Journal of Microbiology

Review

Structural Insights into the Lipopolysaccharide Transport (Lpt) System as a Novel Antibiotic Target: Yurim Yoon, Saemee Song; J. Microbiol. 2024;62(4):261-275. Published online May 31, 2024; DOI: https://doi.org/10.1007/s12275-024-00137-w

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Abstract: Lipopolysaccharide (LPS) is a critical component of the extracellular leaflet within the bacterial outer membrane, forming an effective physical barrier against environmental threats in Gram-negative bacteria. After LPS is synthesized and matured in the bacterial cytoplasm and the inner membrane (IM), LPS is inserted into the outer membrane (OM) through the ATP-driven LPS transport (Lpt) pathway, which is an energy-intensive process. A trans-envelope complex that contains seven Lpt proteins (LptA-LptG) is crucial for extracting LPS from the IM and transporting it across the periplasm to the OM. The last step in LPS transport involves the mediation of the LptDE complex, facilitating the insertion of LPS into the outer leaflet of the OM. As the Lpt system plays an essential role in maintaining the impermeability of the OM via LPS decoration, the interactions between these interconnected subunits, which are meticulously regulated, may be potential targets for the development of new antibiotics to combat multidrug-resistant Gram-negative bacteria. In this review, we aimed to provide an overview of current research concerning the structural interactions within the Lpt system and their implications to clarify the function and regulation of LPS transport in the overall process of OM biogenesis. Additionally, we explored studies on the development of therapeutic inhibitors of LPS transport, the factors that limit success, and future prospects.

Journal Articles

[Protocol] Use of Cas9 Targeting and Red Recombination for Designer Phage Engineering: Shin-Yae Choi , Danitza Xiomara Romero-Calle , Han-Gyu Cho , Hee-Won Bae , You-Hee Cho; J. Microbiol. 2024;62(1):1-10. Published online February 1, 2024; DOI: https://doi.org/10.1007/s12275-024-00107-2

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Bacteriophages (phages) are natural antibiotics and biological nanoparticles, whose application is significantly boosted by recent advances of synthetic biology tools. Designer phages are synthetic phages created by genome engineering in a way to increase the benefits or decrease the drawbacks of natural phages. Here we report the development of a straightforward genome engineering method to efficiently obtain engineered phages in a model bacterial pathogen, Pseudomonas aeruginosa. This was achieved by eliminating the wild type phages based on the Streptococcus pyogenes Cas9 (SpCas9) and facilitating the recombinant generation based on the Red recombination system of the coliphage λ (λRed). The producer (PD) cells of P. aeruginosa strain PAO1 was created by miniTn7-based chromosomal integration of the genes for SpCas9 and λRed under an inducible promoter. To validate the efficiency of the recombinant generation, we created the fluorescent phages from a temperate phage MP29. A plasmid bearing the single guide RNA (sgRNA) gene for selectively targeting the wild type gp35 gene and the editing template for tagging the Gp35 with superfolder green fluorescent protein (sfGFP) was introduced into the PD cells by electroporation. We found that the targeting efficiency was affected by the position and number of sgRNA. The fluorescent phage particles were efficiently recovered from the culture of the PD cells expressing dual sgRNA molecules. This protocol can be used to create designer phages in P. aeruginosa for both application and research purposes.

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Characteristics of bioaerosols under high-ozone periods, haze episodes, dust storms, and normal days in Xi’an, China
Yiming Yang, Liu Yang, Xiaoyan Hu, Zhenxing Shen
Particuology.2024; 90: 140. CrossRef
Airborne desert dust and aeromicrobiology over the Turkish Mediterranean coastline
Dale W. Griffin, Nilgün Kubilay, Mustafa Koçak, Mike A. Gray, Timothy C. Borden, Eugene A. Shinn
Atmospheric Environment.2007; 41(19): 4050. CrossRef

