioactive compounds but have not received as
much attention as the sporulating endophytes (Hussain et al., 2009). The islands of
Andaman have several exclusive mangrove communities in isolated patches represented
by different mangrove and mangrove associates species. Marine sponges of different
species are also found here. It would be worthwhile to study the biodiversity and
metabolites profile of mangrove endophytes and sponge fungi of Andaman islands since:
(1) sponge-associate fungi are prolific producers of bioactive metabolites but have not
been studied adequately for this trait (Suryanarayanan, 2012).
(2) these two classes of fungi (the mangrove endophytes and sponge-associate fungi) are
the least studied in India for bioactive metabolites.
(3) non-sporulating, sterile fungi associated with sponges and mangroves have not been
studied for their species diversity and bioactive metabolites.
16.2 (a) Rationale of the study supported by cited literature
Most of the currently used antibiotics are derived from restricted molecular
frameworks whose effective lives have been stretched by synthetic modifications of the
molecules (Fischbach and Walsh, 2009). Prolonged and indiscriminate use of such
antibiotics with limited structural diversity has resulted in bacteria developing antibiotic
resistance genes (resistomes), culminating in the appearance and spread of multiple drug
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resistant (MDR) pathogens (Wright, 2007; Kumarasamy, 2010). To control MDR
pathogens, it is essential to look for new molecular structures with antibiotic properties.
Here, natural products research is imperative because combinatorial chemistry has fallen
short of our hyped expectations of it (Ortholand and Ganesan, 2004). Indeed, marine
invertebrates harbor in their tissues a series of microorganisms such as bacteria, and fungi
and, in some cases, associated micro-organisms may constitute up to 40% of the biomass,
this bacterial concentration exceeding that of the surrounding sea water by two or three
orders of magnitude. Although the real contribution of the microorganisms to the
secondary metabolism of marine invertebrates has not yet been fully evaluated,
essentially because of the difficulties encountered in culturing sponge-associated
microbes, it is generally accepted that these harbored microorganisms play a significant
role in the biosynthesis of the natural products isolated from the invertebrate (Fattorusso
(Fattorusso and Scafati 2009).
Sorbicillactone A was isolated from a strain of Penicillium chrysogenum cultured
from a sample of the Mediterranean sponge Ircinia fasciculata; it possesses a unique
bicyclic lactone structure, seemingly derived from sorbicillin. The compound exhibits
promising activities in several mammalian and viral test systems, in particular a highly
selective cytostatic activity against murine leukemic lymphoblasts (L5178y) and the
ability to protect human T cells against the cytopathic effects of HIV-1(Bringmann and
Muller 2003) .
Cyclosporin A (CsA) (a hydrophobic cyclic peptide produced by a fungus) is widely used
as an immunosuppressive agent to inhibit the rejection of transplanted organs Azouzi et
al. 2010.
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In this context, screening microbes from unexplored and little-studied ecological
niches for novel metabolites appears to be a promising exercise (Fischbach and Walsh,
2009). Among such organisms marine fungi associated with sponges, corals, seaweeds,
mangroves, marine detritus, tunicates, fish, seagrasses, marine sediments, etc. appear to
be prolific producers of molecules of unprecedented structures with potential
pharmaceutical importance (Bugni and Ireland, 2004; Schulz et al., 2008; Raghukumar,
2008; Kjer et al., 2009; Suryanarayanan, 2010b). In addition, the marine derived fungi,
especially from mangroves and sponges, produce novel metabolites with unusual
bioactivites thus making them choice candidates for bioprospecting (Rateb and Ebel ,
2011).
Marine fungi represent a source of fascinating and significant chemical diversity.
So far, more than 1000 novel compounds have been described from marine fungi and
current studies show that they are still a relatively unexplored group of organisms for
novel biochemicals (Rateb and Ebel, 2011) (Fig. 1).
Fig. 1. Types of novel metabolites so far obtained from marine-derived fungi.
In the recent years, marine derived fungi associated with mangrove trees and
sponges have been the subject of intense study as they produce bioactive metabolites of
M. E. Rateb and R. Ebel . 2011.
Secondary metabolites of fungi
from marine habitats
Nat. Prod. Rep., , 28, 290–344
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novel and even unprecedented molecular architectures (Proksch et al., 2008; Höller et al.,
2000). In a broad-based assay of fungi from sponges, Höller et al. (2000) isolated several
novel compounds with unique bioactivities; that were found to inhibit malarial parasite,
tuberculosis bacterium and HIV-1 reverse transcriptase. They therefore concluded that
sponge-associated fungi are a good source of novel bioactive metabolites. This study was
followed by those of Edrada et al. (2002); Lin et al (2003) which confirmed that the
biosynthetic potential of sponge associated fungi is higher than that of their terrestrial
conspecifics. In the recent years, these findings were bolstered by the studies of Proksch
et al (2008) and Liu et al (2011).
Similarly, endophytic fungi which reside inside the living tissues of mangrove
trees without expressing any disease symptoms are also well known for their heightened
synthetic activity (Kumar and Hyde, 2004; U‟Ren et al., 2012). Kjer et al (2009) isolated
14 different natural products from an Alternaria sp. that was endophytic in the mangrove
tree Sonnretia alba. Aly et al (2008) identified a protein kinase inhibitor from a
mangrove endophyte. Lopanick et al (2004) and Mitchell et al (2008) identified several
novel metabolites from endophytes. Three unique metabolites, namely, phomopsis-H76
A (1), B (2), and C (3), were produced by Phomopsis sp. which is a mangrove endophyte
(Yang et al., 2010). Bugni and Ireland (2004) posit that mangrove endophytes are an
untapped source of novel bioactive chemicals. In this context, sterile endophytic fungi
(those that do not sporulate in culture) are also an unexplored source for novel chemicals
(Vinale et al., 2010). For instance Vinale et al (2010) extracted veratryl alcohol (VA) and
4-(hydroxymethyl)-quinoline which showed antibiotic and antifungal effects from a
sterile endophytic fungus. Similarly, Maria et al (2005) showed that a sterile endophyte
from a mangrove produced novel antibiotics that was active against several bacteria.
However, there are very few studies in the tropics on non-sporulating marine derived
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fungi of sponges and endophytes of mangroves with regard to their diversity and
synthetic ability. There is hardly any study on the bioactive metabolite of these ecological
groups of fungi from India.
(b) Hypothesis:
Fungi associated with marine sponges and endophytic fungi of mangrove trees are
metabolically more innovative than their terrestrial counterparts. Together, these two
sources of fungi account for most of the novel metabolites described from marine habitat
Fig. 2; Rateb and Ebel, 2011).
Fig. 2. Contribution by different ecological groups of marine fungi to the tally of novel
metabolites.
This is perhaps an outcome of their constant need to interact with their host milieu.
Endophyte–plant host interactions are different from pathogen–plant host interactions
since neither associate really „wins‟; neither disease symptoms develop on the plant host
nor is the fungus eliminated by the plant host (Pinto et al., 2000; Schulz et al., 2002;
Stone et al., 2004; Sieber, 2007; Saikkonen, 2007). This situation entails sustained and
prolonged reactions against the defense mechanisms of the host by the endophyte and this
M. E. Rateb and R. Ebel . 2011.
Secondary metabolites of fungi
from marine habitats
Nat. Prod. Rep., , 28, 290–344
10
could act as selection pressure for developing novel metabolic pathways – a potentially
beneficial situation for bioprospectors (Weber et al., 2007; Suryanarayanan et al., 2010a).
We feel that many novel bioactive compounds can be isolated from sponge fungi and
mangrove endophytes of Andaman Islands since they support rare and unique organisms
which evolved thorough millions of years owing to the physical separation between the
islands and from the main lands.
(c) Key questions:
What non-sporulating species of fungi occur as endophytes in the mangroves and
what fungi are associated with the marine sponges of the Andaman Islands ?
What is the chemical diversity of these endophytes and sponge fungi?
What antibiotic, antialgal, antifungal, insecticidal, antioxidant and antimalarial
metabolites are produced by these fungi?
16.5 Current status of research and development in the subject
Filamentous fungi are revealed only by actinomycetes and plants in producing
diverse types of secondary metabolites showing varied biological activities. Among the
fungi, those occurring inside the tissues of sponges which are either symbionts or casual
residents (acquired by the sponge in the process of ingestion - Baker et al., 2009) are
unique in producing novel metabolites (Hirot et al., 2004; Suryanarayanan 2012).
Interestingly, unlike the bacteria associated with marine sponges, the fungi produce
metabolites different from those of their sponge hosts (König et al., 2006). In the recent
years, there has been a considerable increase in the studies on microbes associated with
sponges (Fig 3). Ding et al (2010) isolated strong antimicrobial compounds from
Nigrospora sp. associated with the sponge Clathrina luteoculcitella occurring in the
South China Sea. Aspergillus versicolor, another fungus isolated from South China Sea
sponge elaborates a peptide cyclo (L-Trp-L-Phe) having both agricultural and
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pharmaceutical potential (Chu et al., 2011). Another A. versicolor isolated from a marine
sponge produces a novel aromatic polyketide derivative, two xanthones, and five
anthraquinones (Lee et al. 2010). Rateb et al (2010) reported three hitherto undescribed
dibenzofurans from an ascomycete isolated from a Fijian marine sponge; these
compounds inhibited the growth of many bacterial species and the activity of tyrosine
kinase. Proksch et al (2008) isolated 81 fungal strains belonging to 20 different genera
from the sponge Suberites domuncula; from these a total of 19 different fungal
metabolites, including three new natural products were characterized. Li et al (2011)
reported the production of new hexylitaconic acid derivatives by a marine sponge-derived
Penicillium sp. A sponge associated Acremonium sp. elaborates new sesquiterpenoids
with anti inflammatory properties (Zhang et al., 2009). Hao et al (2008) obtained several
new polyketide-originated metabolites from a mycelia sterilia fungus isolated from
sponge. Aspergillus aculeatus¸asssociated with the sponge Xestospongia testudinaria,
elaborates Aspergillusol A, an r-Glucosidase Inhibitor (Ingavat et al., 2009).
