• Title/Summary/Keyword: sortase A

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Inhibitory Effects of Streptomyces sp. MBTH32 Metabolites on Sortase A and Sortase A-Mediated Cell Clumping of Staphylococcus aureus to Fibrinogen

  • Chung, Beomkoo;Kwon, Oh-Seok;Shin, Jongheon;Oh, Ki-Bong
    • Journal of Microbiology and Biotechnology
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    • v.29 no.10
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    • pp.1603-1606
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    • 2019
  • Sortase A (SrtA), a type of transpeptidase responsible for anchoring surface proteins to the peptidoglycan cell wall, is important in the virulence of gram-positive bacteria. Three compounds were isolated from marine-derived Streptomyces sp. MBTH32 using various chromatography techniques. The structures of these compounds were determined based on spectroscopic data and comparisons with previously reported data. Among the metabolites tested, lumichrome showed strong inhibitory activity against Staphylococcus aureus SrtA without affecting cell viability. The results of cell clumping activity assessment suggest the potential for using this compound to treat S. aureus infection by inhibiting SrtA activity.

Isovitexin, a Potential Candidate Inhibitor of Sortase A of Staphylococcus aureus USA300

  • Mu, Dan;Xiang, Hua;Dong, Haisi;Wang, Dacheng;Wang, Tiedong
    • Journal of Microbiology and Biotechnology
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    • v.28 no.9
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    • pp.1426-1432
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    • 2018
  • Staphylococcus aureus causes a broad variety of diseases. The spread of multidrug-resistant S. aureus highlights the need to develop new ways to combat S. aureus infections. Sortase A (SrtA) can anchor proteins containing LPXTG binding motifs to the bacteria surface and plays a key role in S. aureus infections, making it a promising antivirulence target. In the present study, we used a SrtA activity inhibition assay to discover that isovitexin, a Chinese herbal product, can inhibit SrtA activity with an $IC_{50}$ of $28.98{\mu}g/ml$. Using a fibrinogen-binding assay and a biofilm formation assay, we indirectly proved the SrtA inhibitory activity of isovitexin. Additionally, isovitexin treatment decreased the amount of staphylococcal protein A (SpA) on the surface of the cells. These data suggest that isovitexin has the potential to be an anti-infective drug against S. aureus via the inhibition of sortase activity.

Flavonoid Glycosides Inhibit Sortase A and Sortase A-Mediated Aggregation of Streptococcus mutans, an Oral Bacterium Responsible for Human Dental Caries

  • Yang, Woo-Young;Kim, Chang-Kwon;Ahn, Chan-Hong;Kim, Heegyu;Shin, Jongheon;Oh, Ki-Bong
    • Journal of Microbiology and Biotechnology
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    • v.26 no.9
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    • pp.1566-1569
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    • 2016
  • Three flavonoids were isolated from dried flowers of Sophora japonica using repetitive column chromatography and high-performance liquid chromatography. The flavonoids were identified as rutin (1), quercetin-3'-O-methyl-3-O-α-ʟ-rhamnopyranosyl(1 → 6)-β-ᴅ-glucopyranoside (2), and quercetin (3) on the basis of spectroscopic analysis and comparison of values reported in the literature. These compounds inhibited the action of sortase A (SrtA) from Streptococcus mutans, a primary etiologic agent of human dental caries. The onset and magnitude of inhibition of saliva-induced aggregation of S. mutans treated with compound 1 was comparable to that of untreated S. mutans with a deletion of the srtA gene.

Expression of Sortase, a Transpeptidase for Cell Wall Sorting Reaction, from Staphylococcus aureus ATCC 6538p in Escherichia coli

  • LEE, KI-YOUNG;DONG-SUN SHIN;JUNG-MIN YOON;HEONJOONG KANG;KI-BONG OH
    • Journal of Microbiology and Biotechnology
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    • v.12 no.3
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    • pp.530-533
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    • 2002
  • This paper describes the development of an enzymatic assay system for the identification of specific inhibitors of sortase, a transpeptidase that cleaves surface proteins of Cram-positive bacteria, from Staphylococcus aureus ATCC 6538p for antibacterial drug discovery. The coding region of the enzyme was amplified with the exception of the N-terminal membrane anchor sequence, cloned into a vector providing His-Patch-thioredoxin-tag at the N-terminus, expressed in Escherichia coli, and purified by metal chelate affinity chromatography. The enzyme activity was determined by quantifying increased fluorescence intensity upon cleavage of synthetic Dabcyl-QALPETGEE-Edans peptide. The results suggest that the developed in vitro assay system call be used in the search for sortase inhibitors In a short period of time.

