Microbiology Commons^™

Bioactive Potential Of Microbial Isolates From Yoluja Cave And Jepirramüin Rock Shelter, Cabo De La Vela (Colombia), Juan Sebastián López Caro, Virginia Patricia Roa Angulo, Alex Alberto González Vargas

International Journal of Speleology

Subterranean environments harbor diverse microbial communities with significant potential for the production of bioactive compounds. This study evaluated the antimicrobial and antioxidant potential of microbial isolates obtained from Yoluja Cave and the Jepirramüin rock shelter, located in the coastal region of La Guajira, Colombia. Field sampling was conducted during three campaigns in July, September, and November 2025, including soil and surface samples collected along environmental gradients within each site. Microorganisms were isolated using selective culture media and identified through morphological and biochemical characterization. Selected strains were subjected to liquid fermentation and organic solvent extraction to obtain crude metabolites. Analysis of …

Novel Chitosan-Pyrazole Derivatives: Synthesis, Characterization, And Antimicrobial Potential Assessment, Samar A. Salim, Elrefaie S. Kenawy, Mohamed G. Altamami, Ahmed A. Safaan, Manar M. Emara, Elbadawy A. Kamoun, Atif A. Elgharably

Nanotechnology Research Centre

Double novel chitosan Schiff-base series (I and II) based on heterocyclic moieties were synthesized to improve the pristine antibacterial capabilities of chitosan. Series (I) chitosan Schiff bases were prepared by reacting chitosan with six derivatives of 4-formyl-1H-pyrazole in an acidic medium, whereas series (II) was prepared by reacting chitosan with six derivatives of 1-(4-vinylbenzyl)-3-aryl-1H-pyrazole-4-carbaldehyde in an acidic medium. The chemical configurations of the twelve manufactured Schiff bases were verified using FTIR analysis & TGA measurements. We assessed the antimicrobial efficacy of chitosan and its Schiff base derivatives against four standard bacterial species, Staphylococcus aureus, Enterococcus faecalis, Pseudomonas aeruginosa, and Escherichia …

A Single-Step Plate Method For The Screening Of Xyl-A Mutant Libraries Based On Gene Expression, Bilqees Fatima, Muhammad Mohsin Javed

Journal of Bioresource Management

A single-step plate method is described for the screening of transformants of cloned gene xyl-A encoding xylose isomerase. It is rapid, inexpensive, easy to perform, and more specific, as the selection is based on xyl-A expression level. The crucial composition of the plate medium comprised agar, buffer, nutrients, co-factor, substrate, inducer, antibiotic, and an indicator, i.e., 2, 3, 5-triphenyl tetrazolium chloride (TTC). Transformed cells of E. coli BL21 (DE3) with a plasmid (pET21(a)⁺+xyl-A), were exclusively capable of growing, isomerizing the substrate, and producing deep-red formazan by reducing TTC, whereas the untransformed cells and cells containing a …

The Role Of Seed Mix Characteristics And Soil Amendments During Post-Agricultural Prairie Restoration, Valerie N. Thurston, Michaela J. Woods, Madelaine Gregory, Mary C. Lloyd, Ryan W. Mcewan

Biology Faculty Publications

Introduction: Prairie restoration on degraded lands may be influenced by seed mix characteristics, including species richness and legume establishment. Soil amendments may facilitate restoration; however, the practical usefulness of techniques such as applications of whole soil and leaf mulch remains relatively unclear.

Objectives: We assessed whether increasing species richness and legume content in seed mixes enhances prairie restoration success and examined how soil amendments interact with seed mix composition to influence cover, species richness, diversity, and legume establishment.

Methods: We conducted a field experiment on a 12-ha post-agricultural site in Ohio, U.S.A. Twenty 50 × 50 m plots received one …

In Vivo Expression Profiling Of Listeria Monocytogenes Internalins (Inlf, Inlp, Inlj) And Their Corresponding Host Receptors (Vim, Afdn, Muc2) Using A Rabbit Ligated Intestinal Loop Model, Nawar Al-Janabi, Nadhim Mushtaq Hashim Al-Bderee, Zahraa M. Madania, Mohanad Jawad Kadhim

