• 제목/요약/키워드: 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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    • 제26권3호
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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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    • 제19권1호
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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
    • 한국버섯학회지
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    • 제14권4호
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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.

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

  • 장유신;이영근;김재성;조규성;장병일
    • 미생물학회지
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    • 제39권3호
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    • pp.135-140
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    • 2003
  • Bacillus sp.는 다량의 효소와 기능성 펩타이드들을 세포외로 분비하는 것으로 알려져 있다. Signal recognition particle (SRP)과 SRP receptor는 세포외 분비 단백질의 이동에 있어서 중심적 역할을 담당한다. B. lentimorbus WJ5의 DNA microarray 결과, 감마선 조사로 유도된 항진균 활성 결핍 돌연변이체인 WJ5m12에서 B. subtilis srb homologue (srbL)의 발현이 감소되는 것을 관찰할 수 있었다. SRP receptor와 항진균 활성 사이의 연관성을 고찰하기 위하여 B. lentimorbus WJ5의 srbL을 PCR로 증폭하여 pQE30 vector에 클로닝 하였으며, B. lentimorbus WJ5m12에 형질전환 시켰다. 형질전환된 B. lentimorbus WJ5m12::srbL은 항진균 활성이 복원되었다. 이차원 전기영동 결과, 수 종의 세포외 분비 단백질의 전구체들이 항진균 활성 결핍 돌연변이 균주에서 축적되었고 B.lentimorbus WJ5m12::srbL에서는 야생형 균주의 단백질 수준까지 감소되는 현상을 관찰할 수 있었다. 이는 B.lentimorbus WJ5의 항진균 활성 관련 물질의 이동에 있어서 srbL이 중요한 역할을 한다는 것을 암시한다.

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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    • 제10권2호
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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.

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

  • 이상현;이승규;김재영;이총규;김경희;이용섭
    • 한국미생물·생명공학회지
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    • 제40권1호
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    • pp.66-69
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    • 2012
  • Raffaelea quercus-mongolicae에 대한 항균작용을 하는 미생물을 분리하기 위하여 200개의 균주를 분리하였으며 이 중 SG 1-9, 1-12와 SG 2-8, 2-10, 2-17 5개의 균주에서 항균활성을 확인하였다. 5개의 균주는 확인을 위하여 16S rDNA 염기서열 분석을 하였으며, SG 1-9는 Streptomyces cinnamoneus와 98%의 homology가 SG 1-12는 Burkholderia cepacia와 98% homology가 같은 것으로 나타났다. 또한 SG 2-8은 Streptomyces fradiae와 99%의 homology를 SG 2-10은 Staphylococcus epidermidis와 97%를 SG 2-17은 Staphylococcus epidermidis와 99%의 homology를 나타내었다. 위 5개의 균주 중 활성이 가장 강한 SG 2-17의 유기용매추출물과 단백질추출물을 분리하여 활성을 조사하였으며 유기용매추출물에서 강한 활성을 확인하였다. 3개의 유기용매 중 benzene 추출물이 가장 높은 Raffaelea quercus-mongolicae의 균사성장을 억제하였다.

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
    • 한국생물물리학회:학술대회논문집
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    • 한국생물물리학회 1996년도 정기총회 및 학술발표회
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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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    • 제25권4호
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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.

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

  • 이주희;심진기;한용문
    • 약학회지
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    • 제49권5호
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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.