Alpha‑Hemolysin from Staphylococcus aureus Obstructs Yeast‑Hyphae Switching and Diminishes Pathogenicity in Candida albicans: Xiaoyu Yu , Yinhe Mao , Guangbo Li , Xianwei Wu , Qiankun Xuan , Simin Yang , Xiaoqing Chen , Qi Cao , Jian Guo , Jinhu Guo , Wenjuan Wu; J. Microbiol. 2023;61(2):233-243. Published online February 9, 2023; DOI: https://doi.org/10.1007/s12275-022-00006-4

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Abstract

The use of antibiotics can disrupt the body’s natural balance and increase the susteptibility of patients towards fungal infections. Candida albicans is a dimorphic opportunistic fungal pathogen with niches similar to those of bacteria. Our aim was to study the interaction between this pathogen and bacteria to facilitate the control of C. albicans infection. Alpha-hemolysin (Hla), a protein secreted from Staphylococcus aureus, causes cell wall damage and impedes the yeast–hyphae transition in C. albicans. Mechanistically, Hla stimulation triggered the formation of reactive oxygen species that damaged the cell wall and mitochondria of C. albicans. The cell cycle was arrested in the G0/G1 phase, CDC42 was downregulated, and Ywp1 was upregulated, disrupting yeast hyphae switching. Subsequently, hyphae development was inhibited. In mouse models, C. albicans pretreated with Hla reduced the C. albicans burden in skin and vaginal mucosal infections, suggesting that S. aureus Hla can inhibit hyphal development and reduce the pathogenicity of candidiasis in vivo.

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Candida albicans and Candida glabrata : global priority pathogens
Myrto Katsipoulaki, Mark H. T. Stappers, Dhara Malavia-Jones, Sascha Brunke, Bernhard Hube, Neil A. R. Gow, Joseph Heitman
Microbiology and Molecular Biology Reviews.2024;[Epub] CrossRef

PROTOCOL] Applications of different solvents and conditions for differential extraction of lipopolysaccharide in Gram-negative bacteria: Mai Phuong Nguyen , Le Viet Ha Tran , Hyun Namgoong , Yong-Hak Kim; J. Microbiol. 2019;57(8):644-654. Published online May 23, 2019; DOI: https://doi.org/10.1007/s12275-019-9116-5

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Abstract

Lipopolysaccharide (LPS) is one of the major components in the outer membrane of Gram-negative bacteria. However, its heterogeneity and variability in different bacteria and differentiation conditions make it difficult to extract all of the structural variants. We designed a solution to improve quality and biological activity of LPS extracted from various bacteria with different types of LPS, as compared to conventional
methods
. We introduced a quality index as a simple measure of LPS purity in terms of a degree of polysaccharide content detected by absorbance at 204 nm. Further experiments using gel electrophoresis, endotoxin test, and macrophage activation test were performed to evaluate the performance and reliability of a proposed ‘T-sol’ method and the biological effectiveness and character of the LPS products. We presented that the T-sol method had differential effects on extraction of a RAW 264.7 cell-activating LPS, which was effective in the macrophage activation with similar effects in stimulating the production of TNF-alpha. In conclusion, the T-sol method provides a simple way to improve quality and biological activity of LPS with high yield.

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Effective Modalities of Periodontitis Induction in Rat Model
Fazle Khuda, Badiah Baharin, Nur Najmi Mohamad Anuar, Bellen Sharon Fred Satimin, Nurrul Shaqinah Nasruddin
Journal of Veterinary Dentistry.2024; 41(1): 49. CrossRef
LPS-Induced Mortality in Zebrafish: Preliminary Characterisation of Common Fish Pathogens
Rafaela A. Santos, Cláudia Cardoso, Neide Pedrosa, Gabriela Gonçalves, Jorge Matinha-Cardoso, Filipe Coutinho, António P. Carvalho, Paula Tamagnini, Aires Oliva-Teles, Paulo Oliveira, Cláudia R. Serra
Microorganisms.2023; 11(9): 2205. CrossRef
Heterogeneity of Lipopolysaccharide as Source of Variability in Bioassays and LPS-Binding Proteins as Remedy
Alexandra C. Fux, Cristiane Casonato Melo, Sara Michelini, Benjamin J. Swartzwelter, Andreas Neusch, Paola Italiani, Martin Himly
International Journal of Molecular Sciences.2023; 24(9): 8395. CrossRef
Identification workflow of endotoxins by pyrolysis–gas chromatography–mass spectrometry based on a database and chemometrics
Jackie Jackie, Chun Kiang Chua, Norrapat Shih, Sam Fong Yau Li
Journal of Analytical and Applied Pyrolysis.2022; 165: 105547. CrossRef
Exploring the Lipidome: Current Lipid Extraction Techniques for Mass Spectrometry Analysis
Julian Aldana, Adriana Romero-Otero, Mónica P. Cala
Metabolites.2020; 10(6): 231. CrossRef
The outer membrane glycolipids of bacteria from cold environments: isolation, characterization, and biological activity
Angela Casillo, Ermenegilda Parrilli, Maria Luisa Tutino, Maria Michela Corsaro
FEMS Microbiology Ecology.2019;[Epub] CrossRef