Isovitexin Protects Mice from Methicillin-Resistant Staphylococcus aureus-Induced Pneumonia by Targeting Sortase A

  • Tian, Lili;Wu, Xinliang;Yu, Hangqian;Yang, Fengying;Sun, Jian;Zhou, Tiezhong;Jiang, Hong
    • Journal of Microbiology and Biotechnology
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    • v.32 no.10
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    • pp.1284-1291
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    • 2022
  • The rise of methicillin-resistant Staphylococcus aureus (MRSA) has resulted in significant morbidity and mortality, and clinical treatment of MRSA infections has become extremely difficult. Sortase A (SrtA), a virulence determinant that anchors numerous virulence-related proteins to the cell wall, is a prime druggable target against S. aureus infection due to its crucial role in the pathogenicity of S. aureus. Here, we demonstrate that isovitexin, an active ingredient derived from a variety of traditional Chinese medicines, can reversibly inhibit SrtA activity in vitrowith a low dose (IC50=24.72 ㎍/ml). Fluorescence quenching and molecular simulations proved the interaction between isovitexin and SrtA. Subsequent point mutation experiments further confirmed that the critical amino acid positions for SrtA binding to isovitexin were Ala-92, Ile-182, and Trp-197. In addition, isovitexin treatment dramatically reduced S. aureus invasion of A549 cells. This study shows that treatment with isovitexin could alleviate pathological injury and prolong the life span of mice in an S. aureus pneumonia model. According to our research, isovitexin represents a promising lead molecule for the creation of anti-S. aureus medicines or adjuncts.

Inhibitory Effects of Flavonoids from Spatholobus suberectus on Sortase A and Sortase A-Mediated Aggregation of Streptococcus mutans

  • Park, Wanki;Ahn, Chan-Hong;Cho, Hyunjoo;Kim, Chang-Kwon;Shin, Jongheon;Oh, Ki-Bong
    • Journal of Microbiology and Biotechnology
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    • v.27 no.8
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    • pp.1457-1460
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    • 2017
  • Seven flavonoids were isolated from Spatholobus suberectus via repetitive column chromatography and high-performance liquid chromatography. The chemical structures of these compounds were identified by spectroscopic analysis and comparison with values reported in the literature. Among the flavonoids tested, 7-hydroxy-6-methoxyflavanone (1) and formononetin (4) exhibited strong inhibitory activity against Streptococcus mutans SrtA, with $IC_{50}$ values of 46.1 and $41.8{\mu}M$, respectively, but did not affect cell viability. The onset and magnitude of inhibition of saliva-induced aggregation in S. mutans treated with compounds 1 and 4 were comparable to the behavior of a srtA-deletion mutant without treatment.

Solution-Phase Strategies for the Design, Synthesis, and Screening of Libraries Based on Natural Products

  • Kim, Sang-Hee
    • Proceedings of the PSK Conference
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    • 2003.10a
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    • pp.88-88
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    • 2003
  • The syntheses of different types of stilbenoid libraries have been studied recently. In these courses, the screening of the generated natural product-mimic focused libraries led to the identification of the novel lead compounds for human cytochrome P450 (CYP) lAs, melanin production, and sortase A. A library of trans-stilbene derivatives was prepared through a new efficient solution pahse synthetic pathway and their inhibitory activities were evaluated on human cytochrome P450s(CYP) 1A1, 1A2, and 1B1 to find a potent and selective CYP1 inhibitor. (omitted)

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Genome Sequence of Bacillus cereus FORC_021, a Food-Borne Pathogen Isolated from a Knife at a Sashimi Restaurant

  • Chung, Han Young;Lee, Kyu-Ho;Ryu, Sangryeol;Yoon, Hyunjin;Lee, Ju-Hoon;Kim, Hyeun Bum;Kim, Heebal;Jeong, Hee Gon;Choi, Sang Ho;Kim, Bong-Soo
    • Journal of Microbiology and Biotechnology
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    • v.26 no.12
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    • pp.2030-2035
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    • 2016
  • Bacillus cereus causes food-borne illness through contaminated foods; therefore, its pathogenicity and genome sequences have been analyzed in several studies. We sequenced and analyzed B. cereus strain FORC_021 isolated from a sashimi restaurant. The genome sequence consists of 5,373,294 bp with 35.36% GC contents, 5,350 predicted CDSs, 42 rRNA genes, and 107 tRNA genes. Based on in silico DNA-DNA hybridization values, B. cereus ATCC $14579^T$ was closest to FORC_021 among the complete genome-sequenced strains. Three major enterotoxins were detected in FORC_021. Comparative genomic analysis of FORC_021 with ATCC $14579^T$ revealed that FORC_021 harbored an additional genomic region encoding virulence factors, such as putative ADP-ribosylating toxin, spore germination protein, internalin, and sortase. Furthermore, in vitro cytotoxicity testing showed that FORC_021 exhibited a high level of cytotoxicity toward INT-407 human epithelial cells. This genomic information of FORC_021 will help us to understand its pathogenesis and assist in managing food contamination.