Karbala International Journal of Modern Science

Listeria monocytogenes is a Gram-positive pathogen that causes severe infections, particularly in immunocompromised individuals. Internalin genes are important virulence factors that facilitate the bacterium's invasion of host cells. This research aimed to quantify in vivo expression of selected internalin genes (inlF, inlP, and inlJ) and their corresponding host receptors (VIM, AFDN, and MUC2) during L. monocytogenes infection. To achieve this aim, a rabbit ligated intestinal loop model was constructed comprising two intestinal loops. The first loop was injected with 1 mL of L. monocytogenes (1 × 10⁷ CFU/mL), while the second loop was injected with 1 mL of PBS as …

Effectiveness Of Organic Amendments In Enriching Nitrogen In Compost Produced From Organic Fractions Of Market Waste Generated In Dar Es Salaam, Tanzania, Amani Bilula, Ally Mahadhy, Anthony Manoni Mshandete, Amelia Kivaisi

Tanzania Journal of Science

Low nitrogen (N) content remains a key limitation of compost derived from urban organic waste. This study evaluated the effectiveness of selected plant-based organic amendments in enhancing nitrogen content and improving compost quality. Four plant- based organic amendments, Leucaena leucocephala (BN01), Commelina benghalensis (BN02), Morus alba (BN03), and Helianthus annuus (sunflower) cake (BN04), were incorporated into market waste compost at ratios of 1:0.06:0.04, 1:0.06:0.25, and 1:0.06:0.5 (organic waste: sawdust: amendment). Composting was conducted using a 4 × 3 factorial design with three replicates per treatment (n = 36). Temperature, moisture, and pH were monitored throughout the composting period. Total nitrogen, …

Endophytic Actinobacteria: Deciphering Therapeutic And Industrial Potential, And Prospects Of Genome Mining In Novel Products Discovery, Yusra Waheed, Rabia Shahdin, Imran Sajid

Journal of Bioresource Management

Endophytic actinobacteria are a diverse group of Gram-positive bacteria with high G+C content, which inhabit plants’ internal tissues, and represent a significant yet underexplored reservoir of bioactive secondary metabolites. In host plants, a remarkable repertoire of metabolites, including broad-spectrum antibiotics, anticancer, antiviral, antifungal, antioxidant, and immunomodulatory compounds, is synthesized by these microbial allies, carrying significant medicinal and agricultural potential. These endophytes, in contrast to their soil relatives, have developed unique biosynthetic gene clusters (BGCs) strongly influenced by plant-associated factors, leading to the production of structurally novel metabolites. In recent years, the advances in genome mining, metagenomics, and metabolomics have accelerated …

Discovery Of Bmi-Associated Gut Microbiome Signatures In The Awi-Gen 2 Cohort, Meriem Yagoubi

Theses and Dissertations

Obesity is increasing across sub-Saharan Africa amid rapid dietary, environmental, and urbanization transitions, yet the gut microbiome features associated with adiposity in African populations remain under-characterized. This study investigated whether species-level gut microbiome composition is associated with body mass index (BMI) in the AWI-Gen 2 cohort, and whether BMI-associated microbial signals remain detectable after accounting for the dominant geographic structure of the microbiome. Gut microbiome community structure and diversity were characterized across 1,665 participants, and differential abundance analysis was used to identify taxa associated with BMI after controlling for study site. Machine learning models were then trained to assess how …

A Novel Endolysin Of Phage Persinger Infecting Pseudomonas Aeruginosa: Activity Against Biofilms And Structural Prediction, Layla Abbud

James Madison Undergraduate Research Journal (JMURJ)

Endolysins are proteins that degrade bacterial cell walls. They are encoded on the genome of bacteriophages (viruses that infect and kill bacteria). At the end of the lytic cycle of a bacteriophage, endolysins cut the peptidoglycan layer of the bacterial cell wall, allowing the release of viral progeny. Pseudomonas aeruginosa is a well-known multidrug-resistant human pathogen that causes serious problems, especially in patients diagnosed with cystic fibrosis (CF). One property of P. aeruginosa is the production of biofilms, which is a defense against antibiotics. Previous research has shown that endolysins from bacteriophages can degrade biofilms. The endolysin protein from Persinger, …

Assessment Of Fecal Indicator Bacteria And Vendors' Handling Practices For Safety Of Salads And Raw Leafy Vegetables In Dar Es Salaam, Tanzania, Gloria B. Mrisho, Thomas J. Lyimo, Godwill D. Mrema

