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Item Proximate and heavy metal analysis of carnivorous and herbivorous fish species from Lahore, Pakistan(UMT Lahore, 2023-02-13) Nazish Riaz; Mnahil Baig; Ayaz Khan; Aiman RafiqThis study is done to determine the proximate analysis and metal analysis between selected groups of Herbivore and Carnivore fish. The proximate composition includes the ash, moisture, fat and protein contents and the analysis was done all these proximate compositions and on selected metals in Herbivore and Carnivore fish. The overall contents of nutritional parameters between and within the groups varies. The protein content ranges from 13.39333333% - 16.82% while the overall protein content in Herbivores are greater than Carnivores. The fat content ranges from 1.076666667% - 2.15% whereas the fat level in Herbivore is more than the Carnivores but the difference is very small to notice. The moisture content ranges from 74.18% - 81.03333333% while the overall amount of moisture is higher in Carnivores then the Herbivores whereas the ash content also high in Herbivores then the Carnivores and ranges from 2.553333333% - 6.143333333%. For metal analysis the Iron (Fe) and Copper (Cu) are not detected (ND) in all Herbivore and Carnivore. The levels of Cadmium (Cd) in Herbivores are more than the levels in the Carnivores. The level of Manganese (Mn) in Herbivores are more than the levels in Carnivores, but the difference is not much greater. The levels of Sodium (Na) and Calcium (Ca) are also greater in Herbivores as compared to Carnivores. Despite all the metals the levels of Zinc (Zn) are seen much higher in Carnivores as compared to Herbivore. The profile of these metals and proximate composition of fish will be useful in identifying feed-related indicators of exposure in aquatic creatures in the future.Item Isolation and growth optimization of chromium resistance bacteria from wastewater(UMT Lahore, 2023-02-03) Iqra Sohail; Nimrah Khaliq; Saifullah Malik; Javairia AslamThe current study was aimed at the isolation of bacteria that is resistant to Chromium from industrial wastewater. As industries are increasing day by day, different environmental risks associated with them are also increasing. The major environmental risk associated with industries is the discharge of effluents containing heavy metals. Cr (VI) is one of the most toxic from of chromium and contamination of this metal poses a threat to the environment. In this study strains of different bacteria were isolated from the dye industry effluents. A total of 6-gram positive heavy metals resistant strains were isolated. By biochemical characterization different chromium resistant strains were identified. The bio sorption capacity of these strains for chromium was conducted at a concentration of 0.25mM. At different concentrations of chromium, all the strains showed different MIC and growth range which indicates that all the strains have different Cr resistance. Among all the selected bacterial strains MC3 and MC4 showed maximum chromium reduction ability in a short period of time. At MIC of 6mM, MC3 showed an optical density of 0.2 at a wavelength of 600nm while MC4 showed an optical density of 0.23 at 600nm. The strain MC3 reduced 1.6 to 0.4 of Cr (VI) at an OD of 540nm in a four- hour time period while the strain MC4 reduced Cr (IV) from 1.05 to 0.42 at an OD of 540nm in a four-hour time period. The Cr (VI) reduction ability of these strains showed that the strains MC3 and MC4 can probably be used for bioremediation of Chromium.Item Relationship of body measurements of neoscona theisi with web characteristics in field conditions(UMT Lahore, 2023-02-13) Huzaifa Binte Rashid; Ayesha Zahid Ghuman; Warda Razzaq Cheema; Hina ShahzadiThe goal of the current study was to determine and record the changes in the web's various parameters, including capture thread length (CTL), web height, and number of radii. Only the webs constructed by Neoscona theisi in field conditions were assessed in this study. Relationship of body measurements of N. theisi with web characteristics were studied in this research. According to data, spiders with high body weight and high body length build webs with greater capture thread lengths (CTL) and greater web height. But the body length showed weak or almost no correlation with number of radii. Carapace width also had effect on CTL. Increase in carapace width led to increase in CTL as well. Whereas Carapace width had negative correlation with number of radii. Web height was also influenced by carapace width. Increase in carapace width led to increase in web height as well. Also, the relationship between plant height and web height was also recorded. The data showed that there was positive correlation between pant height and web height. This study showed that web design variation in N. theisi was influenced by spider’s weight, body length and carapace width. This is helpful for determining their potential as biological control in agronomy.Item Isolation and characterization of mercury resistant bacteria from waste water.