Department of Biochemistry – Biotechnology Project Proposals for 2023 - 25

MSc. Dissertation Projects 2023 - 2025

Dear Students,
Every project work has to have sufficient base work before the actual experimental work begins, and
hence there has to be sufficient time to read, gain confidence about the project and then decide the
hypothesis and objectives with methodologies. Dissertation projects are a training to think science,
design experiments, interpret and analyze data and if the work is significant, then publish the work. The
latter is to share the knowledge created, with others in the field. Therefore, the work has to be good and
original. Please do keep in mind that this is a learning process and it is not possible to get breakthrough
results in four to six months and yet your work is significant.
The department having initiated MSc Dissertation has had several thought patterns of how to lead
students into projects. Brainstorming by the faculty led to two important decisions. One is that the
department has to build its research such that there is a direction and that each year the projects
strengthen the idea or topic with sufficient and significant data. Therefore, for the institution and the
department, the projects changing every year does not establish good work. Second, techniques will be
standardized leading to proficiency in work with standard operating procedures that will help students
to take forward a project rather than reinventing the wheel each year.
Each faculty member was asked to prepare a write up on what research they would like to take forward.
These write ups are included below for your kind perusal. You are requested to go through each of the
proposed projects, read a little more on your own, if need be discuss with respective mentors, if you are
not clear of what the proposal is about. Please understand that based on your reading, you will have to
then give your preferences in the order of 1 to 17. The project codes are mentioned as DP – 1, DP – 2
etc. In DP – 5 and DP – 7 etc. Preferably choose only one of the several projects proposed by a mentor.
You will find a table, in which you have to fill up your preferences and submit by September 16, 2023
by uploading the same in the Moodle Page of PBC 1802 for MSc Biochemistry students and PBT
1802 for MSc Biotechnology.
Your listing out of the preferences, will help to allot projects/mentors to students. Please note that it is
not necessary that each student will get his/her first preference. Moreover, every faculty will be allotted
students. Your having spoken to a faculty member is also not an assurance of getting the respective
project. Once you are allotted a project, only after that you have to coordinate with the mentor to build
up your assignment and dissertation project. This semester, the assignment will include writing
literature review on the project allotted to you.
This needs your serious and sincere attention. Moreover, with time in your hands, you are expected to
read and then submit your preferences by the date mentioned above.

Warm regards
Sudeshna Menon
 Department of Biochemistry – Biotechnology Project Proposals for 2023 - 25

DP – 1: Solid waste management - Conversion of organic waste into biofertilizer through

composting

Mentor: Dr. Avni Divatia and Dr. Kinjal Bhatt

Solid waste is one of the most pressing issues of our time. World over, people are generating
greater amounts of garbage without much thought. Solid waste includes municipal,
agricultural, industrial waste etc. Mostly they are disposed by means of incineration which
causes pollution. Hence, there is always a high demand of discovering a waste management
method which is cost effective and ecofriendly. Recoverable materials that are organic in
nature, such as plant material, food scraps, and paper products, can be recovered
through composting. The resulting organic material is then recycled as mulch or compost for
agricultural or landscaping purposes. In addition, waste gas from the process (such as methane)
can be captured and used as biogas for various purposes. The intention of biological processing
in waste management is to control and accelerate the natural process of decomposition of
organic matter.
Recently, many studies and research have been focussed on developing and commercializing
agro-waste-based biofertilizer. Agro-waste is defined as waste which is produced from various
agriculture activities. These agro-wastes include manures, bedding, plant stalks, hulls, leaves,
and vegetable and fruits matter. Agro-waste is usually produced through farming and
household activities. The agro-waste is often useless and will be discarded. The accumulation
of agro-waste may cause health, safety, environmental, and aesthetic concern. Thus, this
represents a problem which requires safe disposal. Such agro-wastes contain insoluble
chemical constituents (e.g., cellulose and lignin) and soluble constituents (e.g., sugar, amino
acids, and organic acids). Other constituents are fats, oil waxes, resins, pigment, protein, and
mineral. The agro-wastes such as decaying part of plants are the primary source of organic
matter in soil. Therefore, agro-wastes are the cheapest source that can be used by farmers to
improve the fertility of soil.
Over the years, there are plenty of interests of using solid-state fermentation (SSF) process in
development of various bioprocesses and products. SSF can be defined as a process that occurs
in the absence of free water using solid substrates or support. Thus, the objective of this
research was to overcome the problems of the agro-waste accumulation in the environment by
utilizing them to produce biofertilizer using SSF method. This study also investigates the
applicability of the biofertilizer in vegetable plantation, isolation of indigenous microbial
strains, enzyme profiling as well as primary and secondary metabolites and above all reactor
designing.
DP – 2: Bioplastic formation

Mentor: Dr. Avni Divatia and Dr. Kinjal Bhatt

Biodegradable plastics are a new generation of polymers emerging on the world market.
Biodegradable plastics have an expanding range of potential applications, and driven by the
growing use of plastics in packaging and the perception that biodegradable plastics are
 Department of Biochemistry – Biotechnology Project Proposals for 2023 - 25

‘environmentally friendly’, their use is predicted to increase. The attempts can be made to
synthesize biodegradable plastic material using banana peels, starch, cellulose and other agro
waste. Biopolymers are the ideal option for replacing synthetic petroleum-based plastics due
to their shorter biodegradation period, superior biocompatibility with other materials, and high
availability. Moreover, the efforts can be made to prepare biodegradable food packaging
material out of it. The biodegradable packaging films are typically made up of natural
biopolymers, including polysaccharides, lipids, proteins, and their complexes, which solve
those root environmental problems caused by plastic waste.
DP – 3: Scaling up cultivation of algae to tap its commercial potential

Mentor: Dr. Kinjal Bhatt

Algae are a group of photosynthetic autotrophs which normally thrives in various types of
water bodies such as lakes, rivers, and sea. They are accounted for producing atmospheric
oxygen via photosynthesis, which is a process that converts water and carbon dioxide into
carbohydrate using solar energy. Algae group is diverse and encompasses numerous different
phyla with their own unique characteristics and properties ranging from prokaryotic single
cellular cyanobacteria to more complex multicellular eukaryotic algae.
Microalgae are capable to grow rapidly. Their high photosynthesis efficiency coupled with the
ability to accumulate a large amount of bioproducts within their cells make them a suitable
candidate to serve as industrial raw material. Besides, cultivation of microalgae does not
require fertile land, a large quantity of freshwater, and herbicides and pesticide when compared
to the other crops and thus will not be competing for resources. Furthermore, cultivation of
microalgae can even be performed using wastewater such as domestic sewage water and palm
oil milling effluents which can assist in bioremediation of wastewater. Apart from wastewater
treatment, cultivation of microalgae can also help with reduction of atmospheric carbon dioxide
through photosynthesis, effectively contributing to the efforts of tackling greenhouse effect and
global warming. Despite the benefits of microalgae cultivation, its developments are still
plagued with various problems. For example, the low biomass production and the small size of
cells when they are cultured in liquid medium render the harvesting process of microalgae very
costly. It is well established that microalgae have tremendous potential as a source of biofuel,
food and high value bio-compounds. However, the limitations in productivity of microalgae
and the drawbacks of bioprocessing technologies render the fully utilization of microalgae
biomass to be impractical.
Therefore, more work needs to be done to further improve the existing technology. For
instance, more advanced culturing technique should be developed to increase the productivity
of microalgae, and novel biotechnology such as gene editing can be attempted to increase the
output of bioactive compounds from the microalgae strain.