Antagonistic effect of peptidoglycan of Streptococcus sanguinis on lipopolysaccharide of major periodontal pathogens: Sung-Hoon Lee; J. Microbiol. 2015;53(8):553-560. Published online July 31, 2015; DOI: https://doi.org/10.1007/s12275-015-5319-6

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Abstract

Streptococcus sanguinis is often found in subgingival biofilm including periodontopathogens, and is correlated with a delay in colonization by periodontopathogens. However, the effect of S. sanguinis on inflammation induced by periodontopathogens is poorly understood. Thus, this study investigated the effect of S. sanguinis peptidoglycan (PGN) on induction of TNF-α, IL-6, and IL-8 expression by lipopolysaccharide (LPS) of periodontal pathogens. LPS was extracted from Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, and Tannerella forsythia, and PGN was isolated from S. sanguinis. THP-1 cells, a monocytic cell-line, were cotreated with LPS of the periodontal pathogens and S. sanguinis PGN, and then the expression of inflammatory cytokines was analyzed by real-time RT-PCR. To analyze the underlying mechanism, the binding assay of the LPS to CD14 or LPS-binding protein (LBP) was performed in the presence or absence of the PGN after coating recombinant human CD14 and LBP on EIA plate. The PGN inhibited the binding of LPS to CD14 and LBP in a dose-dependent manner. Also, THP-1 cells were co-treated with the LPS in the presence of N-acetylmuramic acid and N-acetylglucosamine, as components of PGN, and the competition binding assay to CD14 and LBP was performed. N-acetylmuramic acid inhibited the induction of inflammatory cytokine expression by LPS and the binding of LPS to CD14 or LBP whereas Nacetylglucosamine did not show such effect. Collectively, the
results
suggest that S. sanguinis PGN inhibited the cytokine expression induced by the LPS of periodontopathogens due to the inhibition of LPS binding to LBP and CD14. N-acetylmuramic acid of PGN may play a role in inhibition of the LPS binding of periodontopathogens to CD14 and LBP.

