• Title/Summary/Keyword: Antifungal peptide

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Effective Antibacterial Action of Tat (47-58) by Increased Uptake into Bacterial Cells in the Presence of Trypsin

  • Jung, Hyun-Jun;Jeong, Kyu-Shik;Lee, Dong-Gun
    • Journal of Microbiology and Biotechnology
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    • v.18 no.5
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    • pp.990-996
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    • 2008
  • In a previous study, we found an antifungal effect on human pathogenic fungi by the cell-penetrating peptide Tat (47-58) derived from HIV-1. Tat (47-58) immediately entered into the fungal nucleus and affected some physiological changes on the intracellular condition. In this study, Tat (47-58) showed a broad spectrum of antibacterial activity against pathogenic bacteria including bacterial clinical isolates. To improve resistance against proteases for use in vivo, we synthesized an analog of Tat (47-58) by substituting the L-amino acid for the D-amino acid. The D-enantiomer of Tat (47-58) also exhibited a broad spectrum of antibacterial activity at almost the same level of L-Tat (47-58) concentration. Unlike L-Tat (47-58), D-Tat (47-58) showed a significant proteolytic resistance against all proteases tested and antimicrobial activities in the presence of trypsin. Moreover, D-Tat (47-58) inhibited MRSA infection in human HeLa cells whereas L-Tat (47-58) partially allowed MRSA infection, and the results were due to the proteolytic resistance of D-Tat (47-58).

Purification and characterization of antifungal compounds produced by Bacillus subtilis KS1 (Bacillus subtilis KS1이 생산하는 항진균물질의 정제 및 특성)

  • Ryoo, Sung-Woo;Maeng, Hack-Young;Maeng, Pil-Jae
    • The Korean Journal of Mycology
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    • v.24 no.4 s.79
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    • pp.293-304
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    • 1996
  • A bacterial strain, KSl, possessing strong antifungal activity was isolated from soil samples of ginseng fields and identified as Bacillus subtilis. In greenhouse test, the culture filtrate of B. subtilis KS1 showed strong protective effect against several fungal diseases of agricultural plants such as cucumber gray mold and wheat leaf rust. In addition, the crude butanol fraction of the culture filtrate exhibited antagonistic effect against several fungi including plant or human pathogens, such as Botrytis maydis, Chytridium lagenarium and Candida albicans. The antifungal compound, SW1, produced by B. subtilis KS1 was purified through consecutive chromatographic separations on a pep-RPC column and a ${\mu}$ Bondapak $C_{18}$ reverse phase column. Temperature and pH showed little effect on the stability of the compound in the ranges $-20-121^{\circ}C$ and pH 4.0-10.0, respectively. The composition and structural characteristics of SW1 were analysed by HPLC and by $^1H-,\;^1H-^1H-COSY$, NOESY, COSY-NOESY and HOHAHA NMR spectroscopy, respectively, which revealed that the compound belongs to iturin A, a typical cyclic antifungal compound produced by B. subtilis. In contrast to the previously reported iturin A compounds which have one or no $-CH_3$ side chain in the hydrophobic hydrocarbon chain of ${\beta}-amino$ acids, SW1 was shown to have a ${\beta}-amino$ acid containing 12-carbon skeleton with two $-CH_3$ side chains.

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INTERACTION OF TENECIN FRAGMENTS WITH LIPOSOMES

  • Park, Myeong-Jun;Cho, Hyun-Sook;Hong, Sung-Yu;Yoon, Jeong-Hyeok;Lee, Keun-Hyeong;Moon, Hong-Mo;Cheong, Hong-Seok
    • Proceedings of the Korean Biophysical Society Conference
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    • 1996.07a
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    • pp.37-37
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    • 1996
  • Tenecin fragments are antimicrobial and antifungal peptide from Tenebrio molitor with highly positive charged amino acid residues. To elucidate their membrane selectivity and molecular mechanism, various forms of tenecin fragments were synthesized, and their interaction with acidic phospholipid, Gram (+), fungal and human erythrocyte membrane were investigated by ANTS/DPX leakage, membrane binding and fusion assay. (omitted)

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Immunomodulatory Effects of Hypocrellin A on MHC-restricted Antigen Processing

  • Park, Sun-Im;Im, Sun-A;Kim, Ki-Hyang;Lee, Chong-Kil
    • IMMUNE NETWORK
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    • v.11 no.6
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    • pp.412-415
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    • 2011
  • Hypocrellin A has gained much attention in recent years due to its light-induced antitumor, antifungal and antiviral activities. Here we report that hypocrellin A exerts immunomodulatory effects on MHC-restricted presentation of antigen. Hypocrellin A inhibited class II-MHC restricted presentation of exogenous antigen, but not class I MHC-restricted presentation of exogenous antigen, in dendritic cells. Hypocrellin A also inhibited the cytosolic pathway of endogenous antigen presentation. However, hypocrellin A did not inhibit the expression of class I and class II MHC molecules on dendritic cells (DCs), the phagocytic activity of DCs, or the $H-2K^b$-restricted presentation of a synthetic peptide, SIINFEKL. These results show that hypocrellin A differentially modulates the MHC-restricted antigen presentation pathways.