References
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decolorization. Journal of Environmental Protection, 3, 889.
2. Parmar, N. D., & Shukla, S. R. (2019). Decolourization of dye wastewater by microbial
methods-A review. Indian Journal of Chemical Technology (IJCT), 25(4), 315-323.
 Department of Biochemistry – Biotechnology Project Proposals for 2023 - 25

3. Dong, Q., Chen, Q., Li, Z., & Zeng, S. (2017). Application of Microbial Technology in
Wastewater Treatment. Progress in Applied Microbiology, 23-28.
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screening, and crude oil degradation characteristics of hydrocarbons-degrading bacteria
for treatment of oily wastewater. Water Science and Technology, 78(12), 2626-2638.
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crude oil by microorganisms in soils from four geographic regions of China. Scientific
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biofortified compost using a microbial consortium. International Journal of Recycling
of Organic Waste in Agriculture, 6(4), 321-334.
11. Al-Dhabi, N. A., Esmail, G. A., Mohammed Ghilan, A. K., & Valan Arasu, M. (2019).
Composting of Vegetable Waste Using Microbial Consortium and Biocontrol Efficacy
of Streptomyces Sp. Al-Dhabi 30 Isolated from the Saudi Arabian Environment for
Sustainable Agriculture. Sustainability, 11(23), 6845.
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microbes from Muthupet mangrove soil. J Chem Pharm Res, 5(8), 122-127.
13. Asmita, K., Shubhamsingh, T., & Tejashree, S. (2015). Isolation of plastic degrading
micro-organisms from soil samples collected at various locations in Mumbai, India. Int
Res J Envir Sci, 4(3), 77-85.
14. Indumathi, A., Gayathri, T., Biotechnology, E., Nadu, T., & Nadu, T. (2016). Plastic
Degrading ability of Aspergillus oryzae isolated from the garbage dumping sites of
Thanjavur, India. Int. J. Curr. Microbiol. Appl. Sci, 8(13), 2319-7706.
15. Sangale, M. K., Shahnawaz, M., & Ade, A. B. (2012). A review on biodegradation of
polythene: the microbial approach. J Bioremed Biodeg, 3(10), 1-9.
16. Deepika, S., & Jaya, M. R. (2015). Biodegradation of low density polyethylene by
microorganisms from garbage soil. J Exp Biol Agric Sci, 3, 1-5.
17. Dang, T. C. H., Nguyen, D. T., Thai, H., Nguyen, T. C., Tran, T. T. H., Le, V. H., ... &
Nguyen, Q. T. (2018). Plastic degradation by thermophilic Bacillus sp. BCBT21
isolated from composting agricultural residual in Vietnam. Advances in Natural
Sciences: Nanoscience and Nanotechnology, 9(1), 015014.
18. Ghosh, S., Qureshi, A., & Purohit, H. J. (2019). Microbial degradation of plastics:
Biofilms and degradation pathways. Contaminants in Agriculture and Environment:
Health Risks and Remediation, 1, 184.
19. Hayat, R., Ali, S., Amara, U., Khalid, R., & Ahmed, I. (2010). Soil beneficial bacteria
and their role in plant growth promotion: a review. Annals of Microbiology, 60(4), 579-
598.
20. Souza, R. D., Ambrosini, A., & Passaglia, L. M. (2015). Plant growth-promoting
bacteria as inoculants in agricultural soils. Genetics and molecular biology, 38(4), 401-
419.
 Department of Biochemistry – Biotechnology Project Proposals for 2023 - 25

21. Rashid, M. I., Mujawar, L. H., Shahzad, T., Almeelbi, T., Ismail, I. M., & Oves, M.
(2016). Bacteria and fungi can contribute to nutrients bioavailability and aggregate
formation in degraded soils. Microbiological Research, 183, 26-41.
22. Veerapagu, M., Jeya, K. R., & Priya Rand Vetrikodi, N. (2018). Isolation and screening
of plant growth promoting rhizobacteria from rhizosphere of chilli. Journal of
Pharmacognosy and Phytochemistry, 7(4), 3444-3448.
23. Rodrigues, A. A., Forzani, M. V., Soares, R. D. S., Sibov, S. T., & Vieira, J. D. G.
(2016). Isolation and selection of plant growth-promoting bacteria associated with
sugarcane. Pesquisa Agropecuária Tropical, 46(2), 149-158.
24. Manimaran, D. S., raj Nadaraja, K., peter Vellu, J., Francisco, V., Kanesen, K., &
Yusoff, Z. B. (2016). Production of Biodegradable Plastic from Banana Peel. J.
Petrochemical Engineering, 1(19), 2.
25. Sultan, N. F. K., & Johari, W. L. W. (2017). The development of banana peel/corn
starch bioplastic film: A preliminary study. Bioremediation Science and Technology
Research, 5(1), 12-17.
26. Yaradoddi, J., Patil, V., Ganachari, S., Banapurmath, N., Hunashyal, A., Shettar, A., &
Yaradoddi, J. S. (2016). Biodegradable plastic production from fruit waste material and
its sustainable use for green applications. INTERNATIONAL JOURNAL OF
PHARMACEUTICAL RESEARCH AND ALLIED SCIENCES, 5(4), 56-65.
27. Rizwana Beevi. K, Sameera Fathima. A.R, Thahira Fathima. A.I, Thameemunisa. N,
Noorjahan, C.M, Deepika. T. (2020). Bioplastic Synthesis Using Banana Peels and
Potato Starch and Characterization. International journal of scientific & technology
research, 9(1),1809-1814.
28. Dineshkumar, R., Narendran, R., & Sampathkumar, P. (2016). Cultivation of Spirulina
platensis in different selective media. Indian Journal of Geo Marine Sciences, 45(12),
1749-1754.
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30. Usharani, G., Saranraj, P., & Kanchana, D. (2012). Spirulina cultivation: a review. Int
J Pharm Biol Arch, 3(6).

DP – 4: Production of polyhydroxyalkanoates (PHAs) from waste materials and

by products using submerged and solid state fermentation
Mentor: Dr. Avni Divatia

Polyhydroxyalkanoates are biodegradable polymers produced by prokaryotic organisms from

renewable resources. The production of PHAs by submerged fermentation processes has been
intensively studied over the last 30 years. In recent years, alternative strategies have been
proposed, such as the use of solid-state fermentation or the production of PHAs in transgenic
plants.
This project we make use of submerged and solid-state fermentation processes to produce
PHAs from waste materials and by-products. The use of these low-cost raw materials has the
potential to reduce PHA production costs, because the raw material costs contribute a
significant part of production costs in traditional PHA production processes.
 Department of Biochemistry – Biotechnology Project Proposals for 2023 - 25

DP – 5: Study of anti – stress and anti – ageing effects of plant extracts

using Caenorhabditis elegans as a model
Mentor: Dr. (Fr) Suresh Antony
Summary: Reactive Oxygen Species (ROS) are a free radicals that play a role in aging and
oxidative stress-related conditions. When the generation of ROS is not in balance with
antioxidant activity, it can damage biomolecules, including lipids, proteins and DNA. These
activities can mediate aging as well as other degenerative diseases in humans such as diabetes,
arthritis, cancer, and cardiovascular complications in humans. Secondary metabolites, such as
polyphenols, present in plants exert prevention of such diseases related to oxidative stress
generated by ROS. This study, therefore, aims to examine the in vivo antioxidant and lifespan
extension activities of plant extracts in C. elegans (wild type and mutant worm strains).
Furthermore, the underlying molecular mechanism involved in the management of stress
responses in the worms by plant extracts would be elucidated.

Experiments:
1. Preparation of crude plant extracts and its quantification of polyphenols and
antioxidant property
2. C. elegans strain maintenance and synchronization
3. Lifespan assay
4. Stress resistance assay (Oxidative stress and Thermal stress)
5. Gene expression analysis by Quantitative Reverse Transcription PCR

DP – 6: Optimization and scale up of biosulphidogenesis for treatment of

sulphate containing waste
Mentor: Dr. (Fr) Suresh Antony
Sulphate is known as one of the toxic anions. High amount of sulphate use in many industries
like dye, paper, pulp industries etc. The maximum permissible limit for sulphate in water is
500 ppm. Sulphate concentrations beyond this cause several health problems. It is required
before discharging the sulphate containing effluent into water bodies to convert much sulfate
to non-toxic form. Hot water springs have presence of sulphate and may serve as an excellent
source for the sulphate reducing thermophilic/thermotolerant bacteria (SRB) which can be used
for the sulphidogenesis process. The sulphate reduction through biosulphidogenesis process is
eco-friendly and economically viable.

Objectives:
• Optimization of sulphate reduction with the help of SRB
• Optimization of SRB immobilization for field applications.
 Department of Biochemistry – Biotechnology Project Proposals for 2023 - 25

• Testing of immobilized SRB for sulphidogenesis from selected waste.

To achieve these objectives, we will use sulphate reducing bacteria and also try with other
bacterial species. Immobilization of SRB on suitable support for their use in column reactor
will be carried out and application of the column reactor for the treatment of high sulphate
containing selected waste will follow. At present (there is no) economically viable and
environmentally sustainable comprehensive biotechnological process is not available in the
market to treat the industrial effluent effectively and efficiently to reduce their Sulphur content
before being released into the environment. Chemical processes are expensive and have several
environmental hazards and are time consuming. Considering the present scenario, the idea is
to prepare a column bioreactor in order to cater to the need for biosulphidogenesis in industrial
effluents.