(UMT Lahore, 2023-02-03) Muhammad Ihtasham Mughal; Shahzara IrshadMercury (Hg) is one of the most toxic heavy metals found on earth, and has significantly reported as a contaminant from industrial and agricultural activities. Its release in the environment has led to extreme toxicity to all life forms. Various strategies like physico-chemical techniques have been proposed for Hg removal; bioremediation has proved to be most promising among all. This study focuses on the isolation of Hg resistant bacteria from Ravi River samples. Several biochemical tests were performed like gram staining, catalase, oxidase, sugar fermentation, antibiotic resistance, and motility test. Minimum inhibitory concentration (MIC) was determined to screen the isolate resisting highest selected concentration of Hg. Salt tolerance and pH tolerance tests were also performed. Metal reduction analysis was also run to determine the reduction ability of the isolates for 1100ppm Hg. Hg4, Hg6 and Hg7 were selected due to their higher MIC, salt tolerance, and pH values along with greater potential to reduce Hg2+ to Hg0. In future, these isolates are to be taken towards molecular identification followed by field experiments.Item Integrative network analysis to identify hub genes in metastatic prostate cancer(UMT Lahore, 2023-02-16) Asma Muhammad Mumtaz; Atiqa Iftikhar; Faria Faran; Rimsha Hamid MehmoodMetastatic prostate cancer is the leading cause of prostate cancer related deaths in men around the globe. We performed a detailed bioinformatic analysis to identify the genes involved in the metastasis of the prostate cancer. After identifying the prominent genes that may be involved in metastatic mechanism, we identified purposable drugs, targeting those genes. We identified DEGs using GEO databases. The DEGs thus obtained were validated using TCGA datasets available at GEPIA and TACCO databases. About 14 genes were found to be common in all three databases i.e., GEO, GEPIA and TACCO. Furthermore, we performed GO and KEGG enrichment analyses using DAVID database. PPI-network was constructed using STRINGS software. This network was then analyzed using Cytoscape software. Two plug-ins which were already present in CytoHubba were used for this purpose. The plug-in CytoHubba was used for identifying hub genes and the plug-in MCODE was used for the formation of clusters. Among top 10 hub genes, four genes, ADCYS, PRKG1, ESPR1 and RBFOX2, were present in 2 MCODE clusters. These 4 genes were then used as targets to identify suitable drugs from 3 databases, CMap, DrugBank and TTD. The results identified potential drugs against ADCYS and PRKG1 genes. The effectiveness of the drugs against these two genes were then analyzed by performing Molecular Docking using MOE software.Item Association between three genetic variants in micro-RNAs (rs3746444, rs11614913 & rs2910164) and prostate and gastric cancer risk(UMT Lahore, 2023-03-15) SAMAN RAUF RAMAY; MAHEEN ZAHRA; GULNAZ BIBI; BUSHRA MURTAZAThis study is about "Association between Three Genetic Variants in Micro-RNAs (rs3746444, rs11614913 & rs2910164) and Prostate and Gastric Cancer Risk" show that prostate and gastric cancer is a multifactorial disease, which are leading cause of death in Homo sapiens. These cancers can also be cause by single nucleotide polymorphism. Both types of cancer have three common SNPs in miRNA, which causes cancer. The total number of studies for qualitative and quantitative synthesis for data were n= 27. We retrieve data by using PRISMA guidelines. And we perform meta-analysis of those three SNPs in different ethnicities of Asian and European populations by using SPSS and STATA and it is found that the SNP rs11614913, which is associated with miR196a2, cause prostate cancer only, SNP rs2910164 associated miR146a, do not cause any cancer and SNP rs3746444 associated with miR499 can cause both prostate and gastric