Trichopyrone showing antioxidant activity was isolated from Trichoderma viride
associated with the Caribbean sponge Agelas dispar (Abdel-Lateff et al., 2009). Liu et al
(2009) extracted seven new drimane sesquiterpenoids along with the known compounds
deoxyuvidin B, strobilactone from cultures of the fungus Aspergillus ustus, which was
isolated from the marine sponge Suberites domuncula. The compounds showed cytotoxic
activity against a panel of tumor cell lines, including L5178Y, HeLa, and PC12 cells.
Suryanarayanan (2012) in his recent review states that our knowledge of the diversity and
metabolites of sponge-associated fungi is inchoate.
With reference to mangrove endophytes, although many have been isolated
(Kumaresan et al., 2002), their synthetic potential is being recognized only recently
(Raghukumar, 2008). Huang et al (2010) isolated three new dimeric naphtho-γ-pyrones,
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named rubasperone A, rubasperone B and rubasperone C from the mangrove
endophytic fungus Aspergillus tubingensis; these were active against tyrosinase and α-
glucosidase. Maria et al (2005) showed that the culture extracts of mangrove endophytes
inhibit several Gram positive and Gram negative bacteria. Chaeprasert et al. (2010)
isolated several metabolites from endophytes of 10 mangrove species in Thailand and
reported the following: The antimicrobial potential of 71 endophytic fungi isolated from
mangrove plants towards selected bacteria (Bacillus subtilis, Pseudomonas aeruginosa,
Escherichia coli and Staphylococcus aureus) was tested using ethyl acetate extracts of
fungi cultivated under static conditions. All test bacteria were inhibited by a
Cladosporium sp. isolated from the leaves of T. populneoides and an endophytic Xylaria
sp. 1 isolated from A. ilicifolius leaves caused considerable inhibition to Gram-positive
and Gram-negative bacteria. Additionally, the crude extracts of 84 endophytic fungi were
tested for anticancer activities by the MTT assay against A375 (human malignant
melanoma), SW620 (human colorectal adenocarcinoma), Kato III (human gastric
carcinoma), HepG2 (human liver hepatoblastoma) and Jurkat (human acute T cell
leukemia). Most extracts had cytotoxicity against some cancer cell lines.
Iwatsuki and Omura (2011) isolated five tropolone compounds from the culture broth of
Penicillium sp. FKI-4410. Among them, puberulic acid exhibited potent antimalarial
inhibition, with IC50 values of 0.01μg/ml against chloroquine-sensitive and resistant
Plasmodium falciparum strains with low cytotoxicity (IC50 : 57.2μg/ml) in vitro.
Butyrolactones isolated from the fungus Aspergillus terreus BCC 4651 showed
antiplasmodial and antimycobacterial activity ( Haritakun and Isaka 2010). Kasettrathat
and Kittakoop (2008) showed that tetronic acid and nodulisporacid A isolated from a
marine-derived fungus Nodulisporium sp. CRIF1 from a Thai soft coral, was moderately
antiplasmodial (IC50:1–10 μM) against chloroquine-resistant P. falciparum strain 94.
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Pontius and Konig (2008) isolated a heterocyclic substituted xanthone, chaetoxanthone B
from cultures of a marine derived fungus Chaetomium sp. that showed selective activity
towards P. falciparum K1 strain (IC50: 0.5 μg/mL). Wright and Lang-Unnasch (2005)
reported that pycnidione isolated from the marine fungus Phoma sp., had significant
antiplasmodial activity against three strains of Plasmodium falciparum (IC50: 0.15–0.4
μM)
Increasing research interest in marine sponge-microorganism associations. Number of publications retrieved from the
ISI Web of Science database..
16.6 The relevance and expected outcome of the proposed study
Though marine fungi from different substrates are known to be a good source of
bioactive compounds (Bugni and Ireland,2004; Raghukumar 2008), the symptomless
endophytic fungi associated with mangroves and marine sponges have not been studied
in detail for such chemicals in our country. Hence, this proposal is relevant and
pioneering in nature.
Taylor, M.W., Radax, R., Steger, D. &
Wagner, M. 2007. Sponge-Associated
Microorganisms: Evolution, Ecology,
and Biotechnological Potential.
MICROBIOLOGY AND
MOLECULAR BIOLOGY REVIEWS,
June 2007, p. 295–347.
Fig. 3.
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Expected outcome
Novel metabolites of pharmaceutical importance
Identification of potential producers of antibacterial, antifungal antialgal,
antiinsect, antioxidants and antimalarial chemicals.
Biodiversity and ecology of endophytes of mangroves and fungi of sponges of the
Andaman Islands – this study will be the first of its kind for Andaman Islands.
A collection of cultures of non-sporulating fungi associated with marine sponges
and mangroves of Andaman Islands
16.7 Preliminary work done so far
The Principal Investigator‟s lab is the first to have studied endophytic fungi of mangrove
plants (Suryanarayanan et al., 1998). This paper has been cited by many workers. Hyde
and Soytong (2008) in their review state that our lab has made significicant contribution
to knowledge of tropical endophytes. The principal investigator‟s lab is also the first to
isolate endophytes from seagrasses (Devarajan et al., 2002). The principal investigator‟s
lab is specializing in techniques that help isolating, purifying and identifying fungal
endophytes (Suryanarayanan et al., 2002 – cited in Ganley RJ, Brunsfeld SJ and
Newcombe G (2004) A community of unknown, endophytic fungi in western white pine.
Proceedings of the National Academy of Sciences USA 101: 10107-10112;
Suryanaryanan et al., 2003 – cited in Clay K (2004) Fungi and the food of the Gods.
Nature 427: 401-402). It is also engaged in screening of endophytes for antibacterial,
antialgal and antifungal as well as antimitotic properties (under two Indo-German
projects sanctioned by the DBT). We have isolated several bioactive compounds in the
DBT-funded Indo-German project (Suryanarayanan et al., 2009); these include
cytochalasins, enniatins, apicidin, aphidicolin, etc (Suryanarayanan et al., 2009). In a
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preliminary study, we found that the culture extract of a marine fungus isolated from a
marine sponge off Mandapam coast had metabolites that inhibited cancer cell lines
(Suryanarayanan et al., 2010a).
In a collaborative research by the PI and Co-PI 1 that is funded by the DBT, it has
been shown that the fungi associated with seaweeds produce several novel antibiotics and
antioxidants (Suryanarayanan et al., 2010b). We also reported the production of novel
macrolides by Curvularia lunata, a fungus isolated from a seaweed (Geetha et al., 2011).
In a collaborative research funded by the DBT project “Pharmaceuticals from
marine and marine derived fungi associated with seagrasses and seaweeds of Tamil Nadu
coast”, the PI and co PI have studied the fungal associates of marine sponges occurring
along the coast of Rameswaram Southern India. A paper describing the results of this
study (including the diversity and distribution of sponge fungi and their bioactive
compounds) has been published in Fungal Diversity (Thirunavukkarasu et al., 2012).
The PI has published a paper ("L-asparaginase from marine derived fungal endophytes of
seaweeds", 2011, Mycosphere, Online Journal of Fungal Biology - ISSN 2077 7019)
on the anti neoplastic enzyme of fungi associated with marine algae. Currently, about
100 culture extracts obtained from these fungi by the PI are being screened for anti
malarial activity in Co-PI 2's lab.
Contibution by Co- Principle Investigator – 1
Currently completed a DBT sponsored 3 year project titled “Anti-TB compounds from
marine actinomycetes” in collaboration with Tuberculosis research centre, Chennai. A
novel metabolite was identified and its structure was elucidated using analytical tools. A
patent has been filed for the novel compound which shows anti-TB and anti –HIV
activity (New Antituberculosis antibiotic from marine actinomycetes strain R2,
247/DEL/2011, February 2, 2011). So the co-PI 1 has extensive experience in isolating
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metabolites from natural sources using chromatographic and determining the structures
using several analytical tools.
Contribution by Co- Principle Investigator – 2
The CO-PI 2‟s laboratory at the ICGEB, New Delhi has deep interest and wide
experience in activity guided purification of molecules and methods of screening crude
extracts and purified molecules against Plasmodium falciparum strains in human red
blood cell culture. The screening methods include SYBR Green fluorescence dependent
microtiter plate HTP assay and Giemsa stain based microscopy based assays. The former
is useful for HTP screening and the latter is vital for firm validation and mechanistic
studies (using fluorescent dyes like Hoechst 33342 and JC-1). Both assays are in frequent
use in the laboratory. His laboratory is also equipped with MTT based assays for
quantitative estimation of mammalian cell cytotoxicity. Selectivity of molecules is thus
assessed through Selectivity index which is the ratio of IC50
(P.falciparum)/IC50(Mammalian cells). .