DP – 7: Sustainable and renewable energy – Biodiesel

Mentor: Dr. (Fr) Suresh Antony

Energy drives and sustains almost all human activities and is fundamental to human existence
and development (Sokoto et al., 2013). Use of fossil fuel has resulted in increased carbon
emission, global warming, and climate change and health hazards etc. This calls for a serious
reflection on the search for an alternative, renewable and sustainable energy in the form of
biodiesel and bioethanol.
Jatropha curcas is a drought-resistant plant which is widely recognized as an important source
of biodiesel due to the high oil content constituted in seeds. Oil-rich seeds are generally
sensitive to storage and the reactive oxygen species (ROS) generated during seed storage cause
oxidative stress mounting to oxidative damage in oil seeds. The purpose of this study is to
elucidate the biochemical and physiological changes that occur during the storage of Jatropha
curcas seeds. We subjected Jatropha curcas seeds to natural aging (NA) (naturally stored),
accelerated aging (AA) and saturated salt-accelerated aging (SSAA), and studied the associated
biochemical and physiological changes. The chronological sequence of events of seed
deterioration is an essential knowledge.

Experiments to be carried out:-

1. Biochemical assays for MDA and H2O2.
2. Estimation of total antioxidant capacity, enzymatic and non-enzymatic anti-oxidants
3. Extraction – Isolation of oleosin oil body specific protein through SDS-PAGE.
4. Estimation of gamma tocopherol by HPLC
5. Structural studies of oil bodies using Sudan black lipid staining.
6. Study of cell death using DAPI and PI.
 Department of Biochemistry – Biotechnology Project Proposals for 2023 - 25

DP – 8: Sustainable and renewable energy – Biodiesel

Mentor: Dr. (Fr) Suresh Antony

Bio and agricultural waste is best managed to be utilized as feed stock for conversion of
bioethanol. The real challenge is to remove the lignin portion from bulk source of
lignocellulose. Cellulose then serves as the best nutrient for the microbes that convert glucose
into ethanol. Use of chemical and biocatalyst for the conversion of hemicellulose into
cellulose and cellulose into glucose is crust of the research and calls for innovation/novelty.
Research utilizing agricultural waste is an added value to the project of safe guarding the
mother earth.

Experiments to be conducted:-
1. Removal of Lignin
2. Conversion of hemicellulose into cellulose and then into glucose
3. Upstream process of fermentation using effective microbes
4. Downstream process of extraction of ethanol.
5. Estimation of ethanol by GC.

DP – 9: Preparation of biofertilizer containing enhanced phosphate

solubilizers
Mentor: Dr. (Fr) Suresh Antony

Preparation of bio fertilizer containing enhanced phosphate solubilizers.

 Extraction of phosphate solubilizers from the local soil
 Genomic study of these microbes
 Preparation of bio fertilizer enriched with NPK sources
 Checking the effect of this synthesized phosphate solubilizers enriched bio fertilizer
on the millets.

DP – 10: An exploration into Pectinase

Mentor: Dr. Nandini Ghosh
Pectin
Pectin (from Ancient Greek: pēktikós, "congealed, curdled"[1]) was first isolated and described
in 1825 by Henri Braconnot. Pectin is a polymer with linear structure in which few hundred to
thousand galacturonic acid (a sugar acid derived from galactose) monomer units are linked via
α-(1→4)-glycosidic bond forming a backbone interrupted by occasional l-rhamnose residues
In addition, there are sugars attached through side-chain such as rhamnose, arabinans,
 Department of Biochemistry – Biotechnology Project Proposals for 2023 - 25

galactans, and xylose. The average

molecular weight of pectin ranges
between 50 and 150 kDa, although due
to the presence of several sugar moieties
and different levels of methyl
esterification, pectin does not have
defined molecular weight like other
polysaccharides. Hence, it is a structural
acidic heteropolysaccharide contained
in the primary and middle lamella cell
walls of terrestrial plants, where it helps
to bind cells together, but is also found
in primary cell walls. Pectin is
deposited by exocytosis into the cell
wall via vesicles produced in the golgi.
It is mainly present in fruits, especially
citrus such as apples, oranges, lemons. Citrus fruits contain 0.5%–3.5% pectin which is largely
found in the peel portion of the fruit.
Pectin is a fiber and contains almost no calories or nutrients. The daily intake of pectin from
fruits and vegetables can be estimated to be around 5 g if approximately 500 g of fruits and
vegetables are consumed per day. By weight, 29 grams of liquid pectin contains:
Calories: 3
Protein: 0 grams
Fat: 0 grams
Carbs: 1 gram
Fiber: 1 gram
Pectin was first sold as a liquid extract, but is now most often used as dried powder, which is
easier than a liquid to store and handle. Powdered pectin has a similar nutrient content. Neither
the liquid nor powdered form contain significant amounts of vitamins or minerals, and all of
its carbs and calories come from fiber. In human digestion, pectin binds to cholesterol in the
gastrointestinal tract and slows glucose absorption by trapping carbohydrates. A study found
that after consumption of fruit the concentration of methanol in the human body increased by
as much as an order of magnitude due to the degradation of natural pectin (which is esterified
with methyl alcohol) in the colon. Pectins are reported to have hypo-cholesterolemic properties
but due to its high gelling capacity, it cannot be incorporated in food products at higher
concentration as it negatively affects the product sensory characteristics. Pectin is water soluble
in nature and bypass the enzymatic digestion process of human small intestine but is easily
degraded by the microflora of the colon. In human GI tract, pectin is capable of holding water
and forming gel, which ultimately leads to binding of ions and bile acids. Gel forming ability
of pectin is considered as possible mechanism of its beneficial health effects, such as improved
cholesterol, lipid metabolism, gastric emptying, and glucose metabolism. Pectins are also
reported to have some unique abilities for prevention or treatment of diseases, such as intestinal
infections, atherosclerosis, cancer, and obesity.
There are four types of pectic substances classified by American Chemical Society: A. Pectic
acid: It is the galacturonan’s polymer having a property of solubility and the methoxyl groups
 Department of Biochemistry – Biotechnology Project Proposals for 2023 - 25

are negligible. Pectate is the acidic or normal salt of pectic acid. B. Protopectin: This pectic
substance is insoluble in water. Intact tissues of plants contain protopectin. Due to restricted
hydrolysis protopectin produces pectic acid or pectin. C. Pectinic acids: The chain of
polygalacturon is pectinic acid having methylated units of galacturonate. Normal and acidic
salts of pectinic acid are called pectinates. D. Pectin: Pectin is generally known as polymethyl
galacturonate. In pectin, 75% galacturonate unit’s carboxyl groups are linked with methanol
by esterification. On the basis of methyl esterification, there are two different types of pectin—
high methoxyl and low methoxyl pectin. High methoxyl pectins (HM) are characterized with
more than 50% esterified galacturonic acid residues whereas low methoxyl pectins (LM) are
characterized with less than 50% esterified galacturonic acid residues. The gelation is strongly
affected by whether HM or LM pectins are used. For LM pectins, calcium ions tend to be
involved, located between galacturonic residues on neighboring chains in a way resembling an
“egg-box”.
PECTIN | Food Use
The main use for pectin is as a gelling
agent, thickening agent and stabilizer
in food giving the jelly-like
consistency to jams or marmalades,
which would otherwise be sweet
juices. Pectin displays excellent
swelling and erosion properties and is
accordingly useful for colon targeting
systems, is widely explored for
fabricating the colon-biodegradable
drug delivery systems. Moreover,
pectin can be used as an adsorbent,
emulsion stabilizer, bulk-forming
agent and coating material for tablets,
pellets, microparticles, and beads.
Calcium pectinate tablets exhibit
reduced solubility and have a very
good potential to be used in targeted drug delivery to the colon. Furthermore, cross-linking of
pectin with calcium ions hinders the release of the incorporated drug from pectin tablets by
suppressing both the swelling and dissolution of these systems. Pectin is used as a viscosifier
in beverages and soft drinks, and high-ester pectins may be used as a mouth-feel improver.
This use has been widely developed for juice drinks with a reduced juice content or sugar-free
soft drinks. Pectin is extracted from apples and citrus fruits. On the basis of dry mass, apple
pomace contains 10–15% pectin, whereas citrus peel possesses 20–30% pectin. However,
pectin has also been extracted in higher amount from several other fruits and their by-products,
such as sunflower head, mango peal, soybean hull, passion fruit peel, sugar beet pulp, Akebia
trifoliata peel, peach pomace, banana peel, chickpea husk. But detection and extraction of
pectin in higher concentration is not sufficient to qualify that fruit as a source of commercial
pectin because of the structural variation and modification in side-chain sugars, and also that
pectin from different sources has different gelling properties. Lemon peel is the most common
source for extraction of commercial pectin.