cancer.Item Association between three genetic variants in micro-rnas (rs3746444, rs11614913 & rs2910164) and prostate and gastric cancer risk(UMT Lahore, 2023-02-13) SAMAN RAUF RAMAY; MAHEEN ZAHRA; GULNAZ BIBIThis study is about "Association between Three Genetic Variants in Micro-RNAs (rs3746444, rs11614913 & rs2910164) and Prostate and Gastric Cancer Risk" show that prostate and gastric cancer is a multifactorial disease, which are leading cause of death in Homo sapiens. These cancers can also be cause by single nucleotide polymorphism. Both types of cancer have three common SNPs in miRNA, which causes cancer. The total number of studies for qualitative and quantitative synthesis for data were n= 27. We retrieve data by using PRISMA guidelines. And we perform meta-analysis of those three SNPs in different ethnicities of Asian and European populations by using SPSS and STATA and it is found that the SNP rs11614913, which is associated with miR196a2, cause prostate cancer only, SNP rs2910164 associated miR146a, do not cause any cancer and SNP rs3746444 associated with miR499 can cause both prostate and gastric cancer.Item Level of knowledge, attitude and risk factors of covid-19 among pakistani population(UMT Lahore, 2023-02-14) Iqra Shahbaz; Fiza Nadeem; Mariam Nadeem; NishatCoronavirus is one of the pandemic disease spread worldwide on 20 May 2020 confirmed by the World Health Organization, caused by (SARS-CoV-2). It causes respiratory tract or intestinal infection. Initially, the first case of coronavirus was observed on 19 Dec 2019, in the city of China Wuhan, and later globally that’s why it is called Covid-19. The current study was aimed at the level of knowledge, attitude and risk factors of covid-19 among Pakistani population: a cross sectional study. We conducted a survey on google form. A questionnaire of total 27 questions which include demographics, knowledge, attitude and practices Total 400 responses were collected both online and physical. Among the respondent’s female were outnumbered than male participants. This survey revealed that people had good knowledge, positive attitude and good practices toward COVID-19 pandemic. Respiratory diseases are Major risk factor of COVID-19 infection. Public Education, income status and lifestyle also related to the knowledge and perception about COVID-19. There is further need to educate the public and spread awareness about COVID-19 and to improve the public hospitals and health care services in order to enhance prevention strategies among the general population of Pakistan.Item Synthesis of anti IL-6 polyclonal antibody and its characteristics(UMT Lahore, 2023-10-03) KALSOOM IJAZ; AREEHA ALYAS; MARYAM ZARIFWe present the synthesis and characterization of a polyclonal antibody targeting Interleukin-6 (IL-6), a key cytokine involved in various inflammatory and immune responses. The research involved the development of a novel polyclonal antibody and its subsequent evaluation in a murine model. The primary objective was to establish a reliable and specific antibody against IL-6, which is crucial for understanding its role in various physiological and pathological processes. The synthesis of the anti-IL-6 polyclonal antibody involved immunization of mice with recombinant IL-6, followed by serum collection and antibody purification. The resulting antibody was characterized through immuno dot blot and enzyme-linked immunosorbent assay (ELISA) techniques to confirm its specificity and affinity towards IL-6. Our findings reveal that the synthesized polyclonal antibody effectively recognizes and binds to IL-6 in murine samples, demonstrating its specificity and suitability for further research and diagnostic applications. This research contributes to the development of valuable tools for studying IL-6-related mechanisms in various biological contexts, ultimately enhancing our understanding of its role in health and disease. Additionally, the methodology and results presented in this thesis offer a foundation for future investigations involving IL-6 and related cytokines.Item Computational analysis of variants of uncertain significance of pus7 gene(UMT Lahore, 2023-09-14) GHOSIA NOREEN; SHAHBAZ AHMAD; NIDA NADEEMIntellectual disability is a rare autosomal recessive disorder characterized by mental retardation, limitations in the cognitive abilities and adaptive functioning of an individual. It is caused by mutations in PUS7 gene which is present on chromosome 7 of human genome encoding protein