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- Whole genome and RNA sequencing of oral commensal bacterium Streptococcus anginosus subsp. anginosus with vancomycin tolerance
- Kyu Hwan Kwack , Jae-Hyung Lee , Ji-Hoi Moon
- J. Microbiol. 2022;60(2):167-176. Published online January 7, 2022
- DOI: https://doi.org/10.1007/s12275-022-1425-4
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Abstract
- “Antibiotic tolerance” promotes the rapid subsequent evolution of “antibiotic resistance,” however, it is often overlooked because it is difficult to distinguish between tolerant and susceptible organisms. A commensal bacterium S. anginosus subsp. anginosus strain KHUD_S1, isolated from dental biofilm was found to exhibit a high MBC/MIC ratio of 32 against vancomycin. We observed KHUD_S1 cells exposed to vancomycin did not grow but maintained viability. Transmission electron microscope showed KHUD_S1 cells possessed a dense, thick capsule and maintained the cell wall integrity upon vancomycin exposure. To infer the underlying mechanisms of the vancomycin tolerance in KHUD_S1, we performed whole genome sequencing and RNA sequencing. The KHUD_S1 genome carried three genes encoding branching enzymes that can affect peptidoglycan structure through interpeptide bridge formation. Global gene expression profiling revealed that the vancomycin-induced downregulation of carbohydrate and inorganic ion transport/metabolism as well as translation is less prominent in KHUD_S1 than in the vancomycin susceptible strain KHUD_S3. Based on the transcriptional levels of genes related to peptidoglycan synthesis, KHUD_S1 was determined to have a 3D peptidoglycan architecture distinct from KHUD_S3. It was found that, under vancomycin exposure, the peptidoglycan was remodeled through changes in the interpeptide bridge and transpeptidation reactions. Collectively, these features of S. anginosus KHUD_S1, including a dense capsule and differential gene expression in peptidoglycan synthesis, may contribute to vancomycin tolerance. Our results showing the occurrence of vancomycin tolerance amongst oral commensal bacteria highlight the need for considering future strategies for screening of antibiotic tolerance as an effort to reduce antibiotic resistance.
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Citations
Citations to this article as recorded by
- Gut resistome profiling reveals high diversity and fluctuations in pancreatic cancer cohorts
Xudong Liu, Kexin Li, Yun Yang, Dingyan Cao, Xinjie Xu, Zilong He, Wenming Wu
Frontiers in Cellular and Infection Microbiology.2024;[Epub] CrossRef - The Sexome ‐ A proof of concept study into microbial transfer between heterosexual couples after sexual intercourse
Ruby Dixon, Siobhon Egan, Sheree Hughes, Brendan Chapman
Forensic Science International.2023; 348: 111711. CrossRef
- Gut resistome profiling reveals high diversity and fluctuations in pancreatic cancer cohorts
Research Support, Non-U.S. Gov't
- Expression and Purification of a Recombinant scFv towards the Exotoxin of the Pathogen, Burkholderia pseudomallei
- Kue-Peng Lim , HongBin Li , Sheila Nathan
- J. Microbiol. 2004;42(2):126-132.
- DOI: https://doi.org/2034 [pii]
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Abstract
- A single chain variable fragment (scFv) specific towards B. pseudomallei exotoxin had previously been generated from an existing hybridoma cell line (6E6AF83B) and cloned into the phage display vector pComb3H. In this study, the scFv was subcloned into the pComb3X vector to facilitate the detection and purification of expressed antibodies. Detection was facilitated by the presence of a hemagglutinin (HA) tag, and purification was facilitated by the presence of a histidine tag. The culture was grown at 30^oC until log phase was achieved and then induced with 1 mM IPTG in the absence of any additional carbon source. Induction was continued at 30^oC for five h. The scFv was discerned by dual processes-direct enzyme-linked immunosorbent assays (ELISA), and Western blotting. When compared to E. coli strains ER2537 and HB2151, scFv expression was observed to be highest in the E. coli strain Top10F'. The expressed scFv protein was purified via nickel-mediated affinity chromatography and results indicated that two proteins a 52 kDa protein, and a 30 kDa protein were co-purified. These antibodies, when blotted against immobilized exotoxin, exhibited significant specificity towards the exotoxin, compared to other B. pseudomallei antigens. Thus, these antibodies should serve as suitable reagents for future affinity purification of the exotoxin.
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