Tanzania Journal of Science

Fresh leafy vegetables have gained popularity in Tanzanian cities due to their nutritional benefits and increased promotion by health practitioners. Assessment of their microbiological quality and safety is essential for determination of associated health risks. This study enumerated Fecal Indicator Bacteria (FIB) for determination of microbiological quality and safety of fresh leafy vegetable salads sold by street vendors and raw leafy vegetables obtained from Sinza market in Dar es Salaam. Handling practices, awareness and consumer satisfaction on the safety of leafy vegetable salads was also assessed among salad street vendors and consumers. Samples were collected from four stations and analyzed …

Best Integrated Writing Spring 2026 - Complete Edition, Wright State University School Of Humanities And Cultural Studies, Tracy Smith, Kristie Mckiernan

Best Integrated Writing

Best Integrated Writing includes excellent student writing from Integrated Writing courses taught at Wright State University.

Bacteriophage Therapy For Antibiotic-Resistant Infections, Shiva Pariyar

Best Integrated Writing

Pariyar examines bacteriophage therapy as a promising alternative to antibiotics for treating multidrug-resistant infections, highlighting its targeted mechanism, ability to disrupt biofilms, and synergy with antibiotics. While challenges such as regulation, immune response, and limited clinical trials remain, the study argues that advances in biotechnology and global collaboration position phage therapy as a future cornerstone in combating antimicrobial resistance.

Metagenomic Profiling Of Gut Microbiome Signatures Across Liver Disease Stages And Hcv-Related Hepatocellular Carcinoma In Egyptian Patients, Marwa A. Zahra

Theses and Dissertations

Dysbiosis in the gut microbiome, particularly concerning the synchronous crosstalk between the gut and the liver, has been associated with various diseases. This study profiles the gut microbiome in liver diseases among Egyptian patients, with a focus on the hepatitis C virus (HCV) and hepatocellular carcinoma (HCC), both of which are highly prevalent in Egypt. Utilizing shotgun metagenomic sequencing, we analyzed microbial gene catalogs and taxonomic profiles from 46 Egyptian patients categorized into five groups: healthy individuals, liver disease patients of different etiologies, post-HCV, treated HCV, and HCV-HCC patients. Healthy and treated HCV patients exhibited distinct microbial profiles characterized by …

Environmental Detection And Genetic Profiling Of Staphylococcus Sp. At Taal Volcano Island, Ray Albert E. La Rosa, Shaun Michael C. Basco, Martin Gabriel Z. Dela Cruz, Iluminada A. Ronio

The Lasallian Journal of Health

Taal Volcano Island’s hydrothermal environment supports extremophilic microbial communities. This study isolated, biochemically characterized, and genetically profiled Staphylococcus sp. from shoreline soils. Soil samples were collected near active vents and cultured on acidified gelatin agar (pH 5.0). Biochemical tests—Methyl Red, Spirit Blue Agar, Motility, and Acrolein—were performed. DNA extraction and 16S rRNA sequencing identified Staphylococcus candidatus, with 99 % sequence similarity to GenBank entries JN794602.1 and KX250303.1. Phylogenetic analysis clustered the isolate with S. ureilyticus, S. cohnii, and S. equorum. The organism’s presence in volcanic soils demonstrates adaptation to acidic, thermophilic conditions and suggests potential applications in bioremediation and enzyme …

Combining Crispr And Nanoasv To Uncover The Rare Bacterial Communities Of The Human Oral Microbiome, Gilma Ruth Sevilla Alvarez

Master's Projects

Studying the oral microbiome is vital because its health implications are not contained to the mouth. Cariogenic bacteria cause tooth demineralization and oral microbes that translocate to extra-oral organ systems are linked to conditions like Alzheimer’s disease, heart disease, diabetes, and cancer. Some of these pathogens belong to the most abundant phyla in the oral cavity: Bacillota, Pseudomonadota, Bacteroidota, Actinomycetota, Spirochaetota, and Fusobacteriota, which together comprise 94% of the oral microbiome. This project uses CRISPR-Cas9 to deplete sequences from highly abundant phyla and the Eubacteria domain, enriching the remaining 6% of rarer taxa. Paired with NanoASV for single-nucleotide resolution, this …

Lead Biosorption By Oceanobacillus Profundus Kbz 3-2: Strategies For Bioremediation Systems In Kabwe, Danielle S. Desmarteau

