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Peptide Perfection
No full textThe goal in this research project was to synthesize antimicrobial peptides that are short in length. To facilitate the proper formation of the peptide bonds a protective group was used. These projecting groups ensure that side reactions between free amino and carboxylic acid groups don’t occur. In this project Lysine was used, and since it contains two amine groups, both will be protected by the BOC group. The carboxylic acid will remain reactive. The reaction was then monitored by TLC to determine the reaction has finished. The peptide used, BOC-FF-OMe, will have its carboxylic acid protected with a methyl ester and the BOC group will be removed to free the amine group. In the final step, the peptide bond will be formed with ac coupling reagent, EDC. To ensure the reaction was successful, the synthesized protected amino acids and the final peptide will be analyzed by HNMR and the antimicrobial activity will be studied with growth inhibition assay using optical density measurement at 600nm
A Microbial Goldmine: A Search for New Antibiotics
No full textAntibiotic resistance presents one of the most troubling issues in modern medicine. The overuse of antibiotics has allowed bacteria to evolve and develop resistance, making bacterial infections challenging to treat and increasing the potential threats to public health. The Tiny Earth Project strives to address these problems by encouraging students to research, cultivate, and test bacterial findings against safe relative species, with the hope of potential groundbreaking antibiotic discoveries. This was done by cultivating and serially diluting soil, isolating and screening microbes against the safe ESKAPEs. The following safe ESKAPEs were used in the screening process: Enterococcus raffinosus, Staphylococcus epidermidis, Escherichia coli, Acinetobacter baylyi, Pseudomonas putida, and Enterobacter aerogenes. The first set of microbes showed no signs of inhibition against the safe ESKAPEs. The impact of the research encourages the discovery of new antibiotics and provides a pathway for combating the global health crisis
Antibiotic Discovery From Soil
No full textAntibiotic resistance makes treating bacterial illnesses more difficult and poses a growing threat to world health. New medications are desperately needed as germs develop resistance to existing antibiotic. antibiotics have traditionally been found by separating microorganisms from nature sources. such as soil. However, due to increasing resistance, new techniques are required to find effective antibiotics. This project\u27s objective is to use soil microbes to find new antibiotics. in order to accomplish this, we first gathered soil samples and dug up earth from several location. the germs then spread throughout the agar plates using serial dilution. we separated colonies onto a master plate for additional examination after letting the bacterial develop. we tested the bacterial strains for antimicrobial activity against common infection in order to preform antibiotic screening. to pinpoint the exact bacterial generating the antibiotic chemicals, strains with encouraging outcomes were further separated. this study is significant because it explores the possibility of using soil microbes to produce new antibiotics. our goal is to isolate and screen these bacterial in order to find new compounds that could help in the fight against infections that are resistant to antibiotics. The successful strains will be further isolated, their individual chemicals will be identifies, and their potential for new antibiotics will be tested
Post Storm Water Basin Analysis
No full textAccording to the World Health Organization, at least 1.7 billion people use a contaminated drinking-water source in 2022. Contaminated sources of water can transmit deadly diseases and are estimated to cause roughly 500,000 deaths each year. Monitoring our water is essential to determine the level of contaminants and filtration needed to provide safe drinking water. The natural land around the JCCC campus runs directly into the Indian Creek, which, while not being a main water source, eventually flows into the Missouri River, which is a water source for the Johnson County Area. JCCC has five water sampling sites around campus, we chose to focus on testing the post-basin water sample. Our sample being post-basin means it has already gone through the natural filtration of the native plants. The goal of this research is to determine the effectiveness of JCCC’s stormwater management system and the quality of water, as well as the accuracy of everyday test equipment. The post-basin water was found to be hard water, or water containing high mineral content, and sulfate was the only tested parameter found to be above the recommended range for drinking water. Additional experiments focusing on the calcium ion concentration will be discussed
The Search for Inhibition
No full textThis experiment aims to find antimicrobials from the soil samples collected for class. The sample was taken and used for serial dilutions, which reduced our soil density to a more usable concentration. A master plate is then made by transferring our bacteria onto a petri dish. Different screening plates are then made to test our microbes against different types of safe relatives of ESKAPE pathogens. Those colonies are then used to see which ones create the best zones of inhibition. The safe relatives that produced the largest zones of inhibition were E. faecalis, S. epidermidis, and E. coli. A quadrant streak plate is then made which helps obtain isolated colonies. Finally, metabolic testing, PCR, and gel electrophoresis are used to correctly identify the bacterial colonies grown from the streak plates
The Search for Antimicrobials within Soil