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Inflammasome regulation by the cell surface ecto-5′-nucleotidase of the oral commensal, Streptococcus oralis
Natsuno Nakamura, Hirobumi Morisaki, Momoe Itsumi, Nobuo Okahashi, Haruka Fukamachi, Ayako Sato, Miki Kadena, Mariko Kikuchi, Shohei Matsui, Takahiro Funatsu, Hirotaka Kuwata
Biochemical and Biophysical Research Communications.2025; 744: 151206. CrossRef
New putative periodontopathogens and periodontal health‐associated species: A systematic review and meta‐analysis
Angéline Antezack, Damien Etchecopar‐Etchart, Bernard La Scola, Virginie Monnet‐Corti
Journal of Periodontal Research.2023; 58(5): 893. CrossRef
Correlation and mechanism between cardiac magnetic resonance imaging and oral streptococcus count in patients with primary microvascular angina pectoris
Qi Huang, Shi Sheng Wang, Rong Hua Luo
Medicine.2022; 101(12): e29060. CrossRef
Oral ecological environment modifications by hard-cheese: from pH to microbiome: a prospective cohort study based on 16S rRNA metabarcoding approach
Erna Cecilia Lorenzini, Barbara Lazzari, Gianluca Martino Tartaglia, Giampietro Farronato, Valentina Lanteri, Sara Botti, Filippo Biscarini, Paolo Cozzi, Alessandra Stella
Journal of Translational Medicine.2022;[Epub] CrossRef
Biofilm growth and IL-8 & TNF-α-inducing properties of Candida albicans in the presence of oral gram-positive and gram-negative bacteria
Radhika G. Bhardwaj, Arjuna Ellepolla, Hana Drobiova, Maribasappa Karched
BMC Microbiology.2020;[Epub] CrossRef
Genetics ofsanguinis-Group Streptococci in Health and Disease
Angela Nobbs, Jens Kreth, Vincent A. Fischetti, Richard P. Novick, Joseph J. Ferretti, Daniel A. Portnoy, Miriam Braunstein, Julian I. Rood
Microbiology Spectrum.2019;[Epub] CrossRef
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Daniela Micheline dos Santos, Betina Chiarelo Commar, Emily Vivianne Freitas da Silva, Valentim Adelino Ricardo Barão, Adaias Oliveira Matos, Marcelo Coelho Goiato
Journal of Investigative and Clinical Dentistry.2019;[Epub] CrossRef
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Manlin Qi, Xue Li, Xiaolin Sun, Chunyan Li, Franklin R. Tay, Michael D. Weir, Biao Dong, Yanmin Zhou, Lin Wang, Hockin H.K. Xu
Dental Materials.2019; 35(11): 1665. CrossRef
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Hao Wu, Li Xie, Min He, Ruitao Zhang, Yuan Tian, Suru Liu, Tao Gong, Fangjun Huo, Ting Yang, Qingyuan Zhang, Shujuan Guo, Weidong Tian
Acta Biomaterialia.2019; 97: 597. CrossRef
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Jingjing Xu, Hang Yang, Yongli Bi, Wuyou Li, Hongping Wei, Yuhong Li
Viruses.2018; 10(7): 380. CrossRef
Bacterial Adhesion on Lithium Disilicate Ceramic Surface Exposed to Different Hydrofluoric Solutions
Daniela Micheline dos Santos, Emily Vivianne Freitas da Silva, Adaias Oliveira Matos, Beatriz Cristiane Zuin Monteiro, Rodrigo Antonio de Medeiros, Sandro Basso Bitencourt, Valentim Adelino Ricardo Barão, Elidiane Cipriano Rangel, Marcelo Coelho Goiato
Ceramics.2018; 1(1): 145. CrossRef
Inhibitory effect of Lactococcus lactis on the bioactivity of periodontopathogens
Hyun-Seung Shin, Dong-Heon Baek, Sung-Hoon Lee
The Journal of General and Applied Microbiology.2018; 64(2): 55. CrossRef
The road less traveled – defining molecular commensalism with Streptococcus sanguinis
J. Kreth, R.A. Giacaman, R. Raghavan, J. Merritt
Molecular Oral Microbiology.2017; 32(3): 181. CrossRef
Buckyballs conjugated with nucleic acid sequences identifies microorganisms in live cell assays
Qingsu Cheng, Bahram Parvin
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Antimicrobial activity and regulation of CXCL9 and CXCL10 in oral keratinocytes
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European Journal of Oral Sciences.2016; 124(5): 433. CrossRef

Reducing the Bioactivity of Tannerella forsythia Lipopolysaccharide by Porphyromonas gingivalis: Young-Jae Kim , Sung-Hoon Lee; J. Microbiol. 2014;52(8):702-708. Published online August 1, 2014; DOI: https://doi.org/10.1007/s12275-014-4324-5

