• Title/Summary/Keyword: Antifungal protein

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Antifungal Activity of Salvia miltiorrhiza Against Candida albicans Is Associated with the Alteration of Membrane Permeability and (1,3)-β-D-Glucan Synthase Activity

  • Lee, Heung-Shick;Kim, Younhee
    • Journal of Microbiology and Biotechnology
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    • v.26 no.3
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    • pp.610-617
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    • 2016
  • Candidiasis has posed a serious health risk to immunocompromised patients owing to the increase in resistant yeasts, and Candida albicans is the prominent pathogen of fungal infections. Therefore, there is a critical need for the discovery and characterization of novel antifungals to treat infections caused by C. albicans. In the present study, we report on the antifungal activity of the ethanol extract from Salvia miltiorrhiza against C. albicans and the possible mode of action against C. albicans. The increase in the membrane permeability was evidenced by changes in diphenylhexatriene binding and release of both 260-nm-absorbing intracellular materials and protein. In addition, inhibition of cell wall synthesis was demonstrated by the enhanced minimal inhibitory concentration in the presence of sorbitol and reduced (1,3)-β-D-glucan synthase activity. The above evidence supports the notion that S. miltiorrhiza has antifungal activity against C. albicans by the synergistic activity of targeting the cell membrane and cell wall. These findings indicate that S. miltiorrhiza displays effective activity against C. albicans in vitro and merits further investigation to treat C. albicans-associated infections.

Antifungal Gene (Rs-AFP) Introduction into Rehmannia glutinosa and Gene Expression Mediated by Agrobacterium tumefaciens

  • Lee, Youn-Su;Lim, Jung-Dae;Seong, Eun-Soo;Chae, Young-Am;Yu, Chang-Yeon
    • The Plant Pathology Journal
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    • v.19 no.1
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    • pp.51-56
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    • 2003
  • Rehmannia glutinosa is one of the most important medicinal crops in Korea. However, various plant pathogens, including Fusatium spp., cause great damage on R. glutinosa and result in enormous economic losses. This study was conducted to breed Fusarium-resistant plants by using Agrobacterium tumefaciences and AFP (anti-fungal protein) gene. The plant material used was a native accession of R. glutinosa. The PCR analysis was conducted to verify transgenicity. Based on the PCR analysis, nptII band was observed in transgenic plant genome. Southern blot and AFP protein analyses also showed the expression of this gene in transgenic plants. Expression of AFP in transgenic plants offers the possibility of developing resistance to fungal infection.

Antifungal Effect of Phenyllactic Acid Produced by Lactobacillus casei Isolated from Button Mushroom

  • Yoo, Jeoung Ah;Lee, Chan-Jung;Kim, Yong-Gyun;Lee, Byung-Eui;Yoon, Min-Ho
    • Journal of Mushroom
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    • v.14 no.4
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    • pp.162-167
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    • 2016
  • Lactic acid bacteria (LAB) producing phenyllactic acid (PLA), which is known as antimicrobial compound, was isolated from button mushroom bed and the isolated LAB was identified to Lactobacillus casei by 16 rRNA gene sequence analysis. Cell-free supernatant (CFS) from L. casei was assessed for both the capability to produce the antimicrobial compound PLA and the antifungal activity against three fungal pathogens (Rhizoctonia solani, Botrytis cinerea, and Collectotricum aculatum). PLA concentration was investigated to be 3.23 mM in CFS when L. casei was grown in MRS broth containing 5 mM phenylpyruvic acid as precursor for 16 h. Antifungal activity demonstrated that all fungal pathogens were sensitive to 5% CFS (v/v) of L. casei with average growth inhibitions ranging from 34.58% to 65.15% (p < 0.005), in which R. solani was the most sensitive to 65.15% and followed by C. aculatum, and B. cinerea. The minimum inhibitory concentration (MIC) for commercial PLA was also investigated to show the same trend in the range of 0.35 mg mL-1 (2.11 mM) to 0.7 mg mL-1 (4.21 mM) at pH 4.0. The inhibition ability of CFS against the pathogens were not affected by the heating or protease treatment. However, pH modification in CFS to 6.5 resulted in an extreme reduction in their antifungal activity. These results may indicate that antifungal activities in CFS was caused by acidic compounds like PLA or organic acids rather than protein or peptide molecules.