Scripps Senior Theses

Lead contamination remains a significant global public health concern, particularly in Kabwe, Zambia, the site of a former lead mining operation. Bioremediation, which utilizes biological materials and processes to remove environmental contaminants, represents a promising approach for addressing lead pollution. However, the development of effective bioremediation systems requires a detailed understanding of the mechanisms underlying metal removal and the feasibility of implementation in low-resource settings. Oceanobacillus profundus KBZ 3-2, a bacterium isolated from Kabwe mine waste, has demonstrated the ability to remove lead from aqueous environments. This study proposes to investigate the mechanism of lead biosorption by this organism, …

Interaction Of The Gram-Positive Human Pathogen Staphylococcus Aureus With Host Lipids, Sharanya Paul

Theses and Dissertations

Staphylococcus aureus is a prominent pathogen that is known to incorporate straight-chain unsaturated fatty acids (SCUFAs) into its lipids in addition to the normally biosynthesized branched-chain fatty acids (BCFAs) and straight-chain fatty acids (SCFAs). Furthermore, it has recently been shown that the incorporation of oleic acid into S. aureus lipids significantly enhanced the growth of the organism at low temperatures due to the greater fluidity imparted to the membrane. Together with others in the Wilkinson and Dahl laboratory, I have shown that low-temperature growth is enhanced by various SCUFAs, including the human antimicrobial fatty acids sapienic acid, linoleic acid, and …

Fostering Entrepreneurial Thinking And Practice Through Teaching And Learning Integration, Mohammad G. Sabbir

Learning and Educational Center Virtual Teaching and Learning Conference

This session explores an innovative, evidence-based approach to integrating entrepreneurial thinking into undergraduate neuroscience education through hands-on gene editing experiences. Using the commercially available Bio-Rad CRISPR/Cas9-based “Out of the Blue” kit for bacterial LacZ gene editing, students are guided not only through the scientific principles underlying modern gene editing technologies but also through the entrepreneurial challenge of replicating and manufacturing the kit using existing lab resources. This model empowers students to extend their learning beyond the classroom by engaging in cost-effective, scalable biotechnology practices—particularly valuable for institutions with limited funding. Classroom-based research demonstrates increased student engagement, deeper conceptual understanding, and …

Detection Method For Escherichia Coli Using Real-Time Polymerase Chain Reaction Targeting The Yhav Gene, Muktiningsih Nurjayadi, Anisa Fitriyanti, Royna Rahma Musie, Gusti Angieta Putri, Puan Aqila Azizah, Helzi Angelina, Grace Grace, Ananda Indah Putri Sihombing, Agus Setiawan, Jefferson Lynford Declan, Gladys Indira Putri, Dandy Akbar Juliansyah, Siti Fatimah, Ayu Berkahingrum, Irma Ratna Kartika, Fera Kurniadewi, Vira Saamia, Shyi-Tien Chen, Bassam Aboemolak, Hesham Ali El Enshasy

Makara Journal of Science

Escherichia coli is a foodborne pathogenic bacterium that can cause diarrhea, while yhaV is a virulence-associated gene linked to the toxin–antitoxin system in E. coli. This study was aimed at evaluating the confirmation, specificity, and sensitivity of a yhaV gene primer using real-time polymerase chain reaction. The yhaV-targeting PCR successfully amplified a DNA fragment with an amplicon length of 207 bp (base pairs) under an annealing temperature optimized to a range of 54 °C to 62 °C via gradient PCR. The PCR using the primer pair produced a consistent Ct (cycle threshold) of 14.14 ± 0.05 and showed …

Enhanced Meropenem Activity And Stability Following Load In Polyvinyl Alcohol Nanofiber Scaffolds With Sitagliptin As Quorum Sensing Inhibitor On Pseudomonas Aeruginosa, Samar A. Salim, Abdelrhman Mohsen, Mohamed Emara, Taghrid Elmahdy

Nanotechnology Research Centre

Background The increasing resistance of bacteria to conventional antibiotics poses a significant health challenge. Innovative strategies, such as combining antibiotics with agents like quorum sensing inhibitors (QSIs), have been developed to combat this issue. QSIs enhance antibiotic efficacy without inhibiting bacterial growth, minimizing the risk of resistance. Aims Evaluate the combined effect of Sitagliptin (STG) as a QSI with Meropenem (MER), fabricate drug-loaded Polyvinyl Alcohol (PVA) nanofibers, and investigate their antimicrobial activity against standard Pseudomonas aeruginosa (PAO1) and carbapenem-resistant P. aeruginosa (CRPA). Methods The combinatorial effect was assessed using a checkerboard assay. PVA/STG, PVA/MER, and PVA/STG/ MER nanofibers were fabricated …