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Abstract

Tannerella forsythia is considered a pathogen of periodontitis and forms a biofilm with multi-species bacteria in oral cavity. Lipopolysaccharide is a powerful immunostimulator and induces inflammation and shock. The purpose of this study was to investigate the characteristics of T. forsythia LPS in its co-cultivation with Fusobacterium nucleatum or Porphyromonas gingivalis. T. forsythia was co-cultured in the presence and absence of F. nucleatum and P. gingivalis and then T. forsythia LPS was extracted. The extracts were analyzed by SDS-PAGE and NF-κB reporter CHO cell lines. THP-1 cells were treated with the LPS and evaluated induction of cytokine expression by real-time RT-PCR and ELISA. For analysis of the bioactivity of T. forsythia LPS, the binding assay on LPS-binding protein (LBP) and CD14 was processed. The extracts did not contaminate other molecules except LPS and showed TLR4 agonists. Co-cultured T. forsythia LPS with P. gingivalis exhibited a lower level of induction of TNF-α, IL-1β, and IL-6 expression than singleor co-cultured T. forsythia LPS with F. nucleatum in the conditions of human serum. However, the three T. forsythia LPS did not show difference of cytokine induction in the serum free conditions. Co-cultured T. forsythia LPS with P. gingivalis exhibited a lower affinity to LBP and CD14 as binding site of O-antigen and attached at a lower level to THP-1 cells compared to single- or co-cultured T. forsythia LPS with F. nucleatum. The virulence of T. forsythia LPS was decreased by co-culturing with P. gingivalis and their affinity to LBP and CD14 was reduced, which may due to modification of O-antigen chain by P. gingivalis.

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Anti-Inflammatory Efficacy of Human-Derived Streptococcus salivarius on Periodontopathogen-Induced Inflammation
Dong-Heon Baek, Sung-Hoon Lee
Journal of Microbiology and Biotechnology.2023; 33(8): 998. CrossRef
Virulence of Filifactor alocis lipoteichoic acid on human gingival fibroblast
Hyun-Jun Yoo, Sung-Hoon Lee
Archives of Oral Biology.2022; 135: 105370. CrossRef
Effects of Shiitake mushroom extract on antimicrobial activity against periodontopathogens and inflammatory condition of human gingival fibroblast
Yeol-Mae Jeon
Journal of Dental Rehabilitation and Applied Science.2022; 38(2): 90. CrossRef
Characteristics of Treponema denticola lipooligosaccharide in presence of hemin and quorum-sensing molecule
Dong-Heon Baek, Sung-Hoon Lee
Archives of Oral Biology.2021; 124: 105062. CrossRef
The Associations of Periodontopathic Bacteria and Oral Candida with Periodontal Inflamed Surface Area in Older Adults Receiving Supportive Periodontal Therapy
Hideo Shigeishi, Mariko Nakamura, Iori Oka, Cheng-Yih Su, Kanako Yano, Momoko Ishikawa, Yoshino Kaneyasu, Masaru Sugiyama, Kouji Ohta
Diagnostics.2021; 11(8): 1397. CrossRef
Efficacy of β-caryophyllene for periodontal disease related factors
Hyun-Jun Yoo, Su-Kyung Jwa
Archives of Oral Biology.2019; 100: 113. CrossRef
Inhibitory effect of Lactococcus lactis on the bioactivity of periodontopathogens
Hyun-Seung Shin, Dong-Heon Baek, Sung-Hoon Lee
The Journal of General and Applied Microbiology.2018; 64(2): 55. CrossRef
Consistent and reproducible long-term in vitro growth of health and disease-associated oral subgingival biofilms
Irina M. Velsko, Luciana M. Shaddox
BMC Microbiology.2018;[Epub] CrossRef
Quantification by qPCR of Pathobionts in Chronic Periodontitis: Development of Predictive Models of Disease Severity at Site-Specific Level
Inmaculada Tomás, Alba Regueira-Iglesias, Maria López, Nora Arias-Bujanda, Lourdes Novoa, Carlos Balsa-Castro, Maria Tomás
Frontiers in Microbiology.2017;[Epub] CrossRef
Gingipain‐dependent augmentation by Porphyromonas gingivalis of phagocytosis of Tannerella forsythia
Y.‐J. Jung, H.‐K. Jun, B.‐K. Choi
Molecular Oral Microbiology.2016; 31(6): 457. CrossRef
Development of Filifactor alocis media for single- and co-cultivation with periodontopathogens
조인우, 이성훈
Oral Biology Research.2016; 40(4): 193. CrossRef
Antagonistic effect of peptidoglycan of Streptococcus sanguinis on lipopolysaccharide of major periodontal pathogens
Sung-Hoon Lee
Journal of Microbiology.2015; 53(8): 553. CrossRef
Polymicrobial Oral Infection with Four Periodontal Bacteria Orchestrates a Distinct Inflammatory Response and Atherosclerosis in ApoEnull Mice
Sasanka S. Chukkapalli, Irina M. Velsko, Mercedes F. Rivera-Kweh, Donghang Zheng, Alexandra R. Lucas, Lakshmyya Kesavalu, Salomon Amar
PLOS ONE.2015; 10(11): e0143291. CrossRef