Pectinase
 Department of Biochemistry – Biotechnology Project Proposals for 2023 - 25

During fruit ripening, pectin is broken down by the enzymes pectinase and pectinesterase, in
which process the fruit becomes softer as the middle lamellae break down and cells become
separated from each other. A similar process of cell separation caused by the breakdown of
pectin occurs in the abscission zone of the petioles of deciduous plants at leaf fall. Pectinase
refers to a group of enzymes that catalyze the degradation of pectin. Pectinase is widely
distributed in higher plants and microorganisms, and also found in some protozoa and insects.
In microorganisms, bacteria, actinomycetes, yeasts, and molds can synthesize and metabolize
pectinase. They are also reported to be produced in combination with other industrially
important enzymes by the same microbial isolate.The commercial profit of pectinase industrial
application has promoted its research and development. Many manufacturers around the world
produce pectinase preparations. As technology advances, production costs have gradually
declined and applications of pectinase have become more widespread.
Many molds and small amounts of bacteria and yeasts can produce pectinase, mainly
Aspergillus and Bacillus, others including Penicillium (Penicillium expansum, Penicillium
griseoroseum, etc), Sclerotium rolfsii, Rhizoctonia solani, Mucor pusilus, Mucor mucedo,
Cloctridium thermosaccharolyticum, Rhizopus stolonifer, Aureobasiduim pullulans,
Neurospora rassa, Sporotrichum thermophile, etc. As Aspergillus niger in fungi belongs to
GRAS (General Regarded As Safe), its metabolites are safe, so currently marketed food grade
pectinase is mainly from Aspergillus niger, and the optimum pH value is generally in the acidic
range.
Pectinases can be produced by both submerged (SmF) and solid state fermentation (SSF)
The pectinase enzyme is broadly classified into three types on the basis of their mode of action:
pectin esterase, hydrolases and lyases. Pectin esterase catalyzes the de-esterification of the
methoxyl group of pectin, forming pectic acid. Hydrolases (Polygalacturonases and
Polymethylgalacturonases)—Catalyzes the hydrolytic cleavage of α-1,4-glycosidic linkage in
pectic acid and pectin, respectively, while Lyases (Polygalacturonate Lyase and
Polymethylgalacturonate Lyase)—Catalyzes the cleavage of α-1,4-glycosidic linkage in pectic
acid and pectin, respectively by trans-elimination reaction and forming unsaturated
galacturonates and methyl galacturonates, respectively.
 Department of Biochemistry – Biotechnology Project Proposals for 2023 - 25

Applications
Juices with low viscosity,
high clarity and high in
nutrition are more
desirable by consumers.
Pectinases play a crucial
role to reduce the viscosity,
increase the yield and
clarification of juice by
liquefaction of pulps,
remove off the peels and in
maceration of vegetables to
produce various products
like pastes and purées.
Depectinization depends
upon the composition of
juice, the type of enzyme
used and the time
consumed.
The main functions of pectinolytic enzymes in the wine making process are to support the
extraction process, maximise juice yield, facilitate filtration and intensify the flavour and
colour.
Pectinases are also being used in biorefineries for hydrolyzing pectin present in pectin-rich
agro-industrial wastes. These wastes are processed into simple sugars so that they could be
converted into bioethanol or used as fermentable. Different enzymes such as pectinase,
hemicellulases and cellulases are being used to convert polysaccharides present in the plant
cell wall into simple sugars.
Vegetable oils of olive, sunflower, coconut, palm or canola are obtained by extraction with
organic solvents such as hexane, which is a potential carcinogen. The use of pectolytic
enzymes, in this case preferably alkaline, allows the extraction of vegetable oils in an aqueous
 Department of Biochemistry – Biotechnology Project Proposals for 2023 - 25

process by degradation of cell wall components. Now days, the use of enzyme preparations
containing cellulases, hemicellulases and pectinases has begun for maximum extraction of oil.
Enzymatic treatment resulted not only in increase of oil yield but also increased the
polyphenolic and vitamin E content, thereby also enhanced its organoleptic quality.
In processing of textile material, the conventional scouring process involves the use of harsh
chemicals and is slowly being replaced with the ecofriendly approach by using enzymes. Bio-
scouring is an ecofriendly method for removal of non-cellulosic impurities from the fiber with
specific enzymes. It makes the fibre surface more hydrophilic. Bioscouring also avoids high
energy consumption and severe pollution problems that are associated with conventional
alkaline scouring. Pectinases also prevent fiber damage. Alkaline pectinase has been
considered as the most suitable enzyme for cotton scouring by many researchers, because the
degradation and elimination of pectin facilitates the removal of loosened waxes.
In tea and coffee processing, pectinase treatment accelerates tea fermentation by breaking
down the pectin which is present in the cell walls of tea leaves and also destroys the foam
forming property of instant tea powders by destroying the pectins. The change in colour of tea
during the fermentation also results in the development of characteristic aroma. Pectinolytic
microorganisms are also used in the fermentation of coffee to remove the mucilaginous coat
from the coffee beans.
The use of pectinases in production of ruminant feed decreases the feed viscosity and
increases the absorption of nutrients by ruminants, liberates nutrients by enzymatic action
which also reduces the amount of faeces. The specific enzyme preparations have become a
valuable tool for economically improving the digestive processes in the ruminants
Bio-bleaching of kraft pulp: With the advancement of biotechnology and increased reliance
of paper and pulp industries on the use of enzymes for biobleaching, the use of enzymes like
xylanases, ligninases, mannanase and pectinases is increasing in the paper and pulp industries.
The presence of pectins weakens dewatering during sheet formation due to their high cationic
demand and cause yellowness of paper. The pectinases depolymerize poly-galacturonic acids
and thus decreases the cationic demand in the filtrate from peroxide bleaching of thermo-
mechanical pulp. Pectinases solely and in combination with other enzymes produced by same
or by different microorganisms have been efficiently used for bio-bleaching of mixed
hardwood and bamboo kraft pulps. Reduction in bleaching chemicals would result in reduction
of organochlorine compounds in the effluent.
Recycling of wastepaper: Current deinking process depends upon the use of large amount of
environment damaging chemicals. Deinking using enzymes is less polluting, energy saving,
gives better performance to achieve the desired deinked pulp properties and results in lower
disposal problems. Enzymes being used in deinking process are pectinases, hemicellulases,
cellulases and lignolytic enzymes. Enzymatic deinking alters bonds near the ink particle and
removes the ink from fibre surface. The released ink is then removed by washing or floatation.
Isolation of protoplasts: In plant breeding programmes, many desirable combinations of
characters cannot be transmitted through conventional methods of genetic manipulation. A
process other than the sexual cycle has now become available for higher plants and can lead to
genetic recombination. This non-conventional genetic procedure, involving fusion between
isolated somatic protoplasts (wall less, naked cells) under invitro conditions and subsequent
 Department of Biochemistry – Biotechnology Project Proposals for 2023 - 25

development of their product (heterokaryon) to a hybrid plant, may be an useful tool for the
induction of genetic variability and combination of traits which do not exist in nature. Methods
of protoplast isolation from the plants are classified into three main groups: (a) mechanical
(non-enzymatic), (b) sequential enzymatic (two steps), and (c)mixed enzymatic (simultaneous)
procedures. Mechanical isolation, which includes cutting of plasmolyzed tis-sue with a sharp-
edged knife and release of protoplasts by de-plasmolysis, is now only of historical importance
because of the low yield of the protoplasts obtained by this method. Therefore, enzymatic
processes involving pectinases and cellulases mixture have been tried for this purpose.

As evident from this overview of pectin and pectin degrading enzymes, it is clear that
they have very widespread and diverse applications in all fields of biology. It is possible
that in the near future, an additional plethora of wider applicability of these two natural
biological products will be discovered and commercialized.
However, the production of both, especially of pectinases must be made cost effective
if one has to increase its yield to an industrial scale for large-scale use.
1) Optimization of fermentation conditions for pectinase production by various
microorganisms
2) Physico-chemical characterization of exo-and endo pectinases produced by the
microorganisms and their detailed physico-chemical characterization.
3) Improvement of purification techniques.
4) Being an enzyme, pectinases are subject to denaturation and degradability under
conditions, not conducive for their stability. They are subject to instability low
 Department of Biochemistry – Biotechnology Project Proposals for 2023 - 25

