A5084326072- Bacteriophages and microbial interactions
- Probiotics and Fermented Foods
- Antibiotic Resistance in Bacteria
- Polyamine Metabolism and Applications
- Escherichia coli research studies
- Bacterial Genetics and Biotechnology
- Genomics and Phylogenetic Studies
- Vibrio bacteria research studies
- Enzyme Production and Characterization
- Pharmaceutical and Antibiotic Environmental Impacts
- Bacterial biofilms and quorum sensing
- Protein Hydrolysis and Bioactive Peptides
- Toxin Mechanisms and Immunotoxins
- Monoclonal and Polyclonal Antibodies Research
- Fungal and yeast genetics research
- Gut microbiota and health
- Antibiotics Pharmacokinetics and Efficacy
- Salmonella and Campylobacter epidemiology
- RNA and protein synthesis mechanisms
- Aquaculture disease management and microbiota
- Fermentation and Sensory Analysis
- Transgenic Plants and Applications
- ATP Synthase and ATPases Research
Agricultural Research Service
2020-2024
National Center for Agricultural Utilization Research
2020-2024
Washington State University
2015-2022
Oak Ridge Associated Universities
2022
ORCID
2020
Control of bacterial contamination in bioethanol fermentation facilities has traditionally relied on chemical-based products such as hop acids and use antibiotics. Recent emphasis antibiotic stewardship prompted new research into the development alternative approaches to microbial remediation strategies. We recently described a recombinant peptidoglycan hydrolase, endolysin LysKB317, which inhibited Limosilactobacillus fermentum strains corn mash fermentation. Here, Saccharomyces cerevisiae...
Many pathogenic bacteria of the family Enterobacteriaceae use type III secretion systems to inject virulence proteins, termed "effectors," into host cell cytosol. Although host-cellular activities several effectors have been demonstrated, function and host-targeted pathways most identified date are largely undetermined. To gain insight proteins targeted by bacterial effectors, we performed coaffinity purification from lysates using recombinant intracellular pathogens Salmonella enterica...
The Escherichia coli quorum sensing (QS) signal molecule, autoinducer-2 (AI-2), reaches its maximum concentration during mid-to-late growth phase after which it quickly degrades stationary phase. This pattern of AI-2 coincides with the up- then down-regulation a recently described microcin PDI (mccPDI) effector protein (McpM). To determine if there is functional relationship between these systems, prototypical mccPDI-expressing strain E. 25 was used to generate ΔluxS, ΔlsrACDBFG (Δlsr), and...
Type III secretion systems (T3SSs) contribute to microbial pathogenesis of Vibrio species, but the regulatory mechanisms are complex. We determined if classic ExsACDE protein-protein model from Pseudomonas aeruginosa applies alginolyticus. Deletion mutants in V. alginolyticus demonstrated that, as expected, T3SS is positively regulated by ExsA and ExsC negatively ExsD ExsE. Interestingly, deletion exsE enhanced ability induce host-cell death while cytotoxicity was inhibited trans...
Microcins represent potential alternatives to conventional antibiotics for human and veterinary medicine. For them be applied in this manner, however, we need better understand their spectrum of activity, how these proteins interact with susceptible cells, producer cells are protected against the antimicrobial properties microcins. microcin PDI (MccPDI), report that activity likely includes most E. coli strains due a conserved binding motif found on an outer membrane protein. Shigella has as...
Abstract Microcin PDI inhibits a diversity of pathogenic Escherichia coli through the action an effector protein, McpM. In this study we demonstrated that expression inhibitory phenotype is induced under low osmolarity conditions and primarily controlled by EnvZ/OmpR two-component regulatory system. Functional, mutagenesis complementation experiments were used to empirically demonstrate EnvZ required for regulation mcpM dependent on binding phosphorylated OmpR promoter region. The may...