Antifungal activity of Streptomyces costaricanus HR391 against some plant-pathogenic fungi (여러 식물병원성 진균을 억제하는 Streptomyces costaricanus HR391의 항진균능)

  • Kim, Hae-Ryoung;Song, Hong-Gyu
    • Korean Journal of Microbiology
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    • v.52 no.4
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    • pp.437-443
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    • 2016
  • In this study Streptomyces strains were isolated from soils and their antifungal activities and involved mechanisms were investigated. Among over 400 isolates of actinomycetes, Streptomyces costaricanus HR391 was selected as a potential antagonist to control several plant-pathogenic fungi. S. costaricanus HR391 inhibited mycelial growth of Fusarium oxysporum f. sp. raphani, F. oxysporum f. sp. niveum, F. oxysporum f. sp. lycopersici, and Rhizoctonia solani by 26.5, 26.2, 21.2, and 23.8%, respectively compared to those of uninoculated control after 7-day incubation on PDB medium. S. costaricanus HR391 produced $89{\mu}M$ of siderphore, and showed fungal cell wall-degrading activity including $0.46{\mu}mol/min/mg$ of chitinase and $0.83{\mu}mol/min/mg$ of ${\beta}$-1,3 glucanase. S. costaricanus HR391 secreted 87.49 mg/L of rhamnolipid, and produced 9.49 mg/L and 4.3 mM of lipopeptide, iturin A and surfactin, respectively, all they are membrane-disrupting biosurfactants. It also produced antimicrobial peptide and antibiotics phenazine. In addition to antifungal substances, S. costaricanus HR391 secreted plant growth-promoting phytohormones, zeatin, gibberellins and IAA. These results suggest that S. costaricanus HR391 may be utilized as an environment-friendly biocontrol agent against some important pathogenic fungi.

Molecular Cloning and Characterization of a Flower-specific Thionin in Chinese Cabbage

  • Jung, Bae-Gyo;Choi, Yeon-Ok;Lee, Kyun-Oh;Chi, Yong-Hun;Kang, Soon-Suk;Lee, Seung-Sik;Park, Soo-Kwon;Lee, Jung-Ro;Lim, Chae-Oh;Lee, Sang-Yeol
    • BMB Reports
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    • v.34 no.3
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    • pp.201-205
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    • 2001
  • Thionins are a family of low molecular weight cysteine-rich antimicrobial peptides. We isolated a cDNA encoding thionin gene from a flower bud cDNA library of Chinese cabbage (CFT). The gene contains 611 by nucleotides with 60 bp, and 150 by untranslated regions at its N- and C-terminal, respectively. The deduced amino acid sequence encoded 133 amino acids containing precursor polypeptide. The protein reveals that the precursor has a tripartite structure: a putative signal sequence at the N-terminus, followed by a mature thionin peptide, and a C-terminal acidic domain, which facilitates transport of the mature thionin through membrane. Genomic Southern blot analysis suggests that the CFT gene may be present as a single or two copy gene in the Chinese cabbage genome. Northern blot analysis shows that the gene is specifically expressed in flowers, but not in leaves, stems, or roots. When we analyzed the antifungal activity of the recombinant CFT protein, which was expressed in E. coli using the truncated cDNA region corresponding to the mature protein part, it was not active on fungal growth inhibition.

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Resistance Function of Rice Lipid Transfer Protein LTP110

  • Ge, Xiaochun;Chen, Jichao;Li, Ning;Lin, Yi;Sun, Chongrong;Cao, Kaiming
    • BMB Reports
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    • v.36 no.6
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    • pp.603-607
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    • 2003
  • Abstract Plant lipid transfer proteins (LTPs) are a class of proteins whose functions are still unknown. Some are proposed to have antimicrobial activities. To understand whether LTP110, a rice LTP that we previously identified from rice leaves, plays a role in the protection function against some serious rice pathogens, we investigated the antifungal and antibacterial properties of LTP110. A cDNA sequence, encoding the mature peptide of LTP110, was cloned into the Impact-CN prokaryotic expression system. The purified protein was used for an in vitro inhibition test against rice pathogens, Pyricularia oryzae and Xanthomonas oryzae. The results showed that LTP110 inhibited the germination of Pyricularia oryzae spores, and its inhibitory activity decreased in the presence of a divalent cation. This suggests that the antifungal activity is affected by ions in the media; LTP110 only slightly inhibited the growth of Xanthomonas oryzae. However, the addition of LTP110 to cultured Chinese hamster ovarian cells did not retard growth, suggesting that the toxicity of LTP110 is only restricted to some cell types. Its antimicrobial activity is potentially due to interactions between LTP and microbe-specific structures.