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17. Specific objectives
Determination of the distribution, diversity, host and tissue restriction of non
sporulating fungal endophytes and of non sporulating sponge fungi of Andaman
islands
Creation of a genetic resource of these unique fungi for use in bioprospecting by
other labs
Identification of fungal metabolites which are anti algal, antifungal, antibacterial,
antimalarial and antioxidant in nature
Chemical characterization of bioactive molecule isolated from endophytes and
sponge fungi
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18. Work Plan:
PI’s Lab:
1. Isolation of endophytes from mangroves (various tissues and various species)
2. Isolation of sponge – associated fungi.
3. 3. Identification of the fungi
4. Maintaining pure cultures of the isolated fungi
5. Extraction of metabolites and testing for anti algal and antifungal
metabolites
6. Extracts sent to CoPI 1 and CoPI 2 labs
CoPI 1’s Lab:
1. Test extracts for anti oxidants
2. Test extracts for antibiotics
CoPI 2’s Lab:
1. Test extracts for anti plasmodial activity
2. Antiplasmodial Molecule Isolation
Extracts which show promising activities will be subjected to the following detailed
study. Fungi which produce promising metabolites would be grown in large
quantities (PI 1’s lab) and extracts will be given to CO PI 1 and CO PI 2’s Labs for
separation and isolation. These isolated compounds would be tested again for their
activity and finally chemically catheterized by Co PI 1 and CO PI 2’s labs.
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Identification( by culture criteria &
PCR and gene sequencing, followed
by BLAST search) &
Fig. 2. Bioassays for detecting secondary metabolites from fungi
Antialgal assay
Antifungal assay
Culture filtrate extracted with
hexane & butanolConcentrated
(30 ml in to 1.5 ml fungal extracts)
Sponge/Mangrove
tissue
Isolation &
purification on
PDA
Cultivation in seawater
medium, 21 days
PI’s Lab
Antibiotics Assay
Antioxidants assay
Co PI 1’s Lab
Antimalarial assay
Co PI 2’s Lab
AntialgalAntifungal
Identification( by culture criteria &
PCR and gene sequencing, followed
by BLAST search)
Fig. 2. Bioassays for detecting secondary metabolites from fungi
Antialgal assay
Antifungal assay
Culture filtrate extracted with
hexane & butanolMetabolites will be eluted using methanol,
methanol evaporated in preweighed tubes
and residue subjected to quantitative
potency determination assays listed below
Sponge/Mangrove
tissue
Isolation &
purification on
PDA
Cultivation in seawater
medium (containing the resin
XAD for adsorption of
secondary metabolites), 21
days
PI’s Lab
Antibiotics Assay
Antioxidants assay
Co PI 1’s Lab
Antimalarial assay
Co PI 2’s Lab
XAD samples will be sent by Dr.Suryanarayanan
To Dr. Dhoble and Dr. Sahal
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Promising fungi will be cultivated Broth will be subjected to
on large scale in fermentor water butanol partition
XAD
NO
Antifungal Antibiotic Antioxidant Antialgal Antiplasmodial XAD 1 XAD 2 XAD 3 XAD 4 XAD 5 XAD 6
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Large-scale fermentation of
promising strains on rice medium
(Based on chemical
screening/bioactivity) PI’s lab
Solvent extraction, purification
confirmation of activity by bioassay
(PI’s lab and Co-PI 1 & 2’s lab)
Fractionation and structure elucidation
(CoPI 1 and 2’s labs). The fungal extracts
which show many bioactivities will be further
fractionated in PI 1 and PI 2 labs and tested in
all the 3 labs to identify the active fractions
and metabolites.
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18.1 Work plan
Figure 4 gives the summary of the work plan to be carried out in the PI and CoPI
1 and 2‟s labs.
Work in PI’s Lab:
A. Leaf, bark and root tissues of mangroves and sponge samples occurring along
Rifle man, Manjeri, Wandoor and Plueto Islands of Andaman will be collected
and sampled for isolation of associated endophytes and fungi.
B. The samples will be processed within 48 hours of collection from mangroves will
be isolated using appropriate surface sterilization method (depending on the
texture of the tissue). Sponge samples would be comminuted in sterile sea water
and spread over suitable agar media to isolate associated fungi.
C. Fungi will be isolated, purified and identified based on culture and DNA
sequences. Cultures will be maintained in VINSTROM.
D. Using the data collected, ecological parameters such as species diversity,
colonization frequency, similarity co-efficient, etc. will be completed.
E. This study will throw light on endophyte assemblages of Andaman mangroves
and fungal assemblages of Andaman sponges.
F. Fungi would be grown in sea water amended liquid media and the culture filtrates
will be extracted using suitable solvents. The solvents will be assayed for anti
algal, anti fungal, and anti insect compounds by auto-biogram and maggot assay.
G. A library of extracts of candidate fungi will be maintained. Voucher specimens of
fungi which produce novel and interesting metabolites will be deposited in a
culture collection centre.
Co-PI’1 lab –IIT
Antioxidants assay including (1) DPPH radical scavenging assay, (2) hydroxyl
radical scavenging assay and (3) total reducing power assay will be performed.
Antibiotic activity against Gram positive and negative microorganism will be
performed using Serial dilution MIC method.
The promising compound (s) will be purified using prep TLC, and column
chromatography. Then the structures will be determined with FTIR, GC-MS, 1H
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NMR, and 13C NMR. The novelty of the structures will be ascertained with
SciFinder data base.
ikCo-PI’2 lab - ICGEB
Following work will be done at Malaria Research Lab, ICGEB New Delhi
1) Primary antimalarial screening : Fungal extract provided by PI lab will be screened
for antiplasmodial activity using SYBR Green fluorescence based assay of P. falciparum
growth in microtiter plate wells in vitro. In order to check parasite growth inhibition by
the above extracts, malaria parasite will be grown in vitro (Trager and Jensen1976).
Chloroquine sensitive (3D7) and resistant (INDO) strains of P. falciparum will be used in
culture. The parasite would be cultured in RPMI 1640 supplemented with HEPES,
sodium bicarbonate, gentamycin, 10% human serum or albumax and human erythrocytes.
The parasite will be grown in plates incubated at 370C in carbon dioxide incubator. The
parasitemia would be monitored by microscopic examination of blood smears after
staining with Giemsa stain. For various studies the parasite cultures would be prepared
after Sorbitol mediated synchronization to ring stage.
In vitro drug susceptibility testing in P. falciparum will be determined by a SYBR
Green I-based fluorescence method described previously by Smilkstein and Riscoe
(2004). Stock solutions of each test drug will be prepared in DMSO/ sterile distilled
water. The 50% inhibitory concentration (IC50) will be determined by analysis of dose-
response curves.
2) PURIFICATION OF ANTI-PLASMODIAL MOLECULES
Purification of molecules from active fungal extracts will be done by solvent
extraction followed by reverse phase HPLC using UV dual wavelength and ELSD
24
detection. Antiplasmodial molecules will be identified in the backdrop of bioassay guided
fractionation.
3) Determination of Therapeutic Potential of Purified Antiplasmodial Molecules:
Therapeutic index is a ratio of toxic concentration (TC50) for host to inhibitory
concentration (IC50) for the pathogen. Therapeutic indices will be determined by
measuring the cytotoxicity of test molecules on mammalian cells. Animal cell lines
(HeLa and fibroblast) will be used to determine drug toxicity by using MTT assay for
mammalian cell viability as described by Mosmann (1983).
4) STRUCTURAL ANALYSIS OF ACTIVE MOLECULES:
Structure determination will help in identification and chemical synthesis of nature
derived antiplasmodial molecules. This way we can use nature`s molecules without the
need to perturb ecological equilibrium when the drugs are required in huge amount.
Structural characterization of active molecules will be done using Spectroscopic tools
including Mass Spectrometry, UV-Visible Spectrometry, FT-Infrared Spectrometry, CD
Spectrometry and Nuclear Magnetic Resonance. Our laboratories are well equipped with
these tools that will facilitate structure determination.
5) STUDYING DRUG IN ACTION:
i) In vitro stage dependence of action: Stage of parasite life cycle on which
molecules are acting will be determined by microscopic analysis of drug effect on
different stages (e.g. ring, trophozoite, Schizont) of parasite. Cultures at the ring/troph/
schizont stages will be seeded in 96-well plates. The test compounds will be added to
25
parasite cultures at different stages and drugs will be removed after 6h (Schizonts), 24h
(Trophozoites) , and 48 h (rings) by spinning and washing with complete medium. The
washed cells will be further incubated for 48h under standard drug free culture condition.
Slides will be made and analyzed microscopically. Stage specificity of action will be
assessed by observing the stage transitions in drug treated samples against control.
Parasite invasion inhibition will be examined by action on schizont and formation of ring
as index of invasion in fresh red cells.
ii) Drug Action In-vivo
Plasmodium species that cause human disease are essentially unable to infect non-
primate animal models. So, in vivo evaluation of anti-malarial compounds typically
begins with the use of rodent malaria parasites. In-vitro purified potent antimalarial
molecules will be used to analyze their potential in-vivo using rodent malaria parasite P.
berghei / P. yoelii (Peters and Warhurst, 1977). Briefly, groups of four to five mice will
be intravenously inoculated with 5 x106 P. berghei / P. yoelii infected erythrocytes and
treatment will be initiated when parasitemia has reached 1-3% (Day 0). Drugs will be
administered once-daily on days 0, 1, and 2. Blood drop will be collected 24 h after each
treatment and the smears will be analyzed microscopically. Results will be expressed as
the dose reducing parasitemia to 50% (ED50), 75% (ED75), and 90% (ED90). The
animals will be considered cured if they survived 30 days after the infection without any
detectable parasite.
i) Studying Synergy in Drug Action.