Porphyromonas gingivalis-Derived Lipopolysaccharide-Mediated Activation of MAPK Signaling Regulates Inflammatory Response and Differentiation in Human Periodontal Ligament Fibroblasts: Taegun Seo , Seho Cha , Tae-Il Kim , Hee-Jung Park , Jeong-Soon Lee , Kyung Mi Woo; J. Microbiol. 2012;50(2):311-319. Published online April 27, 2012; DOI: https://doi.org/10.1007/s12275-012-2146-x

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Abstract: Porphyromonas gingivalis (P.g.), which is a potential pathogen for periodontal diseases, contains lipopolysaccharide (LPS), and this endotoxin stimulates a variety of cellular responses. At present, P.g.-derived LPS-induced cellular responses in human periodontal ligament fibroblasts (PDLFs) are not well characterized. Here, we demonstrate that P.gderived LPS regulates inflammatory responses, apoptosis and differentiation in PDLFs. Interleukin-6 (IL-6) and -8 (IL-8) were effectively upregulated by treatment of P.g.-derived LPS, and we confirmed apoptosis markers including elevated cytochrome c levels, active caspase-3 and morphological change in the presence of P.g.-derived LPS. Moreover, when PDLFs were cultured with differentiation media, P.g.- derived LPS reduced the expression of differentiation marker genes, as well as reducing alkaline phosphatase (ALP) activity and mineralization. P.g.-derived LPS-mediated these cellular responses were effectively abolished by treatment of mitogen-activated protein kinase (MAPK) inhibitors. Taken together, our results suggest that P.g.-derived LPS regulates several cellular responses via activation of MAPK signaling pathways in PDLFs.

Research Support, Non-U.S. Gov't

Acinetobacter baumannii Outer Membrane Protein A Modulates the Biogenesis of Outer Membrane Vesicles: Dong Chan Moon , Chul Hee Choi , Jung Hwa Lee , Chi-Won Choi , Hye-Yeon Kim , Jeong Soon Park , Seung Il Kim , Je Chul Lee; J. Microbiol. 2012;50(1):155-160. Published online February 27, 2012; DOI: https://doi.org/10.1007/s12275-012-1589-4

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102 Scopus

Abstract: Acinetobacter baumannii secretes outer membrane vesicles (OMVs) during both in vitro and in vivo growth, but the biogenesis mechanism by which A. baumannii produces OMVs remains undefined. Outer membrane protein A of A. baumannii (AbOmpA) is a major protein in the outer membrane and the C-terminus of AbOmpA interacts with diaminopimelate of peptidoglycan. This study investigated the role of AbOmpA in the biogenesis of A. baumannii OMVs. Quantitative and qualitative approaches were used to analyze OMV biogenesis in A. baumannii ATCC 19606T and an isogenic ΔAbOmpA mutant. OMV production was significantly increased in the ΔAbOmpA mutant compared to wild-type bacteria as demonstrated by quantitation of proteins and lipopolysaccharides (LPS) packaged in OMVs. LPS profiles prepared from OMVs from wild-type bacteria and the ΔAbOmpA mutant had identical patterns, but proteomic analysis showed different protein constituents in OMVs from wild-type bacteria compared to the ΔAbOmpA mutant. In conclusion, AbOmpA influences OMV biogenesis by controlling OMV production and protein composition.

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