activity at higher temperatures and duration of storage. Hence conditions for

stability such as pH and temperature sensitiveness of pectinases produced by
the microorganisms or their mode of action in degrading pectin under these
conditions can be investigated for better optimization of enzyme production and
yield.
5) Immobilization of pectinases using different methods and techniques can be
optimized for long-term storage and later use. The development of thermostable
pectinases is a promising area of investigation that can be explored.
6) Exploring ways to enhance the enzymatic activity is a consideration as the
enzyme with a higher specific activity is likely to achieve any of the above listed
applications at a higher and better efficiency and reduced cost. Molecular
biology techniques coupled with gene cloning technology can be used to
achieve expression of pectinase in other microbial hosts and offers attractive
alternatives.
7) Strain improvement of the enzyme producing microorganisms is an attractive
option for exploration.
8) Screening, isolation and characterization of new microorganisms that produce
the enzyme in combination with others can be researched.
9) Microorganisms growing on substrates other than citrus fruit and apple peels
only can be investigated for the nature and expression of the enzymes produced.
References
1. Chapter: Role of Pectin in Food Processing and Food Packaging in Pectins - Extraction,
Purification, Characterization and Applications. 2019
2. Garg G., Singh A., Kaur A., Singh R., Kaur J., and Mahajan R. Microbial pectinases:
an ecofriendly tool of nature for industries3 Biotech (2016) 6:47 doi: 10.1007/s13205-
016-0371-4
3. Kashyap DR., Vohra PK., Chopra S. and Tewari R. Applications of pectinases in the
commercial sector: a review. Bioresource Technology 77 (2001) 215±227
4. Pedrolli, DB., Monteiro, AC., Gomes, E., & Carmona, EC. (2009). Pectin and
pectinases: production, characterization and industrial application of microbial
pectinolytic enzymes. Open Biotechnology Journal, 9-18. doi:
https://doi.org/10.2174/18744070700903010009
5. Prathyusha, K., & Suneetha, V. (2011). Bacterial Pectinases and their Potent
Biotechnological Application in Fruit Processing/Juice Production Industry: A Review.
Journal of Phytology, 3(6), 16-19.
6. Underkofler, L. A., Barton, R. R., & Rennert, S. S. (1958). Production of microbial
enzymes and their applications. Applied microbiology, 6(3), 212.
7. Edwards,MC., & Doran-Peterson, J. (2012). Pectin-rich biomass as feedstock for fuel
ethanol production. Applied microbiology and biotechnology, 95(3), 565-575. doi:
https://doi.org/10.1007/s00253-012-4173-2
8. Gómez, B., Gullón, B., Remoroza, C., Schols, Henk A., Parajó, JC., Alonso, JL. (2014).
"Purification, Characterization, and Prebiotic Properties of Pectic Oligosaccharides
from Orange Peel Wastes". Journal of Agricultural and Food Chemistry. 62 (40): 9769–
9782. https://doi.org/10.1021%2Fjf503475b
 Department of Biochemistry – Biotechnology Project Proposals for 2023 - 25

DP – 11: Acetylcholinesterase inhibition by plant extracts

Mentor: Dr. Nandini Ghosh
Acetylcholine, a neurotransmitter is found in the postganglionic sympathetic and
parasympathetic neurons and plays an important role in cognition and the activation of muscles.
The neurotransmitter has major implications in brain diseases, mainly dementia, of which
Alzheimer’s Disease is a more severe type which results in a drastic loss of cognitive and
behavioural functions including memory, recall, reasoning and motor functions, severely
impacting daily life. Research has shown that there is both a decrease in the concentration and
the function of acetylcholine in patients with Alzheimer’s disease.
In 1906, Dr. Alois Alzheimer, after whom Alzheimer’s disease is named, had treated a woman
patient for mental disorder, with behaviour matching the now classic clinical symptoms of
Alzheimer’s disease. After her death, he studied her brain tissue and observed several changes
in the tissue such as abnormal clumps, now known as amyloid plaques, and bundles of fibers
which looked tangled, now known as neurofibrillary, or tau, tangles. These plaques and tangles
in the brain are the main features of the disease, along with the loss of connections between
neurons in the brain. However, many other complex changes in the brain also contribute to the
progression of Alzheimer’s.

There are two types of Alzheimer's—early-onset and late-onset. Both types have a genetic
component. Late-onset is the most common type affecting patients in their 60s, and is believed
to be associated with the presence of a specific allelic form of apolipoprotein E (APOE) gene
 Department of Biochemistry – Biotechnology Project Proposals for 2023 - 25

called APOE ɛ4. The early onset is rare and can occur in individuals between the age groups
of 30s to 60s and can be hereditary passed on from one generation to the next by affected
members of the family.
The enzyme, acetylcholinesterase (AChE) breaks down acetylcholine into its inactive
metabolites – choline and acetate, thereby decreasing its concentration and leading to the onset
of the disease, at least partly. Hence, very simply put, if AChE could be prevented from
breaking down acetylcholine, both the level and duration of neurotransmitter action can be
sustained or prolonged and therefore, patients could be prevented from progressing to, at least,
the severe form of the disease.
Compounds called acetylcholinesterase inhibitors inhibit the AChE from breaking down
acetylcholine. AChE inhibitors can be divided into two groups: reversible and irreversible
inhibitors. The reversible inhibitors like Donepezil and Rivastigmine have therapeutic
application in treating diseases like Alzheimer’s and myasthenia gravis. The irreversible
inhibitors include organophosphates which are present in insecticides. Diisopropyl
fluorophosphate (DFP, DIFP, di-isopropyl phosphorofluoridate) is a parasympathomimetic
drug and an irreversible anti-cholinesterase. Another example of an irreversible AChE inhibitor
is Echothiophate (phospholine) which is a parasympathomimetic phosphor-othioate. Some, but
not all patients benefit from these FDA approved drugs only for a limited time.
Patients can experience mild to moderate side effects of these drugs. Diarrhoea, nausea,
headache, drowsiness and dizziness are some of the common ones which can further
incapacitate an already affected person. The more serious side effects which can affect less
than 1 in 100 people are liver problems, severe indigestion, heartburn or stomach pain, severe
or continued vomiting, internal hemorrhage, muscle weakness, pain and cramps in muscles,
high temperature along with stiff muscles, sweating, and hallucinations (called neuroleptic
malignant syndrome).
India is home to 7500 known medicinal plants. The wealth of bioactive compounds present in
them have formed the basis for the development of numerous traditional medicines used since
ancient times for curing a wide variety of diseases and disorders. In recent times, these
medicinal plants have gained wide acceptance because of their fewer side effects when
compared with pharmaceutical products. Science today takes an active interest in investigating
these secondary metabolites of plants for the formulation of compounds that can replace drugs
to improve the general health of the world population. Plants deserving special mention as
having AChE activity are Brassica alba, Brassica nigra, Camellia sinensis, Cinchona
officinalis, Citrus aurantifolia, Citrus aurantium, Ferula assafoetida, Humulus lupulus,
Juglans regia, Juniperus sabina, Myristica fragrans, Pelargonium graveolens, Pistacia vera,
Punica granatum, Rheum officinale, Rosa damascena, Salix alba and Zizyphus vulgaris.
Reports are also available for Centella asiatica, Ginkgo biloba, Withania somnifera, Bacopa
monnieri, Salvia officinalis, Melissa officinalis, Tinospora cordifolia, Glycyrrhiza glabra in
Alzheimer’s disease treatment in animal cell lines and animal models.
The focus of this project is to investigate the secondary metabolite extracts of one or two
specifically selected locally available plant species for their AChE inhibitory properties and
decipher the enzyme kinetics and the dose-response curves of these extracts paving the way for
the development of AChE inhibitory drugs with little or no side effects for arresting the
progression of Alzheimer’s disease and improving the quality of life of the afflicted patients.
 Department of Biochemistry – Biotechnology Project Proposals for 2023 - 25

DP – 12: Isolation and partial purification of azoreductase from microbes

Mentor: Dr. Nandini Ghosh
The use of synthetic dyes extend from leather, textiles, cosmetics to color printing and food
industry. Three most common groups of synthetic dyes which are used extensively are
anthroquinone, phthalocyanine and azo dyes, which are all harmful to living organisms and the
environment.
Azo dyes are organic compounds containing functional group "R−N=N−R", in which R in most
cases is an aryl group. These dyes are both carcinogenic and mutagenic. The complex aromatic
structure of these dyes is the reason behind their stability, toxicity, resistance towards light and
oxidizing agents and intense colour. Azo dyes being arduous to degrade, form layers on the
surface of water bodies. Hence, when they are stagnant on these water bodies, their colours
layer up the whole water surface which in turn is harmful for various hydrophytes lying
underneathThese dyes possess resistance towards oxidizing agents and light which makes their
treatment with conventional methods rather difficult and due to these very reasons (intense
colour, light resistance), azo dyes are used extensively.
Textile industry effluents are harmful not only to humans but also to other living organisms
along with the environment. Major wastes from the textile industries are liquid wastes and
mainly include azo dyes. From the global production of dyes which is 8,00,000 tons every year
world-wide, 10-15% are lost during various processes involved in the industrial stages.
A wide range of methods both mechanical or chemical have been tried in order to treat the
synthetic dyes containing effluents and waste water. Because of the several drawbacks
encountered such as the recalcitrant nature of the resultant solids or liquids which require
further intensive energy and additional treatment for their elimination with these conventional
methods of treatment, biological remediation measures are more suitable.
Bio-transformation is the mechanism in microbes which is responsible for degradation of
contumacious compounds biologically. Azoreductase enzyme, also known as “azobenzene
reductases” as been seen to play the key role in the reduction of azodyes in nature.
Azoreductases involved in degradation of dyes are mainly NADH or NADPH dependent group
of enzymes, some of which may also require flavin as cofactor. As some of the dyes are
carcinogenic in nature, upon their degradation by the azoreductases result into aromatic amines
which also stand to be carcinogenic but the benefit is that these aromatic amines are easily
degraded by the aerobic bacteria and hence the use of azoreductases or in other words the use
of biological methods is preferred.
Thus, bioremediaton or “microbial clean up” is coming up as a good solution for various
hazardous pollutants' treatment including azodye. The biological method or microbial approach
is preferred because microbes have the ability to adapt to their surroundings and hence when
textile effluent containing areas or areas where textile effluents are released are checked for
microbes with dye reducing or decolorizing activities, microorganisms are found which have
such properties and have proved helpful for the more eco-friendly approach for the clean -up
of azo dye containing waste water from various industries. It also assures the complete
detoxification and mineralisation of pollutants. Along with many other benefits of using
azoreductase or biological methods for bioremediation, the economic purpose i.e., it is
 Department of Biochemistry – Biotechnology Project Proposals for 2023 - 25