pseudo uridine Synthase. The enzyme catalyzes the conversion of Uridine into Pseudo uridine during the transcriptional process which stabilizes the RNA as part of post-transcriptional modification. In this study, we applied in-silico analysis to predict the most deleterious missense and splice site mutations in PUS7 gene and their impact on stability, function, post translational modification of pseudouridine synthase. For that purpose, a variety of bioinformatics tools based on different algorithms were utilized. CADD and MetaSNP were used to analyze diseased missense variants. Moreover, PTM analysis was by using ScanProsite which showed three variants lying in those regions. CADD and MetaSNP tools were used to predict pathogenic variants. Out of 217 missense variants, 38 variants were considered pathogenic by both the missense tools. The splice site analysis was performed on 39 variants left after CADD analysis by using spice and splice AI. Moreover, PTM analysis was by using ScanProsite which showed three variants p.Ile352Thr, p.Tyr330Cys and p.Thr299Lys lying in those regions. Out these only one variant is observed on the exposed part of the protein i.e., p. Tyr330Cys. These findings can be very helpful for scientists in the future as they can directly target these mutations in order to produce certain medications and drugs.Item Synthesis of anti-interferon beta 1a antibody in MICE and its characterization(UMT Lahore, 2023-09-15) MUHAMMAD SHOAIB KHAN; RIZWAN ULLAHMultiple Sclerosis (MS) is a disease of nervous system. In this autoimmune disease, the immune system of the body starts attacking the healthy nerves and degenerates the myelin sheath present on them. This results in the formation of scars on the multiple sites on nerves. In the patients of MS, brain to body communication is affected and other symptoms like decline in mobility, blurriness and double vision, Lhermitte sign, and lack of sensation is noticed. Different therapies for multiples sclerosis are used but most commonly used therapy is disease modifying therapy using beta interferons. Considerable clinical problem is the formation of anti-IFN neutralizing antibodies (NABs) against the drug which decrease the therapeutic effectiveness. To confirm this, we immunized the mice with BSA and OAS conjugated activated interferon beta 1a for five weeks, then we took the blood from the mice through cardiac puncture and isolated the serum by centrifugation. We performed ELISA and Immuno Dot Blot tests both of which showed positive test results which confirmed the formation of anti-IFN neutralizing antibody. In future we can further characterize these NABs to exactly know about the mechanism of antibody production against the injected drug, to launch an appropriate treatment mechanism where clinical efficacy of the treatment would sustain.Item Prevalence of pulmonary and extra-pulmonary tuberculosis in patients of the lahore region(UMT Lahore, 2023) ADEEBA ASLAM; RIDA ATHER; SUNDAS AROOJ AFSHANTuberculosis disease is caused by Mycobacterium tuberculosis, a microorganism first discovered by Robert Koch. This bacterium has the ability to interfere the normal functioning of immune system. It causes pulmonary and extra pulmonary Tuberculosis in infected persons. For the Diagnosis of Tuberculosis, different methods are used such as Acid-fast staining, tuberculin skin test, polymerase chain reaction and culture. Mycobacterium tuberculosis has evolved. Mycobacterium tuberculosis complex includes many species that cause disease such as Mycobacterium tuberculosis causes Tuberculosis in humans, Mycobacterium africanum causes TB in African regions only while in domestic animals: Tuberculosis is caused by Mycobacterium caprae, Mycobacterium bovis, etc. In Pakistan, limited data is present on the prevalence of mutated rpoB genes, so Xpert M. tuberculosis/RIF assay is used to study these mutations. Correct detection of mutated rpoB genes is important as it is required for the correct diagnosis of resistance of RIF in M. tuberculosis strains. The recent study is based on 200 patients out of which 173 patients are clinically diagnosed. It is aimed to examine the Prevalence of Pulmonary and Extra-Pulmonary Tuberculosis in Patients of Lahore region. Different tests were performed on the samples taken from 200 patients which include sputum test, GeneXpert test were performed. Furthermore, for the purpose of statistical analysis different software were used in this study which include