Biochemistry And Microbial Sciences - Master Dissertation

Permanent URI for this collectionhttps://kr.cup.edu.in/handle/32116/24

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Start date: 2018End date: 2018

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    Antibacterial and Anticancer property of bioactive secondary metabolites from Actinomycetes isolated from different regions of Himachal Pradesh
    (Central University of Punjab, 2018) Chambiyal, Gourav; Kushawaha Pramod K.
    The prime objective of the present work is to isolate, characterize and purify soil microbes and to know about their antibacterial and anticancer properties. Soil samples were obtained from different locations in Dharamsala, Himachal Pradesh, India. Serial dilutions of the samples were made and plated on starch casein agar. Screening, purification and further tests were performed on the isolates to find the potential colonies. Biochemical and morphological characterization was done as well. Following characterization, isolates were used to extract bioactive secondary metabolites which can be anti-bacterial and anti-cancer. Secondary screening for antibacterial properties of isolates was evaluated by Agar well diffusion method. Further, the isolates were subjected to the GC-MS and MTT cytotoxic assay. Totally 7 isolates of actinomycetes were isolated and out of them only 4 were tested for antagonistic activity against 4 pathogenic microorganisms. Isolates M1,and M3 of dilution 10-1 were active, while M2 and M4 showed less activity against the pathogenic microorganisms. All actinomycetes isolates showed antibacterial activity against S. enterica and P. putida while they showed less activity against S. aureus and E.coli. These isolates showed antibacterial and anti-cancer activities and may be used for the growth of new antibiotics for pharmaceutical or agricultural purposes.
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    Role of Plant Growth Promoting Bacteria in Improving the Quality of Marginal Soil for Multiple Purposes
    (Central University of Punjab, 2018) Kumari, Ritu; Wusirika, Ramakrishna
    Degrading quality of soil by the presence of xenobiotics or change in the natural environment of soil is of great concern. One of them is heavy metal pollution of soil. With increasing population, sustainable food production is a challenge in this climate changing and water deteriorating environment. Marginal soil has been defined from different perspectives. In general, marginal land can be defined as arid and inhospitable land that has low inherent productivity. Marginal land is a good source for the production of biofuels. This can be achieved with the help of Plant Growth Promoting Bacteria (PGPB). This project provides a review of marginal soil and how PGPB can be used to improve its quality, in phytoremediation, and in the production of bioenergy and food.
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    Expression, Purification and Kinetic Characterization of Lactate Dehydrogenase of Lactic Acid Bacteria and ProB of Bacillus subtilis
    (Central University of Punjab, 2018) Gunjan; Verma, Malkhey
    Lactate dehydrogenase is very well known for its role in glucose metabolism in Lactic Acid Bacteria (LAB). These bacteria are widely used in the industrial and research areas. LAB use NADH as a cofactor to produce lactate or lactic acid from glucose through fermentation, whereas ProB is an enzymatic protein which catalyzes the conversion of L-glutamate to ?-glutamyl phosphate at the expense of ATP in proline biosynthesis. Both the recombinant proteins are expressed in the E. coli BL 21 strain using pET plasmids. Proteins are purified using affinity chromatography and purity is established by a single band in SDS Page. Proteins were quantified by Bradford assay. In this study, the proteins selected are Lactate Dehydrogenase and ProB. In the absence of FBP, Pi is an activator of L. lactis LDH at pH 6. This effect can be interpreted by considering the computed binding affinities of Pi to the catalytic and allosteric binding sites of the enzymes modelled in protonation states corresponding to pH 6 and pH 7. In this study, we can find out the delicate interplay among the effects of Pi, FBP, and pH that results in different regulatory effects on the LDH of LAB and ProB of B. subtilis.