Structure-activity relationships of the intramolecular disulfide bonds in coprisin, a defensin from the dung beetle

  • Lee, Jaeho;Lee, Daeun;Choi, Hyemin;Kim, Ha Hyung;Kim, Ho;Hwang, Jae Sam;Lee, Dong Gun;Kim, Jae Il
    • BMB Reports
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    • v.47 no.11
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    • pp.625-630
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    • 2014
  • Defensins, which are small cationic molecules produced by organisms as part of their innate immune response, share a common structural scaffold that is stabilized by three disulfide bridges. Coprisin is a 43-amino acid defensin-like peptide from Copris tripartitus. Here, we report the intramolecular disulfide connectivity of cysteine-rich coprisin, and show that it is the same as in other insect defensins. The disulfide bond pairings of coprisin were determined by combining the enzymatic cleavage and mass analysis. We found that the loss of any single disulfide bond in coprisin eliminated all antibacterial, but not antifungal, activity. Circular dichroism (CD) analysis showed that two disulfide bonds, Cys20-Cys39 and Cys24-Cys41, stabilize coprisin's ${\alpha}$-helical region. Moreover, a BLAST search against UniProtKB database revealed that coprisin's ${\alpha}$-helical region is highly homologous to those of other insect defensins.

Syntheses and Biological Activities of Uridine Nucleoside Derivatives (Uridine Nucleoside 유도체의 합성과 생물 활성)

  • Bong-Hun Lee;Jang-Su Park;Shin-Won Kang
    • Journal of Life Science
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    • v.9 no.1
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    • pp.63-68
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    • 1999
  • Many nucleoside compounds such as 5-halogen substituted uridine, 5'-amino-5'-deoxyuridine conjugates of amino acid, peptide, and penicillin G, 5'-monophosphate uridine derivatives and 5'-monophosphate uridine-fatty acid derivatives were chemically synthesized and their antifungal, antibacterial, and antitumor activities were tested. 5-Bromo-2',3'-O-isopropylideneuridine(6) inhibited the growth of Trichophyton rubrum at $0.2{\mu}$g/ml of MIC. 5'-Amino-5'-deoxyuridine-penicillin G(19), 5'-amino-5'-deoxyuridine-cyclo(Phe-Asp)(20), and 5-iodo-5'-amino-5'-deoxyuridine- penicillin G(22) had antibarterial activity(MIC was $6.25{\mu}$g/ml against S. aureus) and the latter two nucleoside compounds were the most antitumor derivatives(their $IC_{50}$ against L5178Y murine lymphoma cell was $6.5{\mu}$g/ml).

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Three Non-Aspartate Amino Acid Mutations in the ComA Response Regulator Receiver Motif Severely Decrease Surfactin Production, Competence Development, and Spore Formation in Bacillus subtilis

  • Wang, Xiaoyu;Luo, Chuping;Liu, Youzhou;Nie, Yafeng;Liu, Yongfeng;Zhang, Rongsheng;Chen, Zhiyi
    • Journal of Microbiology and Biotechnology
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    • v.20 no.2
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    • pp.301-310
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    • 2010
  • Bacillus subtilis strains produce a broad spectrum of bioactive peptides. The lipopeptide surfactin belongs to one well-known class, which includes amphiphilic membrane-active biosurfactants and peptide antibiotics. Both the srfA promoter and the ComP-ComA signal transduction system are an important part of the factor that results in the production of surfactin. Bs-M49, obtained by means of low-energy ion implantation in wild-type Bs-916, produced significantly lower levels of surfactin, and had no obvious effects against R. solani. Occasionally, we found strain Bs-M49 decreased spore formation and the development of competence. Blast comparison of the sequences from Bs-916 and M49 indicate that there is no difference in the srfA operon promoter PsrfA, but there are differences in the coding sequence of the comA gene. These differences result in three missense mutations within the M49 ComA protein. RT-PCR analyses results showed that the expression levels of selected genes involved in competence and sporulation in both the wild-type Bs-916 and mutant M49 strains were significantly different. When we integrated the comA ORF into the chromosome of M49 at the amyE locus, M49 restored hemolytic activity and antifungal activity. Then, HPLC analyses results also showed the comA-complemented strain had a similar ability to produce surf actin with wild-type strain Bs-916. These data suggested that the mutation of three key amino acids in ComA greatly affected the biological activity of Bacillus subtilis. ComA protein 3D structure prediction and motif search prediction indicated that ComA has two obvious motifs common to response regulator proteins, which are the N-terminal response regulator receiver motif and the C-terminal helix-turn-helix motif. The three residues in the ComA N-terminal portion may be involved in phosphorylation activation mechanism. These structural prediction results implicate that three mutated residues in the ComA protein may play an important role in the formation of a salt-bridge to the phosphoryl group keeping active conformation to subsequent regulation of the expression of downstream genes.