To determine the potential of drugs in combination therapy, action in synergy with
known drugs like chloroquine and artemisinin will be assessed by isobologram analysis
26
using fixed-ratio combination. Briefly, drugs will be diluted in fixed ratios of sub
inhibitory starting concentrations to generate well-defined concentration response
curves. The FICs (Fractional Inhibitory Concentrations) will be calculated by the
following formula: FIC (A) = IC50 of drug A in combination/ IC50 of drug A alone; FIC
(B) = IC50 of drug B in combination/ IC50 of drug B alone; FIC index = FIC (A) +FIC
(B). The isobolograms will be constructed by plotting a pair of FICs for each
combination of drug and the selected compound. An interpretation of a straight diagonal
line (FIC index = 1) on the isobologram indicates an additive effect between the two
drugs. A concave curve below the diagonal line (FIC index < 1) indicates synergy of the
combination, while a convex curve above the diagonal line (FIC index > 1) indicates
antagonism (Kelly and Rescoe, 2009).
18.2 Connectivity of the participating institutions and investigators
The PI and Co-PI ‟1 (IIT, Chennai) have already collaborated in a previous DBT
funded marine biotechnology project to study pharmaceuticals from marine fungi
(BT/PR10169/AAQ/03/376/2007). The PI is already collaborating with Co PI ‟2 (Dr.
Dinkar Sahal, ICGEB, New Delhi) in a preliminary study that screens marine fungal
metabolites for antimicrobial compounds.
18.3 Alternate strategies
Preliminary investigations in the PI‟s lab have shown that sponge fungi and
mangrove endophytes are good sources of novel metabolites. Biodiversity of marine
fungi associated with sponges and mangroves of Andaman Island will be worked out. A
culture collection of this rare fungi will be available for further work. A library of
bioactive metabolites will be available for further development on the path to their
transformations into drugs for clinical use..
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33
19. Timelines:
Period of study Achievable targets
6 months Procuring Instruments, appointing JRF. 2 collection trips, isolation of
fungi; 2 collection trips to Rifle man Islands of Andaman, isolation of
fungi, characterization of fungi using r-DNA sequences
12 months 3 collection trips to Rifle man, Manjeri Islands of Andaman, isolation
of fungi, characterization of fungi, screening for antialgal, antifungal,
antibacterial, insecticidal, antioxidant and antimalarial activities.
Isolation, purification of interesting compounds.
18 months 2 collection trips to Andaman islands (Wandoor Islands). Isolation of
fungi, characterization of fungi, screening for antialgal, antifungal,
antibacterial, insecticidal and antioxidant activities. More isolation
and continuation of search for the above metabolites. Isolation,
purification of interesting compounds. Computation of ecological
parameters of mangrove endophytes. Chemical characterization of
metabolites. Primary Bioassay Guided fractionation of active extracts.
24 months 2 collection trips to Andaman islands (Wandoor and Plueto Islands).
Isolation of fungi, characterization of fungi, screening for antialgal,
antifungal, antibacterial, insecticidal antioxidant and antimalarial
activities. A culture collection as a result of these collections.
Identification of the chemicals that inhibit microbes. Computation of
ecological parameters of sponge associated fungi. Large Scale
purification of identified metabolites
34
30 months 2 collection trips to Andaman islands (Wandoor and Plueto Islands).
Isolation of fungi, characterization of fungi, screening for antialgal,
antifungal, antibacterial, insecticidal antioxidant and antimalarial
activities. Cultures of interesting fungi, and extracted metabolites to
be placed in VINSTROM, IIT and ICGEB. Voucher specimens to be
deposited in a National Culture Collection centre. Large Scale
purification of identified metabolites, Cytotoxicity, In vivo drug
efficacy study of purified molecules, Structural characterization of
active molecules.
36 months Preparation of final report, papers for publication and filing of patents.
Large Scale purification of identified metabolites, Cytotoxicity, In
vivo drug efficacy study of purified molecules, Structural
characterization of active molecules.
20. Name and address of 5 experts in the field:
Sl.
No.
Name Designation Address
1 Prof. V. K. Bhasin Professor Department of Zoology, Delhi
University, India
2 Prof . Santosh K. Kar Professor Centre for Biotechnology,
Jawaharlal Nehru University, New
Delhi, India
3 Prof. Aparna Dixit Professor Centre for Biotechnology,
Jawaharlal Nehru University, New
Delhi, India
4 Dr. Peter Proksch Professor Institute for Pharmaceutical
Biology & Biotechnology,
Heinrich-Heine-Universität
Duesseldorf, Duesseldorf,
Germany
5 Dr. Florenz Sasse Professor Helmholtz Centre for Infection
Research, Department of Chemical
Biology, Braunschweig, Germany
35
Product Development:
Products in the form of drugs or drug leads against Malaria, fungal infections, anti
insects, anti algal and antioxidants are likely to be discovered in the project.
36
PART IV: BUDGET PARTICULARS (Principal Investigator - VINSTROM)
Budget (In Rupees)
A. Non-Recurring
Sl. No. Item Year 1 Year 2 Year 3 Total
1.
2.
Horizontal
autoclave (50 L
capacity)
ELISA Reader
40,000
8,00,000
- - 8,40,000
Sub-Total (A) =8,40,000
B. Recurring
B.1 Manpower
Sl.
No.
Position No. Consolidated
Emolument
Year 1 Year 2 Year 3 Total
1. JRF Rs 12000 pm
+30%
HRA(Rs.
14000 pm
+30% HRA-
third year
only)
1,87,200 1,87,200 2,18,400 5,92,800
Sub-Total (B.1) = 5,92,800
B.2 Consumables
S.
No.
Item
Quantity Year 1 Year 2 Year 3 Total
1. Glassware,
Chemicals for
media, HPLC,
TLC plates, etc.
1,50,000 1,50,000 1,50,000 4,50,000
Sub-Total (B.2) = 4,50,000
Other items Consolidated
Emolument
Year 1 Year 2 Year 3 Total
B.3 Travel
35,000 25,000 35,000 95,000
B.4 Contingency
50,000 50,000 50,000 1,50,000
B.5 Overhead
(20%)
84,440 82,440 90,680 2,57,560
37
Sub-total of B
(B.1+B.2+B.3+B.4+B.5)
15,45,360
Grand Total (A + B) 23,85,360
1. A JRF is needed for collecting samples, isolating endophytes and to culture
them as well as to screen them for metabolites.
2. An ELISA Reader is essential for rapid screening of the hundreds of isolates
that we expect and to identify the candidate fungi for further study.
3. Chemicals (high grade) are needed to obtain pure culture of the fungi;
solvents are needed to extract secondary metabolites from mycelia and
culture extracts.
38
PART IV: BUDGET PARTICULARS (Co-Investigator - 1 & 2 from IIT
Madras and ICGEB)
Budget : IIT
A. Non-Recurring
Sl. No. Item Year 1 Year 2 Year 3 Total
1. - - - - -
2. - - - - -
Sub-Total (A) =
Recurring
B.1 Manpower
Sl.
No.
Position
No.
Consolidated
Emolument
Year 1 Year 2 Year 3 Total
1. JRF Rs 12000 pm
+30% HRA
1,87,200 1,87,200 1,87,200 5,61,600
Sub-Total (B.1) = 5,61,600
B.2 Consumables
Sl. No. Year 1 Year 2 Year 3 Total
Chemicals/solvents/glass
ware/plastic ware
2,00,000 2,00,000 2,00,000 6,00,000
Sub-Total (B.2) = 6,00,000
Other items. Year 1 Year 2 Year 3 Total
B. 3 Travel 30,000 30,000 30,000 90,000
B.4 Contingency 75,000 75,000 75,000 2,25,000
B.5Overhead (20%) 98,440 98,440 98,440 2,95,320
Sub-total of B
39
(B.1+B.2+B.3+B.4+B.5)
Grand Total (A+B) 5,90,640 5,90,640 5,90,640 17,71,920
1. A JRF (with Biochemistry background)is needed to do the activity studies and
structure elucidation
2. We require plenty of solvents for column purification hence the consumable
budget will be high
3. We have to carry out LC-MS and CHN analysis on payment basis, hence require
contingency
Budget : ICGEB
B. Non-Recurring
Sl. No. Item Year 1 Year 2 Year 3 Total
1. SPD-M20A-
PDA Detector
To aid in
purification of
coeluting
molecules that
are spectrally
distinct.
~ 600000 ~ 600000
2. Multiple sample Parallel Evaporation device: Syncore Syncore comprises two instruments and a broad range of accessories suitable for all aspects of multiple sample processing.Its design concept allows one unit to perform as a parallel evaporator (Polyvap) which can elegantly be converted to a parallel concentrator (Analyst). Thus its applications include fast, parallel evaporation of multiple samples, the smooth concentration to predefined residual volumes and sample preparation and purification with SPE.
2200000 2200000
40
Sub-Total (A) = ~ 2800000 (Subject to quotation)
Recurring
B.1 Manpower
Sl.
No.
Position
No.