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    Production and Optimization of Lovastatin by Microbial Fermentation using Aspergillus terreus
    (Central University of Punjab, 2018) Raj, Rohit; Verma, Malkhey
    In the recent times, the fungus Aspergillus (A.) terreus has been highly popularized regarding its domination for the production of the "crackerjack" drugs known as statins, particularly lovastatin. The aim of this research was the production of lovastatin which is a known cholesterol-lowering drug, through microbial fermentation using A. terreus. Besides, it also aimed to analyze certain bioactive chemical products and evaluation of such antibacterial and antifungal products, if any produced. Bioactives (chemical compounds often referred as secondary metabolites) were analyzed using the Gas Chromatography-Mass Spectroscopy technique (GC-MS) technique. A. terreus is known to produce a vast variety of important secondary metabolites with high biological activities. The extraction of the natural statins such as lovastatin or mevastatin from A. terreus is seen as one of the major breakthrough in the field of Industrial Microbiology/Fermentation Technology. Here we report the Aspergillus terreus NBRC (IFO) 31217 (Strain I) and ATCC 11877 (Strain II) don't produce lovastatin but they produce important bioactive compounds of high commercial value like Isovaline (C5H11NO2) and Silane etc.
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    Assessment of Extract of Syzygium cumini Against Doxorubicin Induced Cardiotoxicity
    (Central University of Punjab, 2018) Chayan, Mukherjee; Monisha Dhiman
    For the past four decades, doxorubicin (DOX) has been used to treat cancer, mainly solid tumours and haematological malignancies. However, clinical community is greatly concerned regarding the administration of this as DOX treatment is commonly associated with dose-dependent cardiotoxicity. Attempts at alleviating drug generated cardiac damage using an extract from different parts of plants with radical scavenging property are a promising area of research. Hydroalcoholic extract derived from fruit pulp of Syzygiumcumini which has a significant antiradical scavenging effect. This study aims to assess the effect of parallel administration of SC fruit pulp extract (SC) on mitigating or preventing DOX induced cardiotoxicity in vitro using H9c2 cardiomyoblast cell lines. Addition of SC fruit pulp extract and DOX were performed for both treatment and control sets on H9c2 cells. SC fruit pulp extract showed strong ABTS cation radical scavenging activity in a dose dependent manner. MTT assay was used to study the cytotoxic effect of SC fruit pulp extract and DOX. ROS levels were estimated using NBT assay and DHE assay. The results showed that DOX has significant cytotoxic effect in a dose dependent manner while SC fruit pulp extract did not display any significant cytotoxicity on H9c2 cells. The DOX induced ROS production was found to be significantly reduced in SC fruit pulp extract treated cells. Results of the current study also suggest that the treatment of SC fruit pulp extract along with DOX, displayed cardioprotective potential in H9c2 cells by: 1) reducing lipid peroxidation; 2) decreasing extracellular nitric oxide (NO); 3) decreasing the expression of the protein p47phox and iNOS/NOS-2. These results clearly suggest that treatment of SC fruit pulp extract along with DOX reduces the DOX induced toxicity and hence can be a promising therapeutic intervention in managing DOX mediated cardiotoxicity.
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    Isolation And Determination Of Antibiotic Susceptibility Of Enterococcus Faecalis From Malwa Region
    (Central University of Punjab, 2018) Garg, Dimple; Baranwal,Somesh
    About 20 isolates of Enterococcus faecalis is isolated from six different places of Malwa region. They include isolate from Animal waste soil, Agriculture soil, Sewage soil and public place soil also. Isolation of Enterococcus faecalis from environment causes concern for diseases i.e. Endocarditis, UTI, neonatal infections due to maintaince of unhygienic conditions. The biochemical tests are performed and measure as presumptive results for Enterococcus faecalis. Antibiotic susceptibility test is performed to observe the susceptibility of strains for different antibiotics. Six different types of antibiotics such as Ampicillin, Piperacillin, Tetracycline, Rifampin, Streptomycin, and Gentamycin are used for Antibiotic Susceptibility test. Antibiotic Susceptibility test analyse that Gentamycin and Streptomycin are found more resistant towards Enterococcus faecalis than others in Malwa region. The susceptibility of strains for antibiotics are different due to the IS elements which are horizontal transfer to the other Enterococcus faecalis also formed biofilm which makes it more antibiotic resistant due to slower growth rate, metabolic activities and poor penetration rate.