ANOVA, Chi-square, Graphpad prism, etc. were used. According to study males (age group: 51-60) were significantly infected with Tuberculosis. As well as patients of age group 51-60 were more likely to have pulmonary tuberculosis with significant P-value of 0.016. But regarding to gender WHO and MDR programs are ensuring the prevention and control of TB by applying some strategies to control multi-drug resistance applying the DOT Plus strategies.Item Computational analysis of neem plant (Azadirachta indica) as mosquito repellent(UMT Lahore, 2023-11-23) GHULAM RUBANIThis study was designed for identification of phytochemicals as potential drug candidates targeting five proteins (odorant binding protein, Sterol carrier protein-2 like-3, Carboxypeptidase, Glutathione Transferase, Pyruvate kinase) of Mosquito used in our study. We used online tools to obtain the protein’s structure. Some online servers and tools are RCSB PDB (For getting the 3d structure of the protein), NCBI (For saving protein’s FASTA format), Chimera 1.15 (For the preparation of protein), Discovery Studio Visualizer (Tool used for visualization of protein complexes). Neem plant contain different phytochemicals that can inhabit or kill mosquito. The protein that we use in our study are odorant binding protein, Sterol carrier protein-2 like-3, Carboxypeptidase, Glutathione Transferase, Pyruvate kinase. Maximum phytochemical shows their effect against all five proteins used in our study. Promising results were find out, phytochemicals Nimbaflavone, Gedunin and Rutin show promising result for Odorant binding protein. Phytochemicals Nimolinone, Meliantriol, Quercetin and Rutin show promising result for Sterol carrier protein-2 like-3 chain A. Meldenin, Nimbaflavone, Meliantriol and Rutin show promising result for Carboxipeptidase protein. Similarly, Meliantriol and Rutin show encouraging result for Glutathione Transferase protein. Azadirachtol, Nimbaflavone and rutin show favourable result for Pyruvate Kinase protein. Rutin is the most prominent phytochemical in this study because it shows promising result against all the mosquito proteins.Item Microbiological analysis of coliform bacteria in drinking water from public filtration plants and restaurants in johar town, lahore(UMT Lahore, 2023-10-05) ARIBA NASIR; RANA NABEELThe purpose of the current study was to detect fecal coliform and total coliform in public filtration plants (PFP) and restaurant’s tap water from different areas of Johar Town, Lahore. A total of n=10 water samples were collected including 5 from PFP and 5 from restaurants. Samples were collected from different areas of Johar Town using standard microbiological procedures and were examined for colony forming unit/ml and most probable number of bacteria. The presence of fecal coliform was not detected from both types of water resources. Results showed that 6 out of 10 samples were positive for coliform bacteria including 2 from public filtration plants and the remaining 4 from tap water of restaurants. High MPN value was observed in sample number 6 of restaurant water that was much higher as compared to the value set by the microbial standards of drinking water which is <50/100ml for total coliform and <0.0/100ml for fecal coliform. Our research concluded that restaurant water was more contaminated as compared to public filtration plant water.Item Phenotypic characterization of MBL enzymes in urinary tract infection causing pathogens(UMT Lahore, 2023-09-25) MARWA ABBAS; WANIA MANSOOR; RAHIMA RANA; HARIS ABBASUrinary tract infections (UTIs) are a common bacterial infection. UTIs are one of the most frequent bacterial infections in the entire world, affecting people of all ages and genders. Epidemiological data regarding UTIs can vary depending on the region, population demographics, healthcare access, and reporting practices. Metallo-β-lactamase is an enzyme that is resistant towards most of the antibiotics particularly among gram negative bacteria. In the current research study, the glycerol stocks of bacterial isolates were obtained from Life Science Laboratory, University of management and technology, Lahore and the culture was refreshed and screened on agar plates for MBL production. The antimicrobial resistivity was detected by using disc diffusion methods against meropenem in presence or absence of EDTA. MBL producers was identified and differentiated through biochemical testing. Furthermore, for the purpose of statistical analysis Graph pad prism software was