Expression of Secretion-dedicated Srb Homologue and Antifungal Activity of Bacillus lentimorbus WJ5 (Bacillus lentimorbus WJ5의 분비 전용 Srb Homologue 발현과 항진균 활성)

  • 장유신;이영근;김재성;조규성;장병일
    • Korean Journal of Microbiology
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    • v.39 no.3
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    • pp.135-140
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    • 2003
  • Bacillus sp. secretes high levels of extracellular enzymes into the culture medium. The signal recognition particle (SRP) and the SRP receptor play a central role in targeting pre secretory proteins to the translocase. By the analysis of the DNA microarray of B. lentimorbus WJ5, it was detected that WJ5m12, antifungal activity deficient mutant induced by gamma radiation, had a down-regulated expression of the SRP receptor gene (B. subtitis srb homologue, srbL). To determine the relationship of SRP receptor to antifungal activity, srbL of B. lentimorbus WJ5 was amplified by PCR and ligated into pQE30 vector, and then transferred into WJ5m12. The transformant, WJ5m12::srbL, recovered the antifungal activity. From the 2-DE analysis, the several presecretory proteins accumulated in the mutant cell and decreased to a level of the wild type in WJ5m12::srbL. It seems that the srbL could play an important role in the secretion of the antifungal activity related proteins of B. lentimorbus WJ5.

Effect of method of synthesis on antifungal ability of ZnO nanoparticles: Chemical route vs green route

  • Patino-Portela, Melissa C.;Arciniegas-Grijalba, Paola A.;Mosquera-Sanchez, Lyda P.;Sierra, Beatriz E. Guerra;Munoz-Florez, Jaime E.;Erazo-Castillo, Luis A.;Rodriguez-Paez, Jorge E.
    • Advances in nano research
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    • v.10 no.2
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    • pp.191-210
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    • 2021
  • To compare the antifungal effect of two nanomaterials (NMs), nanoparticles of zinc oxide were synthesized by a chemical route and zinc oxide-based nanobiohybrids were obtained using green synthesis in an extract of garlic (Allium sativum). The techniques of X-Ray Diffraction (XRD), Infrared (IR) and Ultraviolet Visible (UV-Vis) absorption spectroscopies and Scanning (SEM) and Transmission Electron Microscopies (TEM) were used to determine the characteristics of the nanomaterials synthesized. The results showed that the samples obtained were of nanometric size (< 100 nm). To compare their antifungal capacity, their effect on Cercospora sp. was evaluated. Test results showed that both nanomaterials had an antifungal capacity. The nanobiohybrids (green route) gave an inhibition of fungal growth of ~72.4% while with the ZnO-NPs (chemical route), inhibition was ~87.1%. Microstructural studies using High Resolution Optical Microscopy (HROM) and ultra-structural analysis using TEM carried out on the treated strains demonstrated the effect of the nanofungicides on the vegetative and reproductive structures, as well as on their cell wall. To account for the antifungal effect presented by ZnO-NPs and ZnO nanobiohybrids on the fungi tested, effects reported in the literature related to the action of nanomaterials on biological entities were considered. Specifically, we discuss the electrical interaction of the ZnO-NPs with the cell membrane and the biomolecules (proteins) present in the fungi, taking into account the n-type nature of the ZnO semiconductor and the electrical behavior of the fungal cell membrane and that of the proteins that make up the protein crown.

Antifungal Property of Microorganisms against Korea Oak Wilt Pathogen, Raffaelea quercus-mongolicae (참나무시들음 병원균 Raffaelea quercus-mongolicae에 대한 항균미생물 분리)

  • Lee, Sang-Hyun;Lee, Seung-Kyu;Kim, Jae-Young;Lee, Chong-Kyu;Kim, Kyung-Hee;Yi, Yong-Sub
    • Microbiology and Biotechnology Letters
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    • v.40 no.1
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    • pp.66-69
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    • 2012
  • Five strains out of 200 candidate strains (SG 1-9, 1-12, SG 2-8, 2-10, and 2-17) were selected to determine their antifungal activity against Raffaelea quercus-mongolicae. The 16S rDNA sequences of the five strains were determined by sequencing analysis and analyzed by the homology of the blast program at NCBI. The homology search showed that SG 1-9 and 1-12 had a 98% homology with Streptomyces cinnamoneus and 98% homology with Burkholderia cepacia, while SG 2-8, 2-10, and 2-17 had a 99% homology with Streptomyces fradiae, a 97% homology with Staphylococcus epidermidis, and a 99% homology with Staphylococcus epidermidis. Out of the five selected strains, organic extract and protein extracts of SG2-17 strain broth were employed to determine antifungal activity against Raffaelea quercus-mongolicae. The organic extract exhibited antifungal activity, but the protein extracts did not demonstrate such an activity. Three organic solvents, butanol, benzene, and ethyl acetate, were also used for determination of antifungal activities. The activity measurements revealed that benzene extract possessed the greatest inhibitory effect on the growth of Raffaelea quercus-mongolicae, with the next highest being butanol extract, and ethyl acetate extract being the lowest.