The microcin PDI inhibits a diverse group of pathogenic Escherichia coli strains. Coculture single-gene knockout library (BW25113; n=3,985 mutants) against PDI-producing strain (E. 25) identified six mutants that were not susceptible (ΔatpA, ΔatpF, ΔdsbA, ΔdsbB, ΔompF, and ΔompR). Complementation these genes restored susceptibility in all cases, the loss was confirmed through independent gene knockouts E. O157:H7 Sakai. Heterologous expression ompF conferred to Salmonella enterica Yersinia...
Commercial ethanol fermentation facilities traditionally rely on antibiotics for bacterial contamination control. Here we demonstrate an alternative approach to treat using a novel peptidoglycan hydrolase (LysKB317) isolated from bacteriophage, EcoSau. This endolysin was specially selected against Lactobacillus strains that were as contaminants fuel plant. The LysKB317 gene recombinantly expressed in Escherichia coli 33 kDa purified enzyme.In turbidity reduction assays, the recombinant...
Traditional bioethanol fermentation industries are not operated under strict sterile conditions and prone to microbial contamination. Lactic acid bacteria (LAB) often pervasive in tanks, competing for nutrients producing inhibitory acids that have a negative impact on ethanol-producing yeast, resulting decreased yields stuck fermentations. Antibiotics frequently used combat contamination, but antibiotic stewardship has resulted shift alternative antimicrobials.We demonstrate endolysin LysMP,...
We studied the relative fitness benefits of a cephalosporin resistance enzyme (CTX-M-15) that is displacing similar (CMY-2), which extant in E. coli from dairy cattle Washington State. In vitro experiments demonstrated CTX-M-15 provides significant advantage, but only presence very high concentrations antibiotic are found when ampicillin, and to lesser extent ceftiofur, excreted urine treated animals. As such, increasing prevalence bacteria with bla likely occurring ex vivo .
Abstract Clostridium tyrobutyricum strain NRRL B-67062 was previously isolated from an ethanol production facility and shown to produce high yields of butyric acid. In addition, the cell-free supernatant fermentation broth contained antibacterial activity against certain Gram-positive bacteria. To determine source this activity, we report genome mining strain. The complete showed one circular chromosome 3,242,608 nucleotides, 3114 predicted coding sequences, 79 RNA genes, a G+C content...
This study investigated the secretion of endolysin LysKB317 integrated into HO locus Saccharomyces cerevisiae strain NRRL Y-2034 to enable yeast simultaneously perform ethanol fermentation and control bacterial contaminants frequently present in refineries. The cell wall hydrolase gene was expressed using TEF1 NAT5 promoter terminator sequences with α-MF signal an N-terminus poly-histidine tag. detectable by western blot analysis, which showed a molecular weight slightly larger than 33 kDa...
Abstract Background Commercial ethanol fermentation facilities traditionally rely on antibiotics for bacterial contamination control. Here we demonstrate an alternative approach to treat using a novel peptidoglycan hydrolase (LysKB317) isolated from bacteriophage, EcoSau. This endolysin was specially selected against Lactobacillus strains that were as contaminants fuel plant. The LysKB317 gene recombinantly expressed in Escherichia coli 33 kDa purified enzyme. Results In turbidity reduction...
Abstract Background Commercial ethanol fermentation facilities traditionally rely on antibiotics for bacterial contamination control. Here we demonstrate an alternative approach to treat using a novel peptidoglycan hydrolase (LysKB317) isolated from bacteriophage, EcoSau. This endolysin was specially selected against Lactobacillus strains that were as contaminants fuel plant. The LysKB317 gene recombinantly expressed in Escherichia coli 33 kDa purified enzyme. Results In turbidity reduction...
Abstract Background Control of bacterial contamination in bioethanol fermentation facilities has traditionally relied on chemical-based products such as hop acids and the use antibiotics. Recent emphasis antibiotic stewardship prompted new research into development alternative approaches to microbial remediation strategies. We recently described a recombinant peptidoglycan hydrolase, endolysin LysKB317, that inhibited Lactobacillus strains corn mash fermentation. Here, Saccharomyces...