Consolidated
Emolument
Year 1 Year 2 Year 3 Total
1. JRF/SRF 16000/18000
+ 30% HRA
280800 280800 280800 842400
Sub-Total (B1) = 842400
B.2 Consumables
Sl. No. Item Quantity Year 1 Year 2 Year 3 Total
Biological
reagents,
media,
Fluorescent
dyes,
Solvents,
Glass ware
Purchase
and
maintenance
of mice for
in vivo
antimalarial
testing of
lead
compounds
400000 400000 400000 1200000
Sub-Total (B2) = 1200000
Other items. Consolidated
Emolument
Year 1 Year 2 Year 3 Total
B. 3 Travel 0 40000 40000 80000
B.4 Contingency 100000 100000 100000 300000
B.5Overhead (20%) 628480
Sub-total of B
(B.1+B.2+B.3+B.4+B.5)
2292680
Grand Total (A+B) 5092680
41
PART V: EXISTING FACILITIES
Resources and additional information
1. Laboratory: Separate Mycology lab 1300 Sqft. Area.
a. Manpower
b. Equipment
Principal Investigator’s lab:
Air Conditioners, Autoclave, Automatic Shaker, Electrophoresis Unit,
UV Transilluminator, Freeze Dryer (Lyophilizer), Hot air Ovens, Laminar air flow
chambers, Microscopes, NUV lamps, Digital SLR Camera, Zoom lens, Pentax 80-
200 Zoom lens, Computer with Laser Jet printer, Refrigerated Centrifuge, Rotary
Shaker, Rotavapor, Electronic Balance, Vortex Mixer, Magnetic Stirrer, Vacuum
Pump, Growth Chamber, UV-Vis Spectrophotometer, Glass Distillation Unit, Deep
freezer and HPLC.
In Co-PI’1 Lab: IIT
Bacteriological Incubator, Spectrophotometer, Protein Purification System,
Gel Electrophoresis Units, Micro plate Spectrometer, Bench top Freeze Dryer,
Gel Documentation System, High Pressure Liquid Chromatography, Fast
Protein Liquid Chromatography, Membrane ultra-filtration unit , Lyophilizer
Vacuum flash evaporators, UV-Vis spectrophotometer with accessories
UV trans illuminator , Fourier transform Infra Red Spectroscopy,
Ultracentrifuge, pH stat, rotavap, Millipore distillation system, Gel permeation
Chromatography, Sonicator , MALDI-TOF, SEM
Other resources such as clinical material:
Cell culture lab, General Biochemistry Lab
In Co-PI’2 Lab: ICGEB
42
EXISTING FACILITIES
S.
No.
Name of equipment/
accessories
Make Funding
Agency
Year of
Procurement
1 PREP HPLC Gilson DBT 2006
2 Microscope Nikon DBT 2006
3 Coolant circulator Julabo DBT 2006
4 NMR 500 MHz Bruker ICGEB 2009
5 FT-IR Perkin Elmer ICGEB 2006
6 Multiwell florescence
Reader
Perkin Elmer DBT 2007
Other resources such as clinical material, animal house facility, glass house.
Experimental garden, pilot plant facility etc.
Animal House facility on a payment basis is available at the ICGEB, New Delhi
43
PART VI: DECLARATION/CERTIFICATION
It is certified that
a) the research work proposed in the scheme/project does not in any way duplicate the work
already done or being carried out elsewhere on the subject.
b) the same project proposal has not been submitted to any other agency for financial support.
c) the emoluments for the manpower proposed are those admissible to persons of corresponding
status employed in the institute/university or as per the Ministry of Science & Technology
guidelines (Annexure-III)
d) necessary provision for the scheme/project will be made in the Institute/University/State
budget in anticipation of the sanction of the scheme/project.
e) if the project involves the utilisation of genetically engineered organisms, we agree to submit
an application through our Institutional Biosafety Committee. We also declare that while
conducting experiments, the Biosafety Guidelines of the Department of Biotechnology would
be followed in toto.
f) if the project involves field trials/experiments/exchange of specimens, etc. we will ensure that
ethical clearances would be taken from concerned ethical Committees/Competent authorities
and the same would be conveyed to the Department of Biotechnology before implementing
the project.
g) it is agreed that any research outcome or intellectual property right(s) on the invention(s)
arising out of the project shall be taken in accordance with the instructions issued with the
approval of the Ministry of Finance, Department of Expenditure, as contained in Annexure-
V.
h) we agree to accept the terms and conditions as enclosed in Annexure-IV. The same is signed
and enclosed.
44
45
PART VII: PROFORMA FOR BIOGRAPHICAL SKETCH OF
INVESTIGATORS
Name : Dr. T.S. Suryanarayanan
Designation : Director
Department/Institute/University : Vivekananda Institute of Tropical Mycology
(VINSTROM)
Ramakrishna Mission Vidyapith
Chennai 600 004
Date of Birth : 25.07.1951 Sex: Male
Education
Sl. No. Institution Place Degree Awarded Year Field of Study
1. University of Madras M.Sc 1973 Botany
2. University of Madras Post M.Sc.,
Diploma
1974 Mycology and
Plant
Pathology
3. University of Madras Ph.D 1979 Mycology
A. Position and Honors
Position and Employment
Sl. No. Institution Place Position From To
1. Dept. of Botany
RKM Vivekananda
College
Reader and
Head of the
Department
Dec. 1978 July 2009
2. Vivekananda Institute
of Tropical Mycology
(VINSTROM)
RKM Vidyapith
Director August 2009 Till date
Honors/Awards
Fulbright Nehru Senior Fellowship, The Ohio State University, USA, September
2011 – March 2012.
Fulbright visiting faculty, Arizona State University, Tempe, Arizona, USA. May
2004.
Honorary research Associate, University of New Brunswick, St. John, Canada.
46
Invited Lectures
Some strategies to better-utilize fungal endophytes – Intergovernmental Framwork for
European Cooperation in Science & Technology – Symposium on Endophytes in
Biotechnology & Agriculture – Trento, Italy November 2012.
Fungal Endophytes: Ecology and Economic Potential. – Helmholtz Centre for
Infection Research, Braunschweig, Germany. May 2007.
Ecology of fungal endophytes: contribution from VINSTROM. – Plant Pathology
Seminar, University of Arizona, Tucson, Arizona, USA, November 2006.
Tropical Fungal Endophytes: ecological insights gained from forests of Southern
India – 25th
Genetics of Microorganisms Congress, São Paulo, Brazil, March 2006.
Tropical Endophytes: An Ecologist’s Perspective – Technical University of
Braunschweig, Germany, November 2005
Biotechnological Potential of Fungal Endophytes: An Overview – Pipe line Seminar
at the German Research Centre for Biotechnology, Braunschweig, Germany,
November, 2005.
Endophytes of Sonoran Cacti - School of Life Sciences, Arizona State University,
USA. 2004
Fungal endophytes of the mangrove Rhizophora apiculata – Asian Mycological
Congress, Hong Kong, 2000
Diversity of endophytes in tropical forests: the Western Ghats experience – 3rd Asia-
Pacific Mycological Congress on Biodiversity and Biotechnology, 2002, Kunming,
China.
B. Publications
Book chapters 12 Research Papers 88
Selected peer-reviewed publications
1. Suryanarayanan, TS and Hawksworth, DL (2005) Fungi from little-explored and
extreme habitats. In Bio-diversity of fungi: Their role in human life (SK
Deshmukh and MK Rai, eds.): 33-48. Oxford & IBH Publishing, New Delhi.
2. Suryanarayanan, T.S., Ravishankar, J.P., and Muruganandam V. (2010). Drug
discovery: going with the tide. Current Science 99:1308.
3. Kumaresan, V and Suryanarayanan, TS (2001) Occurrence and distribution of
endophytic fungi in a mangrove community. Mycological Research 105: 1388-
1391.
4. Suryanarayanan, TS and Kumaresan, V (2000) Endophytic fungi of some
halophytes from an estuarine mangrove forest. Mycological Research 104: 1465-
1467.
5. Suryanarayanan, TS, Kumaresan, V and Johnson, JA (1998) Foliar fungal
endophytes from two species of the mangrove Rhizophora. Canadian Journal of
Microbiology 44: 1003-1006.
47
6. Suryanarayanan, T.S., Thirunavukkarasu, N., Govindarajulu, M.B., Sasse, F.,
Jansen, R. and Murali, T.S. 2009. Fungal Endophytes and Bioprospecting. Fungal
Biology Reviews 23: 9-19.
7. Ravishankar, JP, Muruganandam, V and Suryanarayanan, TS (1996) Effect of
salinity on amino acid composition of the marine fungus Cirrenalia pygmea.
Current Science 70: 1087-1089.
8. Ravishankar, JP, Muruganandam, V and Suryanarayanan, TS (1995) Isolation and
characterization of melanin from a marine fungus. Botanica Marina 38: 413-416.
9. Ravishankar, JP, Muruganandam, V and Suryanarayanan, TS (1994) Effect of
salinity on fatty acid composition of Cirrenalia pygmea, an obligate marine
fungus. Botanica Marina 37: 479-481.
10. Suryanarayanan, T. S., Thirumalai, E., Prakash, C.P., Govinda rajulu, M.B. and
Thirunavukkarasu, N. 2009. Fungi from two forests of southern India: a
comparative study of endophytes, phellophytes and leaf litter fungi. Canadian
Journal of Microbiology 55: 419-426.
List maximum of five recent publications relevant to the proposed area of work.
1. Suryanarayanan, T.S., Murali, T.S., Thirunavukkarasu, N., Govinda Rajulu, M.B.,
Venkatesan, G., Sukumar, R. (2011). Endophytic fungal communities in woody
perennials of three tropical forest types of the Western Ghats, southern India.
Biodiversity and Conservation 20: 913-928.
2. Govinda Rajulu, M.B., Thirunavukkarasu, N., Suryanarayanan, T.S.,
Ravishankar, J.P., El Gueddari, N.E. and Moerschbacher, B.M. (2011).