Biogenic Calcite Precipitation For Coastal Sand Stabilization: A Synergistic Approach Using Marine Bacteria And Natural Fibers, Aparna Gs

Theses and Dissertations

Coastal erosion, driven by natural forces such as waves, tides, winds, and currents and further intensified by anthropogenic activities like urbanization, climate change, and coastal infrastructure development, poses a significant threat to shoreline stability. Conventional erosion control methods, including jetties, embankments, and artificial reefs, often face limitations in sustainability and effectiveness. In this context, microbially induced calcite precipitation (MICP) has emerged as a promising ecofriendly solution for ground stabilization.

The present study investigates the potential of an indigenous ureolytic marine bacterium, Bacillus sp. N₉, isolated from the coastal sediments of Nagapattinam, Tamil Nadu, for coastal sand stabilization. The strain was …

Genomic Surveillance Of Indian Klebsiella Pneumoniae And Exploration Of Resistance Inhibitors, Abhirami Ks Ms

Theses and Dissertations

Klebsiella pneumoniae is an important ESKAPE pathogen that causes life threatening infections. The strains exhibiting multidrug-resistance (MDR) and hypervirulence are the priority pathogens for which immediate treatment and dissemination prevention strategies are required. To address this, genome profiling was performed to understand the AMR patterns, role of episomes, phages, virulence determinants and its association with sequence types.

The study includes 267 publicly available Indian K. pneumoniae genomes over a period of ten years and four sequenced isolates. Whole genome analysis revealed the widespread presence of diverse β-lactamase genes, with β-lactam resistance being predominant. Genes such as blaCTX-M-15, blaTEM-1B and blaOXA-232 …

Knowledge Of Hiv Transmission And Prevention Among University Students In Pakistan: A Cross-Sectional Study, Muhammad Saleem, Sajid Masood, Shahid Hussain

Journal of Bioresource Management

HIV/AIDS remains a significant public health problem internationally, with young adults being especially at risk because of behavioral and knowledge deficits. In Pakistan, cultural inhibitions and suboptimal health education are the main reasons for limited awareness, particularly among university students. This study had the objectives of determining the knowledge level about HIV transmission and prevention among undergraduate students in Kohsar University Murree (KUM) and to investigate correlations with salient demographic variables. A cross-sectional survey was carried out among 510 students aged 18-27 years through a pre-validated, standardized questionnaire with 12 knowledge-based items. Descriptive statistics and Chi-square tests were utilized to …

Phylogenetic Analysis Of Ftsz In Wolbachia: Implications For Host Specificity, Anwar Rovik, Ayu Rahayu

Makara Journal of Science

Wolbachia are the most prevalent and successful endosymbiont in invertebrates and are categorized into 21 supergroups based on genetic phylogeny. The filamenting temperature-sensitive Z (FtsZ) protein is essential for bacterial cell division and is hypothesized to play a role in Wolbachia's control of host reproduction. However, despite FtsZ's importance, little is known about its diversity and function within Wolbachia. This study investigated the variability of the FtsZ-encoding gene among Wolbachia strains and their hosts. FtsZ gene sequences were retrieved from the National Center for Biotechnology Information database, and representative sequences were randomly selected from each host species and analyzed …

Detection Of Quorum-Sensing Molecules In The Mutualistic Bacteria Vibrio Fischeri Exposed To Simulated Microgravity Using Cell-Based Biosensors., Paulina Slick, Maria C. Tobarra, Alba A. Chavez, Hugo Castillo

Beyond: Undergraduate Research Journal

Bacteria flourish in multiple environments when communicating with each other in a process known as Quorum Sensing. This process is accomplished by the production of small signaling molecules referred to as Autoinducers (AI). This communication allows the bacteria to alter their gene expression in an effort to regulate their cell number, behavior, community formation and virulence. The space environment provides stressful conditions for bacteria as they are exposed to radiation and Microgravity (µG or MG) altering synthesis and concentration of Autoinducers. The purpose of this research was to expose the model system Vibrio fischeri to simulated microgravity using the recently …

Antimicrobial Eats: How Food Additives Are Contributing To Obesity Via The Gut Microbiome, Olivia Mankos

Senior Honors Theses

The typical modern diet is brimming with food additives used in food processing. Additives such as antibiotic residues, GMOs, pesticide residues, anti-caking agents, emulsifiers, artificial sweeteners, colorants, flavor enhancers, and preservatives. Most of these additives seem to shift the gut microbiome towards more pro-inflammatory signals. When all these additives are considered as an amalgam, such as with the modern processed diet, the ramifications on the gut microbiome are concerning. Obesity, which is characterized by low-level chronic inflammation, is likely exacerbated by these food additives. Consequently, most of these additives should be avoided or banned for optimal health.