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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Characterization of a Chitinase Gene Exhibiting Antifungal Activity from a Biocontrol Bacterium Bacillus licheniformis N1

  • Lee, Kwang-Youll;Heo, Kwang-Ryool;Choi, Ki-Hyuck;Kong, Hyun-Gi;Nam, Jae-Sung;Yi, Young-Byung;Park, Seung-Hwan;Lee, Seon-Woo;Moon, Byung-Ju
    • The Plant Pathology Journal
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    • v.25 no.4
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    • pp.344-351
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    • 2009
  • A biocontrol bacterium Bacillus licheniformis N1 grown in nutrient broth showed no chitinolytic activity, while its genome contains a gene which encodes a chitinase. The gene for chitinase from B. licheniformis N1 was amplified by PCR and the deduced amino acid sequence analysis revealed that the chitinase exhibited over 95% identity with chitinases from other B. licheniformis strains. Escherichia coli cells carrying the recombinant plasmid displayed chitinase activity as revealed by the formation of a clear zone on chitin containing media, indicating that the gene could be expressed in E. coli cells. Chitinase gene expression in B. licheniformis N1 was not detected by RT-PCR analysis. The protein was over-expressed in E. coli BL21 (DE3) as a glutathione S-transferase fusion protein. The protein could also be produced in B. subtilis 168 strain carrying the chitinase gene of N1 strain. The crude protein extract from E. coli BL21 carrying GST fusion protein or culture supernatant of B. subtilis carrying the chitinase gene exhibited enzyme activity by hydrolyzing chitin analogs, 4-methylumbelliferyl-$\beta$-D-N,N'-diacetylchitobioside and 4-methylumbelliferyl-$\beta$-D-N,N',N"-triacetylchitotrioside. These results indicated that even though the chitinase gene is not expressed in the N1 strain, the coding region is functional and encodes an active chitinase enzyme. Furthermore, B. subtilis 168 transformants expressing the chitinase gene exhibited antifungal activity against Fulvia fulva by suppressing spore germination. Our results suggest that the proper engineering of the expression of the indigenous chitinase gene, which will lead to its expression in the biocontrol strain B. licheniformis N1, may further enhance its biocontrol activity.

Mode of Action of Coptidis Rhizoma Protein (CRP) and Its Activity Against Subcutaneous Candidiasis due to Candida albicans (황련단백질의 항캔디다 작용기전 및 항피부캔디다증 효과)

  • Lee, Jue-Hee;Shim, Jin Kie;Han, Yongmoon
    • YAKHAK HOEJI
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    • v.49 no.5
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    • pp.422-427
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    • 2005
  • Our previous data showed the protein isolated from Coptidis Rhizoma (CRP) had antifungal activity. In present study, we examined mode of action of the CRP and its activity against subcutaneous candidiasis due to C. albicans yeast cells. Results showed that the CRP blocked hyphal production from yeast form of C. albicans. The CRP also activated RAW 264.7 monocyte/macrophage cell line, which resulted in nitiric oxide (NO) production from the cells. This activation seemed to increase macrophage phagocytosis to destroy the invaders. Like other antimicrobial peptides, CRP was influenced by ionic strength, thus resulting in a decrease of antifungal activity. In murine model of a subcutaneous candidiasis, the sizes of infected areas of the nude mice given the CRP after subcutaneous injection of C. albicans yeast cells to the dorsal skin were $90\%$ less than those of the nude mice groups that received DPBS instead of the CRP. All data indicate that the CRP, which appeared to act like an antimicrobial peptide and to inhibit the morphological transition from blastoconidia, was effec­tive against the subcutaneous disease.