Chitinolytic enzymes from endophytic fungi. Fungal Diversity 47:43-53.
3. Thirunavukkarasu, N., Suryanarayanan, T.S., Murali, T.S., Ravishankar, J.P.,
Gummadi, S.N. (2011). L-asparaginase from marine derived fungal endophytes of
seaweeds. Mycosphere 2:147–155.
4. Suryanarayanan, T.S., Ravishankar, J.P., Muruganandam V. (2010). Drug
discovery: going with the tide. Current Science 99:1308.
5. Suryanarayanan, T.S., Venkatachalam, A., Thirunavukkarasu, N., Ravishankar,
J.P., Doble, M. and Geetha, V. (2010) Internal mycobiota of marine macroalgae
from the Tamilnadu coast: distribution, diversity and biotechnological potential.
Botanica Marina 53:456-468.
B. Research Support
Ongoing research Projects
Sl.
No.
Title of Project Funding
Agency
Amount Date of
sanction and
Duration
1. The potential of fungal
endophytes as biocontrol
organisms
DBT 36,59,000/- February
2011
(3 Years)
48
2. Endophytic fungi of endemic
plants of Arunachal Pradesh:
Diversity and novel metabolites
DBT 14,80,000 May 2012
(3 years)
Completed research Projects
Sl.
No.
Title of Project Funding
Agency
Amount Date of
completion
1. Screening of tropical fungal
endophytes for novel secondary
metabolites
DBT 24,95,000/- 31.08.2008
2. Diversity of microfungi in leaf
litter of the different forests of
Western Ghats
MoEF 12,60,000/- 15.02.2009
3. Enzymes of pharmaceutical
importance from tropical
microfungi
DBT 25,30,000/- 22.05.2010
Place : Chennai
Date : Signature of Investigator
49
PART VII : PROFORMA FOR BIOGRAPHICAL SKETCH OF INVESTIGATORS
Name : Dr. Mukesh Doble
Designation : Professor
Department/Institute/University : Dept. of Biotechnology, Indian Institute of
Technology Madras, Chennai 600 036.
Date of Birth : 16.09.1950 Sex: Male
Education
Sl. No. Institution Place Degree Awarded Year Award
1. IIT, Madras B.Tech 1972 I class
2. IIT, Madras M.Tech 1974 I class
3. University of Aston,
Birmingham
Ph.D 1980
Position and Honors
Position and Employment
Sl. No. Institution Place Position From To
1. Dept. of
Biotechnology, IIT
Chennai.
Professor April 2004 Till Date
Research Experience in various institutions
Professor, IIT Madras, Department of Biotechnology, IIT Madras, (April 2004-
Faculty, Centre for Biotechnology, Anna University, Chennai, (July 2003, April
2004)
Program Leader, Chemical engineering modelling group, GE India Technology
centre, Bangalore (2000 - 2003 )
Process modelling of monomer and polymer reactions
Setting up of chemical engineering lab and polymer reaction lab
Group Manager, ICI India R&T centre , Bombay (1983-88, 1989-2000)
Process development/scale-up, process engineering and technology
development modelling & simulation, reaction kinetics and mechanisms
molecular modelling and QSAR studies
Research Fellow, Texas A&M University, USA (1988-89)
Fischer-Tropsch catalyst design
Research Fellow, University of Cambridge, UK (1980-83)
Car exhaust catalysis design and modelling
Asst Engineer , KCP Ltd, Madras (1974-77)
Pneumatic conveying
sugar processing
50
B. Publications
Books 7 Research Papers 109
Selected peer-reviewed publication
1. Sivakumar, PK and Mukesh Doble, COX-2 Enzyme and its Inhibitors, Current
Bioactive Compounds, 2006 2 (2), 161-179.
2. Mukesh Doble, Biological Treatment of VOCs, Chemical Engineering, 2006, 35-
41.
3. Mukesh Doble and Anil Kumar K, Experimental and modelling studies on
antifungal compounds, Central European Journal of Chemistry 4 (3), 2006, 428-
439.
4. Hemalswarya, Mukesh Doble, Potential synergism of natural products in the
treatment of cancer, Phytotherapy Research, 20, (4)2006.
5. Devarajan, Padma V, Sonavane, Ganeshchandra S and Mukesh Doble, Computer
Aided Molecular Modeling: A Predictive Approach in the Design of
Nanoparticulate Drug delivery System, Journal of Biomedical Nanotechnology,
1(4),2005, 375-383.
6. Bhagwatr, SS, Bevinakatti, Mukesh Doble, Transesterification of substituted
ethanols with ethyl acetate-modelling studies, j. biochem. Engg., 2005, 22(3),
253-360.
7. Mukesh Doble, Putting Six-Sigma processes to work, Chemical Engineering,
2003, 62-67.
8. Mukesh Doble, scale up of chemical reaction, 2002, 48-56.
9. Bhaduri, S, Lahari GK, Munish P and Mukesh Doble (2000) Catalysis Letters,
65,1.
List maximum of five recent publications relevant to the proposed area of work.
1. K. Anil Kumar, Sanjay Kumar Singh, B. Siva Kumar and Mukesh Doble,
Synthesis, anti-fungal activity evaluation and QSAR, studies on podophyllotoxin
derivatives, Central European Journal of Chemistry 5(3) 880-897 (2007).
2. P M Sivakumar, Geetha Babu and Mukesh Doble, QSAR studies on Chalcones
and Flavonoids as Anti-Tuberculosis Agents using genetic function
approximation (GFA) method, Chem. Pharm. Bull. 55(1) 44−49 (2007).
51
3. M. Sudhakar, Priyadarshini, Mukesh Doble,P.Sriyutha Murthy and R.
Venkatesan, Marine Bacteria Mediated Degradation of nylon 66 and 6,
International Biodeterioration and Biodegradation
doi:10.1016/j.ibiod.2007.02.002
4. P. M. Sivakumar, Mukesh Doble, S. Prabu Seenivasan and Vanaja Kumar,
Synthesis, Antimycobacterial activity evaluation and QSAR studies of chalcone
derivatives, Bioorganic & Medicinal Chemistry Letters , 17, (6), 1695-1700
(2007)
5. Puratchikody, Mukesh Doble, Antinociceptive and antiinflammatory activities
and QSAR studies on 2-substituted-4, 5-diphenyl-1H-imidazoles, Bioorganic and
Medicinal Chemistry, Bioorg Med Chem. Jan 15; 15(2):1083-90 (2007).
C. Research Support
Ongoing research Projects
Sl.
No.
Title of Project Funding
Agency
Date of sanction and Duration
1. Enhanced biosurfactant production
for improving biodegradation of
poly propylene
DST May 2007
(3 Years)
2. Synthesis and characterization of
curdlan
DBT August 2006
(3 Years)
Place :
Date : Signature of Investigator
52
PART VII : PROFORMA FOR BIOGRAPHICAL SKETCH OF INVESTIGATORS
Name : DR. DINKAR SAHAL
Designation : SENIOR RESEARCH SCIENTIST
Department/Institute/University : Malaria Research Group
I.C.G.E.B., P.O.Box 10504, Aruna Asaf Ali
Marg,
New Delhi-110 067.
Date of Birth : 18 JAN 1956
Sex (M/F) : Male
Education (Post-Graduation onwards & Professional Career)
SI No. University/Institution Degree awarded Year
1. M.S. university,
Baroda, India
M.Sc (Biochemistry)
1977
2. All India Institute of
Medical Sciences
New Delhi.
Ph.D (Biochemistry) 1984
RESEARCH EXPERIENCE IN VARIOUS INSTITUTIONS:
Professional Experience:
1977 - 1984 : Senior Research Fellow,Department of Biochemistry,All india
institute of Medical sciences, New Delhi, India .
1984 -1986 : Research Associate, Molecular Biophysics Unit, Indian Institute of
Science, Bangalore 560012.
1986 -1988 : Research Fellow, Department of Molecular Genetics, Beckman
Research Institute of the City of Hope, Duarte, California.
1988 -1992 : Lecturer, Department of Biophysics, University of Delhi, South
Campus New Delhi . I was engaged in teaching physical biochemistry to Post graduate students and
conducting research on (a) Peptide models of Protein phosphorylation and
(b)Chromogenic peptides as substrates in testing pyrogens by the amoebocyte lysate
assay from the horse shoe crab system,
1992- till now: Senior Research Scientist, ICGEB, New Delhi I was actively engaged in developing laboratory technologies for production of gene
products both by recombinant and chemical synthesis routes. I have synthesized human
53
Oxytocin by solution phase synthesis in a final yield of 70 grams and human
Adrenocorticotropic hormone by solid phase synthesis in a final yield of 4 grams. By the
recombinant approach, human Insulin was synthesized in milligrams scale following
expression of insulin chains as fusion protein inclusion bodies in E.coli, cyanogens
bromide cleavage , sulfitolysis and A+B chain assembly into chromatographically pure
and biologically active human insulin. At the ICGEB, New Delhi, Sahal made
significant contributions to the study of (a) anomalous mobility of sulfitolysed proteins in
SDS-PAGE and improvising this into a simple method to monitor the kinetics of
sulfitolysis, (b) the invention of a charcoal based method for removal of Iodine used in
the formation of disulfide bonds as exemplified by the synthesis of Oxytocin (c) down
stream processing of recombinantly expressed insulin precursors and (d) peptide cocktails
for application in ELISA based diagnosis of hepatitis C. In more recent times, Sahal has
made a mark by playing a key role in the de novo design and chemical syntheses of
helical hairpin peptides. The first such helical hairpin reported in PNAS (2001) was
designed for proximal orientation of a left handed helix with a right handed helix. Soon
thereafter, Sahal and companions reported in STRUCTURE (2004) the crystal structure
and circular dichroism studies of an analogous homochiral hairpin in which both helices
were right-handed. Most recently, Sahal‟s lab has reported the design, synthesis and
spectroscopic characterization of Trpzip- a de novo designed water soluble mini protein
displaying a motif. Sahal‟s investigations have traversed a large canvas from
structure to function. Thus, one of the major current interests of Sahal‟s laboratory is the
design of potent antibiotic peptides. Mechanistic studies of these peptides are revealing
important clues on the design of the much needed novel antibiotics acting at new targets.