used in this study which Chi-square test. In the current study, the species prevalent in the provided bacterial isolates of the suspected UTI patients were Klebsiella spp. (100%). Out of the total bacterial isolates (n=25), 3 were confirmed to be MBL producers. Percentages of MBL producing isolates were lower in males (8.33%) as compared to females (15.3%) and distribution was statistically non-significant (P=0.587).Item Synthesis and characterization of zinc sulfide nanoparticles and estimation of its anticorrosive and antibacterial activities(UMT Lahore, 2023-10-03) RABIA SHARIF; USMAN; SAHAR SAJIDNanobiotechnology is a rapidly developing sector that is influencing every field of study, from electronics to biology. The core of nanobioctechnology is nanoparticles, because of their incredibly tiny size and large surface area, nanoparticles are capable of bringing about treatments and cures for illnesses that were previously not achievable. In this study zinc sulfide nanoparticles were synthesized by using co-precipitation method, this method produced high yield of nanoparticles with in less time and inexpensive chemicals which was easily available. 0.005mol of zinc acetate and 0.01mol of sodium sulfide were used in this research. Initially clear color of solution was observed which was changed into black color. Centrifuged the solution for 30 minutes at 300 rpm and then dried it at 110 degrees Celsius for 24 hours. UV analysis was done and the maximum absorbance was observed at 500 nm. FTIR analysis also confirmed the synthesis of ZnS nanoparticles. Anticorrosive activity is performed on the iron bar with the aid of nanoparticles which gave the positive results. Agar well diffusion method and culture based method were carried out and significant antibacterial property of the synthesized ZnS nanoparticles was observed against gram negative bacteria E. coli.Item In-silico identification of potential phytochemicals against selected proteins involved in the pancreatic ductal adenocarcinoma(UMT Lahore, 2023-08-11) Aiman Yaseen; Anam Imtiaz; Safa Adrees; Waqar AnwarDue to the highly malignant nature of Pancreatic cancer, it is considered as one of the deadliest cancers. Only 5% of patients survive this gastrointestinal malignancy. This cancer has been diagnosed at later stages, mostly 90% spread and metastasized. There is a tiring need to study the underlying mechanism responsible for progression of the disease. There are many proteins that overexpress during this disease. The proteins which were taken into consideration in this study are LIF, NEK, PPP1CA, KLK8. Common mechanisms shared by these proteins are cell proliferation, differentiation, invasiveness, and metastasis. Existing options include surgical resection of infected areas, chemotherapeutic drugs, some combinational therapies as well. There are many drugs which are in phase I and II, some of which are showing better results and some need deeper consideration. The selected proteins were considered as targets and against them potential anti-cancer drugs have been identified. These drugs will inhibit the function of these proteins through which the cancer progresses. Piroxicam was identified as a novel anti-LIF and anti-KLK8 drug. Boeravinone B was identified as a potential inhibitor for NEK7 and PPP1CA.Item In-silico designing of bi-functional enzyme constructs (cellulase and xylanase) for enhanced hydrolysis of lignocellulosic biomass(UMT Lahore, 2023) MUSHARRAF HUSSAIN SAHAR; ALISHBA SAIF; AYESHA IFTIKHARCurrent biological techniques for breaking down lignocellulosic biomass face challenges due to slow enzymatic degradation. A particular challenge in cellulose degradation is the presence of hemicellulose chains intertwined with cellulose fibrils, hindering overall cellulose hydrolysis. Moreover, hemicellulose is a polymer of various sugars valuable for product synthesis. Efficient hydrolysis of lignocellulosic biomass necessitates breaking down nearly all its components into constituent monosaccharides. This study focuses on computationally designing bifunctional fusion enzymes combining cellulase and xylanase, connected by flexible and rigid linkers while preserving enzyme functionality. Four constructs were designed, and their 3D structures, folding patterns, and molecular interactions with respective substrates were evaluated. The structural characteristics and molecular interactions of individual, unbound enzymes were also analyzed for comparison. Among the four fusion