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    Identification of Novel Natural Inhibitors Of Proteins Involved In Cancer Cell Stemness
    (Central University of Punjab, 2018) Malik, Rebati; Kumar, Shashank
    Cancer stem cells (CSCs) are a small subpopulation of cells identified in a variety of tumors that are capable of self-renewal, differentiation and have the unique property to evade radiotherapy and chemotherapy. CSCs are a very likely cause of resistance to current cancer treatments, as well as relapse in cancer patients. Compared to differentiated tumor cells, CSCs have some important distinguishing feature that confers chemoresistance in these cells. Different proteins such as Bcl-2 (2O21), CXCR4 (3ODU), CHK1 (4FSZ), MTH1 (5ANV), VEGFR2 (1Y6A) and Carbonic anhydrase II (5SZ2) have been reported to involve in cancer cell stemness. Now day's natural products are popular remedies against various diseases including cancer. These products have been reported for their low/non-toxicity and cost-effectiveness. The phytochemical terpenoids, biggest class of naturally occurring compounds derived from five-carbon isoprene unit. They play an important role in binding to the above signaling proteins which are involved in cancer stem cells. Therefore, we studied receptor-based molecular docking of natural terpenoids against target proteins.
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    Role of Helicobacter pylori Enriched Media in Inducing Oxidative Stress in Human Cell lines
    (Central University of Punjab, 2018) Samal, Pallavi; Dhiman, Monisha
    Helicobacter pylori is a gram-negative, helical, microaerophilic bacterium which colonizes the human gastrointestinal tract. Vacuolating cytotoxin A (VacA) is one of the major virulent factors. Reactive oxygen species (ROS) and Reactive nitrogen species (RNS) produced by the immune and epithelial cells damage the host cell thereby resulting in a persistent infection. The prolonged infection results in chronic inflammation, oxidative stress and DNA damage. The microbe affects the major macromolecules of the host tissues lipids, proteins and DNA which leads to lipid peroxidation, protein oxidation and DNA fragmentation hence making the oxidative stress a deleterious damage. Role of H. pylori enriched media (HPEM) in inducing oxidative stress in two human cell lines AGS (human gastric cell line) and THP-1(human monocytic cell line) was studied in present work. The AGS cells and THP-1 cells was treated with various concentrations of HPEM and oxidative stress was evaluated by examining the levels of protein carbonyls, TBARS (thiobarbituric acid reactive species) and nitric oxide by spectophotometric and Western blotting methods. The oxidative stress induced by HPEM showed damaging effects on the cell membrane, protein and produced significantly high nitric oxide (NO) when compared with the untreated controls. From the present work it can be concluded that HPEM exposure to THP-1 and AGS cells enhanced the oxidative stress which leads to cellular damage and is ultimately responsible for the severe H. pylori associated fatal complications during its pathogenesis.
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    Extraction And Identification Of Secondary Metabolites Produced By Soil Actinomycetes And Their Biochemical Characterization
    (Central University of Punjab, 2018) Singh, Jahnavi Kumari; Kushawaha, Pramod
    The worldwide use of antibiotics has rapidly increased since the discovery of penicillin in 1928 by Alexander Fleming, for treating terrible bacterial diseases. But in due course of time, over-consumption of antibiotics has led to an enormous selective pressure on bacteria, which has forced them to adapt to the antibiotics and hence gain development of resistance. In the past years, the excessive use of antibiotics has led to significant evolution in microorganisms where naturally occurring genes resistant to the antibiotic have been spreading fast, thus, the need for new antibiotics has increased. In the stated work, samples were collected from the mountainous sites of Himachal Pradesh and were analyzed for the presence of secondary metabolites. The bacterial species were characterized by biochemical tests and metabolites produced by them were identified.
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    Identification of some novel natural inhibitors of proteins in the life cycle of the malarial parasite
    (Central University of Punjab, 2018) Kumari, Priyanka; Kumar,Shashank
    Antimalarial phytoconstituents were identified and analysed.We studied receptor based molecular docking of protein drug targets( Plasmepsin 2 (1lf2),HSP90 (3k60), PfATPase ( 1u5n), Human orotidine 5 - decarboxylase domain (2v30), and Plasmodium orotidine 5- decarboxylase domain (3n3m) against the natural phytochemical .The dock score were in range of -6.1 to -10.7 for antimalarial targets respectively. Some ligands showed better dock score than standard inhibitors of the respected proteins from the range (-6.9 to -10.7).