Sahal is currently a Principal Investigator of the DBT sponsored project on “Potentiation
of de novo designed conformationally constrained helical peptides for antibiotic action”.
His talk on antibiotic peptides at the Satellite meeting of the joint 15th
IUPAB & 5th
EBSA International Biophysics Congress on “Cell Penetrating Peptides” (Aug 2005)
held at Montpellier, France was well received. Sahal has made important contributions in
proposing new models for our understanding of the antimalarial action of Artemisinin.
He has played a key role in developing validated high through put methods for the
discovery of novel blood stage antimalarials. These methods have been patented,
presented at the World conference on Magic Bullets-to celebrate Paul Ehrlich‟s 150th
birthday, Nurnberg 2004 and published. Using these methods, Sahal‟s laboratory has
identified promising antimalarial hits from chemical combinatorial libraries on the one
hand and marine organisms on the other. The exploration of hits from chemical
combinatorial libraries is a programme in collaboration with CDRI, Lucknow. Sahal is a
Principal Investigator in the DBT sponsored multi institutional project (ICGEB,New
Delhi- Malaria Research Centre, Delhi and Marine Research Lab,University of
Trivandrum) entitled “Discovering antimalarials from marine Organisms”.
RESEARCH SEMINARS PRESENTED
(a)Self-nonself discrimination : a model
(b)An excursion with Peptides
(c)How do Information and Energy play the game of Biology?
(d)Human Chorionic Gonadotrpin: Structure, Function and Immunobilogy
(e)Cryptobiosis
(g) Some significant properties of the peptide bond
(h)Theory of Chromatography
(i) Thermodynamics of Evolution
54
(j) From Ellman‟s reagent to disulfide biochemistry
(k) The chirophilic Glycine Crystal
(j) Origins of biomolecular handedness
(k) In vivo Crystallization and Biomineralization
(l) Protein Electrostatics
(m) Insulin and IGF-I Receptors: a comparative study of isoreceptor Protein
Tyrosine Kinases using exogenous substrates, inhibitors and stimulator
(n) Innovations in Protein Kinase assay
(o) Joys and Travails of Drug discovery
(p) Quality control in Protein Pharmaceuticals
(g) Artemisinin : a colorless drug targets heme a colored cofactor to bring out its
antimlarial action
(h) Making way through the haze of drugs and bugs
(i) Bacterial knock at the SIMIR club
(j) Sensing the color of heme towards discovery of novel antimlarials
REVIEWS
Deciphering ionic interactions in Peptides and Proteins.
Dinkar Sahal in Biophysical Processes in Living Systems. 2001,Editor: P.Pardha
Saradhi, Oxford & IBH Publishing Co.Pvt. Ltd. New Delhi, pp 55-74.
MONOGRAPHS:
1. Production of Human Oxytocin by Solution Phase Chemical Synthesis (1997)
D.SAHAL, ICGEB, New Delhi.
2. Production of Human Adrenocorticotropic Hormone by Solid Phase Chemical
Synthesis (1996) D.SAHAL, ICGEB, New Delhi.
Sahal’s Commentary on the 1992 Nobel Prize in Physiology and Medicine:Key to Life’s
Dynamism- The work of Krebs and Fischer has given a new perspective on controls in
Biological processes. The Economic Times 24th
October 1992, p.11.
Reviewer/Referee/Critic : I have been reviewing manuscripts written by friends, and
papers or proposal received from grant giving agencies like the DBT/DST/CSIR. Also I
am a reviewer of papers sent by various journals like Biochemie, Analytical
Biochemistry, Blood, Current Science, J Photochem. PhotoBiol. etc. I am the guest editor
of the Journal of Amino acids for its special issue entitled “Peptides: Applications from
Drug Discoveries to Materials and Molecular Electronics"
Book review by Sahal
Biophysics reviewed by Dinkar Sahal Current Science 83,(10) 25 Nov 2002, 1267-1269
List of important publications
1. Sahal D, Ramakrishnan,S; Iyer,K.S.N.; Das,C;and Talwar,G.P (1982) Immunobiological properties of a
carboxyterminal 53 aminoacid peptide of the beta subunit of human chorionic gonadotropin. J.
Reproductive Immunology 4,145-156
55
2. Talwar,G.P; Gupta S.K ; Singh,V; Sahal,D; Iyer,K.S.N; and Singh,O. (1985) Bioeffective monoclonal antibody
against the decapeptide gonadotropin release hormone: reacting determinants and action on ovulation and
estrus suppression. Proc. Natl.Acad.Sci.U.S.A 82,1228-1231
3. Iyer,K.S.N.; Sahal,D; and Talwar,G.P. (1986) Chemical synthesis and immunobiological properties of a cyclic
eicosapeptide(Gly )82-101 of the beta subunit of human chorionic gonadotropin. Int.J.Peptide Protein Res.
27,604-612.
4. Sahal,D; and Balaram,P. (1986) Peptide models of electrostatic interactions in proteins:nmr studies on two
beta turn tetra peptides containing Asp---His and Asp---Lys salt bridges. Biochemistry 25, 6004-6013.
5. Sahal,D; and Fujita-Yamaguchi,Y. (1987) Protein kinase assay by paper trichloroacetic acid method:high
performance using phosphocellulose paper and washing an ensemble of samples on flat sheets.
Anal.Biochem 167, 23-30
6. Sahal,D;Ramachandran,J; and Fujita-Yamaguchi.Y. (1988) Specificity of tyrosine protein kinases of the
structurally related receptors for insulin and insulin like growth factor-1:tyr-containing synthetic polymers
as specific inhibitors or substrates. Archives of Biochem. Biophys. 260 , 416-426.
7. Yoko Fujita-Yamaguchi, David B.Sacks, Jay M.Mcdonald, Dinkar Sahal and Satish kathuria. (1989) Effect of
basic polycations and proteins on purified insulin receptor:insulin- independent activation of the receptor
tyrosine-specific protein kinase by poly(l-lysine). Biochem.J. 263, 813-822.
8. Dinkar Sahal and Yoko Fujita-Yamaguchi. (1989) Solid-phase tyrosine specific protein kinase assay in
multiwell substrate-immobilized polyacrylamide gel. Anal.Biochem 182, 37-43.
9. Dinkar Sahal,Shu-Lian Li and Yoko Fujita-Yamaguchi. (1991) Solid-phase protein kinase assay. Methods in
Enzymology 200 , 90-98.
10. Jyoti verma and Dinkar Sahal. ,(1992) Chromatographic and proton nmr study of ion pair interaction
between dodecyl histidine methyl ester and dodecyl hydrogen phosphate in chloroform. Bull.Chem.Soc.Jpn
65, 1719-1721
11. N.Sankar and Dinkar Sahal. (1993) A phosphate staining reagent revisited. Bull.Chem.Soc.Jpn. 66, 2579-
2581.
12. Malhotra,M. and Sahal, D. (1996) Anomalous mobility of sulfitolyzed proteins in SDS- PAGE:Analysis and
Applications. Int.J.Peptide Protein Res 48, 240-248
13. Vijay Kumar , N.Jayasuryan , Honey Reddi , Dinkar Sahal & S.K.Panda. (1998) A monoclonal antibody against
the X protein of hepatitis B virus.Fine mapping of its epitope and application in a quantitative ELISA of the
X protein in sera of Hepatitis B patients. Hybridoma 17 (2) , 157-164.
14. Dinkar Sahal. (1998) Physiology and overcrowding. Current Science 74 (12) 1033.
15. D.Sahal. (1999) Removal of iodine by solid phase adsorption to charcoal following oxidation of
acetamidomethyl-protected peptide precursors to their disulfide bonded products: oxytocin and a Pre-
S1 peptide of hepatitis B virus illustrate the method. J. Peptide Research 53, 91-97.
16. Dinkar Sahal. Deciphering ionic interactions in peptides and proteins. In "Biophysical Processes in Living
Systems", Ed. P.Pardha Saradhi. Oxford & IBH Publishers
17. Udupi A. Ramagopal , Suryanarayanrao Ramakumar, Dinkar Sahal and Virander S.Chauhan. (2001) De novo
design and characterization of an apolar helical hairpin peptide at atomic resolution: Compaction
mediated by weak interactions. Proc.Natl.Acad.Sci (USA) 98 (3), 870 - 874.
56
18. Sahal D, Kannan R, Sinha A, Babbarwal V, Gnana Prakash B, Singh G, Chauhan VS. (2002) Specific and
instantaneous one-step chemodetection of histidine-rich proteins by Pauly's stain. Anal Biochem. Sep
15;308 (2):405-8.