No full textThe overall goal of this research project is to discover antimicrobial species from a soil sample. A soil sample was obtained and diluted using the process of serial dilution. Candidates with inhibition were collected and cultured from the serial dilution plates to form a master plate. The candidates acquired from the master plate were then tested against safe relatives of ESKAPE pathogens to determine if they will prevent growth against those pathogen strains. Metabolic testing, PCR, and gel electrophoresis were used to determine the identity of the candidate
The Great Hunt: Finding a New Cure in the Microscopic World
No full textTest project started with a soil sample taken from a neighborhood lawn. That sample was taken, diluted, and placed on agar plates for bacteria to grow. On those plates, zones of inhibition were visible, areas where the bacteria was inhibiting other species from growing. These zones show specific bacteria that are showing signs of producing antibiotics. 12 bacteria with the best zones of inhibitor were selected and tested them against the safe relatives of medically significant pathogens: Enterobacter aerogenes, Staphylococcus epidermidis, Escherichia coli, Pseudomonas putida, Enteroccous faceilis, Ancinetobacter baylyi. Each safe relative was spread onto an agar plate, fully covering it, with each of the 12 selected bacteria placed on the same plate. From there, one bacteria was found that had shown a zone of inhibition on two of the safe relatives; Staphylococcus epidermidis and Enteroccocus faceilis. From here, metabolic testing, PCR, and gel electrophoresis will be used to identify the species of bacteria
A Comparison of White-Nose Syndrome Treatment Methods for North American Bat Conservation
No full textWhite-nose syndrome (WNS) is a fatal disease affecting North American bats caused by the invasive fungal pathogen Pseudogynoascus destructans, which grows on the skin of bats while they hibernate. Since it was first discovered in 2006 in a cave in New York, likely by human transmission, WNS has killed millions of North American bats, desecrating populations of at least 12 different species. By controlling populations of agricultural pests and mosquitos, and, in turn, reducing the use of pesticides and the incidence of mosquito-borne diseases, bats play a vital role in our ecosystems, economy, and health. With WNS continuing its expansion across North America and no current effective methods of managing it, this presentation is a compilation of previously published data on the most promising treatment methods to assess their applicatory potential in the field and highlight their limitations
Isolation of Antibiotic-Producing Bacteria from Soil in Garnett, KS to Combat Resistant Pathogens
No full textAntibiotic resistance has become an increasing concern for healthcare professionals in recent years, especially with the emergence of pathogens that can resist treatment by common antibiotics. These are known as ESKAPE Pathogens, these complicate the treatment of infections, leading to longer illness durations and increased healthcare costs. As a result, there is an urgent need for the discovery of new antibiotics to effectively combat these resistant strains. To contribute to this effort, I collected soil samples from a garden in Garnett, KS, an area known for its diverse microbial ecosystem. Soil is a valuable source for identifying microorganisms that may produce novel antibiotics, as it contains a wide variety of bacterial species, many of which have the potential to produce antimicrobial compounds. The collected soil was diluted and cultured to isolate microorganisms with antibiotic-producing capabilities. After screening the samples, I identified 12 candidates that showed promising antimicrobial activity. I then focused on two candidates, 8 and 11, both of which demonstrated the ability to inhibit the growth of antibiotic-resistant bacteria, suggesting they could be potential sources of new antibiotics. To verify the purity of the cultures, I performed a streak plate assay, which confirmed that candidate 11 was a pure culture. Microscopic analysis revealed that candidate 11 was a Gram-positive, spore-forming rod, a feature commonly associated with a antibiotic-producing bacteria. These findings suggest that candidate 11 may hold significant potential as a new source of antimicrobial agents
Polypeptide Synthesis Project: Tryptophan
No full textFor this CURE project, a peptide bond will be formed by joining tryptophan with another amino acid, phenylalenine-phenylalenine. Then the synthesized polypeptide will be tested for antimicrobial activity. Current research suggests that some small polypeptides could inhibit microbial growth.
First, to join tryptophan with the phenylalanine-phenylalanine. A protecting-group will be added to the free amine group on tryptophan. This will performed by reacting BOC anhydride, the protection-molecule, with tryptophan under basic conditions using DIPEA as a base, and then water plus acetone for the solvent. The reaction progress will be monitored by Thin Layer Chromatography (TLC). The BOC-protected tryptophan will be isolated by dropping the pH using HCl, washing with ethyl acetate, and using column chromatography.
Then, the protecting group already attached to the free amine group of phenylalanine-phenylalanine will be removed using dichloromethane, THF, and heat for approximately one hour. The carboxylic acid end of phenylalanine-phenylalanine will remain protected and unable to form unwanted side-reactions.
The protecting group on tryptophan and the removal of the protecting group on phenylalanine-phenylalanine is necessary to reduce chances of unwanted side-reactions from occurring but also to allow the coupling of the free amine group of phenylalanine-phenylalanine and the carboxylic acid group of tryptophan. Once tryptophan and phenylalanine-phenylalanine are prepped, they can be joined together with a new peptide bond using EDC as a coupling-reagent. Finally, the protecting groups will be removed from the product and anti-microbial activity will be qualitatively confirmed