comparatively cheaper than the present methods available today for this treatmen. Most
azoreductases are not able to degrade sulphonated azodyes except the azoreductase isoenzyme
isolated from Bacillus sp. B29.
The aim of this project is to explore the microorganisms that can produce azoreductases, their
purification and enzyme kinetics and the establishing optimum enzyme activity at which
azodyes can be effectively degraded.

DP – 13: Herbal Medicine in the Treatment of Non-Alcoholic Fatty Liver

Diseases
Mentor: Dr. Sebastian V A
As per various studies published, non-alcoholic fatty liver disease leads to obesity, diabetes,
liver cirrhosis and fibrosis. When a modification in lifestyle and eating habits is insufficient, it
may be addressed with the help of medicinal plant extracts. Active principles from many
medicinal plants having natural polyphenol has been in use to control the progress to fatty liver.
The research work proposes to use several medicinal plants having such polyphenols and
flavonoids like baicalein to restore the upregulation of genes associated with NAFLD through
wet lab or bioinformatics-based tools.
Students may learn:
1. Survey based epidemiology of NAFLD
2. Identification of relevant medicinal plants available in Gujarat with scope to treat NAFLD
3. Extraction, estimation and identification of active principles
4. Carry out various biochemical/molecular tests to assess the cellular damage
5. Basic tools for docking

Quick reference:
1. DOI: 10.2174/0929867322666150805094940
2. https://www.frontiersin.org/articles/10.3389/fphar.2022.892559/full

DP – 14: Study on Plant pathogen/ plant – fungi interaction and immune

response

Mentor: Dr. Sebastian V A and Sudeshna Menon

Plants respond to pathogens attack either by inducing various elicitors or by activating
resistance genes (R genes). These elicitors are recognized by the host cell, and the signal is
transduced to the nucleus, which leads to activation of the R genes that confer resistance to
 Department of Biochemistry – Biotechnology Project Proposals for 2023 - 25

plants. The R-gene products (Pathogen Related proteins or PR proteins) have various defense
mechanisms which include, 𝛽-1,3- glucanase, chitinase activities to degrade components of
cell wall of pathogens, defensins like endopolygalacturonases and cell wall hydroxyproline-
rich glycoproteins, and lipid transfer proteins which can permeabilize membrane. Moreover
plant storage proteins, like 2S-albumins, lectins, vicilins, glycine-rich proteins etc. accumulate
in storage vacuoles inside plant cells and act as antimicrobial agents in response to pathogen
attack. PR proteins also have roles in seed germination and adapting a plant to environmental
stress.

Five different classes of R genes are reported from various plant species. The most numerous
R gene class is the members of the gene family that code for proteins containing a nucleotide-
binding site (NBS) and leucine-reach repeats (LRRs). It is one of the most numerous gene
families in plants. The NBS domains are conserved and are involved in signaling. There is need
to study the NBS-LRR gene in plants to understand defense mechanism and its activation to
impart resistance to plants from harmful pathogens. This has huge implications in generating
greater yield.

These objectives of the study will be either of the following:

1. Will pre-immunizing plants at a young age help defend themselves efficiently against
future pathogen attacks?
2. Do plants gradually get adapted to the stress as the levels of plant defense molecules
get depleted?
3. Do pathogen treated plants give high phenolic content, and peroxide concentrations?
4. Is there a change in endogenous elicitor content (salicylic acid, jasmonic acid etc.) and in
the activities of antioxidant enzymes after infection with pathogen in the species chosen for
studies?
5. To develop potential genetic markers for marker assisted selection of disease
resistant plants and also for crop improvement.
6. Identify the NBS LRR region using universal primers specific to ITS1, 5.8S, and
ITS2 regions of DNA and its characterisation.
7. Screening and identification of antagonistic fungi against phyto-pathogens through
inhibitory studies.

Reference
Daniela Marone et al., Plant Nucleotide Binding Site–Leucine-Rich Repeat (NBS-LRR).
Genes: Active Guardians in Host Defence Responses, Int. J. Mol. Sci. 14, 7302-7326. 2013.

Deby Anna Augustine and Anu Yamuna Joseph, Isolation and Characterization of NBS-LRR
Resistance Gene in Banana (Musa AAB cv. Nendran), International Journal of Advanced
Biotechnology and Research (IJBR). Vol5, Issue3, pp353-358. 2014.

Eunsook Park et al., Plant–microbe interactions: organelles and the Cytoskeleton in action.
New Phytologist 217: 1012–1028. 2018.
 Department of Biochemistry – Biotechnology Project Proposals for 2023 - 25

Matthias Kretschmer et al., Chloroplasts and Plant Immunity: Where Are the Fungal Effectors?
Pathogens 9, 19. 2020,

Ningxiao Li et. al., Volatile Compound-Mediated Recognition and Inhibition Between

Trichoderma Biocontrol Agents and Fusarium oxysporum, Frontiers in Microbiology Front.
Microbiol. 9:2614. 2018

Parmar, p., Gandhi, and Subramanian, R. B., Isolation and molecular characterization of
avirulence gene from indian isolates of fusarium oxysporum f. Sp. Lycopersici. Journal of Cell
and Tissue Research Vol. 12(1) 3061-3067. 2012.

DP – 15: Evaluation of therapeutic potential of different Bacopa species

under in-vitro, in-vivo and in-silico conditions for neurodegenerative
conditions

Mentor: Sudeshna Menon

Neurodegenerative disorders are mainly caused by protein aggregation, though it is not only
limited to that. Protein aggregations lead to further complications in the nervous system, such
as disruption of protein trafficking, altered energy homeostasis, inflammation, synaptic
dysfunction and cytoskeletal disruption which ultimately leads to neuronal death. These
phenomenon are generally progressive in nature, and as the patients are mostly asymptomatic
during the early stages of these disorders, it is often diagnosed at a late stage when it becomes
even more challenging to treat (Wilson et al. 2023) . Majority of these disorders are caused due
to accumulation of proteins such as Aβ and Tau proteins in Alzheimer’s disease (AD), α-
synuclein in Parkinson’s disease (PD) and TDP-43 in amyotrophic lateral sclerosis (ALS),
hence, the symptoms are often manifested when significant amount of protein aggregation has
occurred in the system. This makes diagnosis of neurodegenerative disorders at early stages
very challenging for physicians around the world (Golde 2009).

Unfortunately, there is no cure for such neurodegenerative disorder regardless of decades

ofresearch. Interestingly, in the time span of 1981-2010, 1355 new drugs were approved out of
which 9.44% were for cancer treatment, 8.7% were antibacterial, 8.11% were antiviral and
5.8% were antihypertension drugs. On the contrary only 12 (0.88%) anti-Parkinsonian, 6
(0.44%) multiple sclerosis and 4 (0.29%) anti-Alzheimer drugs were approved (Newman and
Cragg 2012). Moreover, these anti-Alzheimer drugs are mainly focussed on retarding the rate
of progression of the disease, not to cure it but to delay the inevitable. These drugs are
mainlytargeted to treat some of the symptoms partially. It indicates complexity of the
pathogenesis of these disorders and our failure to understand these diseases despite of decades
long research (prince 2015).

Researchers have been targeting multiple pathways and biomolecules to find a viable treatment
option of neurodegenerative disorders. Some of which are enzymes like secretases, esterases
and proteases, neurotransmitters like acetylcholine, receptor antagonists like NMDA receptor
 Department of Biochemistry – Biotechnology Project Proposals for 2023 - 25

antagonists, protein aggregators, critical molecules involved in inflammation and anti-oxidant

pathways. New targets have also emerged in recent years like asparagine endopeptidase
enzyme for Alzheimer’s disease, which has shown great potential in experimental conditions
(Pohl and Lin 2018) . Stem cell therapy for treatment of neurodegenerative disorders has also
been a promising alternative, though there are many challenges to overcome to translate the
results in large scale clinical trials (Chia et al. 2020).