19. Kannan R, Sahal,D, Chauhan V.S. (2002) Heme-artemisinin adducts are crucial mediators of the ability of
artemisinin to inhibit heme polymerization. Chem Biol. Mar; 9 (3): 321-32
20. Hasan Korkaya, Shahid Jameel, Dinesh Gupta, Shweta Tyagi, Ravinder Kumar, Mohammad Zafrullah, Manjari
Mazumdar, Sunil Kumar Lal, Li Xiaofang, Deepak Sehgal, Suman Ranjan Das, and Dinkar Sahal (2001) The
ORF3 protein of hepatitis E virus binds to Src homology 3 domains and activates MAPK. J Biol Chem. Nov
9;276(45):42389-400.
21. Sahal,D , Kannan R, , Chauhan V.S. (2003) Applying malaria parasite's heme detoxification system for
screening potential antimalarial drugs. Anal Biochem. Jan 15;312(2):258-60.
22. Dinkar Sahal. (2002) Book Review of Biophysics (Vasantha Pattabhi and N.Gautham,Narosa Publishing
House,New Delhi Current Science 83, 1267
23. Sahal,D. (2003) Absence of condensing agent. Current Science 84,978.
24. Rudresh; Ramagopal, U.A.; Inai, Y.; Goel, S.; Sahal, D.; Chauhan, V.S. (2004) De novo design and
characterization of a helical hairpin eicosapeptide: Emergence of an anion receptor in the linker region.
Structure 12, 389-396.
25. Padmashri, R; Chakrabarti, K.S.; Sahal, D; Mahalakshmi,R.; Sarma, S.P.; and Sikdar. S.K. (2004) Functional
characterization of the pentapeptide QYNAD on rNa(v)1.2 channels and its NMR structure. Pflugers Arch.
447, 895-907.
26. Chetal, P.; Chauhan,V.S.; and Sahal, D. (2005) A Meccano set approach of joining trpzip a water soluble β-
hairpin peptide with a didehydrophenylalanine containing hydrophobic helical peptide. J. Peptide Res. 65,
475-484.
27. Kannan, R, Kumar, K, Sahal, D, Kukreti, S, Chauhan, V.S. (2005) Reaction of artemisinin with haemoglobin:
Implications for antimalarial activity. Biochemical Journal 385(2) 409-418
28. Jagdish Rai, S. Raghothama and Sahal D. (2007) De novo design of F containing peptides. Chemical Biology
and Drug Design 69 (2):119-123.
29. Jagdish Rai, S. Raghothama and Sahal D. (2007)Tyrosine-heme ligation in Heme- peptide complex: design
based on conserved motif of catalase. J Pept Sci. 13 (6):406-412.
30. Ipsita Pal-Bhaumick, Ramendra Pati Pandey, Gotam K.Jarori, Santosh Kar, Dinkar Sahal (2007). Structural
and functional studies on Ribonuclease S, Retro S and Retro Inverso S peptides. Biochem Biophys Res
Commun. Oct 18; 17963728
31. Sahal, D. (2009) “Synthetic Life and Artificial Organisms”, chapter in the volume “Life and Organicism”:
Project on History of Indian Science, Philosophy and Culture (PHISPC) Rangaswamy, N.S. (ed)
32. Pooja C.Dewan, Aparna Anantharaman, Virander S Chauhan and Dinkar Sahal (2009) Antimicrobial Action of
Prototypic Amphipathic Cationic Decapeptides and Their Branched Dimers Biochemistry, 48, 5642-5657
33. Anantharaman, Meryam Sardar and Dinkar Sahal (2010) Synergy with rifampicin and kanamycin enhances
potency, kill kinetics and selectivity of de novo designed antimicrobial peptides. Aparna Antimicrobial
Agents and Chemotherapy, 54 ,1693-1699
34. Aparna Anantharaman and Dinkar Sahal (2010) Reverse Engineering Truncations of an antimicrobial peptide
dimer to identify the origins of potency and broad spectrum of action. J Med Chem, 53 (16), 6079–6088
57
35. Wanare G, Aher R, Kawathekar N, Ranjan R, Kaushik NK, Sahal D. (2010) Synthesis of novel alpha-
pyranochalcones and pyrazoline derivatives as Plasmodium falciparum growth inhibitors. Bioorg Med Chem
Lett. 20(15): 4675-8
36. Kumar R, Mohanakrishnan D, Sharma A, Kaushik NK, Kalia K, Sinha AK, Sahal D (2010) Reinvestigation of
structure activity relationship of methoxylated chalcones as antimalarials: Green Synthesis and evaluation
of 2,4,5-trimethoxy substituted patterns as lead candidates derived from abundantly available natural β-
asarone. Eur J Med Chem. 45(11):5292-301
37. Bagavan A, Rahuman AA, Kaushik NK, Sahal D (2011) In vitro antimalarial activity of medicinal plant
extracts against Plasmodium falciparum. Parasitol Res. Jan;108(1):15-22
38. Rao JL, Reddy PS, Mishra RN, Gupta D, Sahal D, Tuteja N, Sopory SK, Reddy MK (2010) Thermo and p H
stable ATP- independent chaperone activity of heat-inducible Hsp 70 from Pennisetum glaucum. Plant
Signal Behav. Feb;5(2):11-121.
39. Bagavan A, Rahuman AA, Kamaraj C, Kaushik NK, Mohanakrishnan D, Sahal D. (2011)Antiplasmodial activity
of botanical extracts against Plasmodium falciparum. Parasitol Res. 108(5):1099-109
40. Aher RB, Wanare G, Kawathekar N, Kumar RR, Kaushik NK, Sahal D, Chauhan VS (2011)
Dibenzylideneacetone analogues as novel Plasmodium falciparum inhibitors. Bioorg Med Chem Lett.
15;21(10):3034-6
41. Tanwar, P.a , Yadav, G.C.a , Jaitley, U.K.b , Kaushik, N.c , Sahal, D (2011) Synthesis and antimalarial activity
of novel N-{2-[2-(2-aminoethoxy) ethoxy] ethyl}-7-chloroquinolin-4-amine and its derivatives Indian Journal
of Chemistry - Section B Organic and Medicinal Chemistry, vol. 50, no. 2, pp. 233–241
42. Sayani Dasgupta, Sharmishtha Samantaray, Dinkar Sahal
and Rajendra P Roy (2011)
Isopeptide Ligation Catalyzed by Quintessential Sortase A: Mechanistic Cues from Cyclic and Branched
Oligomers of Indolicidin. THE JOURNAL OF BIOLOGICAL CHEMISTRY VOL. 286, NO. 27, pp. 23996–24006, July 8,
2011.43.
43. Tandem allylic oxidation–condensation/esterification catalyzed by silica gel:an expeditious approach
towards antimalarial diaryldienones and enones from natural methoxylated phenylpropenes. Abhishek
Sharma, Naina Sharma, Amit Shard, Rakesh Kumar, Dinesh Mohanakrishnan, Saima, Arun K. Sinha and Dinkar
Sahal (2011) Organic & Biomolecular Chemistry 9(14):5211-9.
44. Stilbene-Chalcone Hybrids: Design, Synthesis and Evaluation as a New Class of Antimalarial Scaffolds that
Trigger Cell Death through Stage Specific Apoptosis. Sharma, N, Mohanakrishnan D, Shard A, Sharma A,
Malik S, Sinha, A, Sahal D. (2012) J Med Chem 55,297-311.
45. Antimalarial activities of medicinal plants traditionally used in the villages of Dharmapuri regions of South India Chinnaperumal Kamaraj, Naveen Kumar Kaushik, Abdul Abdul Rahuman
a, Dinesh Mohanakrishnan, Asokan
Bagavan, Gandhi Elango, Abdul Abduz Zahir, Thirunavukkarasu Santhoshkumar, Sampath Marimuthu, Chidambaram Jayaseelan, Arivarasan Vishnu Kirthi, Govindasamy Rajakumar
a, Kanayairam Velayutham
a, Dinkar
Sahal (2012) J Ethnopharmacology
46. Anti-plasmodial action of de novo-designed, cationic, lysine-branched, amphipathic, helical peptides,
Naveen K Kaushik, Jyotsna Sharma and Dinkar Sahal. Malaria Journal (In Press)
PATENTS:
(a) Solid Phase Protein Kinase Assay
58
Dinkar Sahal, U.S.Patent Application Serial No. 321,298
(1989),Date of Filing March 10,1989.
Canadian Patent Application Serial No. 2011768-1, Date of
filing: March 8,1990.
(b) An Improved Process For the Preparation of Alkyl Phosphate.
Dinkar Sahal and Arun Kumar Sinha. Indian Patent Application
: An Improved Process For the Preparation of Alkyl Phosphate.
Dinkar Sahal and Arun Kumar Sinha. Indian Patent Application
Serial Number: 744/Del/91, Date of Filing: 13th
August 1991.
(c) An improved process for the preparation of Chromogenic
Peptides Containing Arginine para Nitroanilide useful for
testing of bacterial pyrogens in parenteral fluids, drugs, food
products, recombinant products, Vaccines, Medical devices,
radiopharmaceutical products, Water supplies etc intended for
human use.
Dinkar Sahal and Subita Srimal
Indian Patent Application Serial Number:317/Del/93/457,
Date of Filing: Dec.1992.
(d) Method of screening a candidate substance as a
potential drug for treatment of malaria. Sahal,D; Kannan,R;
Chauhan,V.S. Indian Patent Application No. 705/DEL/2002.
(e) Antibiotic potential of de novo designed
Didehydrophenylalanine containing
conformationallyconstrained cationic amphipathic peptides and
peptidomimetics
Dinkar Sahal, Pooja Chetal, Aparna Anantharaman and
Virander S.Chauhan.
Indian Patent Application 1343/DEL/2005 filed May 2005.