Metabolite Analysis To Understand Factors Limiting Microbial Fermentation At High Substrate Concentrations, Bishal D. Sharma

Dartmouth College Ph.D Dissertations

Clostridium thermocellum, a thermophilic anaerobic bacterium, holds promise as a biocatalyst for conversion of lignocellulosic biomass into biofuels. This microbe has been subjected to various metabolic engineering strategies, including the deletion of genes encoding secondary fermentation products such as H₂, formate, acetate, and lactate, as well as the expression of heterologous genes to redirect carbon flux toward ethanol production. Despite these efforts, achieving economically viable ethanol titers remains a challenge. We hypothesized that studying how intracellular metabolite concentrations change as fermentation stops could reveal metabolic bottlenecks or regulatory mechanisms limiting ethanol production at high titers. However, metabolomics studies at …

The Design, Construction, And Testing Of Mrna Vaccine Against Equine Herpes Virus, Bhawana Devkota

LSU Master's Theses

Equine Herpesvirus-1 (EHV 1) is a worldwide significant pathogen that causes respiratory illness, abortion, and neurological disorders in equine. Current vaccines, including live attenuated, inactivated, and subunit platforms, do not prevent viral latency, mucosal shedding, or cross-strain immunity, requiring alternative approaches. To address these gaps, this project explores the design, synthesis, and in vitro testing of an mRNA vaccine targeting immunogenic EHV 1 glycoprotein (gB, gC, gD, gG, and gM). Epitopes were computationally predicted using the Immune Epitope Database (IEDB), codon-optimized for Bos taurus, and cloned into pUCIDT vectors using SP6/T7 promoters. In vitro transcription (IVT) used nucleotide modifications …

Development Of A Multi-Epitope Peptide Vaccine Against Monkeypox Virus: Immunoinformatics Analysis For South East Asian Hla Alleles, Nelson Chandra, Mochammad Aqilah Herdiansyah, Viol Dhea Kharisma, Arif Nur Muhammad Ansori, Arli Aditya Parikesit

Makara Journal of Science

The monkeypox virus (MPXV), a DNA virus causing zoonotic disease, poses major global public health challenges, with mortality rates between 3%–6%. Although smallpox vaccines provide partial cross-protection, there is a critical need for a dedicated, effective monkeypox (mpox) vaccine. This study aimed to design a multi-epitope peptide-based vaccine specifically adapted to the HLA allele profiles common in Southeast Asian populations, where MPXV cases are rising. Using immunoinformatics, we screened for and detected B and T cell epitopes from the MPXV cell surface antigen and IFN-alpha/beta receptor proteins. The vaccine design was validated through a rigorous evaluation of its antigenicity, immunogenicity, …

Screening For Penicillin G Acylase (Pga)-Producing Bacteria And Gene Cloning Using Degenerate Oligonucleotide Primed-Pcr, Masdalifah Masdalifah, Sri Rezeki Wulandari, Gabriela Christy Sabbathini, Maria Ulfah, Dini Achnafani, Ahmad Wibisana, Feronika Heppy Sriherfyna, Is Helianti, Niknik Nurhayati

Makara Journal of Science

The growing concern over antibiotic resistance has driven global efforts to explore innovative solutions, including the use of Penicillin G acylase (PGA) to produce semisynthetic β-lactam antibiotics. This study screened four potential in-tracellular PGA-producing bacteria: Alcaligenes faecalis InaCC B444 (AfPGA), Kluyvera cryocrescens InaCC B850 (KcPGA), Providencia rettgeri InaCC B25 (Pr25PGA), and P. rettgeri InaCC B466 (Pr466PGA). Penicillin G Acylase encoding genes (pgas) were isolated from them using a Degenerate Oligonucleotide Primed-PCR (DOP-PCR) approach and sequenced. Microbiological assays confirmed all tested crude extracts to exhibit inhibitory effects. Penicillin G was used for evaluating hydrolytic activity and 6-Amino Penicillanic Acid (6-APA) coupled …