Currently, amyloid-directed antibody, cholinesterase inhibitors and glutamate regulators are

used as drugs for AD, dopamine supplements, decarboxylase inhibitors and dopamine agonist
are used for PD while glutamate-receptor antagonists and free radical scavengers are used for
treatment of ALS (Lamptey et al. 2022) . However, synthetic compounds used as drugs, or
under trials have limitations such as high cytotoxicity leading to severe side effects and large
scale production at affordable pricing leading to high cost of drugs, therefore, researchers have
been exploring plant based compounds for treatment of neurodegenerative disorders.

Several plants have been tested for anti-neurodegenerative properties which have yielded
positive results. For instance some of the plants tested are: Allium sativum, Olea europaea,
Punica granatum and Morus alba for anti-neurotoxicity, Moringa oleifera for cognitive
improvement, Winthania somnifera for anti-paralytic properties, Scutellaria baicalensis for
anti-tremor properties, Berthollletia excelsa, Fibraurea recisa and Zataria multiflora as anti-
inflammatory, Elaeis guineesis for anti-ulcer properties, Origanum vulgare for
immunomodulation, Moringa oleifera, Polygonatum sibricum, Vitis vinifera and Ocimum
tenuflorum for anti-aging properties, and Achyranthes aspera for its anti-epileptic properties.
This list is not exhaustive, since thousands of plants are currently under study for such
properties. Some of these plants like Morus alba and Vitis vinifera have also undergone clinical
trials with positive outcomes (Tyler and Tyler 2023) .

Researchers have used several models to study neurodegenerative diseases and its treatment.
Yeast, Saccharomyces cervisiae, a unicellular organism has around 23% homologous genes
with humans. It has been used widely to study protein misfolding and fundamental cellular
processes involved in neurodegenerative disorders. Yeast has been used to study various
neurodegenerative diseases like Huntington Disease (HD), AD, PD, ALS and prion diseases
(Winderickx et al. 2008). Animal cell culture has contributed greatly to understand the
physiological mechanisms of neurodegenerative disorders, as cell lines can be derived directly
from humans, it has complete genetic similarity with humans unlike animal models used to
study these diseases. Several cell lines like HEK293, SH-SY5Y, PC-12, ST-14A etc. are used
to study neurodegenerative disorders under in vitro conditions (Schlachetzki, Saliba, and de
Oliveira 2013) . Interestingly, Caenorhabditis elegans has 83% homology with human genes.
C. elegans has proved to be a great in vivo model for comprehension of molecular pathways as
well as genetics involved in neurodegenerative disorders. It has also been used extensively to
study protein aggregation (Li and Le 2013). Zebra fish, Danio rerio is an emerging model
organism to study neurodegenerative disorders as it shares 84% human disease-associated
genes. Its easy maintainace and short generation time is making it more attractive as a model
to study such disorders (Kalueff et al. 2015). Fruit flies (Drosophila melanogaster) as well as
 Department of Biochemistry – Biotechnology Project Proposals for 2023 - 25

mice and rats are also used to study neurodegenerative disorders broadly due to high genetics
similarities in disease related genes. However, translating similar results in humans have been
a major limitation (Tello et al. 2022).

DP – 16: Anticancer Potential of Some novel compound derived from plants

against Human Liver Cancer Cells /Human Breast cancer cells/Human
prostate cancer cells
Mentor: Dr. Shital Doshi
The cytotoxic response of the suggested substance will be evaluated by 3-(4, 5-dimethylthiazol-
2-yl)-2, 5-diphenyltetrazolium bromide (MTT), Lactate Dehydrogenase estimation and neutral
red uptake (NRU) assays, and morphological changes. Further, the influence of cytotoxic
concentrations of given compounds on oxidative stress markers, reactive oxygen species (ROS)
generation will be studied by measuring glutathione and lipid peroxidation levels. Additionally,
cell migration and invasion ability will be studied by various assays such as wound healing and
anchorage dependence assay, NA laddering, and fragmentation assay.

DP – 17: Characterization of anti-obesity compounds in C. elegans

Mentor: Dr. Shital Doshi
C. elegans, which have become a popular in vivo model for understanding the regulatory basis
of lipid biosynthesis and deposition will be utilized to screen potential anti-obesity compounds.
Here, the potential compounds will be evaluated in terms of their ability to decrease fat
deposition by Nile red/ Sudan black staining. The ability of compounds to modulate
metabolism will be studied by measuring insulin sensitivity assays and analysis of gene
expression. Total fat content will be evaluated by measuring tri glyceride content. Here, 3T3L1
adipocytes cell lines could also be used along with C. elegans as an additional model to further
support the data.

DP – 18: Biodegradation of plastics using microbial consortia

Mentor: Ms. Sonal Thakur
Worldwide production of plastics is increasing by approximately 5% every year, because
plastics exhibit advantages such as clarity, hardness, processability and lightness together with
price competiveness. However, plastics are mostly made from fossil resources such as
petroleum, coal and natural gas and lots of plastics are discarded after short service life. Waste
plastics lay enormous burden on the environment, because their recalcitrance to degradation
accelerates the accumulation in nature. Waste plastics buried in soil cause the water clogging
phenomena and devastate soil for agricultural cultivation. Lots of animals die of waste plastics
either by being caught in the waste plastics trap or by swallowing the waste plastics debris to
exert ruinous effects on the ecosystem. They are also carcinogenic for humans and affect the
 Department of Biochemistry – Biotechnology Project Proposals for 2023 - 25

human hormones. To curb the problem of plastic pollution, chemical and biological methods
have been used. Use of chemical methods for plastic degradation has further increased
pollution by releasing toxic gases into the atmosphere and so biological methods are preferred
as they are eco-friendly.
Biodegradation of plastic involves the attachment of microorganisms to the surface and use of
polymer as carbon source. In the aerobic process of biodegradation which is eco-friendly,
plastic reacts with oxygen from the air and then the microorganisms, facilitate this degradation
process by secreting plastic degrading enzymes to oxidize or break down the products for its
energy into smaller by-products such as carbon dioxide and water. The limiting factor affecting
the rate and range of hydrocarbon degradation by microorganisms appears to be the lack of
ability of most microbial strains to utilize different components of plastic. Different strains can
degrade different components, but a single strain can usually attack a limited number of
hydrocarbons. Hence, a bacterial consortium is more nutritionally versatile than a single strain
and exhibits considerable competence in utilizing a large number of hydrocarbon components
from plastics. Knowledge regarding the degradation of these compounds is essential to
determination of the fate of these compounds in the environment. In such cases, computational
tools may be used to predict biodegradation pathways for these chemicals.

References
Mahalakshmi, V., & Siddiq, S. A. (2015). Enhanced biodegradation of polyethylene by
development of a consortium. Pelagia Research Library, 6, 183-189.
Yoon, M. G., Jeon, H. J., & Kim, M. N. (2012). Biodegradation of polyethylene by a soil
bacterium and AlkB cloned recombinant cell. J Bioremed Biodegrad, 3(4), 1-8.
Ahmed, T., Shahid, M., Azeem, F., Rasul, I., Shah, A. A., Noman, M., ... & Muhammad, S.
(2018). Biodegradation of plastics: current scenario and future prospects for environmental
safety. Environmental Science and Pollution Research, 25(8), 7287-7298.

DP – 19: Optimization of fermentation conditions to enhance citric acid

production

Mentor: Dr. Yogesh Baravalia

Citric acid is the most important organic acid produced in tonnage and is extensively used in
food (10%) and pharmaceutical industries (60%). It is produced mainly by submerged
fermentation using Aspergillus niger or Candida sp. from different sources of carbohydrates,
such as molasses and starch-based media. Various other agro-industrial residues such as apple
pomace, cassava bagasse, coffee husk, wheat straw, pineapple waste, sugar beet cosset, kiwi
fruit peel, etc. have been investigated with solid state fermentation techniques for their potential
to be used as substrates for citric acid production (Pandey et al. 1999; Pal et al 2016). There is
a need to develop a medium that can ease the production of citric acid. It is known that the
 Department of Biochemistry – Biotechnology Project Proposals for 2023 - 25

factors mainly affecting the citric fermentation are the type and concentration of carbon source,
nitrogen and phosphate limitation, pH, aeration, oligo elements concentration, and morphology
of the producing microorganism. Objective of the present proposal is isolation and evaluation
of citric acid production ability of microbial strain, designing the submerged, solid-state as well
as surface fermentation and comparing the production potential, optimization of production
conditions (physico-chemical), optimizing the extraction method and scale up studies. Output
of the study may have beneficial impact on many industries such as food industry, dairy
industry, pharmaceutical industry, paper industry, textile industry etc.