constructs, (Cell-Xyl)flx, with cellulase at the N-terminus and xylanase at the C-terminus, linked by the flexible linker (GGGS)3, emerged as the optimal choice. It exhibited proper folding, ideal substrate binding to enzymatic domains, maximal molecular interactions with substrates, and higher binding affinity values with both glucan (-7.9kcal/mol) and xylan (-9.2kcal/mol) substrates. Additionally, the nucleotide sequence of the fusion construct was underwent the codon adaptation by JCat tool and in silico cloning of this fusion construct into the pET28a(+) vector resulted in stable expression without altering the amino acid sequence. These findings highlight the effectiveness of computational strategies in designing and molecularly examining various fusion constructs, considering structural parameters, substrate binding, and molecular interactions. Once validated through wet-lab experiments, these ideal constructs hold significant potential for enhancing lignocellulosic biomass hydrolysis into fermentable sugars.Item In-silico designing of Bi-functional enzyme constructs (Cellulase and βGlucosidase) for enhanced hydrolysis of Lignicellulosic biomass(UMT Lahore, 2023-09-12) MUHAMMAD NASIR HUSSAIN; SIDRA NAZIR; BEHZAD ANWARThe rapid depletion of fossil fuels and the escalating environmental concerns have fueled the exploration of renewable energy sources, such as lignocellulosic biomass, for biofuel production. The hydrolysis of lignocellulosic biomass into fermentable sugars is a critical step in this process, and enzymatic approaches using cellulase and beta-glucosidase enzymes have shown promise. However, the suboptimal catalytic efficiency and synergistic interactions between these enzymes limit the hydrolysis yield. This thesis presents an innovative approach to enhance the hydrolysis of lignocellulosic biomass by employing in silico design of bifunctional enzyme constructs. The genetic sequences responsible for cellulase from Bacillus altitudinis and β-glucosidase from Periconia sp. were combined through fusion and subsequently introduced for expression within E. coli K12 bacterial strain. This fusion enzyme was stable at 60 °C for 2 h. Molecular docking simulations were utilized to predict potential binding sites and interactions between the enzymes, enabling the identification of suitable fusion points. Optimum pH for both cellulase and β-glucosidase activities was found to be 6.0. Optimum temperature for cellulase and β-glucosidase activities was found to be 60-65°C, respectively. In future, the docked complexes of stable fusion constructs would be evaluated using molecular dynamic simulations to assess conformational stability in dynamic environment. Combining cellulase and β-glucosidase makes it easier to channel one enzyme's output as a substrate to the other, improving the total efficiency of biomass hydrolysis. Therefore, these fusion enzymes might be useful in industrial applications.Item Isolation and characterization of bacteria from soil samples and their role in plant growth promotion(UMT Lahore, 2023-08-31) Rafiah Kanwal; Maheen Rehan; Eisha Naveed; Muqadas FatimaSoil enormous microbial diversity influences soil fertility and crop health through nutrient recycling and organic matter degradation. Our study aimed to 1) Isolate and purify bacteria from rhizosphere soil samples, 2) Morphological, biochemical characterization, and identification of isolated bacteria and 3) Determining plant growth promoting capability of identified bacterial isolates. Seven bacteria, three from mint soil and four from lettuce rhizospheric soil, were isolated, purified, and characterized. Gram staining, endospore staining, and acid fast staining performed along with ten different biochemical tests for bacterial identification. Plant growth promoting potential of isolates was determine by their growth on Pikovskaya’s agar medium and by plant microbe interaction experiment. Isolate Mnt1 showed phosphate solubilization ability. Isolate Mnt1 and Ltc4 showed the maximum increase in zea mays plant growth compared to the control and other isolates application. Based on the results, isolate 1M has plant growth promoting potential that can be utilized to enhance crop yield at commercial scale. It can be used to produce effective biofertilizers of economic importance. Isolate Mnt1 showed increased plant width by 80% and Ltc4 showed 44% increased weight. Moreover, Isolate Mnt2 showed negative impact and reduced plant growth, reduced plant’s weight by 53%.