Key words: Citric acid fermentation, Aspergillus niger, optimization, organic waste, molasses,
solid state fermentation, liquid state fermentation

DP – 20: Isolation, characterization and biological activity of oil from Citrus

fruit peels and seeds

Mentor: Dr. Yogesh Baravalia

Citrus fruits are one of the most important health beneficial foods to combat the disease and
improve the health. Peels and seeds of these fruit have valuable constituents known as oil /
essential oil. These oils are used in various industries like, food, pharmaceuticals and
cosmetics. Some of the essential oils from citrus have high economic importance mainly in
food and cosmetic industries due to health beneficial and desired flavour properties. Normally
citrus peels and seed are thrown out as waste. It is possible to make best use from waste.
Purpose of this study is to use this waste to isolate oil / essential oil from peels and seeds of
some citrus fruit (lemon, orange, etc.) by various methods, Physicochemical characterization
of isolated oils, assess its antioxidant and antimicrobial properties and to explore other possible
area from the same.

DP – 21: An in-silico and in-vitro approach for management of cancer

Mentor: Dr. Monica Thakur

Cancer remains the most enigmatic of all diseases and despite a plethora of research attempts
to demystify the related etiology, signaling events and resultant pathology, any cure remains
elusive, and cancer remains the leading cause of death worldwide. In addition, the pattern and
mortality rates vary from countries and across different cancer types. Differences in lifestyles,
environment and exposure to risk factors results in the mortality variations. Cancer has always
been a key concern for healthcare in developing as well as under-developed countries. India
also experiencing an increase in the cancer incidence and mortality rate. Despite advances in
the diagnosis and treatment, cancer is still a number one cause for the mortality, chiefly due to
the inability of early detection of the disease. Most of the patients are diagnosed at locally
advanced or at a metastatic stage when the management of the disease is difficult due to
extreme progression of the disease and hence, patients have to face the trauma of side effects
 Department of Biochemistry – Biotechnology Project Proposals for 2023 - 25

and toxicity of the treatment. Asymptomatic progression, distant metastasis, ineffective

treatment, low sensitivity, and specificity of the biomarkers available for detection and
treatment, resistance to the therapy are the major causes of high mortality rate in cancer.
Therefore, a better understanding of the biological processes at an early stage is crucial for the
discovery of further therapeutic interventions. Hence, there is an urgent need for a better
biomarker, combinational or alternative treatment options to improve the prognosis at an early
stage.

Computational or in-silico approaches for protein interaction studies have been developed over
recent years using various tools. These in-silico tools provide significant opportunity to
investigate oncogenesis by detecting protein-ligand and protein-protein interactions. These
interactions are an important part of many signaling pathways related to biological processes
and protein functions. Several structure-based approaches are beneficial today for prediction
of these interactions. Ligand and target based virtual screening are computational methods
widely accepted for the identification of bioactive components for the target of interest and to
estimate the binding affinity of a test component. These approaches can successfully identify
novel bioactive compounds and novel inhibitors for targets.

In India, herbs have proven to be efficacious in the management of various health problems;
the use of medicinal herbs has been escalating in recent times in developing countries, as these
herbs have been reported to be safe and with less adverse effects than synthetic drugs.
Phytocomponents present in these herbs have various therapeutic properties that can be
effectively employed for the control of cancer related events; qualitative analysis of medicinal
herbs can be helpful in identifying the presence of such phytocomponents. Free radicals, the
naturally occurring highly reactive chemicals potentially harm the normal cells of the body
when present in abnormally high concentration and increase the risk of cancer development.
Phytocomponents are rich in antioxidants and can neutralize the free radicals, preventing the
ensuing cell damage. Assessment of free radical scavenging activity of medicinal herbs can be
advantageous in predicting the antioxidant effect of phytocomponents on proliferating cells.
Phytochemicals possess various properties including antioxidant, free radical scavenging and
anti-proliferative; hence phytocomponents can modify tumor behavior resulting in the
reduction of tumor cell proliferation. Hence, the role of potent phytochemicals in preventing
activation of oncogenic signaling pathways by blocking the active sites of specific proteins and
their action as possible inhibitors against target proteins can be investigated. The analysis of
proteins by quantitative methods can detect the effect of medicinal herb on the proteins in tumor
cells, hence these herbs can benefit cancer patients by preventing further growth of tumor and
improving the survival rate without severe side effects and the target proteins can serve as a
marker for early detection and new therapeutic approach.

In this investigation therefore, in-silico analysis will be carried out by employing computational
methods to identify phytomolecules from medicinal herbs. These identified phytomolecules
were then screened to establish binding interactions against potential biomarker protein targets.
The in-vitro studies will be performed for the phytochemical analysis and for evaluation of
antioxidant potential of the prepared extracts of selected herbs. The efficacy of the selected
extracts as potent anti-proliferative agents against the cancer will be investigated through in-
vitro analysis and further, the protein extraction and separation from cultured treated and
untreated cells with extracts of selected plants was performed by PAGE to identify potential
protein markers.
 Department of Biochemistry – Biotechnology Project Proposals for 2023 - 25

References:

 Durgawale, P. P., Patil, M. N., Joshi, S. A., Korabu, K. S., and Datkhile, K. D. (2019).
Studies on phytoconstituents, in vitro antioxidant, antibacterial, antiparasitic,
antimicrobial, and anticancer potential of medicinal plant Lasiosiphon eriocephalus
decne (Family: Thymelaeaceae). Journal of Natural Science, Biology and Medicine,
10(1), 38.
 Ferlay, J., Ervik, M., Lam, F., Colombet, M., Mery, L., Piñeros, M., Znaor, A.,
Soerjomataram, I. and Bray, F. (2020). Global Cancer Observatory: Cancer Today.
Lyon, France: International Agency for Research on Cancer. Available from:
https://gco.iarc.fr.
 GLOBOCAN 2020, International Agency for Research on Cancer, World Health
Organization, 2020. (https://gco.iarc.fr/today/data/factsheets/cancers/15-Lung-fact-
sheet.pdf).
 Jacobs, E. C. (2018). Potential therapeutic effects of phytochemicals and medicinal
herbs for cancer prevention and treatment. Arch Gen Intern Med. 2018; 2 (3): 44-48.
 Pinzi, L., and Rastelli, G. (2019). Molecular docking: Shifting paradigms in drug
discovery. International journal of molecular sciences, 20(18), 4331.
 Sung, H., Ferlay, J., Siegel, R. L., Laversanne, M., Soerjomataram, I., Jemal, A. and
Bray, F. (2021). Global cancer statistics 2020: GLOBOCAN estimates of incidence and
mortality worldwide for 36 cancers in 185 countries. CA: a cancer journal for clinicians,
71(3), 209-249.
 The ClusPro web server for protein-protein docking. Nat Protoc 2017;12:255-78.
Kozakov D, Beglov D, Bohnuud T, Mottarella SE, Xia B, Hall DR, et al. How good is
automated protein docking? Proteins 2013; 81:2159-66.
 Thomsen R, Christensen MH. MolDock: A new technique for high-accuracy molecular
docking. J Med Chem 2006; 49:3315-21. Kozakov D, Hall DR, Xia B, Porter KA,
Padhorny D, Yueh C, et al.
 Umadevi, M., Kumar, K. S., Bhowmik, D., and Duraivel, S. (2013). Traditionally used
anticancer herbs in India. Journal of Medicinal Plants Studies, 1(3), 56-74.
 Wild, C. P., Weiderpass, E., and Stewart, B. W. (2020). World cancer report: cancer
research for cancer prevention. Lyon: International Agency for Research on Cancer.
 YASARA: Krieger, E., and Vriend, G. (2015). New ways to boost molecular dynamics
simulations. Journal of computational chemistry, 36(13), 996-1007.

DP – 22: Herbal extracts improve Glucose tolerance

Mentor: Ms. Srishti Chhabaria
Type 2 Diabetes is characterized by insulin resistance, a reduction in insulin secretion by
pancreatic beta cells and impaired incretin response, is a global epidemic and is still increasing.
More than 300 million people worldwide have been diagnosed .Inadequate T2D management
is a leading cause of cardiovascular diseases, blindness, kidney disease and even death. Owing
to the role of Beta cell dysfunction and improper insulin secretion, major efforts have been
taken to understand the factors and pathways involved in insulin secretion. Medicinal plants
have been explored for their potential to directly stabilize blood glucose levels or to indirectly
help issues related to diabetes such as insulin resistance, obesity etc. Studies in this area will
definitely help in the treatment regimens of diabetic patients.
 Department of Biochemistry – Biotechnology Project Proposals for 2023 - 25

MSc Dissertation Projects 2023 - 25

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